JP2011240328A - Method for treating object to be treated using microorganism - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a simple and effective decomposition method etc. of an object to be treated, such as sewage sludge, using microorganisms by a simple operation.SOLUTION: The method for treating an object to be treated includes: adding and mixing the object to be treated into a bacterial bed containing microorganisms belonging to the genus Bacillus, Ureibacillus and Brevibacillus; and decomposing organic matter contained in the object to be treated by the microorganisms. There is also provided a method for reducing the water content of the object to be treated using heat generated by the fermentation by the microorganisms in the bacterial bed.

Description

  The present invention relates to a method of decomposing (digesting and extinguishing) a processing object such as sewage sludge using microorganisms.

At present, many local governments incinerate sewage sludge discharged from final treatment plants. However, incineration treatment of organic waste containing sewage sludge has a large environmental load such as emission of global warming effect gas, and a review of the treatment method is required.

In recent years, treatment techniques using microorganisms have been researched and developed as an alternative to incineration of organic waste. According to this method, emission of global warming effect gas can be suppressed more than incineration with existing fossil fuels, so it is attracting attention as “organic waste treatment technology with the least burden on the global environment”. .

  In recent years, with the rapid progress of biotechnology, various attempts have been made to effectively use microalgae, and many techniques have been put to practical use. For example, one of the highly feasible biotechnology fields is the medical / health area of pharmaceuticals, physiologically active substances, and functional foods that aims to use functional substances produced by microalgae. The second area mainly uses microalgae itself, and is related to agriculture, water and environmental biotechnology for the purpose of herbivorous livestock and bivalve feed and environmental purification. Furthermore, the area | region of industrial biotechnology including biomass resources and biofuel which is going to utilize a micro algae as a means of industrial production can be mentioned.

In particular, the microalgae energy industry has a high carbon dioxide consumption by photosynthesis that microalgae are much larger than other plants, microalgae can be grown in seawater and sewage, and can be cultured in places where food is difficult to grow. Promote marginal land use, increase food production by utilizing microalgae after energy extraction, enable rapid diffusion without the need for large-scale investment in distribution network development, and the food industry It is attracting a great deal of attention because it can be expected to generate a wide range of job creation power.

JP 2007-44604 A JP 2008-36593 A JP 2008-126169 A JP 2009-125636 A

In Patent Document 1, the water to be treated containing an organic substance and a solid component is first separated and separated in a sedimentation basin, and biologically treated in an activated sludge treatment apparatus, and then subjected to solid-liquid separation in the sedimentation basin. In the activated sludge treatment method, the biochemical oxygen demand derived from solid components in the treated water out of the biochemical oxygen demand in the treated water flowing into the activated sludge treatment apparatus is the treated water. The activated sludge treatment method is characterized in that the water to be treated is caused to flow into the activated sludge treatment apparatus 2 in a state of 60% or more of the total biochemical oxygen demand.

In Patent Document 2, an effective microorganism group consisting of 55% aerobic bacteria group and 45% anaerobic bacteria group is mixed in okara, and the ratio of the number of mixed bacteria is 24 to 72 hours at a temperature of 40 ° C to 200 ° C. First time fermentation, and further mixed with an effective microorganism group consisting of 55% aerobic bacteria group and 45% anaerobic bacteria group, and 24 hours at a temperature of 40 ° C to 200 ° C. Fermentation material for sewage sludge treatment is produced by secondary fermentation for 48 hours, 1000 parts by weight of sewage sludge to be treated is put into the fermentor, and moisture content is added to this to make the moisture content 45 to 55%. Then, 1-2 parts by weight of the fermentation material is added and mixed, and then the temperature in the fermenter is heated to 60 ° C. to 200 ° C., air is injected into the fermenter, and the inside of the fermenter is continuous. Of sewage sludge characterized by fermenting sewage sludge by mechanical stirring The law has been described.

Patent Document 3 discloses a sewage treatment method in which wastewater is sent to a sewage treatment plant via a pipe culvert and a relay pump station, and has a function of decomposing organic matter in the effluent in the pipe culm. A microorganism preparation containing microorganisms is introduced from the relay pump station, and at least a part of the organic matter is decomposed by the microorganisms generated from the microorganism preparation in the pipe tube from the relay pump station to the sewage treatment plant. A sewage treatment method characterized in that is described. Examples of the microorganisms include gram-negative bacteria, or solid microbial preparations containing gram-positive and gram-negative bacteria, and preferable examples include the genus Bacillus, particularly Bacillus toyoi.

Patent Document 4 discloses a sludge treatment method that performs biological treatment with microorganisms such as sewage treatment, human waste treatment, and residue treatment caused by a food processing process, and performs dehydration and drying treatment of sludge after biological treatment. A sludge treatment method is described in which a microorganism killing process for killing microorganisms is performed before the drying process.

The inventor of the present invention uses a microbial bed composed of a combination of specific microorganisms for sewage sludge discharged from a final treatment plant, and further, residues discharged from industrial sites such as food factories and garbage discharged from ordinary households. By adding and mixing, it was found that these can be easily decomposed and extinguished at an extremely high decomposition rate, and the present invention has been achieved.

Furthermore, the present inventors have found that the water content of the object (composition) to which the fungal bed is added and mixed can be significantly reduced by the heat generated by the fermentation by the microorganisms in the fungal bed. did.

The present invention adds a treatment object such as sewage sludge to a bed containing microorganisms belonging to the genus Bacillus, microorganisms belonging to the genus Ureibacillus, and microorganisms belonging to the genus Brevibacillus, and the organic matter contained in the treatment object The present invention relates to a treatment method for sewage sludge and the like, which consists of decomposition by microorganisms.

  Furthermore, the present invention also relates to a method of adding the above-mentioned fungal bed to the treatment object (composition) and mixing, and reducing the water content of the object by heat generated by fermentation by microorganisms in the fungus bed. It is.

According to the method of the present invention, it has been proved that almost 100% of organic substances contained in a processing object such as sewage sludge is decomposed by a very simple operation. Furthermore, the water content in the object to be treated can be significantly reduced, and it is effective in reducing the fossil fuel required for the evaporation process of the treated water.

As representative examples of treatment objects that can be treated by the method of the present invention, sewage sludge, residues discharged from industrial sites such as food factories, and more degradable organic substances such as garbage discharged from general households are collected. Various kinds of waste and waste can be raised.

  The sewage sludge is discharged from, for example, a terminal treatment plant of sewage, and usually contains various decomposable organic substances in addition to about 80 to 90% by weight of water, and has a pH of 7 to 9 The range of the degree.

  In addition, the water content of the object to be treated can be significantly reduced by the heat generated with the fermentation by the microorganisms in the bacterial bed in the method of the present invention. Therefore, as processing objects in this method, for example, natural resources (natural products), biomass, and cultures of various organisms such as microalgae have a high water content (for example, 50 wt% or more, 50 wt% to 98 wt%). %).

Microalgae (microalgae) are unicellular organisms, also called phytoplankton. It has chlorophyll (Chlorophyll), and fixes carbon dioxide (CO ₂ ) in the atmosphere by photosynthesis to produce oxygen (O ₂ ). Currently, it breeds not only in the ocean but also in freshwater systems (lakes, ponds, rivers), and there are 100,000 types. Representative species include, for example, chlorella, spirulina, Dunaliella and Euglena.

In recent years, microalgae have attracted rapid attention as an economically viable biomass that does not compete with food. For the growth and growth of microalgae, it basically requires carbon dioxide, minerals and light, and does not require starch. A method for procuring biofuel by culturing these microalgae has been considered. Biofuel obtained from microalgae is similar to diesel oil in terms of chemical structure, so it is considered that infrastructure such as refineries and storage facilities can be used from existing diesel. Biofuels made from such microalgae are also recently called photosynthetic biofuels or algal biofuels (algal biofuels) to distinguish them from biofuels made from vegetable oils and cellulose raw materials .

  Alternatively, a technique for utilizing the cultured microalgae itself as a fuel material for thermal power generation is being developed. Originally, the oil and fat components produced by microalgae that prospered in the ocean several hundred million years ago are used as oil.

Since microalgae and the like are cultured in an aqueous system, these cultures have a very high water content. Therefore, when these cultures are used as the above biofuels or fuel resources, in the production process thereof, for example, before the extraction process of active ingredients such as hydrocarbons produced by microalgae, etc. It is necessary to reduce the water content of According to the method of the present invention, it is possible to reduce the water content of the culture in such a production process by the heat generated with fermentation by microorganisms in the fungus bed. In this case, since microorganisms can be fermented using various components contained in the fungus bed as nutrient sources, the treatment object itself does not need to be used as a nutrient source by the microorganisms. Therefore, by appropriately adjusting the amount of fungal bed added in the method of the present invention, it is possible to substantially suppress the decomposition / disappearance of active ingredients contained in microalgae and the like. As a result, the utility of the present invention as a biofuel such as microalgae or a fuel resource is not impaired.

The fungus bed used in the method according to the present invention includes simple morphology observation of colony specimens obtained by bacterial isolation, analysis of 16S rDNA partial base sequences, and microorganisms contained in the specimens, as shown in the following examples. By PCR-DGGE (PCR Denaturing Gradinet Gel Electrophoresis) and RT-PCR-DGGE for communities, microorganisms belonging to the genus Bacillus such as Bacillus coagulans and Bacillus licheniformis, Ureibacillus suwonensis, or related species such as Ureibacillus genus It is characterized by containing microorganisms belonging to the genus Brevibacillus such as Brevibacillus thermoruber. All of these are resident bacteria, and the biosafety level (a) is level 1. “Anything that may cause disease in humans or veterinary diseases in animals (including opportunistic infections) It was judged as “Biosafety Guidelines for Japanese Bacteria Society”.

The fungus bed may further contain microorganisms belonging to the genus Tepidimicrobium and / or microorganisms belonging to the genus Pelotomaculum.

  In addition, the fungus bed is, for example, suitable materials such as rapeseed oil squeezed rice cake, charcoal, rice bran, bark, bacus, sugar condensate, and hay on the soil collected around the place where the method of the present invention is carried out Depending on the condition, it can be prepared by appropriately adding natural water such as tap water, spring water or seawater to the return fungus bed (part of the fungus bed previously used in the method of the present invention) as water for water adjustment. . Furthermore, when the decomposition treatment ability of the fungus bed is lowered, the ability can be recovered by adding the above materials as appropriate.

The fungus bed used in the method of the present invention may contain other microorganisms derived from the soil used for the production. Moreover, the content rate of said various microorganisms contained in a microbial bed changes with various conditions, such as the kind of soil used for the production, and there is no restriction | limiting in particular.

The volume ratio between the bacterial bed and the added treatment object during treatment can be selected and maintained by a person skilled in the art depending on the purpose of the method, the type of the treatment object, the moisture content, etc. . For example, as a suitable range, the volume ratio between the fungus bed during treatment and the added treatment target can be 10: 1 to 60: 1, particularly 45: 1 to 55: 1.

  In general, the sewage sludge is divided into multiple times so that the volume ratio between the fungus bed and the added treatment target falls within the above range according to the purpose of the method of the present invention, the type of the treatment target, the moisture content, etc. Add to the bed at appropriate intervals, for example, every day to several days, mix well with the bed using an appropriate means such as a shovel loader, and leave it in the meantime. I can do it. Alternatively, it is also possible to always stir and mix the fungus bed and the object to be treated during the execution of the method of the present invention using any stirring device known to those skilled in the art.

Although depending on the type of treatment object and fungus bed, external environment, etc., the water content of the mixture of the fungus bed and treatment object in the method of the present invention is usually 50% by weight or more, for example, 50% to 75% by weight. The temperature (article temperature) of the mixture of the fungus bed and the treatment target is about 25 to 75 ° C. (measured at a depth of 30 cm from the surface), and the pH is 5.5 to 9.0.

In addition, in the method of the present invention, there is no particular need for deodorizing measures against fermentation odor discharged by fermentation by microorganisms or ammonia generated in an anaerobic state, but when deodorization is necessary, an appropriate deodorant (for example, Probiotics Water (PBW): Manufactured by Kanagawa Furniture Co., Ltd. (including lactic acid bacteria, Bacillus bacteria, and yeast) Propellants can be deodorized by spraying them on the bacterial bed or floor.

  The present invention will be described in detail with reference to the following examples. In addition, an Example is one aspect | mode of this invention, and the technical scope of this invention is not limited to description to an Example.

Demonstration test Experimental method Prepare 17.04 m ³ (ll, 155.2 kg), which is a habitat of the microbial waste decomposing bacteria group, and divide it into 8.52 m ³ (5, 577.6 kg). Put 0.15 m ³ / day of sewage sludge in Amami City every other day in the fungus bed, mix well with the fungus bed with an excavator loader, and let it stand for 48 hours. The product temperature, moisture content, pH ( (Hydrogen ion concentration) was measured, and the digestion and extinction status of the fungus bed was observed. The bacterial flora analysis and bacterial bed component analysis were performed before and after the experiment, and the difference was also verified.

2. Experiment place: 482 Nase Nakakatsu, Amami City, Kagoshima Prefecture 8 In the Amami City Kamesaku Complex Reinforcement Warehouse (about 150 m ² )

3. Sample (sewage sludge) provider 523 Nase Nagahama, Amami-shi, Kagoshima Nase terminal treatment plant

4). Bacterial bed component analysis of inspection organization: Chiba Prefectural Environmental Foundation Chiba Prefectural Governor Registration No. 519 1-11-1 Chuo-ku, Chuo-ku, Chiba
Certificate No. BA21-14-1 BA21-49-1 Bacterial Flora Analysis: Technosuruga Lab Co., Ltd. 330, Nagasaki, Shimizu-ku, Shizuoka, Japan

5). Experiment period and content period: August 20 to October 31, 2009
August 20-25 :
Indigenous bacteria collection (Nakakatsu Prefectural Forest near the experimental site)
August 21 :
Mushroom bed awake back bacteria bed 2m ^3, charcoal 0.25 m ^3, rapeseed marc 0.25 m ^3, rice bran 0.05 m ^3, probiotics TAKES Water (PBW) :( Ltd.) Kanagawa Furniture made 0.02 m ^3, Tohaimitsu 0.02 m ^3. Two sets of fungi beds were prepared using 0.02 m ^{3 of} water and awakening work was performed.
August 25 :
Mass bed culture Awakened bed 2.55 m ³ , Back bed 3 m ³ , Rape seed squeezed 0.5 m ³ , Charcoal 0.25 m ³ , Bark 2 m ³ (from Yamato Village), Shochu waste solution 0.02 m ³ (Amami City) ), Bacus (produced in Setouchi Town), hay (cutting around the experimental area), PW 0.02 m ³ , seawater 0.02 m ³ (taken at Nase Port), water 0.7 m ³ Ome City Okuma), and indigenous bacteria 0.01 m ³ (collected in the vicinity of the experimental site) was mixed, and two sets of bacterial beds were cultured in large quantities.
September 1 :
Start of experiment The fungus bed was divided into two sets of A and B, 0.15 m ³ of sewage sludge was added to the fungus bed A, mixed with a loader, and allowed to stand until 10:00 am (48 hours later) tomorrow. The next day, 2 days, it was put into the fungus bed B, and it was allowed to stand until 10 am. Thereafter, this operation was repeated alternately.
October 7 :
The work was suspended due to the influence of Typhoon No. 18, and the test period was extended by one day.
October 21 :
The sewage sludge was thrown into the fungus bed B and finished. A total of 3.75 m ³ was added to both A and B (7.5 m ³ in total for both).
October 22 :
The bacteria bed was allowed to cool down until 28th.
October 28 :
Bacteria bed volume measurement, 15.735 m ³ (9,445.8 kg) L.0.I. (Lost of Ignition), moisture 48.1% (4,543.4 kg), crude ash 24.4% (2,333.2 kg) Organic matter was 27.2% (2,569.2kg).
October 31 :
End of verification test

Organic matter decomposition rate :
As shown in Table 1 below, due to the above-described decomposition treatment, the organic matter that is the energy source for digestion and extinction treatment is not enough for the organic matter (937.5 kg) contained in the sewage sludge, and the organic matter in the fungus bed is also used as 1,067.4 kg 7500 kg of sewage sludge was treated. Furthermore, with the decomposition treatment of sewage sludge by microorganisms, the water content (83.1%) contained in the sewage sludge decreased to the water content (48.1%) of the final fungus bed.

Method for measuring the amount of fungus bed
(a) Collect samples from the fungus bed and sewage sludge by the quadrant method.
(b) With a 1 l measuring cup, measure the mass per l and average it.
(c) Measure the bed volume with a shovel loader bucket (250 l / cup), 10 l bucket, and 1 l measuring cup.
(d) The total mass was measured with bxc.

Method for measuring composition of fungus bed and sewage sludge
(a) Collect 5g from the sample from the fungus bed and sewage sludge by the quadrant method.
(b) Apply a to the infrared moisture meter to obtain the moisture content, and measure the dry matter as it is. Further, dry matter is treated with L.0.I. and the remaining life is measured as crude ash.
(c) Weighed dry matter minus crude ash as organic matter.
(d) The calculated percentages were multiplied by the amount of fungus bed and the amount of sewage sludge to calculate the water content, the amount of organic matter, and the amount of crude ash.

The reason why the remainder of L.0.I. was not disturbed without “ash” was because the measurement method was processed by a simple method. It is impossible to digest and extinguish ash with this microorganism treatment technology. If ash increases, it is theoretically expected that ash will not be extinguished. There was still no noticeable decline in the ability to decompose the fungus bed during the seven experiments until October of the year.

Bacterial bed component analysis before and after sewage sludge treatment :
Table 2 below shows the results of analysis of the fungal bed components before and after the sewage contamination treatment. Bacteria beds A and B are both lead and chlorine and exceed the environmental standard values by soil analysis. As for phosphorus, the bacterial bed A decreased by 2.2% and the bacterial bed B increased by 0.6%. For the other 24 items, although there was an increase, it was below the reference value, no increase or decrease, or no detection.

The environmental standard value was diverted from the soil component analysis. Samples were collected before the experiment: June 5, 2009 and after the experiment: October 28, 2009. Analysis organization: Chiba Environmental Foundation.

Bacterial bed temperature, water content and pH during sewage sludge treatment :
In measuring the volumetric mixing ratio of the mushroom bed and sewage sludge 56.6: 1 (bacteria bed 8.52 m ^3: Sewage Sludge 0.15 m ³⁾ and mushroom bed sewage sludge for the (average water content 83.1%) water content in the turned on, Bacteria bed A averaged 46.4%, maximum 51.9% and minimum 38.3%, Bacteria bed B averaged 48.5%, maximum 56.7% and minimum 42.4%.

The average temperature of the fungus bed (30 cm deep from the surface) is 56.6 ° C, 70 ° C and 28.3 ° C on average for the fungus bed A, and 55.1 ° C, 70.3 ° C and 27.8 ° C on average for the fungus bed B. It was. In the above demonstration test, the volume mixing ratio of the fungus bed and sewage sludge was 56.6: 1, so the maximum product temperature during the stationary period was input and detected at the first measurement by switching the fungus bed at the time of sludge addition. After that, the case where the temperature gradually decreased and reached the lowest product temperature at the end of standing was 3 times for both B and A (B, 408-454 hours 67-59.6 ° C, 888-934 hours 48.9-48.7 ° C, 1296-1342 hours 3 times at 45.1-42.5 ° C, B for 720-766 hours 43-42.6 ° C 768-814 hours 46.9-40.7 ° C, 1,176-1,222 hours 58.3-44.9 ° C 3 times) It was observed for the first time in a fungal demonstration test.

The pH (hydrogen ion concentration) of the microbial bed was microbial bed A average 8.1, maximum 8.9, and minimum 6.7, and microbial bed B average 7.6, maximum 8.2, and minimum 5.9. These values were generally within the alkaline region where aerobic bacteria were actively active. In addition, when the effect of the cumulative input of sewage sludge on the pH of the fungus bed was investigated, there was almost no effect on either A or B.

The water content of sewage sludge averaged 83.1%, maximum 85.4% and minimum 77.1%. The pH averaged 8.1, maximum 8.8 and minimum 7.5. Similar to pH, there was almost no effect on the relationship between the cumulative input and the water content. Since sewage sludge was added with a polymer-based flocculant, no soup was leached even when left in the stockyard. A glass-like substance that seems to be a crystal of the flocculant was found in the fungus bed after the experiment.

Carbon dioxide equivalent balance of global warming gas :
(1) Carbon dioxide equivalent amount (reduced amount) estimated to be discharged when incinerated
(Sewage sludge incineration amount) x (carbon dioxide emission numerical value) = carbon dioxide equivalent emission kg-CO ₂
7,500kg x 0.84kg-CO ₂ = 6,300 kg-CO ₂
(2) Carbon dioxide emissions (light oil used as excavator loader fuel)
1l per day, 61l of light oil is used for 61 days. Light oil emission factor 2.6
(Light oil consumption) x (Emission factor) = Carbon dioxide equivalent emission kg-CO ₂
61l X 2.6 = 158.6kg-CO ₂
(3) CO2 emission reduction (CO2 emission)-(CO2 emission reduction) = (CO2 emission reduction)
158.6kg-CO ₂ − 6,300 kg-CO ₂ = −6,141.4 kg-CO ₂

This reduction is Nase Clean Center annual emissions (25,589 tons incineration) 21,494.76 tons C02
Equivalent to 0.0286% of In addition, from the same calculation, the total amount of sewage sludge currently incinerated (if 1,191 tons of FY2008 results were treated with microorganisms, the reduction amount was (incinerated sewage sludge amount) x (carbon dioxide emission value) = (dioxide dioxide Carbon equivalent emissions tons-CO ₂ ): 1,191 tons × 0.84 tons-CO ₂ = 1,000.44 tons-CO _2. This is equivalent to the carbon dioxide equivalent emissions from the clean center (incineration amount of 25,589 tons, carbon dioxide It is estimated to be 4.65% of carbon emissions 21,494.76 tons-CO ₂ ).

Sewage sludge (7,500 kg, organic matter content 937.5 kg) that has been digested and extinguished as such absorbs carbon dioxide in the atmosphere during its production process, so even if the sewage sludge itself is incinerated, as a carbon offset, That amount is excluded. Therefore, the global warming effect gas emitted during the incineration process is calculated as the amount of carbon dioxide equivalent due to the use of dinitrogen monoxide (N ₂ 0) derived from nitrogen contained in sewage sludge and fossil fuel as auxiliary fuel. The

In addition, regarding the inorganic components contained in the fungus bed, the fungus bed components before and after the start of the fungus beds A and B were subjected to analysis of soil environmental standard analysis items (26 items) and total phosphorus content test. . As a result, the fungus bed A showed an increase in the contents of cadmium, arsenic, total mercury, fluorine and boron, while the fungus bed B showed an increase in the contents of lead, arsenic and fluorine. Phosphorus increased from 4.7% to 2.5% in Fungus Bed A, minus 2.2%, and in B, B increased from 3.1% to 3.7%, 0.6% (according to Chiba Environmental Foundation).

Measuring equipment used :
(1-1) Product temperature / outside temperature: THERMMODAC EF (automatically measured by Eto Electric Co., Ltd.) 18 thermocouple sensors are prepared and measured every hour.
(1-2) Outside temperature / Inside temperature: Install two thermocouple sensors at 150cm above the ground.
(1-3) Bacterial bed product temperature: Eighteen thermocouple sensors fixed on the support at 30, 60, 100 cm from the surface of the fungus bed are inserted into the fungus bed. In addition to this, a compost sensor (MC-K7108 Φ8 × 900mm Co., Ltd., Sato Weighing Mfg. Co., Ltd.) was used for product temperature measurement.
(2) Moisture content / crude ash: Infrared moisture meter FD-600 (Kett Science Laboratory, Heating Drying / Mass Measurement System)
The fungus bed was measured for 5 g of sample at 110 ° C for 20 minutes. For the crude ash, the moisture content of a 5g sample was measured at 110 ° C for 20 minutes, and then the mass of the remaining dry matter was measured. The residue was measured as crude ash with L.0.I.
(3) pH: Use TEST PAPERS UNIVERSAL pHl-14 KAAGAT LTD. Bacteria bed: Water = 1: 5
(4) Bacteria bed and sewage sludge volume measurement: The fungus bed was measured with a shovel loader (Mitsubishi, WS210) bucket volume squeeze 250 l, a plastic bucket 10 l, and a measuring cup 1 l.

Identification of microorganisms contained in fungus bed First, microorganisms (spore bacteria) contained in each specimen were separated by the following method.
Sample:
Specimen 1: 7861-03 (collected from fungus bed B at maximum activity)
Sample 2: 7861-04 (collected from fungus bed A)
Specimen 3: 7861-05 (collected from fungus bed B)

1. Pretreatment / Suspension adjustment: 1 g of sample is suspended in 10 ml of physiological saline / heat treatment: The suspension is heated at 80 ° C. for 15 minutes.

2. Isolation / medium: Nutrient agar (Oxford, Hampshire, England) + agar, culture temperature: 55 ° C
・ Culture time: 1 day ・ Separation method: smear plate method, 0.1 ml surface smear (Edited by Soil Biology Society; New Soil Microbiology Experiment, Yokendo, 1992; Microbial Separation Method, R & D Planning, 1986)
・ Number of smears: 1 ・ Other conditions: Aerobic culture

3. Simple colony morphology observation (1) Gram staining: Faber G "Nissui" (Nissui Pharmaceutical, Tokyo)
(2) Optical microscope: BX50F4 (Olympus, Tokyo)
(3) Stereo microscope: SZH10 (Olympus, Tokyo)

4. 16S rDNA-500
The operations from extraction to cycle sequence were performed based on the following protocols.
・ DNA extraction: InstaGene Matrix (BIO RAD, CA, USA)
・ PCR: PrimerSTAR HS DNA Polymerase (Takara Bio, Shiga)
Cycle sequence: BigDye Terminator v 3.1 Cycle Sequencing Kit (Applied Biosystems, CA, USA)
・ Primers used: PCR (9F, 926R), Sequence (9F, 536R)
・ Sequence: ABI PRISM 3130 xl Genetic Analyzer System (Applied Biosystems, CA, USA)
・ Sequencing: ChromasPro 1.4 (Technelysium, Pty Ltd., Tewantin, AUS)
・ Homology search and simple molecular phylogenetic analysis software: Apollon 2.0 (Techno Suruga Lab, Shizuoka)
・ Homology search and simple molecular phylogenetic analysis database: Apollon DB-BA 5.0 (Techno Suruga Lab, Shizuoka) International nucleotide sequence database (GenBank / DDBJ / EMBL)

Specimen: SIID 7861-03-B1 (one of the colonies derived from Specimen 1)
As a result of simple morphological observation, SIID7861-03-Bl is positive for Gram staining and is a spore-forming bacilli (0.9-1.0 x 2.0-5.0μm) that shows viability under aerobic conditions. Colony color on Nutrient agar plate medium Had a pale yellow color.
As a result of homology search for Apollon DB-BA 5.0 using BLAST, the 16S rDNA partial sequence of SIID7861-03-Bl showed high homology to 16S rDNA of the genus Bacillus, and B. coagulans (SKERMAN (VBD), McGOWAN (V.) and SNEATH (PHA) (editors): Approved Lists of Bacterial Names. Int. J. Syst. Bacteriol., 1980, 30, 225-420) 98.9% homology to ATCC 7050 16S rDNA Showed the highest homology.
Also in the results of homology search against GenBank / DDBJ / EMBL, 16S rDNA of SIID7861103-Bl shows high homology to 16S rDNA of Bacillus genus, and in the reference strain, 16S rDNA of B. coagulans ATCC 7050 strain, It showed high homology with a homology rate of 98.9%.
As a result of a simple molecular phylogenetic analysis using 16S rDNA of 16S rDNA of SIID7861-03-Bl and 16S rDNA of the top 10 homology search for Apollon DB-BA5.0, SIID7861-03-Bl is 16S of Bacillus sp. It was found to be contained within a cluster formed of rDNA. The specimen also clustered with 16S rDNA of B. coagulans, and both showed the same molecular phylogenetic position.
From the above, it was estimated that SIID7861-03-Bl is contained in the genus Bacillus and belongs to B. coagulans.

Specimen: SIID 7861-03-B2 (one of the colonies derived from Specimen 1)
As a result of simple morphological observation, SIID7861-03-B2 is Gram-staining-positive, a koji mold (0.7-0.8 x 1.5-2.0μm) that shows viability under aerobic conditions, and the colony color on Nutrient agar plate medium is light It was yellow.
As a result of homology search for APOLON DBIBA 5.0 using BLAST, 16S rDNA partial sequence of SIID7861-03-B2 showed high homology to 16S rDNA of Ureibacillus genus, and U. suwonensis (KIM (BY) LEE (SY ), WEON (HY), KWON (SW), GO (SJ), PARK (YK), SCHUMANN (P.) And FRITZE (D.): Ureibacillus suwonesis sp. Nov., isolated from cotton waste composts. Int. J Syst. Evol. Microbiol., 2006, 56, 663-666) It showed the highest homology with 99.4% homology to 16S rDNA of 6T19 strain.
Also in the results of homology search for DDBJ / EMBL, 16S rDNA of SIID7861-03-B2 showed high homology to 16S rDNA of Ureibacillus genus, and the reference strain was 99.4% homologous to 16S rDNA of U. suwonensis 6T19 strain. % Showed the highest homology.
As a result of simple molecular phylogenetic analysis using 16S rDNA of SIID7861-03-B2 and 16S rDNA of the top 10 homology search for Apollon DBIBA5.0, SIID7861-03-B2 is a 16S rDNA of the genus UneibacilIus. It was found to be contained within the cluster formed. The specimen also formed a cluster with U. suwonensis 16S rDNA and was shown to be closely related. However, one base is clearly different between the SllD7861-03-B2 and U.suwonesis 16S rDNA partial sequences.
Based on the above, it is appropriate that SIID7861-03-B2 is included in the genus Ureibacillus and belongs to U. suwonesis or is closely related to Ureibacillus sp.

Specimen: SIID 7861-04-B1 (Colony derived from Specimen 2)
As a result of simple morphological observation, SIID7861-04-Bl is a gonococcus (0.8-0.9 x 1.5-2.5μm) that shows growth stability under aerobic conditions, and the colony color on the Nutrient agar plate medium is It had a cream color.
As a result of homology search against Apollon DB-BA 5.0 using BLAST, the 16S rDNA partial sequence of SIID7861-04-Bl shows high homology to 16S rDNA of Bacillus genus, and 16S rDNA of B. licheniformis DSM 13 strain On the other hand, it showed the highest homology with a homology rate of 99.6%.
As a result of homology search against GenBank / DDBJ / EMBL, 16S rDNA of SIID7861-04-Bl showed high homology to 16S rDNA of Bacillus genus, and B. licheniformis ATCC 14580 strain and DSM 13 It showed the highest homology with the homology of 99.6% for the strain 16SrDNA.
As a result of simple molecular phylogenetic analysis using 16S rDNA of 16S rDNA of SIID7861-04-Bl and 16S rDNA of the top 10 homology search for Apollon DB-BA5.0, SIID7861-04-Bl was obtained from B. licheniformis, B. It was found to be contained in clusters formed by 16S rDNA of aerius and B. stratosphericus. The specimen showed the same molecular phylogenetic position as B. licheniformis 16S rDNA, and there was a distance between the other two species.
From the above, SIID7861-04-Bl was estimated to be a strain belonging to the genus Bacillus and belonging to B. licheniformis.

Specimen: SIID 7861-05-B1 (Colony derived from specimen 3)
As a result of simple morphological observation, SIID7861-05-Bl is gram-stained, gonococci (0.8-1.0 x 2.0-4.0μm) showing viability under aerobic conditions, and the colony color on Nutrient agar plate medium is light It was yellow.
As a result of homology search for Apollon DB-BA5.0 using BLAST, the 16S rDNA partial sequence of SIID7861-05-Bl showed high homology to Brevibacillus genus 16S rDNA, and B. Thermoruber (SHIDA (O.) , TAKAGI (H.), KADOWAKI (K.) and KOMAGATA (K.): Proposal for two new genera, Brevibacillus gen. Nov. And Aneurinibacillus gen. Nov. Int. J. Syst. Bacteriol., 1996, 46, 939 -946) It showed the highest homology of 99.2% with 16S rDNA of DSM 7064 strain.
As a result of homology search against GenBank / DDBJ / EMBL, 16S rDNA of SIID7861-05-Bl showed high homology to 16S rDNA of the genus Brevibacillus, and B. Thermoruber (SKERMAN (VBD), McGOWAN ( (V.) and SNEATH (PHA) (editors): Approved Lists of Bactarial Names. Int. J. Syst. Bacteriol., 1980, 30, 225-420) High homology of 99.2% with 16S rDNA of DSM 7064 strain However, 16S rDNA derived from the reference strain was not searched.
As a result of a simple molecular phylogenetic analysis using 16S rDNA of the top 10 strains of homology search for 16S rDNA of SllD7861-05-Bl and Apollon DB-BA5 ・ 0, SllD7861-05-Bl was found to be 16S of Brevibacillus species It was found to be contained within a cluster formed of rDNA. In addition, the sample formed a cluster with B. thermoruber 16S rDNA, indicating that it was closely related. However, 3 bases are clearly different between SllD7861-05-Bl and B. thermoruber 16S rDNA partial sequences.
From the above, it is appropriate that SIID7861-05lBl is included in the genus Brevibacillus and belongs to B. thermoruber or is closely related to Brevibacillus sp.

PCR-DGGE and RT-PCT-DGGE analysis :
In the following manner, the DNA of the microbial community contained in the sample was analyzed using PCR-DGGE and RT-PCR-DGGE.
(1) DNA extraction / extraction method: ISOIL for Beads Beating (Nippon Gene, Tokyo)
(2) RNA extraction and extraction method: RNA Power Soil Total RNA Isolation Kit (MO-BIO, USA)
(3) PCR
・ Primer used: GC-341f-534r (Muyzer G., et al., Appl. Environ. Microbiol., 1993, 59, 695-700) (for 200bp amplification)
PCR conditions: Touch Down method (Muyzer G., et al. Above)
・ PCR preliminary test: Examination of DNA amount 2.5 ng and 10 ng (4) RT-PCR
-Reverse transcription reaction primer: 907r (Ishi K., et al., J. Gen. Appl. Microbiol., 2000, 46, 85-93)
-RT-PCR primer: GC-341f-534r
・ RT-PCR conditions: Same as above ・ RT-PCR preliminary test: Same as above (5) DGGE
・ DGGE equipment: DcodeDGGE complete system (BIO RAD, CA, USA)
・ Denaturer concentration gradient: 25% → 65%
・ Polyacrylamide gel concentration: 8%
・ Electrophoretic conditions: Voltage 100V, 12 hours ・ DGGE marker: DGGE Marker I (Nippon Gene, Tokyo)

  As a result of performing an international base sequence database BLAST search for the bands obtained from each sample by PCR-DGGE RT-PCT-DGGE, the above-mentioned fungus bed contains microorganisms belonging to the genus Bacillus such as Bacillus coagulans and Bacillus licheniformis, It is confirmed that microorganisms belonging to the genus Ureibacillus such as Ureibacillus suwonensis or related species, and microorganisms belonging to the genus Brevibacillus such as Brevibacillus thermoruber are included, and microorganisms belonging to the genus Tepidimicrobium and / or Pelotomaculum genus It was shown that microorganisms belonging to

By the method of the present invention, the carbon dioxide emission can be reduced as shown in the above demonstration test. Sewage sludge is finally incinerated, but organic matter in the sewage sludge is decomposed and treated by the method of the present invention, and the water content is significantly reduced, so the amount of incineration can be reduced. As a result, it is possible to extend the useful life of incineration facilities such as local governments. Further, various useful components such as phosphorus can be recovered and reused from the incinerated ash after incineration.

  In addition, the method of the present invention can be effectively used for dehydration in an industrial production process of biofuels and fuel materials using various biomass such as microalgae as raw materials.

Claims (13)

A treatment comprising adding a treatment object to a fungus bed containing a microorganism belonging to the genus Bacillus, a microorganism belonging to the genus Ureibacillus, and a microorganism belonging to the genus Brevibacillus, and decomposing the organic matter contained in the treatment object by the microorganism. A method of processing an object. The method according to claim 1, comprising adding a treatment target to a bacterial bed containing a microorganism belonging to the genus Bacillus, a microorganism belonging to the genus Ureibacillus, and a microorganism belonging to the genus Brevibacillus, and then allowing to stand still. The method according to claim 1 or 2, wherein the object to be treated is sewage sludge. A bacterial bed containing a microorganism belonging to the genus Bacillus, a microorganism belonging to the genus Ureibacillus, and a microorganism belonging to the genus Brevibacillus is added to and mixed with the object to be treated, and the object to be treated is generated by heat generated by fermentation by the microorganisms in the bacteria bed. A method for reducing the water content of food. The method of Claim 4 that a process target object is biomass. 6. The method of claim 5, wherein the biomass is a culture of microalgae. The microorganism belonging to the genus Bacillus is Bacillus coagulans or Bacillus licheniformis and / or the microorganism belonging to the genus Ureibacillus is Ureibacillus suwonensis or a closely related species, and / or the microorganism belonging to the genus Brevibacillus is Brevibacillus thermoruber. The method as described in any one of -6. The method according to claim 7, wherein the fungal bed further comprises a microorganism belonging to the genus Tepidimicrobium and / or a microorganism belonging to the genus Pelotomaculum. The method as described in any one of Claims 1-8 whose volume ratio of the microbial bed in a process and the added process target object is 10: 1-60: 1.   The method as described in any one of Claims 1-9 which divides a process target object into multiple times and adds and mixes to a microbial bed every 1 to several days. The method as described in any one of Claims 1-10 whose moisture content of the mixture of a microbial bed and a process target object is 50 to 75 weight% in the decomposition process of a process target object. The temperature of the mixture of a fungus bed and a processing target object during the decomposition process of the processing target object is 25 to 75 ° C (measured at a depth of 30 cm from the surface). the method of. The method as described in any one of Claims 1-12 whose pH of the mixture of a microbial bed and a process target object in the decomposition process of a process target object is 5.5-9.0.