KR100473027B1 - Method for preparing microbial preparation having excellent deodorizing and organic material-degrading capability, and synthetic medium suitable for preparing the same - Google Patents

Abstract

The present invention relates to a method for producing a microbial preparation comprising a microbial community useful for decomposition of various malodorous gases and organic matter and a medium used therein. Specifically, the present invention provides a method for preparing a microorganism preparation for removing odors and useful microorganisms including decomposed microorganisms having excellent decomposition ability of each malodorous component such as hydrogen sulfide, ammonia, and amines generated during the manure treatment process, and separation of useful microorganisms. A synthetic medium suitable for the present invention.

Description

Method for preparing microbial preparation having excellent deodorizing and organic material-degrading capability, and synthetic medium suitable for preparing the same}

The present invention relates to a method for producing a microbial agent excellent in the decomposition of odor gas and organic matter and a synthetic medium suitable for the preparation thereof. Specifically, the present invention provides a method for producing a microorganism preparation for removing organic matter and odor including each of the malodorous components such as hydrogen sulfide, ammonia, amines and the like decomposing microorganisms having excellent resolution of organic matters generated during the manure treatment process and a synthetic medium suitable therefor. It is about.

Odor gas generated in the use of toilet and manure treatment is harmful to the human body and causes air or environmental pollution, as well as causing discomfort or aversion, which is one of the important problems to be solved.

The odor generated in the toilet is caused by the odor-causing gas component generated during the anaerobic decomposition of organic substances, and its composition varies depending on the stage of biological decomposition. The main components of the odorous gas generated are methane and carbon dioxide, and inorganic and low molecular weight organic substances such as ammonia, hydrogen sulfide, mercaptans, amines and acetic acid are generated at the same time to induce odor, and these substances can be regarded as main odor causing substances.

As a method of removing odors, adsorption, absorption, combustion, biological and chemical oxidation are mainly used. Among these methods, adsorption is a method of removing odor causing substances through an adsorbent such as activated carbon, and biological oxidation is a method of removing odor causing substances by microorganisms using a packed bed microbial reaction tank. Chemical oxidation is a method of oxidizing odor causing substances using an oxidizing agent such as a chlorine-based oxidizing agent. Combustion can cause air pollution, and adsorption using activated carbon is expensive and therefore uneconomical.

Until now, most studies have focused on the odor reduction effect according to physicochemical conditions. For example, liquid manure in pigs produces less odor when intermittently aerated than continuous aeration. Gram negative and non-fermentative bacteria It is reported to have been found primarily. Hennlich, W. (1985), Zentralbl. Bakteril. Mikrobiol. Hyg. [B] 181 (1-2), 37-51 and Zentralbl. Bakteril. Mikrobiol. Hyg. [B] 181 (1-2), 52-63.].

There are few studies on biological approaches, but studies on the inhibition of ammonia production using inhibitors of urease (cyclohexylphosphoric triamide and phenyl phosphorodiamidate) have been made by Varel [Varel, V.H. (1998). Agricultural research magazine 46 (10), 22].

In recent years, biological purification to develop and apply special microbial inoculants in various applications such as bioremediation of contaminated soil or wastewater and biofilter for biological removal of odors and volatile organic substances. Research on bioaugmentation is actively conducted [Cho, KS, Hirai, M. and Shoda, M. (1992), Journal of Fermentation and Bioengineering, 73 (1). 46-50; Herrmann. R. F., and J. F Shann (1997), Microbial ecology. 33, 78-85; Tanji, Y., Kanagawa, T. and Mikami, E. (1992), Journal of Fermentation and Bioengineering. 67 (4). 280-285]. The purpose of this study is to maximize the efficiency of biological processes by developing, incubating and inoculating useful microorganisms that are more effective than microorganisms in contaminated soil or wastewater.

In recent years, unlike pure cultures or mixed cultures of a single species of useful microorganisms with regard to biological purification, two or more different microorganisms can coexist closely and have a positive synergistic effect. There are many studies to apply microbial community or microbial consortia to various fields such as biological purification and composting. These studies ultimately aim to secure microbial ecology management techniques to maximize the efficiency of biological processes by regulating the ecology of microorganisms in specific environments.

However, a single microbial species or community of microorganisms reported so far with odor degrading was selected from a medium containing ammonia or sulfur compounds, which are the main components of fecal malodors, as the only energy source, and the presence of high concentrations of organic matter, which is the actual environment of the feces. Underneath, there is a problem in that the growth rate is slow and difficult to be commercialized.

Accordingly, the present inventors have diligently studied to solve the problems of the conventional method as described above, the microorganisms isolated using a synthetic medium useful for accumulating and cultivating a microbial community similar to the actual feces is active in the manure environment, It was found that the odor resolution was excellent, and even when this isolated microbial community was grown in other manures, it could show a synergistic effect as a dominant species, and came to complete the present invention.

Accordingly, an object of the present invention is to decompose each odor component and organic matter, such as hydrogen sulfide, ammonia, and amines, generated in a feces treatment process, such as a manure treatment process or a biological purification system, using a synthetic medium very similar to an actual feces or manure environment. The present invention provides a method for preparing a microbial agent for removing organic substances and odors, which exhibits a synergistic effect as a superior dominant species while coexisting with excellent and indigenous microorganisms, and a synthetic medium used therein.

The present invention has a synergistic effect as a definite dominant species with excellent degradability of each odor component and organic matter such as hydrogen sulfide, ammonia and amines generated in feces treatment process, for example, manure treatment process or biological purification system using synthetic medium. Provided are a method for preparing a microorganism for removing organic matter and odor, and a synthetic medium used therein.

Most of the ammonia oxidizing bacteria reported so far are known as autotrophic bacteria, and pure separation of these microorganisms is performed in a medium using ammonia as an energy source. However, high concentrations of organic matter are present in the manure, and high concentrations of organic matter are known to inhibit growth. Therefore, in the case of ammonia oxidizing bacteria separated from the existing medium, if used as a preparation for the treatment of the actual manure is likely to be unable to comply with the environment containing high concentration of organic matter. Sulfated bacteria, which are known to be excellent in the decomposition of sulfide-based odorous substances, such as Thiobacillus sp., Also belong to autotrophic bacteria.

Therefore, suitable culture methods are most important for the isolation of malodor-decomposing microorganisms to be put into manure treatment. Of Almeida et al. (Almeida MC et al. (1999), Urban Water 1, 49-55) and Choi et al. (Choi E. et al. (1997), Wat. Sci. Tech. 35 (1), 233-240). The characteristics of the manure reported in the literature are shown in Tables 1 and 2 below.

Contaminant concentration of excrement (mg) 1 g of stools 1 ml of urine COD _t 287 17.5 NH ₃ -N 1.5 2.49 NO ₃ -N 0.03 0.012 PO ₄ -P 5.8 5.5 TSS 208 21

In the table, COD _t is total chemical oxygen demand, NH ₃ -N is ammonia nitrogen, NO ₃ -N is nitrate nitrogen, PO ₄ -P is phosphate, TSS is a total rich solid material.

Influent characteristics of manure treatment plant (mg / L except pH) Configuration Sewage Treatment Plant Influent Manure mean SeptageAvg maximum at least Average Standard Deviation pH     8.6     7.2    7.94   0.269    7.9    7.3 TCOD  72,300  16,330  41,050  14,837  48,000  32,000 SCOD  36,200   7,400  18,400   7,363  26,000  12,000 TBOD  29,700   8,100  15,200   4,825  19,000   8,000 SBOD  15,800   5,200   9,300   3,635  12,500   4,300 TSS  34,300  19,500  26,800   8,591  25,000  18,000 VSS  27,600  16,300  21,200   7,282  21,000  12,000 TKN   8,512   1,260   4,424   1,514   6,000   3,000 NH ₄ -N   6,540    940   3,680   1,156   4,800   2,500 T-P   1,317    414    687    226   1,000    500 PO ₄ -P    884    265    515    193    600   9,000 Alkalinity  28,600   9,300  19,800  3,899  24,000     64 COD / TP   59.8    48    10.7 COD / TN    9.3     8

In the table above, TCOD is the total chemical oxygen demand, SCOD is the soluble chemical oxygen demand, TBOD is the total biological oxygen demand, SBOD is the soluble biological oxygen demand, TSS is the total rich solids, VSS is the volatile rich solids, and TKN is Kjeldahl. Nitrogen, NH ₄ -N is ammonia nitrogen, TP is the concentration of total phosphorus, PO ₄ -P is the phosphate phosphorus, COD / TP is the COD / COD ratio per total phosphorus, COD / TN is the COD ratio per total nitrogen.

Based on such manure characteristics, the inventors of the present invention show that the composition is similar to that of general manure, and is a new medium for separating microorganisms capable of efficiently decomposing odor components and high concentrations of organic matter (hereinafter referred to as BH1 medium herein). To synthesize.

BH1 medium according to the present invention is based on water, glucose per liter of medium 4.17g, ammonium sulfate ((NH ₄ ) ₂ SO ₄ ) 1.08g, potassium dihydrogen phosphate (K ₂ HPO ₄ ) 0.37g, urea 0.22g, It contains glucose, ammonium sulfate, potassium dihydrogen phosphate, urea, peptone and sodium thiosulfate at a concentration of 0.17 g peptone and 0.23 g sodium thiosulfate (Na ₂ S ₂ O ₃ ). It is obvious that such ranges are also included within the scope of the present invention, provided that the functionality of the medium as intended in the present invention is the same even if the content of the medium is slightly different from the content given.

Thus, in another aspect, the present invention provides the above-described synthetic medium similar to the manure composition, which is useful for accumulating and separating microorganism communities capable of efficiently decomposing malodorous components and high concentrations of organic matter.

The composition and content of the medium can be verified for its suitability by DGGE or various test methods.

Depending on the DNA sequence difference, the concentration of denaturant required for denaturation of the structure of the double helix will vary. The DGGE method is a molecular biological technique used to analyze the microbial community structure by using the principle that DNA bands denatured at different positions according to DNA sequences during electrophoresis in gels having a denaturation gradient of denaturant. To explain the structural changes of microbial communities in the ecosystem, it has traditionally been based on the count of colonies expressed in the medium, direct counting through fluorescence staining, and the biochemical properties of the microorganisms purely isolated from the sample. Given that only a few of the microorganisms that can be cultivated in natural ecosystems generally exist, approaches based on the separation of microorganisms do not go beyond the limit of incomplete representation of natural state. Thus, molecular biology methods are very useful for understanding the structure and diversity of bacterial communities within a particular ecosystem without microbial culture and isolation. DGGE method extracts DNA directly from environmental samples and amplifies 16S rDNA which can reveal the flexible relationship of bacteria by PCR and analyzes microbial community structure by DGGE method.

In the present invention, the method for obtaining microorganisms that decompose malodor using the BH1 medium is a sample containing a high concentration of organic matter and microorganisms in the soil, manure or foliar soil of a bad malodorous place, BH1 By culturing in a medium to obtain the microorganisms that are actively growing in this medium. The microorganisms actively growing in the medium actively decompose odors or organic substances in the manure environment. In the present invention, in order to formulate microorganisms which are actively growing in the BH1 medium, the microorganisms are fixed to the carrier and the microorganisms are fixed. The carrier is added with amino acids and nutrients, and dried to have a water content of 10% or less. Formulations made in this way are effective because they can minimize the reduction of bacterial counts over a period of time.

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If desired, a specific population of microorganisms may be isolated from the microorganisms obtained by culturing in BH1 medium and used for the preparation of the formulation of the present invention. For example, when microorganisms obtained by culturing in BH1 medium are cultured in a medium that satisfies conditions favorable for growth of ammonia oxidizing bacteria as described below, ammonia oxidizing bacteria can isolate microorganisms that are predominant, and When cultured in a medium that satisfies favorable conditions, sulfur oxidizing bacteria can isolate the predominant microorganisms. As described above, according to the present invention, microorganisms having excellent odor and organic substance decomposition ability can be easily separated from the sample collected in soil, manure or foliar soil of odorous places using the synthetic medium of the present invention.

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Specifically, in order to isolate microorganisms that decompose odorous substances such as ammonia and amines, samples collected from soils in areas where odors are generated are integrated and cultured in the BH1 synthetic medium of the present invention to grow actively even in the presence of high concentration organic matter. The primary integrated culture was obtained, and the culture was inoculated in ammonia oxidizing bacteria separation medium to purely isolate colonies with high growth.

Similarly, the soil samples collected to separate microorganisms that degrade sulfide-based odorous substances such as hydrogen sulfide are integrated and cultured in the BH1 synthetic medium of the present invention to obtain a primary integrated culture medium having high growth in the presence of high concentration of organic matter. Was inoculated in the medium for the separation of sulfated bacteria to isolate pure growth.

As described above, according to the present invention, microorganisms accumulated and cultured in BH1 medium are cultured using ammonia and sulfur compounds as substrates, thereby easily separating microorganisms that efficiently decompose specific substrates.

Preferred media for the cultivation of sulfated bacteria include 5 g sodium thiosulfate pentahydrate, 1.5 g potassium dihydrogen phosphate, 4.5 g dibasic sodium phosphate, 0.3 g ammonium chloride, 0.1 g magnesium sulfate hexahydrate, and Santa rare metals per liter of medium. 5.0 ml of solution. Sintered rare metal solution (pH 6.0) contains 50.0 g of EDTA, 22.0 g of ZnSO ₄ · 7H ₂ O, (NH ₄ ) ₆ Mn ₇ O ₂ · 4H ₂ O 1.10, FeSO ₄ · 7H ₂ O 4.99 g, CaCl · 2H _{2 per} liter O 5.54 g, MnCl ₂ · 4H ₂ O 5.06 g, CuSO ₄ · 5H ₂ O 1.57 g, CoCl ₂ · 6H ₂ O 1.61 g.

The composition of the medium for culturing ammonia oxidizing bacteria according to the present invention is as follows: 0.5 g ammonium sulfate or urea 3.78 mM, potassium monophosphate 1.47 mM, calcium chloride dihydrate 140 mM, magnesium sulfate hexahydrate 160 mM, per liter of medium. HEPES 20 mM, FeNaEDTA 3.8 mg, sodium molybdate 0.1 mg, manganese chloride 0.2 mg, cobalt chloride hexahydrate 0.002 mg, zinc sulfate hexahydrate 0.1 mg, copper sulfate pentahydrate 0.02 mg, sodium bicarbonate 1.5 mM, phenol red 1 mg ( pH 7.5).

In a further aspect, the present invention is adapted to the environment in which the high concentration organic matter is produced by the method described above, can actively grow in the actual excreta and odors such as hydrogen sulfide, ammonia, amines generated in the excreta Provided is a microbial preparation for odor treatment of feces containing a microbial community including microorganisms having excellent resolution of gas and organic matter.

Microorganisms can be identified using a variety of identification methods, such as microbial classification identification methods commonly used in the art, such as Bergey's Manual of Systematic Bacteriology and identification analysis kits (API 20E Strip Kit, Biomerieux).

The preparation obtained by the method of the present invention has the characteristics of effectively decomposing odorous gases such as ammonia, hydrogen sulfide and amines and not being inhibited by high concentration organic matter. As a result of identifying the microorganism to be separated by the method of the present invention, sulfuric acid bacteria and glucose, starch, hemicellulose, soybean oil and degreasing with high decomposition rate of sulfuric acid-based odorous gases such as ammonia oxide bacteria and hydrogen sulfide which efficiently decompose ammonia It was found to contain organic decomposition bacteria having excellent resolution of organic matter including milk powder. Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are intended to illustrate and not limit the invention.

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<Example 1>

Isolation of Microorganisms and Evaluation of Efficacy of Synthetic Media BH1 and Preparation of Microbial Agents

A sample taken from a simple toilet in Daejeon was used as a microbial colony preparation (in situ, B preparation) before treatment, and the sample was used as BH1 medium of the present invention (4.17 g of glucose per liter of medium and ammonium sulfate ((NH ₄ ) ₂ ). SO ₄ ) 1.08 g, potassium dihydrogen phosphate (K ₂ HPO ₄ ) 0.37 g, urea 0.22 g, peptone 0.17 g, sodium thiosulfate (Na ₂ S ₂ O ₃ ) 0.23 g) culture after enrichment (enrichment culture) After the treatment with water, the effect of BH1 medium on the microbial species that dominated the manure in the microbial community preparation (integrated culture, B formulation) was investigated. As a comparative group, a microbial community (A formulation) in a sample collected from a simple toilet in Gwangju, Gyeonggi-do, which was treated with high efficiency without actually generating odors was used.

After inoculating each of the test preparations and the comparative preparations in BH1 medium and incubating at 30 ° C., the cultures were examined for the composition of the microbial community by means of Denaturing Gradient Gel Electrophoresis (DGGE), and the results are shown in FIG. 1. It was. In FIG. 1, the In situ B preparation represents a microbial community configuration before being put into the toilet, and the In situ A preparation represents a microbial community configuration collected in the toilet in which manure treatment is performed without generating an actual odor. As a result of DGGE analysis, it can be seen that microorganisms showing 1, 2, 3, and 4 bands of FIG. As a result of culturing using the medium proposed in the present invention, it can be seen that when cultured using the microorganism of the B preparation as an inoculum, the culture is integrated and cultured in a similar group to the microbial community showing excellent manure treatment ability. The microbial community composition of the B formulation integrated culture in BH1 medium was confirmed to be similar to the microbial community (A formulation) configuration of the toilet was treated with high efficiency unlike before treatment. From these results, it can be seen that the BH1 medium according to the present invention is very suitable for culturing and separating microorganisms for manure treatment. In Example 1, microorganisms of the B agent integrated in BH1 medium are fixed to denatured silt, which is a carrier having excellent deodorizing ability. , Amino acids and nutrients were mixed in a certain amount (0.5-2% for amino acids, 1.5-5% for nutrients) and dried to a water content of 10% or less to make a final product. In the microbial preparations prepared as described above, the final bacterial count was present at 3x10 ⁹ CFU / g at the time of preparation, and after 3 months at room temperature, the degree of bacterial count was decreased and decreased to 8x10 ⁸ CFU / g at 1 month of preservation. There is no significant difference until 3 months thereafter, so it is expected that there will be no problem in commercialization. However, in order to maintain the freshness and efficacy of the product, it is recommended to use it within 2 weeks after manufacture.

<Example 2>

Microbial Identification

To separate microorganisms for decomposition of organic matter, three soil samples were collected, including stalk samples, manure samples from Gwangju, Gyeonggi-do, and oil-polluted soils in Busan, and glucose, starch, hemicellulose, soybean oil, and skim milk powder were diluted by dilution method. Solid medium (Agar 15g / l, (NH ₄ ) ₂ SO ₄ 2g / l, K ₂ HPO ₄ 1g / l, MgSO ₄ · 7H ₂ O 1g / l, CaCl ₂ · 2H ₂ O 0.08g _{/ l, FeCl 2 · 6H 2} O 8.3mg / l, MnCl 2 · 4H 2 O 1.4mg / l, Na 2 MoO 4 · 2H 2 O 1.17mg / l, ZnCl right dropwise after dilution plated for ₂ 1mg / l) Screened colonies. The separated colonies were separated again with a liquid medium ((NH ₄ ) ₂ SO ₄ 2g / l, K ₂ HPO ₄ 1g / l, MgSO ₄ , containing 1% of glucose, starch, hemicellulose, soybean oil, and skim milk powder, respectively). _{7H 2 O 1g / l, CaCl} 2 · 2H 2 O 0.08g / l, FeCl 2 · 6H 2 O 8.3mg / l, MnCl 2 · 4H 2 O 1.4mg / l, Na 2 MoO 4 · 2H 2 O 1.17mg / l, ZnCl ₂ 1mg / l) in order to select a species having excellent growth rate as the final strain, the culture time was limited to 24 hours. As a result, two kinds of strains excellent in specific growth rate (μ) for each substrate were purely separated.

Each strain isolated purely was cultured in large quantities using 5% (v / v) of the medium as a preculture, and identified by examining the morphological, cultural, physiological and chemical taxonomic characteristics with the culture.

As a result, in the present invention, four types of ammonia oxidizing bacterial strains, sulfated bacterial strains and organic substance degrading strains, which were determined to be different in form and biochemical characteristics, were purified.

The colony morphology of some of the selected organic degradation strains showed a milky bulge, and the microscopic observation showed no motility and bacilli. Gram staining showed that both strains were gram positive and catalase positive, and 97% identity with Bacillus bacteria was identified using the API kit. As a result of the resolution analysis by carbon source, one of the strains was excellent in soybean oil but not in the resolution of hemicellulose, while the other strains were good in hemicellulose but not in soybean oil. If you do, it will be complementary to each other.

Similarly, other selected organism degradation strains were identified as the genus Xanthomonas.

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<Example 3>

Degradation Effect of Microorganisms on Organic Matter

In order to investigate the decomposition effect of organic matter according to the microorganism preparation of the present invention, a liquid medium ((NH ₄ ) ₂ SO ₄ 2g / l, K ₂ HPO ₄ 1g / l added glucose, starch, hemicellulose, soybean oil and whole milk powder) , MgSO ₄ · 7H ₂ O 1g / l, CaCl ₂ · 2H ₂ O 0.08g / l, FeCl ₂ · 6H ₂ O 8.3mg / l, MnCl ₂ · 4H ₂ O 1.4mg / l, Na ₂ MoO ₄ · 2H 500 ppm of the microbial preparation of Example 1 was added to 50 ml of ₂ O 1.17 mg / l and 1 mg / l of ZnCl ₂ , and the culture medium was collected by shaking culture at 120 ° C. for 120 hours at 120 ° C. to measure absorbance at 600 nm. The growth rate was presented as a graph (Fig. 2).

As a comparative group, product J (Hipolka-O, Hikokku, Japan) and product A (Custom GT, Custom Biology, USA) were also treated in the same manner as described above, and the growth rates were graphically represented together.

As shown in FIG. 2, the microbial preparation (C) of the present invention showed a faster growth rate in all carbon sources than product J or product A, and particularly, showed high resolution of cellulose and starch in fecal matters in Korea. The applicability of this model is expected to be excellent.

<Example 4>

Microbial Agent Effects by Temperature

In order to compare the growth rate of microbial community with temperature, constant temperature fixed at 4 ° C., 15 ° C., 25 ° C. and 35 ° C. in medium containing 0.2% of glucose, starch, cellulose, soybean oil and whole milk powder respectively as carbon source The growth rate of the microorganisms was measured while shaking incubator for 120 hours. As a comparative group, product J and product A were also treated in the same manner as described above, and the growth rates were shown in the graph of FIG. 3 together.

As can be seen from the graph of Figure 3, the overall microbial preparation of the present invention was shown to be superior to the comparative products in the growth rate according to the temperature, especially the low temperature 15 ℃ and 4 ℃ showed excellent growth in the climate of Korea Due to its characteristics, it is expected to reduce the organic matter in the manure even in winter.

Example 5

The number of microorganisms present in the microbial agent of the present invention

As a result of examining the number of microorganisms present in each product, as shown in Table 3, it was found that the product had various types of organic matter degrading ability and was also excellent in the number of microorganisms.

Number of microorganisms by carbon source for each product (CFU / g-dry) THB SDB CDB LDB PDB AOB SOB Invention (C) 8 X 10 ⁸ 2 X 10 ⁸ 9 X 10 ⁷ 8 X 10 ⁷ 3 X 10 ⁸ 5 X 10 ⁵ 3 X 10 ⁴ Comparative product (J) 3 X 10 ⁸ 5 X 10 ⁷ 6 X 10 ⁵ 9 X 10 ⁷ 4 X 10 ⁷ 2 X 10 ² - Comparative product (A) 8 X 10 ⁸ 9 X 10 ⁷ 3 X 10 ⁷ 4 X 10 ⁷ 2 X 10 ⁸ 4 X 10 ³ - THB: Total heterotrophic bacteria (grown in glucose) SDB: Starch degraded bacteria (grown in soluble starch) CDB: Cellulose degraded bacteria (grown in hemicellulose) LDB: Lipolytic bacteria (grown in soybean oil) Growth) AOB: ammonia oxidizing bacteria (assistance of ammonium salt) SOB: sulfated bacteria (assistance of H ₂ S)

As described above, the microbial preparation prepared by the method and the medium of the present invention is active in a medium containing a high concentration of organic matter, and has an advantage in the actual manure environment, and thus, methane, carbon dioxide and ammonia, amines, hydrogen sulfide, etc. Since the decomposition effect is large, it has an excellent effect in decomposing odor gas generated in the manure treatment process and organic matter in the manure, so it can be effectively used in various manure treatment processes such as biological purification or composting.

1 is a diagram showing the results of Denaturing gradient gel electrophoresis (DGGE) of a microbial community cultured in BH1 medium, which is a synthetic medium according to the present invention.

Figure 2 is a graph showing the measured values obtained by measuring the growth rate for each carbon source of the product by OD ₆₀₀ every 12 hours by selecting one item from the microbial agent and the comparative group presented in the present invention.

Figure 3 is a graph showing the measured value obtained by measuring the growth rate according to the temperature of the product every 12 hours by OD ₆₀₀ with a microbial agent and a comparative group according to the present invention every 12 hours.

Claims (5)

Collecting a sample containing high concentrations of organic matter and microorganisms from soil, manure or foliar soil in a highly malodorous place; The samples were grown on a synthetic medium containing 4.17 g of glucose, 1.08 g of ammonium sulfate, 0.37 g of potassium dihydrogen phosphate, 0.22 g of urea, 0.17 g of peptone, and 0.23 g of sodium thiosulfate per liter of medium. Separating active microorganisms; Fixing the separated microorganisms to a carrier, and mixing the amino acids and nutrients to the carrier to which the microorganisms are immobilized, and drying the microorganisms to a moisture content of 10% or less. Process for the preparation of microbial preparations for The method of claim 1, wherein 0.5 g of ammonium sulfate or urea 3.78 mM, potassium phosphate 1.47 mM, calcium chloride dihydrate 140 mM, magnesium sulfate hexahydrate 160 mM, HEPES 20 mM, FeNaEDTA before fixing the microorganism to the carrier. 3.8 mg, sodium molybdate 0.1 mg, manganese chloride 0.2 mg, cobalt chloride heptahydrate 0.002 mg, zinc sulfate heptahydrate 0.1 mg, copper sulfate pentahydrate 0.02 mg, sodium bicarbonate 1.5 mM, phenol red 1 mg (pH 7.5) 5 g of sodium thiosulfate pentahydrate, 1.5 g of potassium dihydrogen phosphate, 4.5 g of dibasic sodium phosphate, 0.3 g of ammonium chloride, 0.1 g of magnesium sulfate hexahydrate, and sant per liter of ammonia oxidizing bacteria and / or medium And dissociating the microorganism having a rapid growth rate into a medium for sulphating bacteria separation medium containing 5.0 ml of a rare metal solution. delete A synthetic medium based on water, containing 4.17 g of glucose, 1.08 g of ammonium sulfate, 0.37 g of potassium dihydrogen phosphate, 0.22 g of urea, 0.17 g of peptone, and 0.23 g of sodium thiosulfate per liter of medium. delete