JPS5878581A - Cultivation tool for microorganism - Google Patents

Abstract

PURPOSE:To obtain a tool capable of cultivating aerobic and anaerobic microorganisms in the same cultivation tool, by packing a specific culture medium in a vessel, and sealing the opening part with a stopper. CONSTITUTION:A culture medium 2 for cultivating aerobic and anaerobic microorganism consisting of tryptone, soybean peptone, meat extract, hepatic hydrolyzate, glucose, potassium hydrogenphosphate, L-cysteine hydrochloride, P-aminobenzoic acid, sodium polyanetholesulfonate, hemin, agar and gelatin is packed in a glass cultivation tool 1, and a stopper 4 is mounted on the mouth part 3 of the cultivation tool 1 to close the cultivation tool 1. The stopper 4 consists of an integrally formed head part 5 and a drum 6 of somewhat smaller diameter than the head part 5, and a recess 7 is provided at the end of the drum 6 in the interior of the vessel 1.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a microorganism culture device filled with a medium, and particularly relates to a microorganism culture device filled with a medium that can culture both aerobic bacteria and anaerobic bacteria.

Conventionally, when attempting to culture aerobic bacteria and anaerobic bacteria using a culture device, it was necessary to remove the seal from the culture device and inject a medium suitable for culturing each type of bacteria. but,
At this time, bacteria from the outside air may be mixed in, and it may be difficult to distinguish them from the bacteria being cultured. Therefore, it has been considered to enclose a culture medium in advance in a culture device. However, since it is necessary to use a culture device filled with a culture medium suitable for each of aerobic bacteria and anaerobic bacteria, the culture method becomes extremely complicated and is not a rational and reliable culture method. Therefore, there has been a demand for a microorganism culture device filled with a medium capable of cultivating both anaerobic bacteria and aerobic bacteria.

The present letter of invention was created in view of the above circumstances, and its purpose is to have a simple structure, yet be able to completely seal microbial culture equipment before use, and to allow cultivation using the same stopper and culture medium. An object of the present invention is to provide a microorganism culture device that can rationally and reliably culture and handle aerobic bacteria and anaerobic bacteria.

In order to achieve the above object, there is provided a microbial culture device comprising a container filled with a culture medium and a stopper that fits into the opening of the container to seal the opening. Yeast extract, liver lysate, glucose, potassium hydrogen phosphate □, L-cystine hydrochloride,
P-aminobenzoic acid, sodium polyanethole sulfonate.

The present invention provides a microorganism culture device characterized by containing hemin, kuten, and gelatin.

Furthermore, the stopper is made of an elastic material that can be penetrated by a sample injection needle, and has a body portion that fits into the opening of the container to seal the opening, further comprising: The present invention provides a microorganism culturing device according to claim 1, wherein an internal atmosphere surrounded by the container and the stopper is in a reduced pressure state.

Next, a specific configuration of an embodiment of the present invention will be explained with reference to the drawings.

Microorganism culture device 1 fitted with rubber stopper 4 in Fig. 1
0 Indicates the assembled state. The inside of the glass culture device 1 is filled with a medium 2 for culturing aerobic bacteria and anaerobic bacteria. A stopper 4 is attached to the opening 3 of the culture device 1, and the culture device 1 is sealed tightly. The plug body 4 consists of a head 5 integrally formed, a body 6 having a slightly smaller diameter than the head 5, and a recess 7 on the end surface of the body inside the container.
have. Further, a cover 8 is fitted over the stopper 4 attached to the mouth portion 3 of the culture device 1.

This cover 8 is made of plastic or a malt/stopper.

Even an aluminum cap or aluminum foil can be fitted onto the bottom end 9 of the head 5 of the stopper 4 with a small gap maintained.

A recess 12 is formed on the outer end surface of the container of the head 5 of the stopper 4, and serves as a trap to facilitate penetration of a sample injection needle and a bacteria collection needle during bacterial transplantation.

On the other hand, when collecting a sample, if the inside of the culture device 1 is brought into a reduced pressure state and then the sample injection needle is inserted, the injection can be performed automatically. In this way, samples can be collected while maintaining an airtight state even in anaerobic culture and aerobic culture. In the above example, a test tube-shaped culture device was used, but the present invention The medium is not limited to this, and may be flask-shaped or bottle-shaped, with only the mouth portion having a relatively long cylindrical shape.Next, the culture medium will be explained. The same medium can be used for both anaerobic and aerobic culture,
The following media are suitable.Table 1, margins below 0.Medium compositionMedium is prepared by the ash method. Among the nine medium components shown in Table 1, weigh each component other than gelatin, and dissolve them in 500-g of distilled water. However, L-cysti/
Hydrochloride and hemin are dissolved in advance and added. 9. Separately, weigh gelatin and warm it with 500ml of distilled water to completely dissolve it. After mixing both well, cool to room temperature and adjust the pH to 7.3±0 with alkaline solution.
．． 1, and then conduct a preliminary sieve using absorbent cotton.゛The R liquid thus obtained is further filtered through a glass filter under reduced pressure, and the resulting R liquid serves as a culture medium.The plug body is manufactured by the following method. A plug is obtained by pouring the heated and melted rubber-like material into a mold and solidifying it. Next, if necessary, heat a needle having a predetermined shape and size, and punch a hole in the plug body using the needle, or use a tool such as a punch or the like to punch a hole.

After washing the culture device stopper obtained above, it is naturally dried or heated and coated with silicone oil. Next, combine the culture device and the stopper. The method is to use a 20-sized tube (15,5 φ x 165
) or a specified amount of culture medium in a suitable flask-shaped or bottle-shaped container (20sg tube is lam, flask-shaped or bottle-shaped container, etc.), and then gently add a mixed gas (nitrogen: carbon dioxide = 9: 1) is pumped in.Next, the pressure is reduced to the specified amount of water absorption, and the stopper is pushed in under reduced pressure to seal it.Then, the culture equipment is sterilized at 115℃ for 15 minutes in a high-pressure steam sterilizer. Sterilize.

Next, the effect of the present invention will be explained in the case where a holder for vacuum blood collection tubes (manufactured by Chil-Nana Co., Ltd.) is used. After disinfecting the stopper part of the culture device with alcohol, iodine tincture, etc., insert the culture device into the holder with the sample injection needle.Next, after puncturing the sample injection needle into the patient's vein, insert the culture device into the holder. When the culture device is inserted deeply enough, a predetermined amount of sample is injected into the culture device because the pressure inside the culture device is reduced. In addition, the present invention can also be practiced using a syringe.

After collecting the sample with a syringe, insert the syringe needle into the center of the specimen in the culture device whose stopper part has been sterilized with alcohol and iodine tincture. At this time, since the inside of the culture device is under reduced pressure, a predetermined amount of sample is automatically aspirated through the syringe and injected.

After this, culturing is carried out as follows. In the case of anaerobic culture, a predetermined amount of sample is aspirated using the sample strain counting method, and then
Cultivation is carried out for 1 to 14 days at 27'' to 37°C.Culture can be continued further if necessary.Incidentally, if the stopper is rotated once just before the start of culturing, degassing will be carried out well. In the case of aerobic culture, samples are collected in the same manner as in anaerobic culture, and after mixing, the stopper is lifted so that part or all of the hole is exposed to the outside of the mouth of the culture device, and then culture is performed.

Experimental Example 1 First, in an anaerobic culture, various gas-producing bacteria were suspended in an appropriate amount of sterile distilled water, adjusted to a concentration of 10" - 10" bacteria/-, collected, and then cultured at 37°C. did. If you observe this every day and check the growth status of bacteria inside the culture equipment,
Number 21 below! Table 2 Experimental Example 2 On the other hand, in aerobic culture, various bacterial strains were suspended in an appropriate amount of sterile distilled water, so that the number of microorganisms was 1 x 10''/Wd. After adjusting the bacteria concentration, collect the bacteria, lift the stopper so that the hole in the stopper is above the 4th part of the culture device, and then
Culture was started at ℃, and two days later, the number of viable bacteria in the culture solution was measured according to a conventional method.
It was growing well. 9. In a control culture device in which bacteria were not collected, they were similarly cultured at 37°C under aeration conditions for 21 days. No contamination was observed 〇 Below is the margin. Please note that the tube-shaped culture equipment used above was not used.
The same results as those obtained above were obtained when culturing was carried out using a flask-like or bottle-like container with only a relatively long cylindrical mouth.

As described above, according to the present invention, it is possible to completely seal a microorganism culture device before use, and it is possible to rationally and reliably maintain aerobic culture with a culture device using the same stopper and medium. A bacterial flora and seeds for culturing anaerobic bacteria can be created.

Furthermore, if the internal air surrounding the stopper is kept in a reduced pressure state, a sample can be automatically collected by piercing the sample injection needle.

[Brief explanation of the drawing]

FIG. 1 is a partially cutaway sectional view of the assembly of the plug and the culture device before use, and FIG. 2 is a partially sectional view of the plug. DESCRIPTION OF SYMBOLS 1... Culture device, 2... Culture medium, 3... Mouth part of culture device, 4... Stopper, 5... Stopper head, 6... Stopper body, 7... ... Recessed part of body, 10... Hole, 11...
・Circular groove〇Applicant Terumo Corporation District 1

Claims (2)

[Claims] (1) A microbial culture device consisting of a container filled with a medium and a stopper that fits into the opening of the container to seal the opening, in which the medium contains tryptone, soybean peptone, meat extract,
Yeast extract, liver hydrolyzate, glucose, potassium hydrogen phosphate, L-cystine hydrochloride, P-aminobenzoic acid,
Sodium borianethole sulfonate, hemin. Microbial culture equipment containing agar and gelatin 0% (2) The stopper is made of an elastic material that can be penetrated by a sample injection needle, has a body that fits into the opening of the container to seal the opening, and further has a body that is larger than the inner diameter of the opening. 2. The microorganism culturing device according to claim 1, wherein the stopper has a head having a large outer diameter, the container being surrounded by the stopper and the atmosphere in the strike portion being in a reduced pressure state.