JPH06104075B2 - Method for analyzing properties of mold - Google Patents

Description

Detailed Description of the Invention [Industrial applications]

The present invention relates to a method and apparatus for analyzing the properties of mold and the like.

[Prior art]

Conventionally, the following method has been used for observing the activity or inactivity of fungal hyphae of mold, analysis for identifying mold, and the like. (1) First, the medium is placed in a container such as a dish and the mold is planted on the surface of the medium. At this time, a chemical substance is allowed to coexist if necessary. (2) By observing the growth degree of colonies, which are aggregates of hyphae, by culturing the molds while keeping them at a constant temperature and humidity, the properties of the molds are analyzed, the mold-killing properties of the chemical substances, and the anti-molding properties of the molds. Make a judgment or identify mold. here,
The growth degree of colonies is judged mainly based on the radius and the number of colonies. To judge by these criteria, it is necessary to grow the colonies to a certain extent. It takes days.

[Problems to be Solved by the Invention]

As described above, in the colony observation method, since it takes a long time to culture the mold, the following points are problems. That is, rapid analysis is not possible, and it is not suitable for a screening test that requires simultaneous measurement of the fungicidal or antifungal properties of many specimens or many chemical substances. In addition, the concentration and composition of chemical substances often change from the start to the end of the test, and it is difficult to keep the test conditions constant. Furthermore, we have observed only the average macroscopic changes of the mycelium as an aggregate, and no information on the activity or inactivity of individual hyphae is obtained, and as a result, the growth of mold and the effect of chemical substances on the growth It is difficult to obtain knowledge about the mechanism of. In view of the problems of the conventional ones, the present invention observes the growth rate and morphological change of one or a specific number of hyphae, not the aggregate of hyphae as in the colony observation method, and thus the properties of mold, etc. It is an object of the present invention to provide a method and an apparatus capable of easily and quickly analyzing.

[Means for Solving the Problems]

In order to achieve the object, the present invention sequentially exposes a target mold to air and then to air containing a vapor of a drug, a drug, or the like, and selects one or a mold of the target mold under the above-mentioned different environments. Cultivate a number of mycelia and observe the growth rate and morphological changes at the same site in the mycelium using visible light or infrared light.
A method for analyzing the properties of mold, which is characterized by obtaining information on the activity or inactivity of the mold by analysis, is provided. At this time, it is possible to determine the antifungal property or antifungal property of the chemical substance in the drug. Further, it is considered that the information on the growth rate and the morphological change of the hyphae is stored and the fungal hyphae to be compared and the known hyphae of the fungus are compared and observed to identify the fungus. Further, it is conceivable to measure the environmental conditions in the room by sequentially exposing specific hyphae to the outside air, then to the room air, and then to the outside air and comparing each growth rate and morphological change. Further, the present invention is characterized by comprising a mold culture cell and an image processing computer for processing an image signal captured by an optical microscope and an image pickup device for observing mold hyphae in the mold culture cell. It also provides a device for analyzing the properties of mold. At this time, it is considered that the mold culture cell is equipped with a drug control device for supplying air, saturated vapor of the drug, or air containing the vapor of the drug. Further, it is considered that the mold culture cell is equipped with a temperature control device.
Further, it is conceivable that the image processing computer is additionally provided with a memory for storing the data of the processed image and a monitor for observing the processed image.

[Action]

In the present invention, by paying attention to the same mycelium, measuring the growth rate thereof and observing the morphological change thereof, the properties of the mold can be easily and rapidly analyzed. Furthermore, when the present invention is used, the fungicidal and antifungal properties of chemical substances can be detected easily, rapidly and with high sensitivity, as compared with conventional methods.

【Example】

Next, the present invention will be described in detail with reference to the drawings. FIG. 1 (A) shows an example of an apparatus for carrying out the method for analyzing the properties of mold according to the present invention. In this figure, 1 is a mold culturing cell, 2 is an optical microscope, 3 is an image sensor, and the mycelium cultured in the mold culturing cell 1 is observed by the optical microscope 2, and the observation signal is sent by the image sensor 3. It is supposed to be done. Reference numeral 4 denotes an image processing computer, and this image processing computer 4 processes the image signal captured by the image pickup device 3.
A video deck 5 and a monitor 6 are connected to the image processing computer 4. The mold cell 1 has a temperature control chamber 7
Inside the temperature control chamber 7, a temperature control device 8 is provided. Reference numeral 9 is a drug control device for mixing a volatile fungicide or antifungal agent with air and circulating the mixture in the mold culturing cell 1. That is, from the medicine supply port a, the medicine fixed quantity supply pump 10, the solenoid valve
11, chemical flow meter 12, saturated vapor generation pipe 13 and admixture adjustment valve 14
A chemical supply path 16 is formed from the air supply port b to the admixture container 15, and an air constant amount supply pump 17 and a solenoid valve 1 are provided.
8. An air supply path 20 is formed to reach the mixing container 15 through the air flow meter 19 and the mixing adjustment valve 14. The mixing container 15 and the mold culturing cell 1 are connected by a supply pipe 21, and a flow path is formed from the mold culturing cell 1 to the outside air via the exhaust pipe 22. FIG. 1 (B) shows an example of the structure of the mold culturing cell 1. In the figure, 31 is a culture sem body, 32 is a medium, 33 is a mold,
34 is a holder, 35 is a chemical vapor inlet, and 36 is a chemical vapor outlet. In the apparatus configured as described above, the analysis of the activity or inactivity of the mold 33 is performed by introducing air into the mold culture cell 1 and then introducing air containing the vapor of the drug or the vapor of the drug into the mold culture cell 1. The growth state of books or a specific number of hyphae is observed by the optical microscope 2, and the growth rate and morphological changes of hyphae are processed by the image processing computer 4 and then observed by the monitor 6. Note that this state is recorded by the video deck 5. FIG. 2 shows an example of analysis of the fungicidal characteristics using the above device. Here, Aspergillus niger was selected as the fungus body of mold 33, and cineole was used as the drug.
In the state in which pure air is circulated in the mold culture cell 1, as shown in the range from 0, mycelia grow linearly in sequence, but when saturated vapor of cineole is introduced, The mycelium stops growing, as indicated by the range. Then, even if pure air is circulated thereafter, the hyphae are not activated again as shown in the range of 1, and it is analyzed that the cineole has excellent fungicidal activity. FIG. 3 likewise shows the antifungal property when ethyl salicylate is used as a drug. In this illustrated example, when the mold culture cell 1 is filled with saturated vapor of ethyl salicylate, the growth of mycelium stops. However, unlike the case of cineole, when ethyl salicylate vapor is removed, the mycelium begins to grow after a few hours. As is clear from this characteristic, it is revealed that ethyl salicylate has an antifungal property against ampergiris niger but not a fungicide. FIG. 4 shows the characteristics when isoamyl salicylate was used as a drug. In this case, even if isoamyl salicylate was introduced into the mold culture cell 1, the growth rate of mycelium was slightly smaller than that before introduction, and never reached zero in some cases. This indicates that isoamyl salicylate does not show very effective antifungal properties. Fig. 5 shows an example of the fungicidal and antifungal properties of natural substances. In this example, finely chopped onion is filled in a saturated steam generating tube 13 to increase the growth rate of Rhizoptonia solani. The anti-mildew effect of onion aroma is shown. In the above analysis example, the morphology of mycelium also shows changes peculiar to each drug. That is, when mycelia were exposed to cineole vapor, the intracellular protoplasm appeared as if it had gelled, and the cracking phenomenon of the current trait was observed. In addition, when exposed to ethyl salicylate vapor, the current traits appear to fly out in a spherical shape from the tip of the hyphae. Was seen. No significant changes were observed with isoamyl salicylate. As is clear from these results, by observing the growth rate of the same part of the same hypha and observing the morphological change, the effectiveness of the drug as a fungicidal and antifungal property can be accurately determined. can do. Further, according to this method, it is possible to perform the observation in an extremely short time (within 10 hours) as compared with the conventional colony observation method. Therefore, for example, even with isoamyl salicylate, which is said to be ineffective in the colony observation method, it is possible to determine the antifungal effect of the drug with high sensitivity so that the antifungal effect can be observed to some extent. In the above analysis example, a vaporizable chemical substance was used as a fungicide and an antifungal agent, but this was used to determine the fungicidal property and antifungal property of a chemical substance of catalytic type and other modes of action. Is also effective.

[Modification]

Another embodiment of the present invention will be described below. [Modification 1] The present invention is also effective for determining the fungicidal and antifungal properties of natural substances. The effects of the invention will be explained by taking the scent component emitted from the onion. The system used is the same as in FIG. The finely chopped onion is put in the saturated steam generating tube 13 shown in FIG.
Figure 5 shows the effect of onion aroma components on the growth rate of Rhizoptonia solani. Initially, hyphae grew linearly in pure air environment. Next, when the aroma component of onion was introduced into the culture cell, the growth of hypha stopped. When pure air was introduced again, the hypha started to grow. The effect of onion on Rhizoptonia solani was examined by the colony observation method.
Mycelia were planted on the medium put in a petri dish, and a chopped onion was attached to the lid of the petri dish so that the aroma component of the onion was transferred to the mycelia. 30 this petri dish for 5 days
No growth of colonies was observed when kept at ℃. Next, the minced onion was removed and cultured for another 5 days at 30 ° C., and growth of colonies was observed. As is clear from the above, the results of the method of the present invention and the colony observing method are in agreement for the fragrance component of onion. That is, using the present invention, the fungi killing of natural substances such as onions
It becomes possible to detect the antifungal property. [Modification 2] The present invention is not limited to the detection of antifungal and antifungal characteristics of chemical substances,
It is also possible to identify the mold. The identification of mold is based on the morphology of spores in addition to the morphology of hyphae, and information on the morphology of spores is especially necessary for the identification. Spores are not always formed in the medium in which the hyphae grow. If spores do not form, try changing the medium. Examples of the mold medium include Gabro Agar medium, Waxman agar medium, potato-glucose agar medium, chlorotiphenier and Glucose peptone agar medium. Therefore, the number of culture cells is the number of required media. FIG. 6 illustrates a device for identifying the mold 33. The basic configuration of this device is the same as that shown in FIG. 1, except that the mold culture cell 1 is arranged in a constant temperature and humidity chamber 23, and a medium moving device 24 inside the same is provided. The image processing computer 4 is equipped with a memory 25. In the figure, 30 is a rotary stage. In this device, in order to identify the mold 33, each medium 32 is exposed in synchronization with the synchronization signal for positioning by the medium moving device 24.
The memory 25 stores the change of the mycelium in. And
The changes in the hyphae of each substratum stored in the memory 25 are frame-fed and displayed on the monitor 6, and the observer visually compares the spore morphology with the morphology of the spores already identified and fixes the unknown mold. To do. Further, in the case of automatically identifying the mold 33, the operation of the flowchart shown in FIG. 7 is performed. That is, first, the culture medium 32 in FIG. 1 (B) is positioned so that the image and the culture medium 32 correspond to each other, and a specific mycelium is brought into focus by fine adjustment of the holder 34. Next, the hyphae are observed K ₀ times for each medium 32 for a certain period of time T ₀ , and the area of the hyphae is calculated. When the area of the hyphae is calculated each time, the time differentiation of the area is taken to calculate the growth rate of the hyphae. Then, when the hyphae are growing, they are continuously observed until the formation of spores. When the growth rate is zero, the next mold 33 of the medium 32 is observed. When such a cycle is performed M ₀ times, it ends, but if an image of spores is detected during this time, it is compared with the image of the already identified mold 33 stored in the memory 25, and the mold 33 Identification is performed. [Modification 3] By the way, in a precision products factory such as a semiconductor manufacturing factory or in a food factory where it is necessary to maintain high hygiene, contamination by microorganisms such as mold in the room is a problem. In many cases, fungicides and antifungal agents are used. When using the chemicals, after sampling the microorganisms floating in the air or the microorganisms adhering to the wall or floor, culturing them on a medium, the effectiveness of the chemical substances used depending on the degree of colony growth I confirmed the sex. These methods have problems that they are limited to the judgment of the contamination status of the building and that it takes time to culture the bacteria. The foodstuffs that are stored or stored have some bacteria or their spores attached to them. Therefore, it is important to quickly detect whether the fungus or its spores are in storage or in a growing environment within the building during storage. The present invention is also effective as an environment measuring device from the viewpoint of mycelial growth. The present invention is also suitable for rapidly determining the above environment in a building. That is, FIG. 8 shows a device therefor. The basic configuration of this device is almost the same as that shown in FIG. 1, but an air suction pump 26 and a solenoid valve 27 in the building are provided to introduce the air inside and outside the building into the mold culturing cell 1, respectively. Further, the air suction pump 28 and the solenoid valve 29 outside the building are provided. A signal indicating whether the air inside the building is being sucked or the air outside the building is being sucked is input to the image processing computer 4. The mold culture cell 1 is movable by a rotating stage 30. The image processing computer 4 also has a memory 25 in this apparatus. With this device, a mold in the target building or a specific mold that propagates in the target food is selected and planted in the medium in the mold culture cell 1. Then, first, the growth rate and the morphological change are observed by focusing on one hyphae under the environmental conditions of the outside air in which the air outside the building is introduced. Then, the air in the building was introduced to observe the growth rate and morphological deformation of the same part in the same hyphae mentioned above, and the environment in the building was judged by comparing the observation results of both and the effect in this environment It is possible to select a chemical substance having a typical antifungal property and an antifungal property, and to promote its proper use.

【The invention's effect】

Therefore, according to the present invention, it is possible to observe the growth rate and morphological change of one or a specific number of mycelia, which is not an aggregate of hyphae as in the conventional colony observation method, and the properties of mold can be easily evaluated. It has an effect that analysis can be performed quickly and with high sensitivity.

[Brief description of drawings]

The drawings show an embodiment of the present invention, FIG. 1 (A) is a configuration diagram showing an example of the apparatus, and FIG. 1 (B) is a perspective view showing an example of the configuration of a mold culture cell, FIG. 2 and FIG. , FIG. 4 and FIG. 5 are characteristic diagrams showing analysis results respectively, FIG. 6 is a configuration diagram of an apparatus showing another embodiment, FIG. 7 is a flow chart showing the operation mode of the same as above, and FIG. It is a block diagram of the apparatus which shows the Example. In the figure, 1 is a mold culture cell, 2 is an optical microscope, 3 is an image sensor, 4 is an image processing computer, 5 is a video deck, and 6
Is a monitor, 7 is a temperature control room, 8 is a temperature control device, 9 is a drug control device, 10 is a drug fixed amount supply pump, 13 is a saturated vapor generation pipe, 15 is a mixing container, 17 is an air fixed amount supply pump, and 23 is a constant temperature. Humidity tank, 24 medium moving device, 25 memory, 26 air suction pump inside building, 28 air suction pump outside building, 30 rotation stage, 31 culture cell body, 32 culture medium, 33 medium Mold, 34 is a holder, 35 is a chemical vapor inlet, and 36 is a chemical vapor outlet.

Claims (4)

[Claims] 1. A target mold is sequentially exposed to air and then to air containing a vapor of a drug having a fungicidal / antifungal property or a drug composed of a substance having a fungicidal / antifungal property. , Culturing one or a specific number of mycelia of the target mold under the above different environments and observing / analyzing the growth rate and morphological change of the same site in the mycelium using visible light or infrared light A method for analyzing the properties of mold, characterized by obtaining information on the activity or inactivity of the target mold. 2. The method for analyzing the properties of mold and the like according to claim 1, wherein the mold-killing property or anti-mold property of the chemical substance in the drug is determined. 3. A method for identifying a mold by storing information on the growth rate and morphological change of the mycelium, and comparing and observing the mold mycelia to be compared with the known mold mycelia. Method for analyzing the properties of the described mold, etc. 4. The indoor environmental conditions are measured by sequentially exposing specific hyphae to the outside air, then indoor air, and then to the outside air and comparing each growth rate and morphological change. Method for analyzing properties of mold, etc.