JPH09289889A - How to store microorganisms - Google Patents

Abstract

(57) Abstract: [PROBLEMS] To provide a method for preserving microorganisms in an active state, which are excellent in initial start-up ability and can be used in industrial fields such as wastewater treatment and food waste treatment. SOLUTION: After supporting microorganisms on a wood strip in which the cross section of the conduit or the temporary conduit is not substantially destroyed on the wood mouth surface, the water content of the wood strip is adjusted to 5% or more and the fiber saturation point or less. A method for preserving microorganisms which is characterized by maintaining.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for preserving microorganisms.

[0002]

2. Description of the Related Art Conventionally, there are known methods for preserving various kinds of microorganisms using a protective substance such as skim milk or glutamic acid or an inorganic substance such as talc as a carrier.

[0003]

However, the microorganisms preserved by these methods for preserving microorganisms are extremely inferior in the initial rising ability and cannot be used as they are. Therefore, it is necessary to perform preincubation in order to bring the microorganism into an active state.

[0004]

In view of the above situation, the present inventors have conducted intensive studies to find a better method for preserving microorganisms, and arrived at the present invention. The present invention is as follows: 1) After supporting microorganisms on wood chips whose cross section of the conduit or the temporary conduit is not substantially destroyed on the wood mouth surface, the water content of the wood chips is 5% or more at the fiber saturation point. A method for preserving microorganisms characterized by maintaining the following (hereinafter referred to as the method of the present invention), 2) A wood strip in which the cross section of the conduit or the temporary conduit is not substantially destroyed at the mouth end thereof. A method for preserving microorganisms, characterized in that, after supporting the microorganisms, the water content of the wood chips is maintained at 10% or more and 20% or less, 3) The cross section appearing at the mouth of the temporary conduit is substantially destroyed. The present invention provides a method for preserving microorganisms, which comprises maintaining a water content of 5% or more and a fiber saturation point or less after supporting microorganisms on wood chips derived from non-coniferous trees.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION As the microorganism used in the present invention, for example, when it is used for wastewater treatment or food waste treatment, plural kinds of aerobic microorganisms obtained from soil are suitable. If necessary, facultative anaerobic microorganisms or anaerobic microorganisms can be used together with the above microorganisms. In addition, when these microorganisms are used as a group of microorganisms obtained by proliferating and culturing under artificial environmental conditions, they exhibit excellent processing ability that appropriately responds to changes in various types of objects to be treated, and that ability is maintained. Will be possible. These microorganisms are, for example, soil collected from forests, fields, seafood processing plants, pig houses, cow houses, poultry houses, fishing ports, fish markets, fruit orchards, fruit and vegetable markets, manure treatment plants, food processing plants, chemical plants, etc. , Microorganisms collected by conventional methods from soil where many microbial groups continue to decompose organic matter for many years, for example, liquid culture using nutrients of the same quality as the treated material under aeration conditions, Proliferation culture under artificial environmental conditions such as solid or solid culture, or
Each microorganism can be selectively grown and cultured, and efficiently prepared. Furthermore, among the microbial group, thermophilic bacteria (5
Existence rate of those that can reproduce in the temperature range of 5 ° C or higher is 3
It is preferable to adjust the ratio to be not more than.

The microbial carrier used in the present invention is a woody piece (hereinafter referred to as a woody piece) in which the cross section of the conduit or the temporary conduit is not substantially destroyed on the wood mouth surface.

Here, the wood strip "in which the cross section of the conduit or the temporary conduit is not substantially destroyed at the mouth end" means
For example, it means that the surface of the wood strip is in a state as shown in FIG. 1, and the liquid is relatively easily introduced without substantially closing the cross section appearing at the mouth of the conduit or the temporary conduit. It has a function that can. The size of each cross section that appears on the ostium of the conduit or the temporary conduit is, for example, about 10 μm to 500 μm in diameter, preferably about 10 μm.
Those having a thickness of about 100 μm are suitable. The specific gravity of the wood chips may be about 0.3 to 0.7 (g / cm ³ ), and preferably about 0.3 to 0.5 (g / cm ³ ) for better effect. Can be obtained. The wood chips are produced by cutting wood with a cutting machine having a plurality of blades so as to have a grain size of about 0.2 mm to about 10 mm so that the cross section appearing on the mouth of the conduit or the temporary conduit is not substantially destroyed. It may be produced by any method as long as it is manufactured. For example, wood made of a wood part made by sawing trees such as cedar, hiba, oak, etc. A suitable manufacturing method is a method of cutting at a high speed of about 1800 rpm to about 2000 rpm using a circular saw having a plurality of blades set in a range of about 60 °. Incidentally, as a particularly preferable tree species of the wood strip, there can be mentioned, for example, a conifer, which is a tissue (temporary conduit) in which most of the woody part has a function of allowing liquid to pass relatively easily.

In order to support microorganisms on the above-mentioned wood chips, for example, about 10 L to about 100 L of wood chips impregnated with a sufficient amount of water in advance are added with a liquid containing microorganisms at a ratio of about 1 L. Add and mix well. If necessary, a nutrient source for microorganisms may be added. As a nutrient source of microorganisms, those normally used for culturing microorganisms, for example, rice bran, bran, oil cake, fish meal, fish cake,
Bone meal, beer dregs, soy sauce dregs, yeast extract, peptone,
Amino acids, vitamins, sugars such as glucose and molasses,
Alternatively, fertilizer components such as ammonium sulfate, urea, lime superphosphate, molten phosphorus, potassium chloride, liquid fertilizer, compound fertilizer and the like can be used. After mixing, the mixture is deposited and left for about 3 days to about 1 month to grow the microorganisms. During this time,
The center temperature of the deposit is, for example, about 30 ° C. to about 50 ° C.
It is advisable to carry out the stacking (returning) several times so as to maintain the temperature at about 30 ° C., preferably about 30 ° C. to about 40 ° C. The degree of growth of the microorganisms is judged by the temperature of the central part of the deposit, and it may be terminated when the degree of heat generation becomes low. In this way, the microorganisms can be carried on the wood chips by sufficiently propagating the microorganisms in the pores of the wood chips.

Next, water is removed from the wood chips supporting the microorganisms, and the water content of the wood chips is adjusted to be in the range of 5% or more and the fiber saturation point or less, and then maintained within the range. To do. Preferably, the water content of the wood chips is 10% or more 2
It is better to keep it at 0% or less. Here, the water content means the weight% of water with respect to the dry weight of the wood chips. The water content can be measured using an electric resistance type moisture meter. As a method of removing water from the wood chips supporting microorganisms, for example, air drying, heating, a method utilizing centrifugal force,
And the method etc. which combined them can be mentioned.

The microorganism preserved by the method of the present invention (hereinafter referred to as the present microorganism) has an excellent initial rising ability and can be used, for example, in the industrial field of wastewater treatment as described below. For example, the wastewater can be treated by contacting the wastewater with the microorganisms stored together as described above (hereinafter referred to as the present microbial preparation together with the wood pieces carrying the microorganisms) under aeration conditions. Specifically, for example, the capacity is about 60 m ³ to about 1000 m ³ ,
Moreover, about 15 L to about 80 L, preferably about 40 L to about 60 L is filled per 100 L volume as the treatment medium in the treatment tank having an aeration means. About 3 to about 7 parts by weight of waste water (for example, BOD: 1000 mg / L to 2000 parts) is added to about 1 part by weight of the processing medium in this processing tank.
(about mg / L) and let it stay for about 12 hours to about 2 days. The treated wastewater is removed from the drain of the treatment tank. The application rate of this microbial preparation varies depending on various conditions such as the activity strength of the microorganism used, the type of wastewater, the treatment temperature, the treatment time, etc., and can be appropriately increased or decreased without regard to the above range. it can.

[0011] Examples of the use of the present microbial preparation include industrial use such as treatment for reducing the volume of food waste, treatment for composting food waste, and treatment for preventing malodor, and promotion of plant growth. Treatment, soil improvement treatment,
It can also be used in the agricultural field such as treatment for controlling harmful microflora.

[0012] Specifically, in the case of treatment for reducing the volume of raw garbage, for example, the present microbial preparation may be applied to about 3 kg to about 10 kg per 1 kg of raw garbage.

[0013]

EXAMPLES The present invention will be described in more detail with reference to Reference Production Examples, Examples and Test Examples, but the present invention is not limited to these Reference Production Examples, Examples and Test Examples.

Reference Production Example (Preparation of Wood Piece) A wood piece in which the cross section of the conduit or the temporary conduit is not substantially destroyed at the mouth end of the wood is a wood obtained by cutting a cedar log. Use a circular saw with 30 blades set at 60 ° with respect to the axis of the original tree to rotate at high speed (200
It was manufactured by cutting at 0 revolutions / minute.
Sampling a part of the wood pieces produced in this way and observing the surface of the sampling ostium with an inverted optical microscope revealed that the majority of the cross-section appeared on the ostium of the temporary conduit. It was confirmed that the wood pieces were not destroyed. Furthermore, when observing some of the samples with an electron microscope, it was found that the wood chips were in an orderly and closely aligned state as shown in Fig. 1 without substantially destroying the cross-section appearing at the mouth of the temporary conduit. Existed.
The size of each cross-section appearing at the mouth of the temporary conduit is about 20 μm to 50 μm in diameter, and the specific gravity of the wood strip is about 0.35 to 0.40 (g / cm ³ ). It was

Example (Preservation of microorganisms by the method of the present invention) 20 L of soil collected from a waste treatment plant that decomposed organic substances for many years was sieved to 10 mesh or less.
Next, about 785 L of the classified soil, 4 kg of a nutrient source consisting of rice bran, fish meal and steamed bone meal, 100 L of the wood strip obtained in Production Example 1 impregnated with a sufficient amount of well water in advance, and 150 L of well water It was added to a container (diameter 1 m, depth 1 m) and mixed well. The mixture is heated from 15 ° C to 25
The microorganisms were propagated by leaving it in the atmosphere in the range of ° C. During the standing, 60 L / min of air was blown into the mixture. After standing for 3 days, only the liquid was selectively collected from the outlet at the bottom of the container. Next, to a wood strip 1000 L obtained in Production Example 1 in which a sufficient amount of well water was impregnated beforehand, 30 L of the liquid (containing microorganisms) recovered above and a nutrient source consisting of bran, fish meal and sugar 1
2 kg was added and mixed well. After mixing, the mixture was deposited and left for 3 days to grow the microorganisms. During this time, the deposits (returns) were changed to 5 so that the center temperature of the deposit was maintained at about 30 ° C to about 40 ° C.
Performed times. After that, the degree of heat generation in the deposit decreased, and when the temperature of the central portion of the deposit became stable in the range of about 30 ° C. to about 40 ° C., the standing was stopped. In this way, the microorganisms were sufficiently propagated in the pores of the wood chips. Next,
The obtained wood chips were transferred to a stirring and mixing tank, and while being slowly stirred, dry air was blown thereto, and dried at room temperature for 48 hours. After the completion of drying, the water content of the obtained wood chips was measured with an electric resistance moisture meter and found to be 15%. The wood chips thus obtained were stored in a constant temperature and humidity chamber for 20 days under cool and dark conditions. In addition, when the wood pieces stored were sampled and the water content thereof was measured using an electric resistance type moisture meter, it was maintained in the range of 10% to 20%.

Comparative Example 20 L of soil collected from a waste treatment plant that decomposed organic substances for many years was sieved to 10 mesh or less.
Next, about 785 L of the classified soil, 4 kg of a nutrient source consisting of rice bran, fish meal and steamed bone meal, 100 L of the wood strip obtained in Production Example 1 impregnated with a sufficient amount of well water in advance, and 150 L of well water It was added to a container (diameter 1 m, depth 1 m) and mixed well. The mixture is heated from 15 ° C to 25
The microorganisms were propagated by leaving it in the atmosphere in the range of ° C. During the standing, 60 L / min of air was blown into the mixture. After standing for 3 days, only the liquid was selectively collected from the outlet at the bottom of the container. Next, to a wood strip 1000 L obtained in Production Example 1 in which a sufficient amount of well water was impregnated beforehand, 30 L of the liquid (containing microorganisms) recovered above and a nutrient source consisting of bran, fish meal and sugar 1
2 kg was added and mixed well. After mixing, the mixture was deposited and left for 3 days to grow the microorganisms. During this time, the deposits (returns) were changed to 5 so that the center temperature of the deposit was maintained at about 30 ° C to about 40 ° C.
Performed times. After that, the degree of heat generation in the deposit decreased, and when the temperature of the central portion of the deposit became stable in the range of about 30 ° C. to about 40 ° C., the standing was stopped. In this way, the microorganisms were sufficiently propagated in the pores of the wood chips. Next,
The obtained wood chips were stored as they were in a thermo-hygrostat for 20 days under cool and dark conditions. In addition, when the wood particles in storage were sampled and the water content thereof was measured using an electric resistance type moisture meter, it was found to be maintained in the range of 50% to 60%.

Test Example 1 Microorganisms preserved according to the examples, that is, 6 L of wood chips carrying the microorganisms were put into a 40 L acrylic treatment tank together with 30 L of factory wastewater (BOD: 1000 mg / L), and the mixture was treated from the bottom of the treatment tank. Air was supplied at 30 L / min for 24 hours, and the treatment was performed at room temperature. When the liquid in the treatment tank was observed, the color of the wastewater began to decrease from the initial rising point and became thin in a short time. In addition, the odor was also reduced. After 24 hours, the BOD dropped to 20 mg / L and the odor disappeared. Next, after removing only the treated wastewater from the treatment tank, the same wastewater (BOD: 1000 mg /
(L) 30 L was put in, and air was blown from the lower part of the treatment tank at 30 L per minute for 24 hours in the same manner as above, and the treatment was carried out at room temperature. Further, the same operation was repeated 10 times every 24 hours, but no phenomenon such as bulking or rancidity was observed in the treatment tank.

Test Example 2 After preliminarily adding water to the microorganism preserved in Example, ie, 20 L of wood particles carrying the microorganism, this was equipped with a stirrer at the bottom, and the length was 60 cm, width 50 cm, depth 50 cm.
The container with a plastic lid was filled. Into this container, 1.0 kg of household garbage, which had been lightly drained, was charged every day and treated with stirring at a speed of 1 rotation per minute. When observing the garbage in the plastic container,
After 7 days from the start of the test, the amount of food waste was remarkably reduced and almost no residue was present. Almost no bad odor was observed.
Even after 6 months from the start of the test, there was almost no residue of food waste and the generation of foul odor was prevented.

Test Example 3 Microorganisms preserved according to the examples, that is, wood chips carrying the microorganisms, were stored for 3 months under cool and dark conditions while maintaining the water content in the range of 10% to 20%. did.
The water content during storage was monitored by periodically sampling the wood strips and measuring the water content using an electric resistance moisture meter. A test was conducted by using the wood strips stored for a long period of time in the same manner as in Test Example 2 and the food waste treatment capacity was confirmed. I was not able to admit.

Comparative Test Example Microorganisms preserved according to the comparative example, that is, wood chips carrying the microorganisms, had a water content of 50 for an additional 3 months.
It was stored under cool dark conditions while maintaining the range of 60% to 60%. The water content during storage was monitored by periodically sampling the wood strips and measuring the water content using an electric resistance moisture meter. A test was conducted by using the wood strips stored for a long period of time in the same manner as in Test Example 2 and the food waste treatment capacity was confirmed. Occurred.

[0021]

The microorganisms preserved by the method of the present invention are
It has an excellent initial start-up ability and is suitable for use in wastewater treatment and food waste treatment, for example.

[Brief description of drawings]

FIG. 1 is an electron micrograph of a wood piece (tree species: cedar) in which the cross section of the conduit or the temporary conduit is not substantially destroyed on the mouth side thereof.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Akihiko Mine 5-3-15 Yamatonishi, Kawanishi City, Hyogo Prefecture

Claims (3)

[Claims] 1. After supporting microorganisms on a woody piece whose cross section of a conduit or a temporary conduit is not substantially destroyed on its wood mouth surface, the woody piece has a water content of 5% or more and a fiber saturation point or less. A method for preserving microorganisms, which is characterized in that 2. The water content of the wood strip is increased to 10% or more and 20% or less after the microorganisms are loaded on the wood strip whose cross section of the conduit or the temporary conduit is not substantially destroyed on the wood mouth surface. A method for preserving microorganisms which is characterized by maintaining. 3. After supporting microorganisms on wood particles derived from coniferous wood whose cross-section appearing on the ostium of a temporary conduit is not substantially destroyed, the water content of the wood particles is reduced to 5% or more and below the fiber saturation point. A method for preserving microorganisms which is characterized by maintaining.