WO2008146961A1 - Bacillus cereus bc3 kccm 10859p reducing nasty odor and heavy metal and clarification methods for resource recovery from food waste or livestock waste water by using it - Google Patents
Bacillus cereus bc3 kccm 10859p reducing nasty odor and heavy metal and clarification methods for resource recovery from food waste or livestock waste water by using it Download PDFInfo
- Publication number
- WO2008146961A1 WO2008146961A1 PCT/KR2007/002224 KR2007002224W WO2008146961A1 WO 2008146961 A1 WO2008146961 A1 WO 2008146961A1 KR 2007002224 W KR2007002224 W KR 2007002224W WO 2008146961 A1 WO2008146961 A1 WO 2008146961A1
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- kccm
- bacillus
- bacillus cereus
- food waste
- cereus
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- 239000011368 organic material Substances 0.000 description 1
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- 229910052760 oxygen Inorganic materials 0.000 description 1
- 235000020636 oyster Nutrition 0.000 description 1
- 238000010422 painting Methods 0.000 description 1
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- 229960000292 pectin Drugs 0.000 description 1
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 230000001699 photocatalysis Effects 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 108010064470 polyaspartate Proteins 0.000 description 1
- 229920000656 polylysine Polymers 0.000 description 1
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- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000004094 preconcentration Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
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- 238000010298 pulverizing process Methods 0.000 description 1
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- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
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- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- NESLWCLHZZISNB-UHFFFAOYSA-M sodium phenolate Chemical compound [Na+].[O-]C1=CC=CC=C1 NESLWCLHZZISNB-UHFFFAOYSA-M 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
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- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
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- 230000004083 survival effect Effects 0.000 description 1
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- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/345—Biological treatment of water, waste water, or sewage characterised by the microorganisms used for biological oxidation or reduction of sulfur compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE NOT OTHERWISE PROVIDED FOR
- B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/40—Treatment of liquids or slurries
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/32—Nature of the water, waste water, sewage or sludge to be treated from the food or foodstuff industry, e.g. brewery waste waters
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2303/00—Specific treatment goals
- C02F2303/02—Odour removal or prevention of malodour
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/085—Bacillus cereus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/10—Bacillus licheniformis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/11—Bacillus megaterium
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/141—Feedstock
- Y02P20/145—Feedstock the feedstock being materials of biological origin
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Definitions
- 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.
- 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.
- 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.
- the nasty odor generated from animal stalls such as pigsties, cattle pens, chicken farms, etc. is contributed mainly by hydrogen sulfide (H 2 S) while ammonia
- Oxygenation oxygenates and degrades malodor ingredients 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 aieas, 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 deadly odor, can solve the nasty odor problem temporarily, but cannot pi ovide a fundamental solution thereto.
- ozone or UV radiation is used alone or in combination as a deodorizing agent. Ozone, although useful for sterilization and deodo ⁇ zation, may itself act as a pollutant.
- a photocatalyst such as TiO 2
- ozone a method in which a photocatalyst, such as TiO 2 , is used, in combination with ozone, to degrade odor sources.
- a limitation is imposed on an increase in the surface area of photocatalysts, such as TiO 2 .
- the photocatalysts, such as TiO2 are readily poisoned thereby, subsequently suffering from the disadvantage of a rapidly decreasing catalyst efficiency.
- BTXs including benzenes, toluene and xylene
- VOCs volatile organic compounds
- VOCs volatile organic chemicals
- An incineration method produces secondary air pollution while an absorption method is economically disadvantageous due to the use of active carbon.
- chemical methods require a low expense for the construction of facilities therefor, but a high expense for the operation thereof.
- 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 deodorization, uses a bioreactor loaded with a carrier onto which deodorizing microorganisms are immobilized. The lowest operation cost is found m the biofilter system. Operation costs increase in the order of bioflltration, chemical cleaning, catalytic incineration, and direct combustion.
- Korean Patent Application No. 10-1996-0025606 discloses a microbial composition for the biological treatment of food waste, comprising thermoresistant filamentous fungi adapted to high temperatures, thermophilic bacillus bacteria which degrade piotems 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 producing 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.
- the use of mi croorganism agents offers an improvement in growth conditions for livestock and living environments for stock farmers .
- Korean Patent No. 378667 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., Flavobacterium 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 discloses a microbial composition for the biological treatment of malodorants and a method for the preparation thereof.
- the microbial composition comprises deodorizing microorganisms 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.
- 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 s ' pp.
- thermoresistant and thermophilic bacillus capable of growing on a broad range of substrates may be Bacillus subtilis CH-IO, Bacillus mesentericus, Bacillus r ⁇ egaterium, Bacillus licheniformmis 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.
- 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
- Korean Patent Application No. 10-2004-0011349 suggests a Geotrichum-immobilized biofliter 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 oi game compounds.
- the present invention provides a novel strain Bacillus cereus BC3 KCCM 10859P, which has the ability to deodorize livestock wastewater and food waste faster and more effectively than conventional strains, and to remove heavy metals.
- the present invention provides a microorganism composition for removing nasty odor and heavy metals, comprising the novel strain Bacillus cereus BC3 KCCM 10859P as an active ingredient.
- the novel strain Bacillus cereus BC3 KCCM 10859P 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 piesent 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 .
- FIG. 1 is a schematic diagram showing a device used in the experiments to determine the ability to deodorize livestock wastewater.
- FIG. 2 is a schematic diagram showing a device used m the experiments to determine the ability to deodorize food waste.
- the present invention provides a novel bacterial strain, identified as Bacillus cereus BC3 KCCM 10859P, which is useful in deodo ⁇ zation and heavy metal removal.
- the present invention provides a microbial composition for the removal of malodor and heavy metals, comprising an effective amount of the novel strain Bacillus cereus BC3
- the bacterial strain may be formulated into a solid preparation, 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.
- 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 in terms o ⁇ preservability.
- 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 cereus BC3 KCCM 10859P culture into a 0.5-3.0 wt% solution containing a metal salt such as Ca 2+ , Cu 2+ , Zn 2+ , Ni 2+ , Co 2+ , Mn 2+ , Al 2+ , Fe 2+ , or Mg 2+ 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 .
- a positively charged biodegradable polymer such as chitosan, a chitosan derivative or polylys
- 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.
- 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
- 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 cereus BC3 KCCM 10859P in a sufficiently effective amount to conduct the above-mentioned functions, m combination with one or more Bacillus strain selected from the group consisting of Bacillus megatenum BCl-I KCCM 10856P, Bacillus mega tenum.
- BCl-2 KCCM 10857P Bacillus megatenum BC2-1 KCCM 10858P, 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.
- the present invention provides a method for reducing malodor or heavy metals of food waste or livestock wastewater, comprising culturing the novel strain Bacillus cereus BC3 KCCM 10859P with malodorous food waste or livestock wastewater.
- 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 cereus BC3 KCCM 10859P, in combination with one or more Bacillus strain selected fi om the group consisting of Bacillus megatenum BCl-I KCCM 10856P, Bacillus megatenum BCl-2 KCCM 10857P, Bacillus megatenum BC2-1 KCCM 10858P, 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 firr ⁇ us BC9 KCCM 10865P and Bacillus cereus BClO KCCM
- 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 cereus BC3 KCCM 10859P.
- organic liquefied fertilizer from livestock wastewater and animal feeds from food waste may be achieved using a conventional process without the use of the novel Bacillus cereus BC3 KCCM 10859P.
- organic liquefied fertilizer for example, it may be prepared from livestock wastewater using peat.
- a conventional method comprises pulverizing and mixing peat, organic 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 deodo ⁇ zation, extracting organic liquefied fertilizer, neutralizing the extracted organic liquefied fertilizer with dj luted hydrochloric acid, and aerating the neutralized organic liquid to decrease the chemical oxygen demand thereof.
- a dry fermentation method On the basis of the novel strain Bacillus cereus BC3 KCCM 10859P, a dry fermentation method, a mixed fermentation method, a combination thereof, or a liquid- phase fermentation method may be provided for the preparation of animal feeds from food waste.
- the dry fermentation method greatly reduces the food waste in mass, but demands an inordinate amount of energy for drying and produces 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 high 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 in 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 85%, has an advantage over the other methods in terms of pioduction cost, but the products are inconvenient to transport .
- 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 cereus BC3 KCCM 10859P 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 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, and a combination thereof.
- Bacillus cereus BC3 KCCM 10859P the principle of the present invention, was taken from mushrooms and hyphae grown on fallen leaves in a forest located in Samgi, Iksan,
- 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 - 30 0 C. 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.
- Bacillus cereus BC3 KCCM 10859P 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 livestock wastewater (see FIG. 1) and the gas of the
- 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 F]G.1) .
- 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.
- 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 ⁇ 3O 0 C, followed by measuring absorbance at 640 nm. 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.
- III phenol pentacyanonitrosyl ferrate
- a sample gas was adsorbed and concentrated at -170 0 C with an SPT (Sample Preconcentration Trap) , desorbed by heating, and injected into gas chromatography equipped with a PFPD
- Table 1 were summarized the analysis results obtained after culturing Bacillus cereus BC3 KCCM 10859P alone and in combination with other Bacillus strains for 4 days in wastewater.
- Bacillus cereus BC3 KCCM 10859P 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 cereus BC3 KCCM 10859P Alone or in Combination with other Bacillus Strains on Food Waste
- Bacillus cereus BC3 KCCM 10859P 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 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 to microorganism reaction vessels containing food waste (see FIG. 2) and the gas of the vessels
- 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.
- Bacillus cereus BC3 KCCM 10859P showed excellent ability to remove ammonia, methyl mercaptan, dimethyl disulfide, trimethyl amine and acetaldehyde . However, this bacteria was found to be unable to effectively remove dimethyl sulfide and, as similar concentrations of these chemicals were detected between the samples and the control.
- Bacillus cereus BC3 KCCM 10859P In order to evaluate the ability of Bacillus cereus BC3 KCCM 10859P 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 cereus BC3 KCCM 10859P 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
- Bacillus cereus BC3 KCCM 10859P 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, zinc and lead from food waste. Nickel and chromium, 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 chromium-removing ability of Bacillus cereus BC3 KCCM 10859P unclear, [industrial Applicability] As described above, Bacillus cereus BC3 KCCM 10859P according to the present invention is very useful in deodorizing livestock wastewater and food waste.
- the novel bacterial strain of the present invention is highly effective at removing the nasty odor of livestock wastewater and food waste.
- the strain of the present invention is capable of very effectively removing heavy metals from livestock wastewater or food waste.
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Abstract
The present invention relates to a novel strain Bacillus cereus BC3 KCCM 10859P 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] [inventxon Title]
BACILLUS CEREUS BC3 KCCM 10859P 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 gJ obal 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 raalodor 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, mghtsoil 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 ((CH3J3N), 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 malodor ingredients 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 aieas, 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 pi ovide 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 deodoπzation, 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 ((CH3J2S2), the photocatalysts, such as TiO2, 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 deodorization, uses a bioreactor loaded with a carrier onto which deodorizing microorganisms are immobilized. The lowest operation cost is found m the biofilter system. Operation costs increase in 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 piotems 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 producing 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 mi 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., Flavobacterium 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) discloses a microbial composition for the biological treatment of malodorants and a method for the preparation thereof. The microbial composition comprises deodorizing microorganisms 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 s'pp. The thermoresistant and thermophilic bacillus capable of growing on a broad range of substrates may be Bacillus subtilis CH-IO, Bacillus mesentericus, Bacillus rαegaterium, Bacillus licheniformmis 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 biofliter.
Korean Patent Application No. 10-2004-0011349 (filed Feb. 2, 2004) suggests a Geotrichum-immobilized biofliter 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 oi game compounds.
[Disclosure] [Technical Problem]
It is an object of the present invention to provide a novel bacterial strain, identified as Bacillus cereus BC3 KCCM 10859P, 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 cereus BC3 KCCM 10859P
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 cereus BC3 KCCM 10859P, 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 cereus BC3 KCCM 10859P as an active ingredient.
The novel strain Bacillus cereus BC3 KCCM 10859P 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 piesent 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 in the experiments to determine the ability to deodorize livestock wastewater.
FIG. 2 is a schematic diagram showing a device used
m 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 cereus BC3 KCCM 10859P, 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 cereus BC3
KCCM 10859P.
Conventional techniques may be applied for the formulation of the bacterial strain Bacillus cereus BC3 KCCM 10859P according to the present invention. The bacterial strain may be formulated into a solid preparation, 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 in terms oϊ 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 cereus BC3 KCCM 10859P 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 cereus BC3 KCCM 10859P in a sufficiently effective amount to conduct the above-mentioned functions, m combination with
one or more Bacillus strain selected from the group consisting of Bacillus megatenum BCl-I KCCM 10856P, Bacillus mega tenum. BCl-2 KCCM 10857P, Bacillus megatenum BC2-1 KCCM 10858P, 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.
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 cereus BC3 KCCM 10859P 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 cereus BC3 KCCM 10859P, in combination with one or more Bacillus strain selected fi om the group consisting of Bacillus megatenum BCl-I KCCM 10856P, Bacillus megatenum BCl-2 KCCM 10857P, Bacillus megatenum BC2-1 KCCM 10858P, 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 firrαus 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 cereus BC3 KCCM 10859P.
Also, the production of organic liquefied fertilizer from livestock wastewater and animal feeds from food waste may be achieved using a conventional process without the use of the novel Bacillus cereus BC3 KCCM 10859P. As for organic liquefied fertilizer, for example, it may be prepared from livestock wastewater using peat. In detail, a conventional method comprises pulverizing and mixing peat, organic 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 dj luted hydrochloric acid, and aerating the neutralized organic liquid to decrease the chemical oxygen demand thereof.
On the basis of the novel strain Bacillus cereus BC3
KCCM 10859P, a dry fermentation method, a mixed fermentation method, a combination thereof, or a liquid- phase fermentation method may be provided for the preparation of animal feeds from food waste. The dry fermentation method greatly reduces the food waste in mass, but demands an inordinate amount of energy for drying and produces 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 high 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 in 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 85%, has an advantage over the other methods in terms of pioduction 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 cereus BC3 KCCM 10859P 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 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, 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 cereus BC3 KCCM 10859P
Bacillus cereus BC3 KCCM 10859P, 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 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 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 cereus BC3 KCCM 10859P Alone or in Combination with other Bacillus Strains on Livestock Wastewater
In order to evaluate the ability of Bacillus cereus BC3 KCCM 10859P to reduce the malodor of livestock wastewater, Bacillus cereus BC3 KCCM 10859P 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 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 F]G.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 cereus BC3 KCCM 10859P and 5 ml of a mixture containing equal amounts of Bacillus megateπum BCl-I KCCM 10856P, Bacillus megateπum 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 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 ni troprusside solution and a sodium hydrochlorite solution, both of which react with ammonium ions to produce indophenols 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 ~ 3O0C, followed by measuring absorbance at 640 nm. 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 tπmethyl 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 700C 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 Lm Instrument Co. Ltd.) and analyzed.
(2) Analysis Results
Table 1 were summarized the analysis results obtained after culturing Bacillus cereus BC3 KCCM 10859P alone and in combination with other Bacillus strains for 4 days in wastewater.
< TABLE 1 > Deodorizing Effect of Bacillus cereus BC3 KCCM 10859P on Livestock Wastewater (unit ppm)
As is apparent from the data of Table 1, Bacillus cereus BC3 KCCM 10859P 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 cereus BC3 KCCM 10859P Alone or in Combination with other Bacillus Strains on Food Waste
In order to evaluate the ability of Bacillus cereus BC3 KCCM 10859P to reduce the malodor of food waste, Bacillus cereus BC3 KCCM 10859P 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 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 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 cereus BC3 KCCM 10859P 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 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 cereus BC3 KCCM 10859P on Food Waste (unit ppm)
As is apparent from the data of Table 1, Bacillus cereus BC3 KCCM 10859P showed excellent ability to remove ammonia, methyl mercaptan, dimethyl disulfide, trimethyl amine and acetaldehyde . However, this bacteria was found to be unable to effectively remove dimethyl sulfide and, 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 cereus BC3 KCCM 10859P 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 cereus BC3 KCCM 10859P 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 HCIO4 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 m 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 cereus BC3 KCCM 10859P to Reduce Heavy Metals of Livestock Wastewater (Units of ppm)
< TABLE 4 > Ability of Bacillus cereus BC3 KCCM 10859P to Reduce Heavy Metals of Food Waste (Units of ppm)
Bacillus cereus BC3 KCCM 10859P 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, zinc and lead from food waste. Nickel and chromium, 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 chromium-removing ability of Bacillus cereus BC3 KCCM 10859P unclear, [industrial Applicability] As described above, Bacillus cereus BC3 KCCM 10859P 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 deodorization (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
[Claim 1]
A novel strain Bacillus cereus BC3 KCCM 10859P 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 cereus BC3 KCCM 10859P.
[Claim 3] A microorganism composition for reducing nasty odors and heavy metals, comprising an effective amount of a novel strain Bacillus cereus BC3 KCCM 10859P 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 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, 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 cereus BC3 KCCM 10859P 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 cereus BC3 KCCM 10859P and one or more Bacillus species selected from the group consisting of Bacillus megatenum BCl-I KCCM 10856P, Bacillus megatenum BCl-2 KCCM 10857P, Bacillus megatenum BC2-1 KCCM 10858P, 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, 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 cereus BC3 KCCM 10859P m livestock wastewater.
[Claim 7]
A method for producing organic liquefied fertilizer, using an effective amount of novel strain Bacillus cereus BC3 KCCM 10859P and one or more Bacillus species selected fiom the group consisting of Bacillus megatenum BCl-I KCCM 10856P, Bacillus megatenum BCl-2 KCCM 10857P, Bacillus megatenum BC2-1 KCCM 10858P, 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, 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 cereus BC3 KCCM 10859P. [Claim 9]
A method for preparing livestock feed from food waste, using a microorganism composition comprising an effective -amount of a novel strain Bacillus cereus BC3 KCCM 10859P 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 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, and a combination thereof.
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