JP2011031180A - Method of making heavy metal, dioxin, nitrate and agricultural chemical harmless - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To decompose heavy metals, dioxins, nitrates and agricultural chemicals effectively at low costs to make them harmless. <P>SOLUTION: A method of making the harmful substances harmless comprises mixing soy pulp or rice bran with a group of effective microorganisms comprising alkalophilic Bacillus 'tateyama-turugi' FERM BP-10691, alkalophilic Bacillus 'tateyama-yakusi' FERM BP-10692, halophilic Bacillus 'tateyama-jodo' FERM BP-10693, anaerobic Atopostipes 'tateyama-jojo' FERM BP-10690 and anaerobic and halophilic Clostridium 'tateyama-ryuoh' FERM BP-10694, carrying out primary fermentation at 50-100°C for 24 h to form a bed material, mixing a target material with the bed material, mixing the mixture of the target material and the bed material with the group of the effective microorganisms and carrying out secondary fermentation at 60-80°C for 48-72 h. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a method for detoxifying heavy metals containing cadmium, dioxins, nitrates and agricultural chemicals.

  That is, the present invention relates to a method for decomposing and detoxifying heavy metals, dioxins, nitrates and agricultural chemicals contained in soil, fertilizer, water, incinerated ash, livestock manure, industrial wastewater, sludge, sewage and the like.

  In the present application, soil, fertilizer, water, incinerated ash, livestock manure, factory wastewater, sludge, sewage, etc. containing heavy metals, dioxins, nitrates or agricultural chemicals to be rendered harmless are referred to as objects to be treated.

  In Japanese Patent No. 3710424, the present inventors provide a method for decomposing heavy metals, dioxins and agricultural chemicals (hereinafter referred to as [conventional method]).

In the conventional method, an effective microorganism group composed of 55% aerobic bacteria group and 45% anaerobic bacteria group is mixed in organic waste, and primary fermentation is performed at a temperature of 40 to 200 ° C. By mixing this with an effective microbial group consisting of 55% aerobic bacteria group and 45% anaerobic bacteria group, and subjecting it to secondary fermentation at a temperature of 100 to 200 ° C. together with the material to be treated. A method for decomposing heavy metals, dioxins and agricultural chemicals, characterized in that heavy metals, dioxins or agricultural chemicals contained in the object to be treated are decomposed.

Japanese Patent No. 3710424

  Although the above conventional method is capable of decomposing heavy metals, dioxins and agricultural chemicals, the present invention is a method for effectively decomposing and detoxifying heavy metals, dioxins, nitrates and agricultural chemicals at low cost. It was made to offer.

  In order to solve the above problems, the present invention provides a method for detoxifying the following heavy metals, dioxins, nitrates and agricultural chemicals.

(1) The alkalophilic Bacillus genus Tateyama sword FERM BP-10691, the alkalophilic Bacillus genus Tateyama pharmacist FERM BP-10682, the halophilic Bacillus genus Tateyama pure soil FERM BP-10893, and the anaerobic fungus Atopostipes An effective microorganism group consisting of the genus Tateyama female FERM BP-10690 and the anaerobic and halophilic Clostridium genus Tateyama Ryuo FERM BP-10694 is mixed in okara or rice bran at a temperature of 50 to 100 ° C. 24 hours primary fermentation to form flooring,
Mixing the object to be treated with the flooring;
To the mixture of the object to be treated and the flooring, an alkalophilic Bacillus genus sword FERM BP-10691, an alkalophilic Bacillus genus Tateyama pharmacist FERM BP-10692, and a halophilic Bacillus genus Tateyama pure soil FERM BP- An effective microorganism group consisting of 10893, an anaerobic bacterium, Atostipes genus TATEYAMA FERM BP-10690, and anaerobic and halophilic Clostridial genus TATEYAMA Ryuo FERM BP-10694 are mixed. A method of detoxifying heavy metals, dioxins, nitrates and agricultural chemicals, characterized by subjecting to secondary fermentation at a temperature of 60 to 80 ° C. for 48 to 72 hours.

(2) The alkalophilic Bacillus Tateyama sword FERM BP-10691, the alkalophilic Bacillus genus Tateyama pharmacist FERM BP-10692, the halophilic Bacillus genus Tateyama pure soil FERM BP-10893, and the anaerobic fungus Atopostipes An effective microorganism group consisting of the genus Tateyama female FERM BP-10690 and the anaerobic and halophilic Clostridium genus Tateyama Ryuo FERM BP-10694 is mixed in okara or rice bran at a temperature of 50 to 100 ° C. 24 hours primary fermentation to form flooring,
Mixing the object to be treated with the flooring;
To the mixture of the object to be treated and the flooring, an alkalophilic Bacillus genus sword FERM BP-10691, an alkalophilic Bacillus genus Tateyama pharmacist FERM BP-10692, and a halophilic Bacillus genus Tateyama pure soil FERM BP- An effective microorganism group consisting of 10893, an anaerobic bacterium, Atostipes genus TATEYAMA FERM BP-10690, and anaerobic and halophilic Clostridial genus TATEYAMA Ryuo FERM BP-10694 are mixed. A method for detoxifying heavy metals, dioxins, nitrates and agricultural chemicals, characterized by natural fermentation for 1 to 2 months without heating (Claim 2).

  According to the present invention, heavy metals, dioxins, nitrates and pesticides contained in soil, fertilizer, water, incinerated ash, livestock manure, industrial wastewater, sludge, sewage, etc. can be effectively decomposed and rendered harmless. Moreover, since the effective microorganism group and okara or rice bran are used, it is extremely advantageous in terms of cost, and the okara or rice bran that is normally discarded can be effectively used. Furthermore, useful compost can also be obtained by carrying out the fermentation process of the to-be-processed object.

FIG. 1 is a molecular phylogenetic tree of a Bacillus sp. Tateyama sword and a Bacillus sp. Tateyama pharmacist. FIG. 2 is a molecular phylogenetic tree of Bacillus sp. FIG. 3 is a molecular phylogenetic tree of Atopostipes sp. Tateyama. FIG. 4 is a molecular phylogenetic tree of Clostridium sp.

The method for detoxifying heavy metals, dioxins, nitrates and agricultural chemicals according to the present invention is as follows.

[Formation of flooring]
First, the alkalophilic Bacillus sp. Tateyama sword FERM BP-10691, the alkalophilic Bacillus sp. Tateyama pharmacist FERM BP-10682, and the halophilic Bacillus genus (Bacillus sp.).
sp.) Tateyama Pure Land FERM BP-10893, anaerobic bacteria Atopostipes sp. Tateyama Lady FERM BP-10690, anaerobic and halophilic Clostridium sp. An effective microorganism group consisting of Tateyama Ryuo FERM BP-10694 is mixed in okara or rice bran and subjected to primary fermentation at a temperature of 50 to 100 ° C. for 24 hours to form a flooring material.

As an example, 2 parts by weight of the effective microorganism group is mixed in 35 parts by weight of Okara or rice bran.

[Mixed material to be treated with flooring]
To-be-treated material such as heavy metals, dioxins, nitrates or agricultural chemicals containing soil, fertilizer, water, incinerated ash, livestock manure, industrial wastewater, sludge, sewage, etc. are mixed with the flooring.

As an example, 58 parts by weight of the workpiece is mixed with 37 parts by weight of a flooring material.

[Secondary fermentation]
To the mixture of the material to be treated and the flooring, an alkalophilic Bacillus sp. Tateyama sword FERM BP-10691, an alkalophilic Bacillus sp. Tateyama pharmacist FERM BP-10682, Salty Bacillus
sp.) Tateyama Pure Land FERM BP-10893, anaerobic bacteria Atopostipes sp. Tateyama Lady FERM BP-10690, anaerobic and halophilic Clostridium sp. An effective microorganism group consisting of Tateyama Ryuo FERM BP-10694 is mixed and subjected to secondary fermentation at a temperature of 60 to 80 ° C. for 48 to 72 hours.

  As an example, 5 parts by weight of the effective microorganism group is mixed in 95 parts by weight of a mixture of 58 parts by weight of the workpiece and 37 parts by weight of the flooring material.

[Natural fermentation]
About the said secondary fermentation, the said effective microorganism group may be mixed in the mixture of this to-be-processed object and this flooring, and you may ferment naturally for 1 to 2 months, without heating this.

It should be noted that the anaerobic bacteria Atopostipes sp. Tateyama woman (FERM BP-10690), the alkalophilic Bacillus sp. Genus (Bacillus sp.) Tateyama pharmacist (FERM BP-10692), halophilic Bacillus sp. Tateyama pure soil (FERM BP-10893), anaerobic and halophilic clostridium genus ( Clostridium sp.) Tateyama Ryuo (FERM BP-10694) is deposited at the National Institute of Advanced Industrial Science and Technology Patent Biological Depositary.

An alkalophilic Bacillus sp. Tateyama sword and an alkalophilic Bacillus
sp.) Tateyama pharmacist, halophilic Bacillus sp. Tateyama pure soil, anaerobic Atopostipes sp. Tateyama woman, and anaerobic and halophilic Clostridium Identification was conducted with Clostridium sp. Hereinafter, an identification method and a microbial test result are shown.

As a bacterial first stage test, cell morphology, Gram staining, presence of spores and motility by flagella were observed with an optical microscope U-LH1,000 (Olympus, Japan). Colony morphology was observed on NUTRIENT AGAR (OXOID, HAMPSHIRE, ENGLAND) (hereinafter referred to as “NT”) or GAM agar medium (Nissui, Tokyo) (hereinafter referred to as “GAM”). Tests were made for catalase reaction, oxidase reaction, acid / gas production from glucose and glucose oxidation / fermentation (O / F).
Table 1 shows the results of the bacteriological property test.

Next, gene analysis of 16S rDNA base sequence was performed. INSTAGENE MATRIX (BIO RAD, CA, USA) was used for extraction of genomic DNA. Using the extracted genomic DNA as a template, a region having a total base sequence of about 1,500 to 1,600 base pairs was amplified by PCR. Thereafter, the amplified 16S rDNA was sequenced to obtain a base sequence. For purification and cycle sequencing of PCR products, BIGDYE TERMINATOR V3.1 CYCLE SEQUENCING KIT (APPLIED BIOSYSTEMS, CA, USA) was used. PRIMESTAR HS DNA POLYMERASE (Takara Bio, Shiga) for thermal cyclers and ABI PRISM 3100 GENETIC ANALYZER SYSTEM (APPLIED BIOSYSTEMS, for DNA sequencers)
CA, USA). The primers were according to the document “Identification and Classification of Actinomycetes, p88-117, Japan Society for Business Administration, 2001”.

The analysis results of the base sequence of 16S rDNA are shown in SEQ ID NO: 1 for Bacillus sp. Tateyama sword, SEQ ID NO: 2 for Bacillus sp. Tateyama pharmacist, and Bacillus sp.
sp.) Tateyama Pure Land, SEQ ID NO: 3, and Atopostipes sp. Tateyama Maiden, SEQ ID NO: 4, and Clostridium.
sp.) Tateyama Ryuo is shown in SEQ ID NO: 5, respectively.

The above base sequence is searched for homology using the Apollon DB bacterial reference strain database (Techno Suruga, Shizuoka). From the obtained 16S rDNA base sequence, a homology search for species considered to be closely related to the sample is performed. Five strains were determined.
The homology search results are shown in Table 2 for Bacillus sp. Tateyama swords, in Table 3 for Bacillus sp. Tateyama pharmacists, and Bacillus sp.
sp.) Table 4 for Tateyama Pure Land, Table 5 for Atopostipes sp. Tateyama, and Clostridium
sp.) Tateyama Ryuo is shown in Table 6, respectively.

Based on information obtained so far, CLUSTAL W and MEGA VER3.1 were used to construct a molecular phylogenetic tree using the 16S rDNA base sequence and the 16S rDNA base sequence of the bacterial group considered to be closely related thereto. . The neighbor linking method was used to estimate the molecular phylogenetic tree, and a bootstrap showing the validity of the tree type was generated 1000 times.
CLUSTAL W is a document “Nucleic Acids Research, 1994, 22: 4673-4680.” MEGA is a document “Briefings in
Bioinformatics, 2004, 5, 150-163. "
The results of molecular phylogenetic analysis are shown in FIG. 1 for the Bacillus sp. Tateyama sword and the Bacillus sp. Tateyama pharmacist.
sp.) The Tateyama Pure Land is shown in Figure 2, and the Atopostipes sp. Tateyama Lady is shown in Figure 3, Clostridium.
sp.) is shown in FIG.

The alkalophilic Bacillus sp. Tateyama sword FERM BP-10691, the alkalophilic Bacillus sp. Tateyama pharmacist FERM BP-10682, and the halophilic Bacillus genus (Bacillus sp.)
sp.) Tateyama Pure Land FERM BP-10893, anaerobic bacteria Atopostipes sp. Tateyama Lady FERM BP-10690, anaerobic and halophilic Clostridium sp. 2 parts by weight of an effective microorganism group consisting of Tateyama Ryuo FERM BP-10694 was mixed in 35 parts by weight of Okara and subjected to primary fermentation at a temperature of 70 to 80 ° C. for 24 hours to form a flooring material. 58 parts by weight of soil contaminated with cadmium was mixed with 37 parts by weight of flooring. To 95 parts by weight of a mixture of 58 parts by weight of the soil and 37 parts by weight of the flooring material, an alkalophilic Bacillus sp. Tateyama sword FERM BP-10691 and an alkalophilic Bacillus sp. Tateyama pharmacist FERM BP-10692 and halophilic Bacillus
sp.) Tateyama Pure Land FERM BP-10893, anaerobic bacteria Atopostipes sp. Tateyama Lady FERM BP-10690, anaerobic and halophilic Clostridium sp. 5 parts by weight of an effective microorganism group consisting of Tateyama Ryuo FERM BP-10694 was mixed, and this was subjected to secondary fermentation at a temperature of 60 to 80 ° C. for 60 hours.

  As a result, the soil contaminated with cadmium was detoxified and became safe soil for agricultural products.

  58 parts by weight of soil contaminated with cadmium was mixed with 37 parts by weight of the flooring material of Example 1, and 95 parts by weight of the mixture was mixed with 5 parts by weight of the effective microorganism group. Container bag) and naturally fermented for 1 month without heating.

  As a result, the soil contaminated with cadmium was detoxified and became safe soil for agricultural products.

  The same method as in Example 1 was performed on the soil contaminated with dioxin. As a result, the soil contaminated with dioxin was detoxified and became safe soil for crops.

  The same method as in Example 2 was performed on the soil contaminated with dioxin. As a result, the soil contaminated with dioxin was detoxified and became safe soil for crops.

  The same method as in Example 1 was performed on soil contaminated with agricultural chemicals and nitrates. As a result, soil contaminated with pesticides and nitrates was rendered harmless and became safe soil for agricultural crops.

  The same method as in Example 2 was performed on soil contaminated with agricultural chemicals and nitrates. As a result, soil contaminated with pesticides and nitrates was rendered harmless and became safe soil for agricultural crops.

Claims (2)

Alkalophilic Bacillus genus sword FERM BP-10691, alkalophilic Bacillus genus Tateyama pharmacist FERM BP-10692, halophilic bacillus genus Tateyama pure soil FERM BP-10893, anaerobic bacteria Apostostis genus Tateyama有効 FERM BP-10690 and anaerobic and halophilic Clostridium genus Tateyama Ryuo FERM BP-10694 are mixed with okara or rice bran and primary at a temperature of 50-100 ° C. for 24 hours. Fermented to form flooring,
Mixing the object to be treated with the flooring;
To the mixture of the object to be treated and the flooring, an alkalophilic Bacillus genus sword FERM BP-10691, an alkalophilic Bacillus genus Tateyama pharmacist FERM BP-10692, and a halophilic Bacillus genus Tateyama pure soil FERM BP- An effective microorganism group consisting of 10893, an anaerobic bacterium, Atostipes genus TATEYAMA FERM BP-10690, and anaerobic and halophilic Clostridial genus TATEYAMA Ryuo FERM BP-10694 are mixed. A method for detoxifying heavy metals, dioxins, nitrates, and agricultural chemicals, wherein secondary fermentation is performed at a temperature of 60 to 80 ° C. for 48 to 72 hours. Alkalophilic Bacillus genus sword FERM BP-10691, alkalophilic Bacillus genus Tateyama pharmacist FERM BP-10692, halophilic bacillus genus Tateyama pure soil FERM BP-10893, anaerobic bacteria Apostostis genus Tateyama有効 FERM BP-10690 and anaerobic and halophilic Clostridium genus Tateyama Ryuo FERM BP-10694 are mixed with okara or rice bran and primary at a temperature of 50-100 ° C. for 24 hours. Fermented to form flooring,
Mixing the object to be treated with the flooring;
To the mixture of the object to be treated and the flooring, an alkalophilic Bacillus genus sword FERM BP-10691, an alkalophilic Bacillus genus Tateyama pharmacist FERM BP-10692, and a halophilic Bacillus genus Tateyama pure soil FERM BP- An effective microorganism group consisting of 10893, an anaerobic bacterium, Atostipes genus TATEYAMA FERM BP-10690, and anaerobic and halophilic Clostridial genus TATEYAMA Ryuo FERM BP-10694 are mixed. A method of detoxifying heavy metals, dioxins, nitrates and agricultural chemicals, characterized by natural fermentation for 1 to 2 months without heating.

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