WO2010044613A2

WO2010044613A2 - Microorganism culture vessel that can be opened or closed selectively and method for preparing novel microorganism culture medium containing moisturizing additive

Info

Publication number: WO2010044613A2
Application number: PCT/KR2009/005927
Authority: WO
Inventors: 오정균; 이상수; 이진성
Original assignee: (주)나래바이오테크
Priority date: 2008-10-15
Filing date: 2009-10-15
Publication date: 2010-04-22
Also published as: KR20100042237A; WO2010044613A3; KR101151297B1

Abstract

The present invention relates to a microorganism culture vessel that can be selectively opened and closed and to a method for preparing a microorganism culture medium that is injected into the culture vessel. More specifically, the present invention relates to a culture vessel which suppresses water evaporation from a microorganism culture medium by allowing or preventing supply of air into the vessel by selectively opening or closing the vessel, to an additive for a microorganism culture medium that effectively suppresses water evaporation, and to a method for preparing a medium using same, which enable long-term preservation of the quality (or performance) of a microorganism medium.

Description

Method for producing a novel microbial culture medium comprising a microorganism culture vessel and a moisturizing additive that can be selectively opened and closed

The present invention relates to the production of a microorganism culture vessel capable of selective opening and closing and a medium to which a moisturizing agent inhibits water evaporation is added. More specifically, it is possible to supply or block air into a container by selective opening and closing of the culture medium for microbial culture. Microbial culture vessel that can be selectively opened and closed to maintain the performance of the long-term, moisturizing additives to maintain the long-term quality of the microbial medium by inhibiting evaporation of moisture in the microbial culture medium and a method for producing a microbial culture medium using these additives It is about.

In general, the microbial culture vessel consists of a cylindrical container body for receiving the culture medium for microorganisms, and a lid coupled to the upper portion of the container body.

The microbial culture vessel having such a structure cannot prevent the evaporation of water, which accounts for more than 90% of the components of the microorganism culture medium, and the degeneration of components that lose its function due to oxidation when contacted with air. A period of reliable performance is recognized as less than two weeks, and many inconveniences due to frequent media production are expected.

In fact, due to the disadvantage that the medium performance does not last for a long time, scientists and technicians who perform actual microbial culture in industries, hospitals, laboratories, and the like have no choice but to use and use the microbial culture as needed. It causes cost inefficiency.

In addition, the conventional microbial culture vessel has a problem that is difficult to store and manage, due to the unstable structure when stacked a plurality of, the working space is limited.

On the other hand, some of the microbial culture medium is dispensed in a conventional microbial culture vessel to commercialize the previously prepared medium, but the conventional microbial culture vessel that is easy to evaporate moisture due to disadvantages, the performance preservation capacity of the microbial culture medium is not significantly improved have.

Since the water molecules in the microbial medium are adsorbed on the surface of the solute or solid, the more these substances, the less the amount of water the microorganism can use, resulting in an extremely poor microbial culture. Osmotic concentration determined by the amount of water affects the physiological characteristics of microorganisms. The degree of availability of water available to microorganisms is indicated as an indicator of water activity. It can be seen that the optimum water activity is different.

Table 1

Minimum water activity (Aw) required for microbial growth <sup> * </ sup>

Water activity	Germ	Environment
0.95	Most Gram-negative bacteria	Blood, vegetables, fruit
0.95	Most Gram-positive bacilli	bread
0.90	Most Aureus and Bacillus	ham
0.85	Staphylococcus aureus	salami
0.75	Basophils, pressure-resistant osmotic yeast	Pickled Sugar

^* ADBrown, "Microbial water stress, in Bacteriological Reviews, 40 (4): 803-846, 1976.

As a result, the water activity can maintain the optimal culture environment of the bacteria when minimizing or minimizing the evaporation of free water other than the solute and bound water attached or bound to the culture vessel surface in consideration of the microbial medium. .

However, the conventional microbial medium does not contain a component that inhibits moisture evaporation or moisturizing function, so that the moisture of the medium is easily evaporated, and drying proceeds rapidly. In addition, the importance of water in cultivating microorganisms is very important because the water content of the microbial culture medium is significantly reduced, as described in the test operation section of the Food Code. It is an important factor.

Therefore, the method of minimizing or suppressing a series of processes or a process in which the microbial culture medium evaporates moisture and becomes dry may have a significant improvement in time, labor and cost for many people who culture, examine, diagnose or study microorganisms. will be.

The present invention has been made to solve the above problems, through the functional characteristics of the structure to realize the selective sealing and opening of the container to ensure that the inside of the container is sealed during storage and the inside and outside air of the container when opening Improves the problem of lack of medium performance preservation ability by simultaneously communicating with limited or smooth communication, and at the same time to meet anaerobic and aerobic culture conditions in microbial culture, improving inefficiency of time, labor and cost of microbial culture The first objective is to provide a microbial culture vessel that can be selectively opened and closed, and to facilitate the storage and management by allowing the safe stacking of multiple culture vessels.

The second purpose is to delay the evaporation of moisture essential for the growth of microorganisms through the addition of a new moisturizing agent that inhibits the evaporation of water to the microbial medium injected into the container to prolong the optimum quality of the medium to occur during frequent production of the medium. To improve time and money.

Finally, the present invention relates to a method for preparing a microbial culture medium using the additive for inhibiting or delaying water vaporization, and relates to a method for producing a solid medium using the additive having a novel water vaporization inhibitory effect. .

The present invention is the top is open and the outer circumferential surface is formed inclined in the form of upper and lower light, a plurality of cutting grooves are formed in the outer peripheral surface of the medium therein, including a lid for opening and closing the inlet of the container body according to the user's selective opening and closing Provides a microorganism culture vessel that can be selectively opened and closed.

The present invention also provides a method for preparing a solid medium for microbial culture by injecting a liquid medium into the microbial culture vessel of the present invention.

The microorganism culture vessel capable of selectively opening and closing according to the present invention and the additive having an effect of inhibiting water evaporation are characterized by minimizing contact with air, ie, oxygen, introduced into the microorganism culture medium injected into the container, thereby preventing the oxidation of the culture medium. By prolonging the optimum quality maintenance period and providing the effect of selectively implementing anaerobic and aerobic culture environments and the growth environment of the culture medium for culturing microorganisms injected into the container by additives that suppress water evaporation, By increasing the period of storage and distribution while maintaining the effectiveness of the effect is expected to commercialize the culture medium prepared through a series of manufacturing processes.

In addition, it is easy to store and manage a plurality of culture vessels to be safely stacked, there is an effect that can overcome the spatial constraints in performing microbial culture.

1 is an exploded perspective view showing a microbial culture vessel capable of selectively opening and closing in accordance with the present invention.

Figure 2 is a perspective view showing a combined state of the microbial culture vessel capable of selectively opening and closing in accordance with the present invention.

3 is a cross-sectional view taken along the line A-A of FIG.

Figure 4 is a view showing the inflow of air when using a microbial culture vessel capable of selectively opening and closing in accordance with the present invention.

1: container body

11: protrusion

12: partition

13: base plate

14: outer wall

141: jamming

2: lid

21: cutting groove

22: recessed groove

23: hanging groove

24: top plate

25: outer wall

B: air inflow passage

A microorganism culture vessel capable of selectively opening and closing according to the present invention includes a container body having an upper portion open and an outer circumferential surface inclined in the form of upper and lower light beams to receive a medium therein, and coupled to an upper portion of the container body and having an outer circumferential surface A plurality of cutting grooves are formed in and characterized in that it comprises a lid for opening and closing the inlet of the container body according to the user's selective opening and closing.

In addition, the lower portion of the container body is characterized in that the projection is formed downwardly protruding along the circumferential direction, the recessed groove corresponding to the projection is formed on the upper surface of the lid.

In addition, a grid-shaped partition is formed on the inner bottom of the container body.

The present invention also provides a method for preparing a solid medium for culturing microorganisms by introducing a liquid medium into the microorganism culture vessel according to the present invention.

In the method according to an embodiment of the present invention, the liquid medium may contain an additive having a moisture evaporation inhibiting effect. The solid medium for culturing the microorganism prepared by the present invention refers to a solid medium solidified by appropriately cooling the liquid medium containing or not containing an additive having a water evaporation inhibiting effect and then adding the appropriate amount to the microbial culture container according to the present invention.

In the method according to an embodiment of the present invention, the additive having a moisture evaporation inhibiting effect is propylene glycol (propylene glycol), trihalose (trehalose), sodium lactate (sodium lactate), chondroitin (chondroitin), hyaluronic acid (hyaluronic acid), meso erythritol and one or more selected from the group consisting of glycerol, or a derivative of the additive or a complex including the additive, but is not limited thereto.

Derivatives of the additives or the composite containing the additives Phosphatidylcholine, Palmitoyl pentapeptide, Sodium Pyrrolidone Carboxylate, Pseudodoceramide (PC104), Cholesterol, Linolelic Acid, Pythoping ), Stearic acid, propylene glycol, hyaluronic acid triethanolamine, polypeptides and sodium lactate (PSL), sphingolipid, cerebromide (cerebroside), dipropylene glycol, sodium stearoyl glutamate, squalene, dimethicone, cetaryl isononanoate, sodium polyacrylate ( sodium polyacrylate), PEG-40 hydrogenated caster oil, PEG-60 hydrogenated caster oil, methyl Methyl paraben, propyl paraben, phenyl trimethicone, octyldodeceth 16, hydrogenated lecithin, triethanolamine, trisodium EDTA And the like.

In the method according to an embodiment of the present invention, the medium is Plate Count Agar, Endo Agar, Eosine Methylene Blue Agar, Nutrient Agar, Desoxycholate Lactose Agar, Plate Count Agar with Brom Cresol Purple, Potato Dextrose Agar, Mannitol Salt Agar with Egg Yolk Agar) Thiosulfate Citrate Bile Salt Sucrose Agar, Liver Agar, Clostridium perfringens Agar, Salmonella Shigella Agar ), MacConkey Agar, Desoxycholate Citrate Agar, Oxford Agar, Lithium Cla Lithium Chloride Phenylethanol Moxalactam Agar, Tryptic Soy Agar, Tryptose-Sulfite-Cycloserine Agar, MacConkey Sorbitol Agar Agar, Cefsulodin Irgasan Novobiocin Agar, Mannitol Egg Yolk Polymyxin agar, Liver-Veal Egg Yolk Agar, Anaerobic agar Agar, Xylose Lysine Desoxycholate Agar, Chromogenic Enterobacter sakazakii Agar, Violet Red Bile Glucose Agar, Violet Red Bile Agar Red Bile Agar, Bidd-Parker Agar, Bismuth Sulfite Agar or Polymyxin Acriflavin LiCl Septa Polymyxin Acriflavin LiCl Ceftazidime Esculin Mannitol Agar, but is not limited thereto.

In the method according to an embodiment of the present invention, it is preferable to add the additive having the moisture evaporation inhibiting effect to the liquid medium after sterilizing the liquid medium, which can be modified by heat when sterilized with the liquid medium. Because there is. Therefore, the additive is preferably added to the sterilized liquid medium after sterilization separately through a sterilization apparatus such as a syringe filter.

In the method according to an embodiment of the present invention, the propylene glycol, trihalose, sodium lactate, hyaluronic acid, mesoerythritol, glycerol, derivatives or complexes thereof are added to the liquid medium at a concentration of 5% or less of the final concentration. It is preferable to add to the liquid medium more preferably at a concentration of 0.1-5% of the final concentration.

In the method according to one embodiment of the present invention, it is preferable to add the chondroitin, derivatives or complexes thereof to the liquid medium at a concentration of 0.1% or less of the final concentration, more preferably at a concentration of 0.01 to 0.05% of the final concentration. To the liquid medium.

In the method according to an embodiment of the present invention, the microbial culture vessel is characterized in that it has a circular or square or a shape that is regularly arranged with a plurality of circles and squares. The microbial culture vessel is characterized by the Sanghyup light, which is the patented technology, and means a shape in which a plurality of microbial culture spaces (1 or more) are formed, such as 6-well plates, 12-well plates, and 96-well plates. The user can culture one kind of bacteria in one or more different media in one container, and can test the culture properties of the bacteria by various media types. In addition, by making a plurality of medium of one kind and cultivating different kinds of bacteria in one container has the advantage that the culture properties of a plurality of bacteria on one medium can be tested in one container at the same time.

An embodiment is described with reference to the accompanying drawings.

1 is an exploded perspective view showing a microorganism culture vessel capable of selectively opening and closing according to the present invention, Figure 2 is a perspective view showing a coupling state of the microorganism culture vessel capable of selectively opening and closing according to the present invention, Figure 3 is a line AA of Figure 2 4 is a cross-sectional view showing the inflow of air when using a microbial culture vessel capable of selectively opening and closing in accordance with the present invention.

As shown in Figures 1 to 3, the microorganism culture vessel that can be selectively opened and closed according to the present invention is configured to include a container body (1), and a lid (2) coupled to the container body (1).

The microorganism culture vessel capable of selectively opening and closing (hereinafter referred to as 'microbial culture vessel' for convenience) may completely seal the container body 1 or allow external air to be circulated according to the selective opening and closing of the lid 2. .

More specifically, the lid 2 and the container body 1 are combined in a closed state in which communication between the inside and the outside of the container is completely blocked, and an anaerobic culture state in which communication between the inside and the outside of the container is limited, In order to be made selectively in aerobic culture state that the communication between the inside and the outside of the container is made smoothly. This extends the medium performance storage period accommodated inside the container body 1 and enables selective implementation of anaerobic and aerobic culture environments.

First, the container body 1 is open at the top and the outer circumferential surface is formed to be inclined in the form of upper and lower light and accommodate the medium therein. The container body 1 is composed of a disk-shaped base plate 13 on which a microorganism culture medium is placed, and an outer wall 14 protruding upward from the outer periphery of the base plate 13.

A locking protrusion 141 is formed at an upper end of the outer wall 14 of the container body 1 so as to be caught by a locking groove 23 formed at an inner upper portion of the lid 2 when the lid 2 is coupled with the lid 2.

The outer diameter of the locking projection 141 is formed larger than the inner diameter of the lid 2, the outer wall 14 of the container body 1 is formed to be inclined. Therefore, in the storage and distribution stage of the microbial culture vessel, the lid 2 is combined with the lid 2 to prevent the microbial culture medium to be stored or to prevent evaporation of water contained in the microbial culture medium generated during the distribution process. As a result, the water content of the microorganism culture medium is prevented from being reduced, thereby improving the storage capacity of the microorganism culture medium.

Meanwhile, a partition 12 having a lattice structure is formed at an inner bottom of the container main body 1, and according to the formation of the partition 12, a flow of the microorganism culture medium accommodated inside the container main body 1 is prevented. And facilitates counting of colonies.

The protrusion 11 is formed downwardly protruding downward along the circumferential direction on the lower surface of the container body 1, the recessed groove 22 corresponding to the protrusion 11 is formed on the upper surface of the lid (2).

Through the depression structure of the lid (2) it is possible to minimize the amount of air remaining in the culture medium in the culture medium. In other words, by minimizing the amount of air remaining in the microbial culture vessel, by minimizing the amount of oxygen in the air to prevent the degeneration by oxidation of the microorganism culture medium components accommodated to improve the performance preservation capacity of the microorganism culture medium.

In addition, the formation of the protrusions 11 and the recessed grooves 22 ensures stability when stacking and storing a plurality of microbial culture vessels, thereby facilitating storage and management.

On the other hand, the lid 2 is coupled to the upper portion of the container body 1, a plurality of cutting grooves 21 are formed on the outer circumference circumference and open and close the inlet of the container body 1 according to the user's selective opening and closing . The cutting groove 21 is preferably formed in a semi-elliptic shape, not limited to this, and can be deformed as much as possible.

The lid 2 includes a circular upper plate 24 and an outer wall 25 protruding perpendicularly to the lower portion of the upper plate 24.

As shown in Figure 4, when culturing the microorganisms using the microbial culture vessel according to the present invention, according to the oxygen demand of the microorganisms present in the sample sample, that is, in the case of aerobic microorganisms, the lid of the microbial culture vessel Push up and open towards the top to increase the inflow of oxygen into the container, in the case of anaerobic microorganisms can be controlled by the user to block the inflow of oxygen into the container by closing the lid of the microbial culture vessel to the lower side. In other words, by adjusting the lid (top) of the container and through this can increase or decrease the height of the internal space, the user can control the growth conditions of aerobic and anaerobic microorganisms.

A plurality of cutting grooves 21 formed in the outer wall 25 of the lid 2 and the outer wall 14 of the container body 1 by slightly placing the lid 2 coupled to the container body 1 without pressing them. The air inflow passage (B) is finely formed between the top. Therefore, the air is introduced into the microbial culture vessel through the air inlet passage (B) according to the culture environment.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

실시예 1: 신규한 미생물 배양용기의 보습 효과Example 1 Moisturizing Effect of a Novel Microbial Culture Container

Microbial culture vessel (LOP, Lab On a Plate) designed by the present invention as a control group Petri dishes currently used the most as a control to evaluate the water evaporation inhibiting ability, that is, the moisturizing effect. First, LB agar medium (Luria Bertani agar, Casein enzymic hydrolysate 10g, yeast extract 5g, sterilized for 15 minutes at 121 ° C. and 1 atm) for the preparation of a medium for use in the conventional petri dish and the microorganism culture container of the present invention as a control group. , sodium chloride 10g / 1000ml) was used as the standard medium, and 17ml was inoculated and solidified by drying for 30 minutes in aseptic mass. Weigh the weights of the dried medium, the Petri dish containing the same, and the microbial culture vessel of the present invention using an electronic balance, and 4 ° C, 25 ° C, 32 ° C and 37 ° C as conditions for evaporation of water. Into the thermostat set to an artificial set temperature was weighed the control group and the experimental group containing the medium every 24 hours. In this study, water evaporation was defined as the difference between the remaining net medium volume weighed every four days, the net medium weight of T0, which is the initial time (when the medium was first injected), minus the Petri dish weight. The net medium weight was 100%, and then every 4 days (T1 to T8), the net medium weight was weighed and described as a percentage compared to the time point T0. All tests weighed every four days weighed (3 replicates) each of three test samples made under the same conditions to yield an average value.

As a result of the remaining medium weight according to each set temperature (Table 2), at all four set temperatures, the moisturizing effect of the microbial culture vessel of the present invention was higher than that of the conventional Petri dish.

TABLE 2

Remaining medium weight for each set temperature

Exam conditions		T0	T1	T2	T3	T4	T5	T6	T7	T8
4 ℃	Petri Dish	100	98.65	98.28	95.48	94.31	93.4	91.15	89.21	89.03
4 ℃	Microbial culture vessel of the present invention	100	100.00	100.00	99.84	99.83	99.68	99.68	99.29	98.8
25 ℃	Petri Dish	100	93.08	86.60	91.90	88.20	58.73	26.33	23.59	23.04
25 ℃	Microbial culture vessel of the present invention	100	99.92	98.26	99.83	99.58	99.59	99.36	99.28	98.69
32 ℃	Petri Dish	100	78.01	56.78	30.14	4.66	4.28	3.69	3.65	3.63
32 ℃	Microbial culture vessel of the present invention	100	99.13	97.90	96.50	95.47	94.61	93.89	92.24	89.26
37 ℃	Petri Dish	100	59.29	19.44	4.56	4.29	4.23	4.07	3.57	3.34
37 ℃	Microbial culture vessel of the present invention	100	98.63	96.26	94.99	92.38	90.6	86.94	82.85	82.16

In addition, the data on the water evaporation amount (water evaporation amount = 100-remaining medium amount at each time point) is applied to the SPSS statistical program at 4 ° C., which is a general storage temperature of ready-to-use among the test temperatures obtained above. When the moisture in the medium evaporated 5% (5% water evaporation amount) was set as the limit quality indicator and the safety factor was 0.7, the shelf life of the microorganism culture vessel and the petri dish was compared. As a result (Table 3), the microbial culture vessel of the present invention was calculated to have a shelf life of 110 days (110.8121212 days), which is about 100 days longer than conventional petri dishes (10.04092141 days). Compared to Petri dishes, the product showed more than 11 times better shelf life.

From the above results, it can be confirmed that the microbial culture vessel of the present invention, which is one of the most important environmental factors when culturing microorganisms, has a superior effect than the conventional microbial culture vessel Petri dish in preserving and maintaining the moisture content for the longest time there was.

TABLE 3

Comparison of shelf life of petri dishes with microbial culture vessel of the present invention

	Regression equation	Expiration date (days)	Shelf life with safety factor in days
Microbial culture vessel of the present invention	y = -0.033x + 100.224	158.3030303	110.8121212
Petri Dish	y = -0.369x + 100.293	14.34417344	10.04092141

실시예 2: 미생물 배양 배지에 신규한 첨가제(보습제)의 첨가에 따른 보습 효과Example 2: Moisturizing effect of the addition of a novel additive (humidant) to the microbial culture medium

Seven kinds of propylene glycol, trihalose, sodium lactate, chondroitin, hyaluronic acid, meso erythritol and glycerol In order to test whether the additive can inhibit water evaporation of the microbial culture medium, a petri dish, which is a conventional microbial culture medium, was carried out as follows. First, the weight of Petri dishes was weighed for the preparation of the medium to be used for the control group without additives and the control group with the experimental group, and then sterilized for 15 minutes at 121 ° C. and 1 atm, and cooled to 55 ° C. And as shown in Table 4 was added to the different concentrations of each additive as the experimental group medium in 17ml metered dose and dried for 30 minutes in aseptic mass. The weight of the dried medium and the petri dish containing the same was measured using an electronic balance, and placed in an incubator set at 30 ° C. to accelerate water evaporation, and the weight of the petri dish containing the medium was measured every 24 hours. In this test, water evaporation was defined as the difference between the remaining net medium volume weighed every 24 hours, the net medium weight of T0, the initial time (when the medium was first injected), minus the weight of the Petri dish. The net weight of the medium was 100%, and then the net medium weight was weighed every 24 hours (T1 to T9) and described as a percentage compared to the time point T0. All tests weighed every 24 hours weighed each of the three test samples made under the same conditions to yield an average value. In addition, in order to evaluate whether the additive used in the test has an effect of inhibiting water evaporation in other media, the test was conducted in the same manner as described above on PCA, PDA (Potato dextrose agar), and Coliform medium using glycerol as an additive. It was. In order to know whether the additives added to the medium had a significant water evaporation inhibitory effect, the paired samples T test of the statistical program SPSS (ver. 13.0) was used. Concentrations were analyzed (Table 7).

Table 4

Concentration Conditions of Additives Used in Moisture Inhibition Testing

	Additives to inhibit water evaporation (moisturizer)
	Propylene glycol	Trehalose	Sodium lactate	Chondroitin	Glycerol	Mesoerythritol	Hyaluronic acid
Test concentration (%)	0, 1, 3, 5	0, 1, 3, 5	0, 1, 3, 5	0, 0.01, 0.05, 0.1	0, 1, 5, 10	0, 0.1, 1	0.01, 0.02

Table 5

Table 6

Statistical analysis of the seven types of additives used in the test to evaluate the effect of inhibiting the water vaporization of the microbial culture medium (Table 7) shows that, in the PCA medium, all seven additives suppress the water evaporation. It was confirmed to be effective. In detail, it was analyzed that propylene glycol had the effect of inhibiting water evaporation at 1.35% and 0.67% at 1% and 5% concentrations (P = 0.001, P = 0.002), respectively. Trehalose and sodium lactate inhibited water evaporation at all test concentrations of 1, 3 and 5%. In the case of chondroitin, significant evaporation inhibitory effect was observed in 0.01% and 0.05%, and in hyaluronic acid and meso erythritol, 0.02% and 0.1%, respectively. In the case of glycerol, significant moisture evaporation inhibitory effects were observed at concentrations of 3% and 5%. Second, Trehalose, sodium lactate and chondroitin were found to inhibit water evaporation proportionally with increasing concentration. Finally, as the effect of inhibiting water evaporation of glycerol in PDA medium was significant at 1% and 3%, all of the above additives could be applied to all microbial culture medium regardless of the medium type, and different at the optimum concentration. As can be seen, it has been found to have a significant characteristic of inhibiting water evaporation.

TABLE 7

In addition, propylene glycol, trehalose, chondroitin, meso erythritol, and glycerol were tested at 16 concentrations in the test of 16 kinds of microorganisms according to the concentration of each additive whose significant effects were recognized through the statistical verification of the above water vapor suppression effect. It was confirmed that it did not inhibit the growth of microorganisms. However, sodium lactate did not inhibit the growth of microorganisms only at a concentration of 1% and hyaluronic acid was found to inhibit bacterial culture at a concentration of 0.02% (Table 8).

Table 8

Bacterial Culture Results According to the Optimal Concentration of Additives with Inhibiting Water Evaporation

Standard strain name

Propylene glycol

Trehalose

Sodium lactate

Chondrotin

Hyaluronic acid

Mesoerythritol

Glycerol

One%

5%

One%

3%

5%

One%

3%

5%

0.01%

0.05%

0.1%

0.02%

0.1%

One%

3%

5%

Bacillus cereus (ATCC 11778)

L

P

L

P

L

Bacillus subtilis (ATCC6633)

L

P

L

P

L

Pseudomonas aeruginosa (ATCC 27853)

L

P

L

P

L

Yersinia enterocolitica (ATCC 23715)

L

I

L

P

L

Enterococcus faecalis (ATCC 29212)

L

I

L

P

L

Klebsiella pneumoniae (ATCC 13883)

L

P

L

P

L

Citrobacter freundii (ATCC 8090)

L

P

L

P

L

Escherichia coli (ATCC 25922)

L

P

L

P

L

Listeria monocytogenes (ATCC 19111)

L

P

L

P

L

Serratia marcescens (ATCC 14756)

L

I

L

P

L

Staphylococcus aureus (ATCC 25923)

L

I

L

P

L

Proteus mirabilis (ATCC 25933)

L

I

L

P

L

Candida albicans (ATCC 10231)

L

P

L

P

L

Enterobacter aerogenes (ATCC 13048)

L

I

L

P

L

Shigella sonnei (ATCC 25931)

L

P

L

P

L

Salmonella enterica (ATCC 43971)

L

P

L

P

L

L: luxuriant growth, P: partial growth, I: Inhibition or no growth

While the present invention has been described with reference to the accompanying drawings, it will be apparent to those skilled in the art that many various obvious modifications are possible without departing from the scope of the invention from this description. Therefore, the scope of the invention should be construed by the claims described to include examples of many such variations.

Claims

A container body 1 having an upper portion open and having an outer circumferential surface inclined in the form of upper and lower light beams to receive the medium therein;

Is coupled to the upper portion of the container body 1 is formed with a plurality of cutting grooves 21 on the outer circumference circumference and includes a lid (2) for opening and closing the inlet of the container body 1 according to the user's selective opening and closing A microbial culture vessel that can be selectively opened and closed.
According to claim 1, wherein the projection 11 is formed in the lower surface of the container body 1 in the circumferential direction downwardly, the recessed groove corresponding to the projection 11 on the upper surface of the lid ( 22) microorganism culture vessel capable of selectively opening and closing, characterized in that is formed.
The microorganism culture vessel of claim 1 or 2, wherein a partition 12 of a lattice structure is formed on an inner bottom of the vessel body (1).
Liquid badges A method for preparing a solid medium for culturing microorganisms by adding to the microorganism culture vessel according to claim 1.
5. The method according to claim 4, wherein the liquid medium contains an additive having a moisture evaporation inhibiting effect.
The method of claim 5, wherein the additive having a water evaporation inhibiting effect is at least one selected from the group consisting of propylene glycol, trihalose, sodium lactate, chondroitin, hyaluronic acid, mesoerythritol and glycerol or derivatives or complexes thereof. How to feature.
The method according to claim 5, wherein the additive having the effect of inhibiting water vaporization is added to the liquid medium after sterilizing the liquid medium.
The method of claim 6, wherein the propylene glycol, trihalose, sodium lactate, hyaluronic acid, mesoerythritol, glycerol, derivatives or complexes thereof are added to the liquid medium at a concentration of 5% or less of the final concentration. .
7. The method of claim 6, wherein said chondroitin, derivatives or complexes thereof are added to the liquid medium at a concentration of 0.1% or less of the final concentration.
The method according to claim 4, wherein the microbial culture vessel has a shape in which a circle or a square or a plurality of circles and rectangles are regularly arranged.