JPH08301712A - Antifungal agent and its usage - Google Patents

Abstract

PURPOSE: To obtain an antifungal agent, containing Bacillus subtilis natto capable of strongly proliferating under the same environment as compared as that for molds and suppressing the proliferation an propagation of molds as an active ingredient and applicable to the molds in a bathroom, a kitchen, a closet, a shoe box, etc. CONSTITUTION: This antifungal agent uses readily available and commercial Bacillus subtilis natto, suspended in water and regulated to 10<8> to 10<9> bacterial cells/ml number of the Bacillus subtilis natto. The dispersion of the Bacillus subtilis natto is sprayed in a volume of 10-20ml/m<2> at a time on a place where molds are grown and proliferated or a site with the possibility for growing and proliferating thereof. The spraying may be carried out once or several times. The molds can be prevented from growing or proliferating by directly allowing the dispersion to stand. When the molds are preremoved by a simple means in spraying, effects thereof can more be enhanced. A means such as coating of the dispersion of the Bacillus subtilis natto is also included in the spraying.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an antifungal agent or an antifungal method utilizing the antifungal action of natto bacteria.
INDUSTRIAL APPLICABILITY The antibacterial agent or the antibacterial method of the present invention can be used for the prevention of fungus in places such as a bathroom, kitchen, push-in, shoe box, where mold is likely to be generated or propagated.

[0002]

2. Description of the Related Art If the temperature and humidity conditions are the same in a general household throughout the year, dark brown or black molds are often found everywhere in bathrooms, kitchens, shoe boxes and the like. Also,
In the push-in room, there were many experiences that "moldy odor" snorted when ventilation was insufficient. The generation and reproduction of such molds not only spoils the aesthetics of the living environment, but also causes problems such as the health of the inhabitants when mold spores (mold fungi) float in the living / indoor space. .

On the other hand, conventionally, as a means for sterilizing / controlling fungi or preventing the occurrence of fungi, it has been routinely practiced to use chemicals as an anti-fungal agent. Specifically, some kind of commercially available chlorine-based chemicals are applied or sprayed, and then wiped off. However, during the moldy season,
Since this single operation is not sustainable and it is impossible to completely prevent the generation of mold, the actual situation is that the application and spraying are repeated regularly.

Chlorine chemicals which have been conventionally used for sterilizing or preventing the growth of mold bacillus require cleaning after spraying or application due to problems with odor. However, there was a problem that the chemicals had to be repeatedly used as an anti-fungal agent. On the other hand, when spraying the drug, there is a health problem to the user,
Also, due to lack of knowledge and misuse of the user, when several chemicals are used in combination, the chlorine component that should have been dissolved dissociates and gasifies, causing poisoning to the person in use or death in some cases. Accidents such as accidents may occur. It was necessary to pay close attention to the use of these chemicals against mildew, and it was necessary to carry out post-treatment such as thorough washing after use.

[0005]

DISCLOSURE OF THE INVENTION The present inventor has earnestly studied to solve these problems of the conventional means for preventing or suppressing the formation of mold by chemicals. That is, an object of the present invention is to provide an antifungal agent which can be easily used and post-treated after use, and a method of using the antifungal agent.

[0006]

In the present invention, in order to solve such a problem, that is, means for easily and safely preventing or suppressing the generation of mold has been studied. In particular,
Anti-bacterial means using bacteria, specifically, whether or not it is possible to prevent the growth of fungi by allowing coexistence of fungi that are stronger than fungi even in the presence of fungi and are harmless to the human body. In pursuit of possibility, we considered mold vs. first.

[0007] That is, fungi and their microorganisms were made to coexist by using various microorganisms, and the state of their growth was examined. As a result of such a coexistence test, the natto fungus ( Bacillus subtilis Natto), which had been usually used for food in the diet, proliferates more strongly in the same environment as compared with mold, and grows and propagates in molds. The inventors have found that spraying can suppress the generation and growth of mold, and completed the present invention.

The present invention relates to an antifungal agent containing natto bacteria as an active ingredient. Further, the present invention relates to an anti-fungal method comprising applying a suspension of natto bacteria to a place where mold may be generated or propagated or where mold is generated or propagated. In the present invention, the background of focusing on the natto bacterium is that no mold has been generated in the natto manufacturing plant since ancient times. Focusing on such a background, the present inventor considered that natto bacteria may have an action of destroying the mold survival environment, and examined the mold growth inhibitory action of natto bacteria.

The present invention was tested as follows.

[Test example]

(1) Separated culture of mold 1) Peeled potatoes cut into 1 cm pieces 200
g was put in 1000 ml of distilled water, boiled in boiling water for about 20 minutes, and potatoes were removed with gauze. Then glucose 20
g and agar 15g, then add distilled water and re-add 100
0 ml was used to prepare a potato-glucose agar medium (culture medium (A)) used for fungal separation culture. 2) Separation of test mold Black mold was collected from a bathroom of a general household, and this was cultured for 1 week at 25 ° C using the culture medium (A) as a separation medium. In order to further isolate the isolated strain into a single strain, the culture was repeated several times by the same method to perform pure isolation. The isolate showed good growth on culture medium (A) (see Figure 1). 3) Identification of the separated molds The culture medium (A) is smeared with the test molds and cultured for 3 days at 25 ° C, observed under an optical microscope, and as a result of a morphological search, it belongs to the genus Cladosporium. It was identified as mold (see FIGS. 2 and 3). This mold is
It is a typical mold that breeds in bathrooms, kitchens, closets, shoe boxes, etc.

(2) Culture of Natto Fungus Meat extract 5 g, peptone 10 g, NaCl 5 g, agar 1
5 g was dissolved in 1000 ml of distilled water and sterilized by heating to prepare an agar medium (culture medium (B)) used for culturing natto. The pH was 7.0.

(3) Antifungal Proliferation Inhibitory Effect of Natto Fungus 1) Co-Cultivation of Fungus and Natto Fungus with Culture Medium (A) A plate mold (A) is smeared with a test fungus, and a culture solution of Natto fungus is applied from above. Drop a small amount in 4 places, 25 ℃, humidity 7
The culture was performed at 0% for 1 week and at 18 ° C. and 70% humidity for 1 week. After 1 week, the state of proliferation was observed. The results are shown in FIGS. 4 and 5 (photograph). FIG. 4 shows the culture at 25 ° C., and FIG. 5 shows the culture at 18 ° C. It can be seen that in any case, the mold growth is extremely small in the white-growing region of the natto fungus. On the other hand, in the case of culturing at a low temperature, since the growth conditions of natto were bad, the growth of mold was observed on the entire surface of the plate, and the effect of natto to suppress the growth of mold was not sufficient.

2) Co-cultivation of mold and natto fungus with culture medium (B) Since the culture medium (A) was suitable for mold, the culture medium (B) suitable for natto was used for the same culture as in 1) above. When the test was carried out, the mold growth inhibitory effect was sufficient even at a low temperature of 18 ° C.

3) Mixed culture of mold and natto A mixed medium containing 50% each of the culture medium (A) and the culture medium (B) was prepared to prepare a plate medium (C). Next, 50% of both fungi and natto bacteria were mixed and smeared on the bacteria. It was cultured at 37 ° C. and 70% humidity and at 25 ° C. and 70% humidity for 1 week each. The state of proliferation after 1 week is shown in FIG. 6 (photograph). FIGS. 6 (a) and 6 (b) are comparative photographs of the one cultured at 37 ° C. (a) and the one cultured at 25 ° C. (b). At 37 ° C, the growth of mold is completely suppressed. In the case of 25 ° C., although mold growth is partially observed, it can be clearly seen from comparison with FIG. 4 that the natto bacteria suppress mold growth.

From the above-mentioned test example, it was revealed that the growth condition of natto is relatively high temperature and humidity, but this condition is similar to the environment in summer season, in which mold is easily generated in Japan. It clearly shows that natto can be used for the development of mold. Moreover,
By utilizing natto bacteria as a fungicide, it is possible to prevent the harmful effects of chemicals on the human body.

As can be seen from the above-mentioned test results, the present inventor has a strong effect of preventing natto fungus from developing and reproducing, and the present invention was made based on this finding. Especially under high temperature and high humidity conditions of 25 ° C or higher, the effect of inhibiting the growth and reproduction of mold is strong.
It is possible to prevent the growth and growth of mold in kitchens, closets, shoe boxes and the like.

The natto bacterium in the present invention is commercially available and can be easily obtained by a third party. This commercially available natto bacterium may be suspended in water before use. Natto bacteria number 10 ⁸ to 10
It is preferable to use ⁹ pieces / ml. However, this concentration can be appropriately changed in consideration of various conditions such as the mold growth condition, season, and growth period. As the diluting water, distilled water, well water, tap water, a buffer solution, or other appropriate water can be used.

The dispersion of the natto fungus dispersion liquid according to the present invention is applied to the above-mentioned dispersion liquid in an amount of 10 to 20 ml per time at a place where the mold may be generated or propagated, or at a place where the mold is generated or propagated. / M ² spray. The spraying may be carried out once or several times. By leaving it as it is, it is possible to prevent the growth and reproduction of mold. Also,
The effect can be further enhanced by removing the mold beforehand by a simple means at the time of spraying, and the spraying also includes means for applying the natto bacteria dispersion liquid.

Next, the present invention will be specifically described with reference to examples.

[Examples] 10 g of a commercially available natto bacterium (Sawamura Chemical Co., Ltd.) was dissolved in 1 liter of tap water to give a natto bacterium concentration of 10 ⁸ / ml.
Was prepared. It is used as an anti-fungal agent for this, and it is used in areas with dark black mold in the bathroom.
10 to ²⁰ ml was sprayed per m ² . After spraying, it was left for about 2 to 3 days and then washed with water. This application was repeated twice.
The results were as follows. In addition, as a control, the one sprayed only with tap water and the one using a chlorine-based antifungal agent were used. The effect is shown in Table 1.

[0019]

[Table 1]

[0020]

EFFECTS OF THE INVENTION Since the antifungal agent of the present invention contains natto bacteria as an active ingredient, it is harmless to humans and can safely exhibit antifungal effects without being damaged by misuse. In particular, since natto fungus grows at high temperature and high humidity, its property is effective for the prevention of fungus such as household baths, kitchens, indentations where molds are easily generated in summer, shoe boxes and the like.

[Brief description of drawings]

FIG. 1 shows a black mold isolated from a bathroom used in the present invention.

[Fig. 2] The above black mold [Cladosporiu
m)] is a micrograph (magnification of FIG. 2 is 200 times, FIG.
Is 600 times)

[Fig. 3] The black mold [Cladosporiu
m)] is a micrograph (magnification of FIG. 2 is 200 times, FIG.
Is 600 times)

[Fig. 4] Fig. 4 shows the antifungal effect of natto bacteria in the above black mold culture medium (A). Culture at 18 ° C is shown.

FIG. 5 shows the antifungal effect of natto bacteria in the above black mold culture medium (A). Culture at 25 ° C. is shown.

FIG. 6 shows a state in which a natto bacterium and a black mold are symbioticly cultured in a mixed medium of the culture medium (A) and the culture medium (B). (A) shows cultivation at 25 ° C, and (b) shows cultivation at 37 ° C.

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[Procedure amendment]

[Submission date] September 7, 1995

[Procedure Amendment 1]

[Document name to be amended] Statement

[Name of item to be corrected] Figure 1

[Correction method] Change

[Correction content]

[Figure 1] The present invention of black mold that was away from the bathroom, which was used single in
The morphology is shown.

[Procedure Amendment 2]

[Document name to be amended] Statement

[Name of item to be corrected] Fig. 4

[Correction method] Change

[Correction content]

[4] form of Natto in culture medium of the black mold (A)
State . Culture at 18 ° C is shown.

[Procedure 3]

[Document name to be amended] Statement

[Name of item to be corrected] Figure 5

[Correction method] Change

[Correction content]

[5] the form of Natto in culture medium of the black mold (A)
State . Culture at 25 ° C. is shown.

[Procedure amendment 4]

[Document name to be amended] Statement

[Name of item to be corrected] Figure 6

[Correction method] Change

[Correction content]

FIG. 6 shows the morphology of natto and black mold co-cultivated in a mixed medium of the culture medium (A) and the culture medium (B).
(A) shows cultivation at 25 ° C, and (b) shows cultivation at 37 ° C.

Claims (3)

[Claims] 1. Bacillus subtilis Natt
Antifungal agent containing o) as an active ingredient. 2. A method of preventing rust, which comprises spraying a suspension of natto fungi on a place where mold generation and reproduction are likely to occur or where they are occurring. 3. The method according to claim 2, wherein the place where mold development and reproduction may occur or is occurring is a bathroom, kitchen, closet or shoe box.