JPH08205854A - Medium - Google Patents

Abstract

PURPOSE: To omit various labors for preparing a medium and to obtain the medium preservable for a long period of time by packing a bag or a container made of a plastic with medium components and a given amount of water, sealing and sterilizing under heating. CONSTITUTION: This medium is obtained by packing a bag or a container made of a plastic with a powdery medium, etc., then a required amount of water or preparing a liquid medium or an agar medium and packing the medium. Then the bag or the container is completely sealed, sterilized under heating, e.g. at 120 deg.C for >=15 minutes. The amount of the medium 2 is large and the material of the bag 1 is required to have sufficient strength and heat resistance and to form a sealing part 3 by thermal fusing. A laminate film is preferable as the material.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a culture medium. More specifically, the present invention relates to a sterilized medium that does not require weighing of the medium or sterilization of the medium when producing the medium.

[0002]

2. Description of the Related Art A typical medium for culturing microorganisms, cells, plants and the like is a liquid medium used in a liquid state and an agar medium obtained by solidifying the liquid medium with agar. Generally, these media are commercially available as powdered media in metal containers, plastic containers or bags, although these media differ depending on the type of the media. Then, at the time of use, the required amount of powder medium according to the amount of liquid medium or agar medium required is weighed, weighed, dissolved in a predetermined amount of water, most of the medium must be further sterilized Currently, these operations require a great deal of time.

Further, since the powder medium is fine particles, there is a problem that the powder medium scatters at the time of weighing, stains the scale and the laboratory bench, and also scatters when the powder medium is put into a container for dissolving the powder medium. This problem has been solved by changing the powder medium from fine particles to particles having a large particle size in some culture media, but many culture media remain conventional fine particles. Furthermore, if the container for dissolving the powdered medium is a container with a narrow opening such as an Erlenmeyer flask, the powdered medium adheres to the opening at the time of charging or spills out of the container, which may cause a problem that the container cannot be charged well. is there.

That is, it is considered that the above problems can be solved if the medium can be obtained in the state of liquid medium or agar medium instead of powder. However, a liquid medium is not commercially available at present, and there are only agar media commercially available in the state of being dispensed in a petri dish, which is the final use form. The medium dispensed in this petri dish has a remarkable evaporation of water from the medium, and if it is stored for a long period of time, the medium becomes dry and unusable.

[0005]

SUMMARY OF THE INVENTION The present invention has been made in view of the above problems, and provides a liquid and an agar medium which can save various kinds of preparation of the medium and can be stored for a long period of time. With the goal.

[0006]

In order to solve the above problems, the present invention fills a plastic bag or container with a liquid medium or agar medium, seals it, and then sterilizes it.
Alternatively, the medium is characterized in that the medium is filled and sealed in a plastic bag or container by aseptic filling.

The present invention will be described in detail below.

First, a method of sterilizing the medium after filling the medium into a plastic bag or container will be described.

The bag or container is filled with the medium by filling the bag or container with a powdery medium or various necessary medium components and then filling the bag with a required amount of water, or by using a liquid medium or an agar medium. The medium may be filled after preparation.
However, in the latter case, when the agar medium is filled, it goes without saying that it is necessary to dissolve the agar by heating so that the agar is completely dissolved, and at a temperature at which the agar does not solidify, that is, at a temperature of 45 ° C or higher. Must be filled. After filling, the bag or container is completely sealed and sterilized.
The sterilization condition may be any condition as long as the microorganisms in the medium can be completely sterilized, but sterilization treatment at 120 ° C. for 15 minutes or more is preferable. It is preferable to use a retort sterilizer for the sterilization treatment. An autoclave can be used, but since the sterilization target is a sealed bag or container, there is a risk of breaking the bag or damaging the container. Further, in the case of retort sterilization, a rotary method is preferable. In particular, when the medium components and water are separately filled, the rotary type can easily and uniformly dissolve the medium components, which is particularly effective in the agar medium. After the sterilization treatment is completed, it is necessary to quickly cool it and prevent the deterioration of the medium components as much as possible, but separately fill the medium components and water,
When the rotary retort sterilization is not performed, it is necessary to stir the content after the sterilization treatment to make the content uniform, and in the agar medium, it must be performed before the agar solidifies by cooling.

When a bag is used in the above method, the material of the bag has heat resistance sufficient to withstand sterilization, and
Any material may be used as long as it can be sealed by heat fusion. However, a laminated film is preferable because it has a large amount of content and therefore needs to have sufficient strength. For example, a bag made of a film in which polyethylene terephthalate (PET), stretched nylon (ONy) or the like is used as a substrate and non-stretched polypropylene (CPP) is laminated as a sealant can be used. Further, in order to impart a barrier property against oxygen and water vapor, a film in which an aluminum foil, a saponified ethylene-vinylacetic acid copolymer (EVOH), polyvinylidene chloride (PVDC), or the like is laminated on the intermediate layer can be used. A film coated with PVDC or a film deposited with aluminum (AL) or a metal oxide can also be used as a substrate or an intermediate layer. However, in the case of the present invention, even if the material does not have a barrier property, there is no fear that the medium will become unusable during storage. Therefore, considering the cost and the like, it is not necessary to intentionally use a material having a barrier property. Furthermore, in order to avoid misuse during use, it is preferable to use a transparent bag that allows the contents to be confirmed. Furthermore, in order to improve the operability, a bag having a self-supporting shape as shown in FIG. 1 is preferable.

On the other hand, although a container such as a bottle can be used, it is necessary to have heat resistance as a material and it is necessary to provide means for preventing leakage of contents. For example, a method of sealing the opening of the container body with a cap having packing, a method of sealing the opening with a lid material by a heat fusion method, and the like are conceivable. However, in terms of suitability for retort sterilization, the heat fusion method is more preferable than the cap method. A container completely sealed with is preferable.

Next, a method for aseptically filling the medium after sterilization will be described.

First, a liquid medium or a molten agar medium is prepared, and the medium is completely sterilized. As the sterilizer, a heat exchange plate type sterilizer can be used.
The sterilization condition may be any condition that allows microorganisms in the medium to be completely sterilized. After the sterilization treatment, the aseptic bag or container may be aseptically filled with the medium using an aseptic filling and packaging machine. In this case, the filling temperature may be room temperature or a high temperature of about 80 ° C as long as it is lower than the heat resistance of the bag or container to be used, but the condition is that the agar is not solidified in the agar medium. The aseptic filling and packaging machine may be any device as long as it is a model capable of filling a bag or container to be used. Further, the material of the bag and the container may be any material as long as it is a material for the aseptic filling device, and there is no limitation. For example, a paper container can be used as long as it is a liquid medium. However, the agar medium needs to be reheated in order to melt the agar at the time of use, and it is necessary to use a material having heat resistance of 100 ° C. or higher. Moreover, since the reheating is often performed in a hot water bath, the paper container is not suitable for the agar medium.

If the medium prepared by the above method is a liquid medium, it may be opened at the time of use, and the required amount may be aseptically dispensed into a sterile container such as an Erlenmeyer flask to be actually used. If the medium is an agar medium, it may be dissolved by heating in a sealed state, dispensed in a sterile petri dish or the like and used. The method of heating and dissolving is generally to heat in a hot water bath at 90 ° C or higher, but if the agar dissolves and there is no problem with the bag or container, use any method such as microwave oven or autoclave. It may be dissolved.

In order to facilitate the work of dispensing the medium, it is possible to provide a spout or a container with a spout to facilitate the dispensing work.

The type of the target medium is not limited, and the present invention can be applied to a liquid medium or an agar medium that requires sterilization.

[0017]

In the present invention, a liquid medium or agar medium is filled in a package such as a bag or a container and sealed, whereby an amount of powder medium according to the production amount is weighed, dissolved in water, and then sterilized. The labor of preparing the culture medium can be omitted, and the culture medium of the present invention obtained can be used as it is by opening the bag or the like, or simply by transferring to a petri dish or the like, and is sealed in a sterile state before use. Therefore, the problem of storage stability of the agar medium is solved.

If a self-supporting bag is used as the package, it is not necessary to take a fall prevention measure when the work is interrupted at the time of dispensing after opening, and if a plastic container is used, it should be transparent. For example, the color of the medium can be seen and misuse can be prevented. Furthermore, if a paper container is used, the shape of the container is maintained at the time of use, and it is easy to hold by hand, so it is easy to dispense into the final use container and when the work is interrupted during dispensing as in a self-supporting bag, It is not necessary to take a fall prevention measure, and advantageous properties in terms of the material and structure of the package can be utilized depending on the purpose.

[0019]

EXAMPLES Examples will be described below. Example 1 18.0 g of powdered normal broth medium and 1 part of water
000 ml was hermetically packed in a 150 mm × 250 mm self-standing bag of the form shown in FIG. 1 consisting of ONy / adhesive / CPP, and sterilized at 120 ° C. for 20 minutes with a retort sterilizer. After the sterilization was completed, the mixture was cooled to room temperature, and the bag was shaken well to homogenize the liquid medium. This medium at room temperature for 6
After being stored for a month, no microbial growth was observed. After that, when used for culturing E. coli, E. coli grew without any problem.

Example 2 Powdered standard agar medium 23.5
g and 1000 ml of water were hermetically packaged in a 150 mm × 250 mm self-standing bag made of ONy / adhesive / CPP shown in FIG. 1, and sterilized at 120 ° C. for 20 minutes by a retort sterilizer. After sterilization, cool to 50 ℃,
The bag was shaken well to homogenize the agar medium, and then left at room temperature to solidify the agar. This medium was stored at room temperature for 6 months, no microbial growth was observed, and then 9
After dissolving the medium in a hot water bath at 5 ° C., it was dispensed into a sterile petri dish and used for counting the number of E. coli cells, but no difference was observed between the freshly prepared medium and the results.

Example 3 A potato dextrose agar medium was prepared at 90 ° C., and 1000 times before the medium solidified.
1 ml was sealed and packaged in a 150 mm × 250 mm self-standing bag composed of PET / adhesive / AL / adhesive / CPP shown in FIG. 1 and sterilized at 120 ° C. for 15 minutes by a retort sterilizer. . After the sterilization was completed, the agar was solidified by leaving it at room temperature. Although this medium was stored at room temperature for 6 months, no growth of microorganisms was observed. After that, the medium was dissolved in a hot water bath at 95 ° C. and then dispensed into a sterile petri dish to try to culture Black Aspergillus. No difference was found in growth between the freshly made medium.

<Example 4> An ordinary broth medium was prepared,
1000 ml of PET / adhesive / AL / adhesive / C
150 mm × 250 m made of PP in the form shown in FIG.
Sealed in a self-supporting bag of m. 12 with retort sterilizer
Sterilization was performed at 0 ° C for 15 minutes. After the sterilization was completed, it was cooled to room temperature. This medium was stored at room temperature for 6 months, but no growth of microorganisms was observed. After that, an attempt was made to culture Escherichia coli, but no difference was found in the growth from the freshly made medium.

<Example 5> The prepared molten agar medium was sterilized by a plate sterilizer at 130 ° C for 15 seconds, cooled to 70 ° C, and stored in a sterile tank kept at 60 ° C. . Using the aseptic filling and packaging machine for the pouch, the medium was aseptically filled in 500 mm × 300 mm flat bags made of stretched polypropylene / polyethylene (PE) by 500 ml each. Although this medium was stored at room temperature for 6 months, no growth of microorganisms was observed. After that, the medium was dissolved in a hot water bath at 95 ° C. and then dispensed into a sterile petri dish and used for counting the number of E. coli. , No difference was found in the results from the freshly prepared medium.

Example 6 The prepared ordinary broth medium was sterilized by a plate type sterilizer at 130 ° C. for 15 seconds, cooled to 30 ° C., and stored in a sterile tank. Using the aseptic filling and packaging machine for paper containers, the medium was aseptically filled into a gable-top type (roof type) paper container with a spout having a capacity of 1000 ml of PE / paper / PE. This medium was stored at room temperature for 6 months, but no growth of microorganisms was observed. After that, an attempt was made to culture Escherichia coli, but no difference was found in the growth from the freshly made medium.

Example 7 The prepared ordinary broth medium was sterilized by a plate sterilizer at 130 ° C. for 15 seconds, cooled to 30 ° C., and stored in a sterile tank. The medium was aseptically filled into a polypropylene bottle having a capacity of 1000 ml using an aseptic filling and packaging machine for bottles. This medium was stored at room temperature for 6 months, but no growth of microorganisms was observed. After that, an attempt was made to culture Escherichia coli, but no difference was found in the growth from the freshly made medium.

[0026]

EFFECTS OF THE INVENTION The medium of the present invention is already filled with water and heat-sterilized. Therefore, in order to prepare the medium, the medium components are weighed, water is added, and the prepared medium is sterilized. No need, no hassle, easy to use. In addition, since the medium is hermetically sealed, evaporation of water during storage is small, and the medium can be stored for a long period of time.

Further, sterilized liquid medium or molten agar medium is filled in a sterile package in a sterile environment,
By sealing, the sterilization time is shorter than the heat sterilization treatment such as retort, and there is an advantage that the deterioration of the medium components is small.

The structure of the package can be selected from bags, containers, bottles, etc. according to the type of culture medium and the purpose of use, and the material is a simple substance of plastic, a laminate, or a laminate of paper and these. It has a wide range of applications because it can be used by selecting from among.

As described above, the medium of the present invention is a medium that is much easier to handle than the conventional powder medium or agar medium dispensed in a petri dish.

[0030]

[Brief description of drawings]

FIG. 1 is a cross-sectional view showing an example of a culture medium of the present invention.

[Explanation of symbols]

 1 Self-supporting bag 2 Medium 3 Seal part

Continued Front Page (72) Inventor Toshimi Yamamoto 1-5-1 Taito, Taito-ku, Tokyo Toppan Printing Co., Ltd.

Claims (7)

[Claims] 1. A package is filled with a medium component and a predetermined amount of water,
A medium that has been sealed and then heat-sterilized. 2. A medium characterized in that the package contains a liquid medium or an agar medium dissolved by heating. 3. A medium comprising a sterilized liquid medium or a molten agar medium filled and sealed in a sterile package under a sterile environment. 4. The culture medium according to claim 1, wherein the package is a plastic bag. 5. The culture medium according to claim 1, wherein the package is a plastic container. 6. The culture medium according to claim 1, wherein the package is a paper container. 7. The culture medium according to claim 4, wherein the plastic bag used is a self-supporting bag.