WO2008136544A1

WO2008136544A1 - Bacillus sphaericus bc6 kccm 10862p reducing nasty odor and heavy metal and clarification methods for resource recovery from food waste or livestock waste water by using it

Info

Publication number: WO2008136544A1
Application number: PCT/KR2007/002222
Authority: WO
Inventors: Gun Ho Choi; Kil Ho Choi
Original assignee: Pyung Kang Special Vehicle Co., Ltd.
Priority date: 2007-05-04
Filing date: 2007-05-04
Publication date: 2008-11-13
Also published as: KR101106034B1; KR20100019475A

Abstract

The present invention relates to a novel strain Bacillus sphaericus BC6 KCCM 10862P which is highly able to remove nasty odors and heavy metals of livestock wastewater and food waste. This novel strain may be used alone or in combination with other Bacillus species in removing nasty odors and heavy metals. It can also be used to convert livestock wastewater or food waste into useful resources.

Description

[Description] [invention Title]

BACILLUS SPHAERICUS BC6 KCCM 10862P REDUCING NASTY ODOR AND HEAVY METAL AND CLARIFICATION METHODS FOR RESOURCE RECOVERY FROM FOOD WASTE OR LIVESTOCK WASTE WATER BY USING IT [Technical Field]

The present invention relates to a novel microorganism species useful for reducing nasty odors and heavy metals in livestock wastewater and food waste. Also, the present invention is concerned with the use of this novel microorganism in converting livestock wastewater and food waste into useful resources. In detail, the present invention relates to a novel microorganism species capable of removing nasty odors and heavy metals from livestock wastewater and food waste, and to a method for the production of organic liquid fertilizer from livestock wastewater, and to a method for the production of animal feeds from food waste. [Background Art]

The excretions from a body of livestock generate nasty odors, which also permeate the stalls where they are bred. In addition to acting as air pollutants incurring global warming, the nasty odor is harmful to humans and animals. The nasty odor is attributed, for the most part, to livestock feces, which can be divided, by microorganisms, into easily degradable matter such as carbohydrates, starch, proteins and cellulose, and sparingly degradable matter such as lipids, lignin, etc.

To the malodor generated upon the degradation of livestock feces under anaerobic conditions, various compounds including ammonia, methyl mercaptan, hydrogen sulfide, methyl sulfide, methyl disulfide, trimethyl amine, acetaldehyde, styrene, propionic acid, lactic acid, and valeric acid make a contribution. For example, methyl mercaptan provides an odor of rotten onions . Hydrogen sulfide is an irritative, suffocative and colorless gas, which also gives off the smell of rotten onions. Ammonia, an irritative and colorless gas, is responsible for the characteristic urine smell. In combination, they give out a complex foul smell, disgusting persons and inhibiting the giowth of the livestock.

There is a need for regulating the generation of the nasty smell because public grievances attributed to the nasty smell tend to increase. In fact, strict legal regulations are enacted against the generation of such a nasty smell. Examples of the facilities which produce intensive and extensive nasty odors include livestock wastewater disposal plants, food waste disposal plants, n.ghtsoil treatment plants, waste burial sites, and animal feed manufacturing plants. In such facilities, generally, odor is produced not from certain processes but rather from the processes taken as a whole, which makes it greatly difficult to design process management, modify the facilities, and change the construction to reduce the nasty odor. Further, because such processing facilities are, in most cases, present in residential districts, a highly efficient method for reducing nasty odors is required in order to minimize negative effects on the surroundings.

Now, legal regulations are enacted against the generation of eight malodor compounds: hydrogen sulfide

(H₂S), methyl mercaptan (CH₃SH), methyl sulfide ( (CH₃) ₂S), methyl disulfide ( (CH₃) ₂S₂), ammonia (NH₃), trimethyl amine ( (CH₃) ₃N), acetaldehyde (CH₃CHO), and styrene (C₆H₅CH=CH₂).

These compounds, unlike other organics, are known to produce a strong stink even at low concentrations .

Particularly, the nasty odor generated from animal stalls, such as pigsties, cattle pens, chicken farms, etc. is contributed mainly by hydrogen sulfide (H₂S) while ammonia

(NH₃), methyl mercaptan (CH₃SH), trimethyl amine ( (CH₃) ₃N) and lower fatty acids make it worse.

On the whole, the malodor of a matter results from a combination of the individual smells of the component compounds. Hydrogen sulfide is responsible for the generation of a smell similar to rotten eggs. Mercaptan is a main ingredient of the stink of rotten vegetables. Amines contribute to the bad smell of rotten fish.

Methods for reducing nasty odor are largely divided into two categories: separation or destruction of malodor ingredients by, for example, oxygenation, enzymatic degradation or absorption; and simple suppression such as masking. Oxygenation oxygenates and degrades raalodor ingredient^'s and sterilizes bacteria, fungi and viruses so as to fundamentally remove odor resources. Many methods have been suggested for oxygenation. Most chemical oxygenation methods use chlorine dioxide, sodium hypochlorite, or chlorate dioxide. However, these compounds, containing chlorine may be injurious to humans and animals. Enzymatic degradation methods are known to use deodorizers prepared from vegetable extracts, but their ingredients and operation principles are not disclosed. Absorption methods, based on absorbents with large surface areas, such as active carbon, decrease in efficiency with time because available surface areas of the absorbents drop after being in use for a period of time. Masking methods in which natural or synthetic odorous compounds are dissolved in highly volatile solvents, such as methyl alcohol, to generate a strong odor which conceals a nasty odor, can solve the nasty odor problem temporarily, but cannot provide a fundamental solution thereto. Instead of such chemical materials, ozone or UV radiation is used alone or in combination as a deodorizing agent. Ozone, although useful for sterilization and deodorization, may itself act as a pollutant. Also, there has been suggested a method in which a photocatalyst, such as TiO₂, is used, in combination with ozone, to degrade odor sources. However, a limitation is imposed on an increase in the surface area of photocatalysts, such as TiO₂. Further, when applied to the treatment of sulfides, such as hydrogen sulfide (H₂S) , methyl mercaptan (CH₃SH) , methyl sulfide

( (CH₃) ₂S), and methyl disulfide ( (CH₃) ₂S₂) , the photocatalysts, such as Ti02, are readily poisoned thereby, subsequently suffering from the disadvantage of a rapidly decreasing catalyst efficiency.

In general, various industry plants, such as petrochemical plants, sewage/excretion disposal plants, rubber manufacturing plants, painting plants, food processing plants, livestock treatment plants, etc. generate BTXs including benzenes, toluene and xylene, and volatile organic compounds (VOCs) including organic acids, aldehydes, ketones, aromatic compounds, etc. in addition to giving off nasty odors. Further, the public grievance raised due to these plants prevents or delays the construction of such industrial operations.

The malodor and volatile organic chemicals (VOCs) generated from industrial plants are generally removed using chemical or biological methods. An incineration method produces secondary air pollution while an absorption method is economically disadvantageous due to the use of active carbon. These chemical methods require a low expense for the construction of facilities therefor, but a high expense for the operation thereof. In contrast, biological methods, although suffering from the disadvantage of requiring high expenses for the construction of facilities therefor and complicated conditions for the operation thereof, are advantageous in that they are easy to maintain and manage and can be operated at low cost. A biofliter method, typical of biological deodoπzation, uses a bioreactor loaded with a carrier onto which deodorizing microorganisms are immobilized. The lowest operation cost is found in the biofilter system. Operation costs increase m the order of bioflltration, chemical cleaning, catalytic incineration, and direct combustion.

Korean Patent Application No. 10-1996-0025606 (filed on June 29, 1996) discloses a microbial composition for the biological treatment of food waste, comprising thermoresistant filamentous fungi adapted to high temperatures, thermophilic bacillus bacteria which degrade proteins and lipids, and thermoresistant actinomyces; and a method for composting food waste.

Korean Patent No. 292879 (filed on April 7, 1999) pertains to a microorganism composition and a method for the treatment of livestock excretions using the same. The microorganism composition comprises a bacillus species (Bacillus subtilis KCTC 1028) which is highly capable of pioducmg an enzyme useful in the degradation of carbohydrates and proteins, and a photosynthetic microorganism (Rhodopseudomonas spp., deposited in the Korean Collection for Type Cultures with the accession number of KCTC 8937P) which can grow on the lysates and deodorize livestock excretions. Thereby, the use of m. croorganism agents offers an improvement in growth conditions for livestock and living environments for stock farmers .

Korean Patent No. 378667 (filed on Aug. 31, 2000) discloses a composition and a method for deodorizing and composting livestock excretions. The composition is used to fermentatively degrade livestock excretions or food waste into organic manure, thereby removing the nasty odor of livestock excretions and the smell generated at an early putrefaction stage of food waste. The deodorizing composition is prepared by aerobically culturing at least one species selected from the group consisting of St reptomyces spp., Bacillus spp., Micrococcus spp., Achromobacter spp., Flavobacteπum spp., Escherichia spp. and Pseudomonas spp. in a general broth such as nutrient broth or TSB broth, adsorbing the culture onto a medium selected from among limestone, bentonite and humus soil, and drying it.

Korean Patent No. 433268 (filed on Oct. 5, 2000) dαscloses a microbial composition for the biological tieatment of malodorants and a method for the preparation thereof. The microbial composition comprises deodorizing πu croorganisms composed of a thermoresistant filamentous fungal species adapted to organic matter and high temperatures; another filamentous fungal species adapted to substrate change; thermoresistant and thermophilic bacillus capable of growing on a broad range of substrates; and thermoresistant Actinomyces adapted to a high concentration of organic substances. The thermoresistant filamentous fungal species is at least one microorganism selected from the group consisting of Aspergillus niger, Aspergillus oryzae, Penicillium spp., Mucor spp., Trichoderma viride, Cephallosporium acremonium, and Issatchenkia orientalis . The filamentous fungal species adapted to substrate change is selected from among Candida spp., Arthrobacter spp., and Nocardia spp. The thermoresistant and thermophilic bacillus capable of growing on a broad range of substrates may be Bacillus subtilis CH-IO, Bacillus mesentericus, Bacillus megaterium, Bacillus licheniforwmis NLRI-33, Bacillus thuringiensis, Bacillus circulans or a combination thereof. The thermophilic actinomyces adapted to high concentrations of organic substances is Thermoactinomyces spp. and/or Thermomonspora spp. Preferably, the thermoresistant actinomyces adaptable to high concentrations of organic substances is selected from among Streptomyces griseus, Streptomyces globisporius, Streptomyces flavus and a combination thereof. The bacterial species able to grow on a broad range of substrates is described to be preferably selected from the group consisting of Pseudomonas putida NLRI-X50, Corynebacterium spp., Staphylococcus lentus, Micrococcus lylae, and a combination thereof.

Korean Patent Application No. 10-2005-0044120 (filed on May 25, 2005) presents an air cleaning apparatus for removing stench and sand dust from inside a barn, with the application of Bacillus subtilis for deodorization. Korean Patent Application No. 10-2006-0032543 (filed

April 11, 2006) suggests a method and an apparatus for efficiently treating malodorous waste air using a hybrid system composed of a photo-catalytic reactor, fluidized aerobic and anaerobic reactors and a biofilter.

Korean Patent Application No. 10-2004-0011349 (filed Feb. 2, 2004) suggests a Geotrichum-immobilized biofilter and a method for removing methylethylketone using the same, disclosing that the removal of methylethylketone with the Geotrichum-immobilized biofilter requires a lower cost than do conventional methods using, for example, active carbon or incineration, and does not produce nitrogen oxides or secondary pollutants.

Korean Patent Application No. 10-2003-0078402 (filed Nov. 6, 2003) provides a biofilter system for the removal of malodor gases and volatile organic compounds, with the application of Pseudomonas spp. to the removal of volatile organic compounds .

[Disclosure] [Technical Problem]

It is an object of the present invention to provide a novel bacterial strain, identified as Bacillus sphaericus BC6 KCCM 10862P, capable of reducing malodor and heavy- metals in livestock wastewater and food waste. It is another object of the present invention to provide a microbial composition for the removal of malodor and heavy metals, comprising Bacillus sphaericus BC6 KCCM 10862P as an active ingredient, and a method for converting livestock wastewater or food waste into a useful resource using the same .

[Technical Solution] In accordance with an aspect thereof, the present invention provides a novel strain Bacillus sphaericus BC6 KCCM 10862P, which has the ability to deodorize livestock wastewater and food waste faster and more effectively than conventional strains, and to remove heavy metals. In accordance with another aspect, the present invention provides a microorganism composition for removing nasty odor and heavy metals, comprising the novel strain Bacillus sphaericus BC6 KCCM 10862P as an active ingredient.

The novel strain Bacillus sphaericus BC6 KCCM 10862P can greatly reduce the nasty odor of livestock wastewater or food waste. Further, this microorganism can remove heavy metals from livestock wastewater or food waste. Having such abilities, the microorganism composition according to the present invention may be applied for the clarification of livestock wastewater and food waste. Further, it can be used to prepare organic liquefied fertilizer from livestock wastewater and prepare animal feed from food waste .

[Description of Drawings]

FIG. 1 is a schematic diagram showing a device used m the experiments to determine the ability to deodorize 1 Lvestock wastewater.

FIG. 2 is a schematic diagram showing a device used in the experiments to determine the ability to deodorize food waste. [Best Mode]

In accordance with an embodiment thereof, the present invention provides a novel bacterial strain, identified as Bacillus sphaericus BC6 KCCM 10862P, which is useful in deodoπzation and heavy metal removal.

In accordance with another embodiment thereof, the present invention provides a microbial composition for the removal of malodor and heavy metals, comprising an effective amount of the novel strain Bacillus sphaericus BC6

KCCM 10862P.

Conventional techniques may be applied for the formulation of the bacterial strain Bacillus sphaericus BC6 KCCM 10862P according to the present invention. The bacterial strain may be formulated into a solid pieparation, such as beads, or a liquid preparation. Solid preparations suffer from the disadvantage of causing an inhomogeneous distribution of the active ingredient therein, decreasing microbial activity and leading to a reduction in the applicability thereof, and generating dust during the manufacture thereof. On the other hand, liquid preparations have advantages over the solid preparations in that they can elicit microbial activity faster and are convenient for treating, but are inferior to solid preparations m terms of preservability .

For preparing the bacterial strain into a bead formulation may be used a method comprising: (a) dropping a mixture of a 0.1-5.0 wt% solution of a negatively charged biodegradable polymer, such as alginate, pectin, carrageenan or polyaspartic acid, and a Bacillus sphaericus BC6 KCCM 10862P culture into a 0.5-3.0 wt% solution containing a metal salt such as Ca²⁺, Cu²⁺, Zn²⁺, Ni²⁺, Co²⁺, Mn²⁺, Al²⁺, Fe²⁺, or Mg²⁺ to form beads; and (b) electrostatistically coating the beads with a 0.1-3.0 wt% solution of a positively charged biodegradable polymer such as chitosan, a chitosan derivative or polylysine by stirring the beads within the solution for 0.5 ~ 2 hrs . After freeze drying, the coated beads may be used. Alternatively, they may be subjected to additional processes for improving the survival rate and activity of the microorganism, including (c) washing the beads with 50 mM sodium acetate (pH 5.5) to remove an excess of the positively charged biodegradable polymer; (d) stirring the bead within a 0.1 - 2.0 wt% solution of a negatively charged polymer; and (e) washing the beads with saline and stirring them within a 0.1 - 2.0 wt% solution of a positively charged biodegradable polymer to form a thicker outer film thereon.

The microbial composition for the removal of malodor and heavy metals in accordance with another embodiment of the present invention comprises the novel strain Bacillus sphaericus BC6 KCCM 10862P in a sufficiently effective amount to conduct the above-mentioned functions, in combination with one or more Bacillus strain selected from the group consisting of Bacillus megaterium BCl-I KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P.

In accordance with a further embodiment thereof, the present invention provides a method for reducing malodor or heavy metals of food waste or livestock wastewater, comprising culturing the novel strain Bacillus sphaericus BC6 KCCM 10862P with malodorous food waste or livestock wastewater .

In still another embodiment of the present invention, food waste and/or livestock wastewater are deodorized and/or depossessed of heavy metals using a method in which a microbial composition comprising a sufficiently effective amount of the novel strain Bacillus sphaericus BC6 KCCM 10862P, in combination with one or more Bacillus strain selected from the group consisting of Bacillus megaterium BCl-I KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 10860P, Bacillus cereus BCS KCCM 10861P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM 10864P, Bacillus fjrmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P, and a combination thereof, is cultured with malodorous food waste and/or livestock wastewater.

In still a further embodiment thereof, the present invention provides a method for composting livestock wastewater into organic liquefied fertilizer and transforming food waste into animal feeds using a sufficiently effective amount of the novel strain Bacillus sphaericus BC6 KCCM 10862P.

Also, the production of organic liquefied fertilizer fiom livestock wastewater and animal feeds from food waste may be achieved using a conventional process without the use of the novel Bacillus sphaericus BC6 KCCM 10862P. As for oiganic liquefied fertilizer, for example, it may be prepared from livestock wastewater using peat. In detail, a conventional method comprises pulverizing and mixing peat, oi game materials, oyster and coal ash at a proper ratio, adding sodium hydrogen carbonate, calcium oxide and calcium chloride to the mixture in effective amounts to achieve deodoπzation, extracting organic liquefied fertilizer, neutralizing the extracted organic liquefied fertilizer with diluted hydrochloric acid, and aerating the neutralized organic liquid to decrease the chemical oxygen demand thereof . On the basis of the novel strain Bacillus sphaericus BC6 KCCM 10862P, a dry fermentation method, a mixed fermentation method, a combination thereof, or a liquid- phase fermentation method may be provided for the pieparation of animal feeds from food waste. The dry fermentation method greatly reduces the food waste m mass, but demands an inordinate amount of energy for drying and pioduces nutritionally insufficient feeds which thus cannot be used per se. Further, the resulting feeds are difficult to pack and apply to an automatic feeding line due to the hα gh water content (around 40%) thereof. The mixed fermentation method allows the animal feeds to be balanced nutritionally, but is unable to produce various animal feeds meeting the demand for different nutrient contents according to age. The use of mixed and dry fermentation methods m combination provides ease in maintaining a nutritional balance and adjusting water content in the resulting animal feed, but requires high production cost because expense must be paid for drying and water content adjustment. The liquid-phase fermentation method, typically resulting m an animal feed with a water content of around 8b%, has an advantage over the other methods in terms of production cost, but the products are inconvenient to transport .

In yet another embodiment thereof, the present invention provides a method for producing organic liquefied fertilizer, from livestock wastewater, and animal feeds, from food waste, using a microbial composition comprising an effective amount of Bacillus sphaericus BC6 KCCM 10862P and one or more Bacillus strain selected from the group consisting of Bacillus megaterium BCl-I KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P, and a combination thereof.

Hereinafter, the present invention will be described in further detail with reference to examples . It is to be understood, however, that these examples are for illustrative purposes only and are not to be construed to limit the scope of the present invention.

[Mode for Invention]

[Example 1] Isolation and Identification of Bacillus sphaericus BC6 KCCM 10862P

Bacillus sphaericus BC6 KCCM 10862P, the principle of the present invention, was taken from mushrooms and hyphae grown on fallen leaves in a forest located in Samgi, Iksan, Korea. The mushrooms and hyphae were immersed in a culture medium comprising 5 g of black sugar and 300 ml of water in a 500 ml beaker before incubation at 28 - 30⁰C. It was found that the nasty odor ceased in 24 hours after the application of a portion of the culture to a sewage treatment tank, showing the deodorizing ability of the culture. When the culture was maintained in molasses and applied to food waste, it was also found to be highly effective in deodorizing food waste. From the culture, useful microorganisms were isolated.

In a microbiology lab of Kunsan National University, 12 microorganism strains were isolated in total from the culture using an NA medium. All of them were found to belong to Bacillus. After being sub-cultured to purity in an NA medium, each of the twelve strains was stored individually. Analysis for 16S rDNA in the Korea Research Institute of Bioscience and Biotechnology, Taejeon, Korea identified the 12 isolated Bacillus strains as Bacillus megaterium BCl-I KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus lichenifoimis BC4 KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus sphaericus BC6 KCCM 10862P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P, respectively, which were all deposited in the Korean culture center of microorganisms on April 27, 2007, with the respective accession numbers.

[Example 2] Deodorizing Effect of Bacillus sphaericus BC6 KCCM 10862P Alone or in Combination with other Bacillus Strains on Livestock Wastewater

In order to evaluate the ability of Bacillus sphaericus BC6 KCCM 10862P to reduce the malodor of livestock wastewater, Bacillus clausii BC7 KCCM 10863P was added alone or in combination with Bacillus megaterium BCl- 1 KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus sphaericus BC6 KCCM 10862P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P to microorganism reaction vessels containing livestock wastewater (see FIG. 1) and the gas of the vessels was collected 4 days after the addition and then analyzed.

Conditions for guaranteeing the microorganisms to actively grow in the reaction vessels included aeration with aerator and temperature maintenance at 37.5°C with a heating band and an automatic temperature controller (see FEG.1) .

Before the addition of microorganisms, the malodor of raw livestock wastewater was analyzed for ingredient concentrations to be used as monitoring reference. Two liters of wastewater was placed in each reaction vessel. 5 ml of a culture of Bacillus sphaericus BC6 KCCM 10862P and 5 mJ of a mixture containing equal amounts of Bacillus megatenum BCl-I KCCM 10856P, Bacillus megatenum BCl-2 KCCM 10857P, Bacillus megatenum BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 1086OP, Bacillus cereus BC5 KCCM 10861P, Bacillus sphaericus BC6 KCCM 10862P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P were added to respective vessels while 5 ml of the wastewater was added to another reaction vessel, serving as a control .

(1) Assay Method

1) Quantification of Ammonia

To an assay sample were added a phenol-sodium nitroprusside solution and a sodium hydrochlorite solution, both of which react with ammonium ions to produce mdophenols which are suitable for spectrophotometry to quantify ammonia. A. Reagents - Collection solution

5 g of boric acid was dissolved in distilled water to produce a total volume of 1 L. Phenol, disodium pentacyanonitrosyl ferrate (III) solution

500 mL of a solution of 5 g of phenol and 25 mg of disodium pentacyanonitrosyl ferrate (III) dehydrate in distilled water was prepared. This solution was stored in a cold and dark place and discarded if it was prepared one month ago or earlier.

- Sodium hypochlorite solution

60/CmL (effective chlorine content 3 ~ 10%) of sodium hypochlorite [C represents an effective chlorine concentration (unit %) of sodium hypochlorite as measured upon preparation], 10 g of sodium hydroxide, and 35.8 g of sodium hydrogen phosphate dodecahydrate were dissolved in distilled water to produce a total volume of 1 L. This solution was freshly prepared just before each use. B. Solution absorption

An absorption bottle equipped with a glass frit impactor having a capacity of 20 ml or greater was used. Two absorption bottles, each containing 20 ml of a collection solution in the mid thereof, were connected in series. After being installed onto the absorption bottles, a suction pump was operated to aspirate air at a rate of 10 L/min or higher. After the air collection, the collection solutions contained in the respective bottles were pooled and transferred into a 50 mL flask. The bottles were washed with a collection solution which was then added to the flask to give a total volume of 50 mL. 10 mL of this solution was aliquoted into a test tube and used as a test sample solution.

C. Measurement of absorbance of test sample solution The test sample solution was mixed with 5 ml of phenol pentacyanonitrosyl ferrate (III) solution by agitation and then with 5 ml of a sodium hypochlorite solution and was allowed to stand for 1 hour at 25 ~ 30⁰C, followed by measuring absorbance at 640 run. As a blank test sample, 10 mL of the collected solution (absorption solution) was used in the same manner as in the test sample solution .

2) Quantitative Assay for methylmercaptan, hydrogen sulfide, dimethyl sulfide and dimethyl disulfide In order to quantitatively assay sulfide, a sample gas was adsorbed and concentrated at -170⁰C with an SPT (Sample Preconcentration Trap) , desorbed by heating, and injected into gas chromatography equipped with a PFPD (Pulse Flame Photometric Detector) .

3) Quantitative Assay for trimethyl amine

A test sample solution prepared by absorbing a sample into an aqueous sulfate solution (359+1) was partially taken with a volumetric syringe and injected into a silicon stopper-plugged degradation bottle containing a potassium hydroxide solution. The solution was bubbled by injecting 2 ~ 3 L of nitrogen at a rate of 0.2 ~ 0.3 L/min to generate trimethyl amine which was then concentrated at a low temperature in a condenser. Trimethyl amine was desorbed by heating the concentration to a temperature of 7O⁰C and introduced into a column for FID analysis.

4) Quantitative Assay for aldehyde

A sample was passed through a sample collection tube filled with 2.4-dinitrophenyl hydrazine-coated octadecylsilylated silica gel. After the evaporation of acetonitrile from the sample collection tube, aldehydes captured by the collection tube were dissolved in ethyl acetate and a portion thereof was introduced into HPLC (Young Lin Instrument Co. Ltd.) and analyzed.

(2) Analysis Results

Table 1 were summarized the analysis results obtained after culturing Bacillus sphaericus BC6 KCCM 10862P alone and in combination with other Bacillus strains for 4 days in wastewater.

< TABLE 1 > Deodorizing Effect of Bacillus sphaericus BC6 KCCM 10862P on Livestock Wastewater (unit ppm)

As is apparent from the data of Table 1, Bacillus sphaericus BC6 KCCM 10862P showed excellent ability to remove ammonia, hydrogen sulfide, methyl mercaptan, dimethyl sulfide, dimethyl disulfide and trimethyl amine. However, this bacteria was found to be unable to remove acetaldehyde as similar concentrations were detected for acetaldehyde between the samples and the control .

[Example 3] Deodorizing Effect of Bacillus sphaericus BC6 KCCM 10862P Alone or in Combination with other Bacillus Strains on Food Waste

In order to evaluate the ability of Bacillus sphaericus BC6 KCCM 10862P to reduce the malodor of food waste, Bacillus sphaericus BC6 KCCM 10862P was added alone or in combination with Bacillus megaterium BCl-I KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus sphaericus BC6 KCCM 10862P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P to microorganism reaction vessels containing food waste (see FIG. 2) and the gas of the vessels was collected 4 days after the addition and then analyzed.

Conditions for guaranteeing the microorganisms to actively grow in the reaction vessels included temperature maintenance at 37.5°C with a heating band and use of an automatic temperature controller (see FIG.2).

Before the addition of microorganisms, the ingredient concentrations of the malodor of raw food waste were analyzed to be used as a monitoring reference. Two liters of food waste was placed in each reaction vessel. 5 ml of a culture of Bacillus sphaericus BC6 KCCM 10862P and 5 ml of a mixture containing equal amounts of Bacillus megaterium BCl-I KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus sphaericus BC6 KCCM 10862P, Bacillus clausii BC7 KCCM 10863P, Bacillus Ijcheniformis BC8 KCCM 10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P were added to respective vessels while 5 ml of the food waste was added to another reaction vessel, serving as a control.

The malodor of food waste was analyzed in a similar manner to Example 2, the analysis of livestock wastewater. The results are summarized in Table 2, below.

TABLE 2 > Deodorizing Effect of Bacillus sphaericus BC6 KCCM 10862P on Food Waste (unit ppm)

As is apparent from the data of Table 1, Bacillus sphaericus BC6 KCCM 10862P showed excellent ability to remove ammonia, hydrogen sulfide, methyl mercaptan, dimethyl sulfide, dimethyl disulfide and acetaldehyde. However, this bacteria was found to be unable to effectively remove trimethyl amine, as similar concentrations of these chemicals were detected between the samples and the control.

[Example 4] Analysis for Reducing Heavy Metals from Livestock Wastewater and Food Waste

In order to evaluate the ability of Bacillus sphaericus BC6 KCCM 10862P to reduce heavy metals of food waste, two liters of food waste was placed in each reaction vessel and 5 ml of a culture of Bacillus sphaericus BC6 KCCM 10862P and 5 ml of a mixture containing equal amounts of Bacillus megaterium BCl-I KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus sphaericus BC6 KCCM 10862P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P were added to the respective vessels while 5 ml of the food waste was added to another reaction vessel, serving as a control. The same above conditions were applied for the analysis of livestock wastewater. The food waste and the livestock wastewater were quantitatively analyzed for heavy metals before and after the incubation of the microorganisms for 4 days. The analysis was performed in the following manner. (1) Preparation of reagent

- Ternary solution: HNO₃, H₂SO₄, and HClO₄ were mixed at a ratio of 10:1:4 according to the quantity needed. - concentrated HNO₃

(2) Decomposition

10 mL of a sampled specimen was placed in a 25OmL Erlenmeyer flask, added with 10 ~ 30 mL of concentrated HNO₃, and left overnight. Then, while being slowly heated on a hot plate, the sample was dried until a white precipitates were formed. After cooling, 10 ~ 50 mL of the ternary solution was added to the precipitates . By heating on a hot plate, H₂SO₄ and HClO₄ were evaporated off as a white gas. When the solution turned white or transparent brown, heating was stopped. The solution was cooled and poured through a No. 6 filter into a 50 mL or 100 mL flask. The Erlenmeyer flask was rinsed with hot distilled water, and the residue was scraped off with rubber policemen and filtered through the filter.

(3) Measurement

A portion of the filtrate was taken and diluted. Calibration curves were constructed for K, Ca, Mg, Na, Fe, Mn, Cu, Zn, Cr, Cd, Pb, and Ni using respective standard solutions . in an Atomic Absorption Flame Emission Spectrometer or ICP (Inductively Coupled Plasma) , and used to determine the levels of the metals in the dilution. < TABLE 3 > Ability of Bacillus sphaericus BC6 KCCM 10862P to Reduce Heavy Metals of Livestock Wastewater (Units of ppm)

< TABLE 4 > Ability of Bacillus sphaericus BC6 KCCM 10862P to Reduce Heavy Metals of Food Waste (Units of ppm)

Bacillus sphaericus BCβ KCCM 10862P was found to greatly remove cadmium, chromium, copper, zinc and lead from livestock wastewater. Also, the bacteria showed a high ability to remove cadmium, copper and zinc from food waste. Nickel and lead, however, was detected to be of a higher concentration in food waste, which was contrary to the result of livestock wastewater, leaving the nickel and lead-removing ability of Bacillus sphaericus BC6 KCCM 10862P unclear . [industrial Applicability] As described above, Bacillus sphaericus BC6 KCCM 10862P according to the present invention is very useful in deodorizing livestock wastewater and food waste. As compared to conventional microbial mixtures which typically require a period of 30 days for deodoπzation (data not shown) , the novel bacterial strain of the present invention is highly effective at removing the nasty odor of livestock wastewater and food waste. In addition, the strain of the present invention is capable of very effectively removing heavy metals from livestock wastewater or food waste.

Claims

[CLAIMS]

[Claim l]

A novel strain Bacillus sphaericus BC6 KCCM 10862P for reducing nasty odors and heavy metals.

[Claim 2]

A microorganism composition for reducing nasty odors and heavy metals, comprising an effective amount of a novel strain Bacillus sphaericus BC6 KCCM 10862P.

[Claim 3] A microorganism composition for reducing nasty odors and heavy metals, comprising an effective amount of a novel strain Bacillus sphaericus BC6 KCCM 10862P and one or more

Bacillus species selected from the group consisting of

Bacillus megaterium BCl-I KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P,

Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4

KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM

10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P, and a combination thereof.

[Claim 4]

A method for reducing nasty odors or heavy metals from livestock wastewater or food waste, comprising culturing a novel strain Bacillus sphaericus BC6 KCCM 10862P with the livestock wastewater or food waste.

[Claim 5]

A method for reducing nasty odors or heavy metals from livestock wastewater or food waste by culturing a microorganism composition, comprising an effective amount of a novel strain Bacillus sphaericus BC6 KCCM 10862P and one or more Bacillus species selected from the group consisting of Bacillus megateπum BCl-I KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P, and a combination thereof with the livestock wastewater or food waste .

[Claim 6] A method for producing organic liquefied fertilizer, using an effective amount of novel strain Bacillus sphaericus BC6 KCCM 10862P m livestock wastewater.

[Claim 7]

A method for producing organic liquefied fertilizer, using an effective amount of novel strain Bacillus sphaericus BC6 KCCM 10862P and one or more Bacillus species selected from the group consisting of Bacillus megaterium BCl-I KCCM 10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus clausii BC7 KCCM 10863P, Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9 KCCM 10865P and Bacillus cereus BClO KCCM 10866P, and a combination thereof in livestock wastewater.

[Claim 8]

A method for preparing livestock feed from food waste, using an effective amount of a novel strain Bacillus sphaericus BC6 KCCM 10862P. [Claim 9]

A method for preparing livestock feed from food waste, using a microorganism composition comprising an effective amount of a novel strain Bacillus sphaericus BC6

KCCM 10862P and one or more Bacillus species selected from the group consisting of Bacillus megaterium BCl-I KCCM

10856P, Bacillus megaterium BCl-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus cereus BC3 KCCM 10859P, Bacillus licheniformis BC4 KCCM 10860P, Bacillus cereus BC5 KCCM 10861P, Bacillus clausii BC7 KCCM 10863P,

Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9

KCCM 10865P and Bacillus cereus BClO KCCM 10866P, and a combination thereof.