KR20160019019A - Microbial agent for deodorant for decreasing bad smell, and the method using the same - Google Patents

Abstract

The present invention provides a microbial agent for deodorization, comprising Bacillus thuringiensis (JLRI 02 KCCM 11490P), Pseudomonas caeni (JLRI 03 KCCM 11491P), and Rhodobacter sphaeroides (JLRI 04 KCCM 11492P).

Description

TECHNICAL FIELD The present invention relates to a microbial agent for odor reduction and a method for reducing odor using the microbial agent,

The present invention relates to a microbial agent for reducing odor and a method for reducing odor using the same, more particularly, it is safe and less harmful to humans and animals, and there is no need for a separate treatment facility for removing or reducing odor in an animal husbandry farm The present invention relates to a microorganism preparation for odor reduction in a housing or manure having a low utility cost and a reduced effective amount of gas induced by odor, and a method for reducing odor using the same.

In general, odors from livestock farming or livestock manure are not only harmful to nearby residents, but also contribute to environmental pollution. For this reason, deodorants have been introduced to remove various types of odors generated from these odor sources.

In general, a deodorant is an agent that removes or weakens odors, and most of them use activated carbon having an adsorptive function of sucking odor, chemical oxidation or chemical cleaning.

However, the method of adsorbing odor using activated carbon has a disadvantage in that not only the deodorization efficiency is low but also the service life is short. The chemical oxidation method requires only a high-level treatment technique because only the component that is a main cause of the odor among the odor components is required There is a problem of secondary contamination through the treatment of the compound, and the cleaning method using the chemical solution is a method of absorbing the gas generating odor into the solution and treating it, so that when the odorous gas of various kinds is mixed, And the application range thereof is limited.

In addition, since the chemical oxidation method and the chemical liquid cleaning method all use chemical chemicals, when they are sprayed in the place where the odor is generated, they are not environmentally friendly because of the problems such as the removal of the odor and the secondary environmental pollution.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made in view of the above problems, and it is an object of the present invention to provide a method and a device for preventing harmful substances from harmful to humans and animals, Which is low in cost and fundamentally reducing the offensive odor gas, thereby providing a microorganism preparation for odor reduction in a poultry or manure having a high utility value and a method for reducing odor using the same.

The technical problem of the present invention as described above is achieved by the following means.

(1) a malodor reducing microorganism preparation comprising Bacillus thuringiensis JLRI 02 KCCM 11490P, Pseudomonas caeni JLRI 03 KCCM 11491P, and Rhodobacter sphaeroides JLRI 04 KCCM 11492P .

(2) a malodor reducing microorganism preparation comprising Bacillus thuringiensis JLRI 02 KCCM 11490P, Pseudomonas caeni JLRI 03 KCCM 11491P, and Rhodobacter sphaeroides JLRI 04 KCCM 11492P To a poultry or manure in powder or liquid form.

(3) Bacillus thuringiensis JLRI 02 KCCM 11490P ( Bacillus thuringiensis JLRI 02 KCCM 11490P) having an odor reducing activity.

(4) Pseudomonas aeruginosa JLRI 03 KCCM 11491P ( Pseudomonas caeni JLRI 03 KCCM 11491P) having an odor reducing activity.

(5) Rhodobacter sphaeroides JLRI 04 KCCM 11492P ( Rhodobacter sphaeroides JLRI 04 KCCM 11492P)

As described above, the present invention is safe and less harmful to humans and animals, and does not require a separate treatment facility to remove or reduce odor in the livestock farm, etc., which is inexpensive and fundamentally reduces odor-induced gases Especially, there is a large effect of removing odor generated due to poultry or manure.

Hereinafter, the contents of the present invention will be described in more detail.

The three microorganisms constituting the microorganism preparation according to the present invention are Bacillus thuringiensis JLRI 02 KCCM 11490P, Pseudomonas caeni JLRI 03 KCCM 11491P, and Rhodobacter sphaeroides JLRI 04 KCCM 11492P), which were isolated from bilge enzyme, skipjack, and compost, respectively, were deposited at the Korea Microorganism Conservation Center, Korea Institute of Microbiology, on November 27, 2013.

The microorganism preparation according to the present invention is not particularly limited, but each microorganism strain may be mixed at a ratio of 1 to 30% by weight. The microorganism preparation according to the present invention mixes the three kinds of microorganisms, thereby exerting a synergistic effect and directly injecting the powder into a powdery or liquid form in a manure or cattle manure, so that the malodor can be eco-friendly.

Each microorganism preparation may be provided as a microorganism preparation in the form of a liquid, a powder, or a solid in combination with each separated microorganism.

The method of injecting the microorganism preparation of the present invention as described above can directly spray the microorganism preparation according to the present invention to a housing or livestock manure using a sprayer or the like in a liquid or powder form, the mixed microbial agent microorganism of the species ^{(1 × 10 8 cfu / ml} ) may be subject to dilution appropriate multiples to spray the spray, in the case of housing a mixed microbial agent microorganism of the three species ^{(1 × 10 8 cfu / ml} ) Can be diluted about 100 times and sprayed into the house every day for one week and every two days for the remaining one week.

The microbial formulation according to the present invention has a high removal efficiency of odor by directly putting it in a place where odor is generated, and has a very high removal efficiency of hydrogen sulfide and ammonia which is a cause of odor and a main cause of air pollution .

Hereinafter, the present invention will be described in more detail with reference to examples. It should be understood, however, that the invention is not limited to the disclosed embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

[Experimental Example 1] Selection and identification of odor-reducing microorganisms

1. Experimental process

In order to isolate the odor - reducing microorganisms, the samples used for isolating strains were collected from microbiological samples such as compost, soil, kimchi, and red sea bream.

(1) Selection of ammonia and sulfur oxidizing bacteria

The growth medium was prepared by adding 0.5 g of ammonium sulfate [(NH ₄ ) ₂ SO ₄ ] and 5 g of sodium thiosulfate (Na ₂ S ₂ O ₃ ) to the nutrient broth and shaking for 30 minutes at 30 ° C for 1 week. 1 ml of the proliferated culture was taken, diluted in PBS (phosphate buffered saline) (10 ^-4 to 10 ^-8 ), and plated on ammonia and sulfur-oxidizing plate medium. After culturing at 30 ° C for 2 to 3 days, the first strain was selected, and the first selected strain was plated on an agar medium (3 g of beef extract, 5 g of peptone, 15 g of agar) For several times.

Ammonia oxidizing bacteria medium composition Furtherance Molecular formula Content (g / L) Potassium monohydrogen phosphate K ₂ HPO ₄ 0.5 Magnesium sulphate heptahydrate MgSO ₄ .7H ₂ O 0.1 Ammonium sulfate (NH _{4) 2} SO ₄ 2 Ferric chloride hexahydrate FeCl ₃ .6H ₂ O 0.014 Calcium chloride dihydrate CaCl ₂ .2H ₂ O 0.004

Sulfur oxidizing bacteria medium composition Furtherance Molecular formula Content (g / L) Potassium dihydrogen phosphate KH ₂ PO ₄ 2.0 Potassium monohydrogen phosphate K ₂ HPO ₄ 2.0 광동기 NH ₄ Cl ₂ 0.4 Magnesium dichloride MgCl ₂ 0.2 Iron sulfate heptahydrate FeSO ₄ .7H ₂ O  0.018 Sodium thiosulfate Na ₂ S ₂ O ₃ 5.0

(2) Selection of photosynthetic bacteria

Samples diluted with PBS (10 ^-4 to 10 ^-6 ) were plated on mineral salts-succinate agar and incubated for 2 to 4 days at 30 ° C under 4,000 lux conditions.

Mineral salts-succinate medium composition Furtherance Molecular formula Content (g / L) Magnesium sulphate heptahydrate MgSO ₄ .7H ₂ O 0.2 Potassium monohydrogen phosphate K ₂ HPO ₄ One Sodium chloride NaCl 0.5 Iron sulfate heptahydrate FeSO ₄ .7H ₂ O 0.01 Calcium chloride CaCl ₂ 0.02 Manganese chloride tetrahydrate MnCl ₂ .4H ₂ O 0.002 Sodium molybate dihydrate Na ₂ MoO ₄ .2H ₂ O 0.001 Sodium succinate dibasic C ₄ H ₄ Na ₂ O ₄ 10 Yeast extract One Ammonium chloride NH ₄ Cl One

(3) Identification of strains using molecular biological methods

DNA extraction

DNA extraction of isolated strains was performed by adding 100 μl 5% Chelex (Biorad) to the E-tube after collecting colonies of the bacteria grown in the solid medium. After heat treatment at 95 ° C for 10 minutes, the supernatant was used as PCR amplified DNA.

PCR reaction

The isolated strains were amplified and sequenced using 16S ribosomal DNA (rRNA). The universal PCR primers used for amplification were 27F (5'-AGA GTT TGA TCM TGG CTC AG-3 ') and 1492R (5'-TAC GGY TAC CTT GTT ACG ACT T-3'). The PCR conditions were 1 μl of DNA, 2 μl of 10 μM forward and reverse primers, 1 μl of each 2.5 mM dNTPs, 0.25 μl of Taq polymerase (Takara), 0.25 μl of PCR buffer (15 mM MgCl 2, 100 mM Tris-Hydrochloric acid pH 8.3, 500 mM KCl), denatured at 94 ° C for 5 minutes, denatured at 94 ° C for 45 seconds, attached at 55 ° C for 45 seconds, and incubated at 72 ° C for 1 minute. Amplified, and finally elongated at 72 ° C for 10 minutes. The final reaction was confirmed by electrophoresis on 1.2% agarose gel to which 1 gel gel red was added for 50 minutes at 100 V, and the amplified product was about 1500 bp in size.

(4) Comparison of isolated strains by ARDRA (Amplified Ribosomal DNA Restriction Analysis, amplified rDNA restriction analysis)

The amplified PCR products were reacted with restriction enzymes HaeIII and HaI (Takara, Japan) at 37 ° C for 5 hours. The obtained products were separated by comparing the band patterns shown by electrophoresis in a 2.5% agarose gel for 70 minutes.

2. Experimental results

The 16S rDNA analysis results are as shown in SEQ ID NOS: 1 to 3. The results of identification of the odor-reducing microorganisms isolated as described above are shown in the following table.

Isolate Closest relative to the closest genetic distance Similarity IRG-2 Paenibacillus macquariensis subsp. macquariensis NCTC 10419 (T) (X60625) 99% Su-1 Bacillus simplex NBRC 15720 (T) (AB363738) 99% Su-4 Bacillus simplex NBRC 15720 (T) (AB363738) 98% Su-8 Bacillus thuringiensis  ATCC 10792 (T) (ACNF01000156) 99% Hong-5 Pseudomonas caeni  HY-14 (T) (EU620679) 99% IRG-7 Bacillus megaterium IAM 13418 (T) (D16273) 99% Hong-2 Bacillus nealsonii DSM 15077 (T) (EU656111) 99% Muk-7 Bacillus methylotrophicus CBMB205 (T) (EU194897) 99% Milk-3 Pseudoclavibacter soli KPO2 (T) (AB329630) 97% IRG-18 Bacillus thioparans BMP-1 (T) (DQ371431) 99% Su-26 Paenibacillus amylolyticus NRRL NRS-290 (T) (D85396) 99% Hong-21 Microbacterium esteraromaticum DSM 8609 (T) (Y17231) 99% M-1 Rhodobacter sphaeroides  ATH 2.4.1 (T) (CP000143) 99%

[Experimental Example 2] Characterization of odor-reducing microorganisms

1. Growth characteristics

After incubation at low temperature (-4, -10, -20 ° C), the optical density values were measured using a UV-visible spectrophotometer in the time zones (0, 4, 8, 12, 24, Were measured.

As a result, growth of Bacillus subtilis Su-8, Nitrogen-degrading bacteria Hong-5 and Photosynthetic bacteria M-1 was excellent as shown in the following table.

-4 ° C culture culture
time
(h) Bacterium IRG-2 Su-1 Su-4 Su-8 Hong-5 IRG-7 Hong-2 Muk-7 Milk-3 M-1 0 0 0 0 0 0 0 0 0 0 0 4 0.028 0.051 0.023 0.021 0.04 0.039 0.031 0.01 0.034 0.033 8 0.032 0.054 0.053 0.047 0.041 0.058 0.039 0.024 0.061 0.054 12 0.066 0.084 0.078 0.054 0.084 0.078 0.04 0.035 0.083 0.068 24 0.106 0.096 0.082 0.064 0.099 0.097 0.074 0.05 0.098 0.096 48 0.189 0.114 0.098 0.153 0.299 0.115 0.098 0.082 0.145 0.118 72 0.422 0.129 0.099 0.17 0.524 0.119 0.1 0.07 0.162 0.169

-10 ° C culture culture
time
(h) Bacterium IRG-2 Su-1 Su-4 Su-8 Hong-5 IRG-7 Hong-2 Muk-7 Milk-3 M-1 0 0 0 0 0 0 0 0 0 0 0 4 0.01 0.016 0.02 0.049 0.026 0.026 0.024 0.012 0.064 0.036 8 0.056 0.038 0.048 0.067 0.062 0.108 0.05 0.034 0.102 0.118 12 0.074 0.22 0.107 0.108 0.16 0.278 0.076 0.17 0.276 0.298 24 0.302 0.344 0.388 0.155 0.262 0.42 0.09 0.344 0.27 0.62 48 0.872 0.556 0.584 0.338 1.576 0.72 0.126 0.5 0.366 0.92 72 1.018 0.792 0.998 0.64 1.598 0.93 0.296 0.56 0.4 1.43

-20 ° C culture culture
time
(h) Bacterium IRG-2 Su-1 Su-4 Su-8 Hong-5 IRG-7 Hong-2 Muk-7 Milk-3 M-1 0 0 0 0 0 0 0 0 0 0 0 4 0.012 0.024 0.02 0.101 0.124 0.029 0.05 0.02 0.031 0.029 8 0.098 0.058 0.072 0.272 0.42 0.05 0.132 0.031 0.048 0.05 12 0.31 0.222 0.19 0.838 1.384 0.468 0.591 0.113 0.114 0.468 24 0.71 0.339 0.212 1.071 1.897 0.521 0.613 0.196 0.153 0.521 48 1.01 0.617 0.927 1.136 2.086 0.58 0.319 0.534 0.576 0.85 72 0.956 0.68 1.068 1.033 2.165 0.775 0.255 0.337 1.049 1.775

2. Antimicrobial activity

Was determined by agar diffusion method (paper disc diffusion method) for: (H7, Staphylococcus aureus subsp aureus ATCC 13301, Salmonella typhimurium ATCC 13311 Bacillus cereus ATCC 27348, E. coli O157.) Total of four kinds of pathogenic bacteria. That is, each strain was cultured in a 20 ml nutrient medium for 18 hours, the pathogenic bacterial culture was adjusted with a spectrophotometer at an optical density value of 0.5 at 620 nm, and the bacteria were evenly spread on the entire surface of the medium while being rotated with a sterilized cotton swab . The cultured odor-reducing microorganisms were absorbed by 100 μl into a sterilized paper disk (8 mm in thickness; Advantec Co.), adhered to a paper disk on a plate medium, cultured in a 37 ° C. incubator for 24 hours, Diameter was measured and the antibacterial activity was compared.

As a result, Su-8, Hong-5 and M-1 had antibacterial activity against all pathogenic bacteria. Among them, Su-8 showed excellent antibacterial activity and showed similar results as the addition of 1 ppm chloramphenicol.

Antimicrobial activity of chloramphenicol density Clear zone (mm) B. cereus
ATCC 27348 E. coli
O157: H7 S. aureus subsp. aureus ATCC 13301 Sal. typhimurium ATCC 13311 500 ppb 12 12 12 8 1 ppm 15 15 14 15 10 ppm 20 30 24 22

Odor-reducing microbial antimicrobial activity Isolate Transparency ( clear zone , mm ) B. cereus
ATCC 27348 E. coli
O157: H7 S. aureus subsp. aureus ATCC 13301 Sal . typhimurium ATCC 13311 Muk-7 19 20 19 19 Water -8 16 16 14 16 Su-1 15 16 14 15 Hong -5 15 15 14 15 M-1 14 15 15 14 IRG-7 14 14 15 14 Su-4 15 14 13 13 Hong-21 13 12 13 13 Milk-3 12 12 10 - Hong-2 - 9 - - Su-26 - 9 - - IRG-18 - 8 - - IRG-2 - - - -

8 ( Bacillus thuringiensis ATCC 10792, 99%), Nitrogen-degrading bacteria Hong-5 ( Pseudomonas caeni HY-1, 99%), photosynthetic bacteria M-1 ( Rhodobacter sphaeroides ATH 2.4.1, 99%) were selected as the malodor reducing strains of the present invention and Bacillus thuringiensis JLRI 02 KCCM 11490P, Pseudomonas aeruginosa, caeni JLRI 03 KCCM 11491P), and Rhodobacter sphaeroides JLRI 04 KCCM 11492P), deposited at Korea Microorganism Conservation Center, Korea Institute of Microbiology, Korea, on November 27, 2013.

[Experimental Example 2] Removal of odor substances by chemical method

(NB + Na ₂ S ₂ O ₃ + NH ₄ Cl) containing nitrogen compound and sulfur compound were added to each 30 ml cap tube, Deg.] C, 120 rpm for 48 hours. The growth of the strain was measured by taking 3 ml of the cultured microorganism and measuring absorbance at 660 nm using a spectrophotometer. The remaining culture was centrifuged at 4000 rpm for 20 minutes, and then only the supernatant was recovered, sterilized by filter, and then used for hydrogen sulfide and ammonia nitrogen removal efficiency.

The supernatant of the strain in which the pellet was removed by the above method was examined for ammonia reduction effect by a chemical method. Ammonia nitrogen (NH ₃ -N) was obtained by diluting, homogenizing and centrifuging the sample according to the Indo-phenol Blue method of Chaney and Marbach (1962) in the presence of phenol and sodium hypochlorite. Sodium nitroprusside was used as a catalyst to produce indophenol (blue). The absorbance (630 nm) was measured and the concentration was calculated according to a standard calibration curve obtained with NH ₄ Cl.

Ammonia nitrogen (NH ₃ -N) removal ability analysis (unit: ppm) Treatment NH ₃ conc. (630 nm) Con (-) 159.64 Con (+, Alcaligenes aquatilis LMG 22996 (T)) 123.51 T1 ( B. simplex, Su-4) 132.68 T2 ( B. thuringiensis, Su-8) 117.52 T3 ( Pseudomonas caeni, Hong-5) 108.02 T4 ( Rhodobacter sphaeroides, M-1) 105.33

※ Con (-): No treatment, Con (+): Excellent nitrogen removal ability strain

As shown in the above results, according to the present invention, ammonia removal ability was superior to that of Alcaligenes aquatilis , which has a known nitrogen removal ability, in T2, T3 and T4 treatments.

Sulfate was added to 1ml of Conditioning regimen, 1ml of sample and 60mg of BaCl _{2 in} 30ml according to the method described by Kolmert, A., P. Wikstr, et al. (2000) And the absorbance was measured at 420 nm using a spectrophotometer.

Analysis of hydrogen sulfide removal (unit: ppm) Treatment SO ₄ conc. (420 nm) Con (-) 1052.12 Con (+, Bacillus licheniformis ) 1659.98 T1 ( B. simplex, Su-4) 1125.52 T2 ( B. thuringiensis, Su-8) 1689.37 T3 ( Pseudomonas caeni, Hong-5) 1437.61 T4 ( Rhodobacter sphaeroides, M-1) 1746.83

※ Con (-): untreated, Con (+): existing odor reduction patented strain

As shown above, according to the present invention, the removal ability of hydrogen sulfide was excellent in the treatments of T2, T3 and T4. In particular, it was found that the photosynthetic bacteria T4 was most efficiently converted into the sulfate by hydrogen sulfide during inoculation.

[Experimental Example 3] Removal of odorous substances in pigs by using a single strain

(1) ammonia

Ammonia gas contents were measured at 0, 24, 48, and 72 hours after the inoculation of the strains. As a result, ammonia gas content was lowest at all time points in T4 treatment, and ammonia gas was effectively reduced.

Ammonia gas reduction (unit: ppm) Treatment 0 24 48 72 Con (-) 55 30 15.5 10 Con (+, Bacillus licheniformis ) 55 10 5 4.25 T1 ( B. simplex, Su-4) 55 7.5 5 3.25 T2 ( B. thuringiensis, Su-8) 55 8 5.5 4.5 T3 ( Pseudomonas caeni, Hong-5) 55 10 7.5 5 T4 ( Rhodobacter sphaeroides, M-1) 55 6 4 2

※ Con (-): untreated, Con (+): existing odor reduction patented strain

(2) hydrogen sulfide

As a result of measurement of hydrogen sulfide gas contents at 0, 24, 48 and 72 hours after strain inoculation, hydrogen sulfide gas was not effectively detected at T3 and T4 treatments at all times and it was observed to effectively reduce hydrogen sulfide gas.

Reduction of hydrogen sulfide gas (unit: ppm) Treatment 0 24 48 72 Con (-) 0 12 14 110 Con (+, Bacillus licheniformis ) 0 0 2.5 20 T1 ( B. simplex, Su-4) 0 0 One 15 T2 ( B. thuringiensis, Su-8) 0 0 0 0.5 T3 ( Pseudomonas caeni, Hong-5) 0 0 0 0 T4 ( Rhodobacter sphaeroides, M-1) 0 0 0 0

※ Con (-): untreated, Con (+): existing odor reduction patented strain

(3) Total mercaptan

Total mercaptan gas contents were measured at 0, 24, 48 and 72 hours after strain inoculation, and it was observed that total mercaptan gas was not detected at T2, T3 and T4 treatments at all times, effectively reducing mercaptan gas.

Total mercaptan gas reduction (unit: ppm) Treatment 0 24 48 72 Con (-) 0 0 2.5 2 Con (+, Bacillus licheniformis ) 0 0 0 3 T1 ( B. simplex, Su-4) 0 0 0 One T2 ( B. thuringiensis, Su-8) 0 0 0 0 T3 ( Pseudomonas caeni, Hong-5) 0 0 0 0 T4 ( Rhodobacter sphaeroides, M-1) 0 0 0 0

※ Con (-): untreated, Con (+): existing odor reduction patented strain

[Experimental Example 4] Removal of odorous substances from pigs of complex microorganisms

The slaughtered sows were kept in a constant temperature water bath at 30 ° C within 3 hours and then mixed with 1: 1 ratio. %, And the mixture was allowed to stand in the room (room temperature 23 to 26 ° C) for 1 hour, and then the experiment was conducted.

(1) ammonia

Ammonia gas contents were measured at 0, 24, 48 and 72 hours after inoculation of the strain. As a result, T2 + T3 + T4 ( B. thuringiensis , Su-8 + Pseudomonas Caeni , Hong-5 + Rhodobacter sphaeroides, M-1) showed the lowest ammonia gas content, indicating that ammonia gas was effectively reduced.

Ammonia gas reduction (unit: ppm) Treatment 0 24 48 72 Con (-) 30 7.5 4.5 4 Con (+, Bacillus licheniformis ) 30 5.5 4.25 3.5 T1 + T4 30 4 3.75 2.75 T2 + T4 30 4.25 3 2.5 T3 + T4 30 4 2.5 2.5 T1 + T2 + T4 30 3 2 3 T1 + T3 + T4 30 5 2.5 2 T2 + T3 + T4 30 3 2.5 1.75 T1 + T2 + T3 + T4 30 4.95 5 1.5

※ Con (-): untreated, Con (+): existing odor reduction patented strain

(2) hydrogen sulfide

As a result of measurement of hydrogen sulfide gas contents at 0, 24, 48 and 72 hours after the strain inoculation, hydrogen sulfide gas was not effectively detected at the treatment time in all treatments, unlike the case where the hydrogen sulfide concentration was increased in the control Respectively.

Reduction of hydrogen sulfide gas (unit: ppm) Treatment 0 24 48 72 Con (-) 0 0 10 90 Con (+, Bacillus licheniformis ) 0 0 2 15 T1 + T4 0 0 0 10 T2 + T4 0 0 0 20 T3 + T4 0 0 0 0 T1 + T2 + T4 0 0 0 10 T1 + T3 + T4 0 0 0 0 T2 + T3 + T4 0 0 0 0 T1 + T2 + T3 + T4 0 0 0 0

※ Con (-): untreated, Con (+): existing odor reduction patented strain

(3) Total mercaptan

The total mercaptan gas contents were measured at 0, 24, 48 and 72 hours after strain inoculation. As a result, the total mercaptan was not detected in the treatments of bacterial strain combined at all times, unlike the increase in the total mercaptan concentration in the control It was observed that the total mercaptan gas was effectively reduced.

Total mercaptan gas reduction (unit: ppm) Treatment 0 24 48 72 Con (-) 0 0 1.5 3 Con (+, Bacillus licheniformis ) 0 0 0 1.5 T1 + T4 0 0 0 1.5 T2 + T4 0 0 0 2 T3 + T4 0 0 0 0 T1 + T2 + T4 0 0 0 0 T1 + T3 + T4 0 0 0 0 T2 + T3 + T4 0 0 0 0 T1 + T2 + T3 + T4 0 0 0 0

※ Con (-): untreated, Con (+): existing odor reduction patented strain

The results showed that ammonia, ammonia, 100% hydrogen sulfide and 100% total mercaptan were reduced in the mixed microbial treatment according to the present invention.

[Experimental Example 5] Effect of complex microorganisms on the removal of odor substances from pigs

The complex microorganism preparation (1 × 10 ⁸ cfu / ml) according to Experimental Example 4 of the present invention was diluted 100-fold in a livestock farm in Jeollanam-do, and sprayed into the pigs at intervals of two days for one week and every two days for one week As a result of measurement according to the disclosed method, as shown in Table 5 below, it was confirmed that the total amount of odor substances decreased until 8 days after spraying, and that ammonia was reduced by 65.5% and hydrogen sulfide by 42.9%.

Result of the odor elimination test in nursery pigs (Unit: ppm) Measurement date Before processing 2 days 4 days 6 days 8 days 10 days 12th 14 days ammonia 14.5 15.75 10.25 8.75 6.25 5.25 5.5 5 Hydrogen sulfide 0.7 0.5 0.3 0.35 0.3 0.4 0.45 0.4

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. It can be understood that

Korea Microorganism Conservation Center (overseas) KCCM11490 20131127 Korea Microorganism Conservation Center (overseas) KCCM11491 20131127 Korea Microorganism Conservation Center (overseas) KCCM11492 20131127

<110> JeollaNamdo <120> MICROBIAL AGENT FOR DEODORANT FOR DECREASING BAD SMELL, AND THE          METHOD USING THE SAME <160> 3 <170> Kopatentin 1.71 <210> 1 <211> 1345 <212> DNA <213> Bacillus thuringiensis JLRI 02 <220> <221> gene &Lt; 222 > (1) .. (1345) <223> 16S rDNA sequence <400> 1 ttgctctcaa gaagttagcg gcggacgggt gagtaacacg tgggtaacct gcccataaga 60 ctgggataac tccgggaaac cggggctaat accggataac attttgaacc gcatggttcg 120 aaattgaaag gcggcttcgg ctgtcactta tggatggacc cgcgtcgcat tagctagttg 180 gtgaggtaac ggctcaccaa ggcaacgatg cgtagccgac ctgagagggt gatcggccac 240 actgggactg agacacggcc cagactccta cgggaggcag cagtagggaa tcttccgcaa 300 tggacgaaag tctgacggag caacgccgcg tgagtgatga aggctttcgg gtcgtaaaac 360 tctgttgtta gggaagaaca agtgctagtt gaataagctg gcaccttgac ggtacctaac 420 cagaaagcca cggctaacta cgtgccagca gccgcggtaa tacgtaggtg gcaagcgtta 480 tccggaatta ttgggcgtaa agcgcgcgca ggtggtttct taagtctgat gtgaaagccc 540 acggctcaac cgtggagggt cattggaaac tgggagactt gagtgcagaa gaggaaagtg 600 gaattccatg tgtagcggtg aaatgcgtag agatatggag gaacaccagt ggcgaaggcg 660 actttctggt ctgtaactga cactgaggcg cgaaagcgtg gggagcaaac aggattagat 720 accctggtag tccacgccgt aaacgatgag tgctaagtgt tagagggttt ccgcccttta 780 gtgctgaagt taacgcatta agcactccgc ctggggagta cggccgcaag gctgaaactc 840 aaaggaattg acgggggccc gcacaagcgg tggagcatgt ggtttaattc gaagcaacgc 900 gaagaacctt accaggtctt gacatcctct gaaaacccta gagatagggc ttctccttcg 960 ggagcagagt gacaggtggt gcatggttgt cgtcagctcg tgtcgtgaga tggttggggt 1020 taagtcccgc aacgagcgca acccttgatt ttagttgcca tcattaagtt gggcactcta 1080 aggtgactgc cggtgacaaa ccggaggaag gtggggatga cgtcaaatca tcatgcccct 1140 tatgacctgg gctacacacg tgctacaatg gacggtacaa agagctgcaa gaccgcgagg 1200 tggagctaat ctcataaaac cgttctcagt tcggattgta ggctgcaact cgcctacatg 1260 aagctggaat cgctagtaat cgcggatcag catgccgcgg tgaatacgtt cccgggcctt 1320 gtacacaccg cccgtcacac cacga 1345 <210> 2 <211> 1327 <212> DNA <213> Pseudomonas caeni JLRI 03 <220> <221> gene &Lt; 222 > (1) .. 1327 <223> 16S rDNA sequence <400> 2 gattgaacgc tggcggcagg cttaacacat gcaagtcgag cggtaacaga gagtagcttg 60 ctactctgct gacgagcggc ggacgggtga gtaatgccta ggaatctgcc tagtagtggg 120 ggataacttg ccgaaaggta agctaatacc gcatacgtcc tacgggagaa agcaggggac 180 tcttcggagc cttgcgctat tagatgagcc taggtcggat tagttagttg gtgaggtaat 240 ggctcaccaa gaccgcgatc cgtaactggt ctgagaggat gatcagtcac actggaactg 300 agacacggtc cagactccta cgggaggcag cagtggggaa tattggacaa tgggggaaac 360 cctgatccag ccatgccgcg tgtgtgaaga aggtcttagg attgtaaagc actttaagtt 420 gggaggaagg gcagttaact aatatttgac tgttttgacg ttaccgacag aataagcacc 480 ggctaacttc gtgccagcag ccgcggtaat acgaagggtg caagcgttaa tcggaattac 540 tgggcgtaaa gcgcgcgtag gtggttcagt aagttaggag tgaaagcccc gggctcaacc 600 tgggaattgc ttctaaaact gctgggctag agtacagtag agggtggtgg aatttcctgt 660 gtagcggtga aatgcgtaga tataggaagg aacatcagtg gcgaaggcga ccacctggac 720 tgatactgac actgaggtgc gaaagcgtgg ggagcaaaca ggattagata ccctggtagt 780 ccacgccgta aacgatgtca actagttgtt gggttccttg aggacttagt aacgcagcta 840 acgcattaag ttgaccgcct ggggagtacg gccgcaaggt taaaactcaa atgaattgac 900 gggggcccgc acaagcggtg gagcatgtgg tttaattcga agcaacgcga agaaccttac 960 ctggccttga catgctgaga actttctaga gatagattgg tgccttcggg aactcagaca 1020 caggtgctgc atggctgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgtaacg 1080 agcgcaaccc ttgtccttat ttaccagcac gtaatggtgg gcactctaag gagactgccg 1140 gtgacaaacc ggaggaaggt gggggatgac gtcaagtcat catggccctt acggccaggg 1200 ctacacacgt gctacaatgg ttggtacaac gggctgccaa gtcgcgagac ggagctaatc 1260 ccataaaacc aatcgtagtc cggatcgcag tctgcaactc gactgcgtga agtcggaatc 1320 gctagta 1327 <210> 3 <211> 1171 <212> DNA <213> Rhodobacter sphaeroides JLRI 04 <220> <221> gene &Lt; 222 > (1) .. (1171) <223> 16S rDNA sequence <400> 3 tttgagagga tgatcagcca cactgggact gagacacggc ccaaactcct acgggaggca 60 gcagtgggga atcttagaca atgggcgcaa gcctgatcta gccatgccgc gtgatcgatg 120 aaggccttag ggttgtaaag atctttcagg tgggaagata atgacggtac caccagaaga 180 agccccggct aactccgtgc cagcagccgc ggtaatacgg agggggctag cgttattcgg 240 aattactggg cgtaaagcgc acgtaggcgg atcggaaagt cagaggtgaa atcccagggc 300 tcaaccctgg aactgccttt gaaactcccg atcttgaggt cgagagaggt gagtggaatt 360 ccgagtgtag aggtgaaatt cgtagatatt cggaggaaca ccagtggcga aggcggctca 420 ctggctcgat actgacgctg aggtgcgaaa gcgtggggag caaacaggat tagataccct 480 ggtagtccac gccgtaaacg atgaatgcca gtcgtcgggc agcatgctgt tcggtgacac 540 acctaacgga ttaagcattc cgcctgggga gtacggccgc aaggttaaaa ctcaaaggaa 600 ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgcagaac 660 cttaccaacc cttgacatgg cgatcgcggt tccagagatg gttccttcag ttcggctgga 720 tcgcacacag gtgctgcatg gctgtcgtca gctcgtgtcg tgagatgttc ggttaagtcc 780 ggcaacgagc gcaacccacg tccttagttg ccagcattca gttgggcact ctagggaaac 840 tgccggtgat aagccggagg aaggtgtgga tgacgtcaag tcctcatggc ccttacgggt 900 tgggctacac acgtgctaca atggcagtga caatgggtta atcccaaaaa gctgtctcag 960 ttcggattgg ggtctgcaac tcgaccccat gaagtcggaa tcgctagtaa tcgcgtaaca 1020 gcatgacgcg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca ccatgggaat 1080 tggttctacc cgaaggcggt gcgccaacct cgcaagagga ggcagccgac cacggtagga 1140 tcagtgactg gggtgaagtc gtaacaaggt a 1171

Claims (5)

A malodor reducing microorganism preparation comprising Bacillus thuringiensis JLRI 02 KCCM 11490P, Pseudomonas caeni JLRI 03 KCCM 11491P, and Rhodobacter sphaeroides JLRI 04 KCCM 11492P. A malodor reducing microorganism preparation comprising Bacillus thuringiensis JLRI 02 KCCM 11490P, Pseudomonas caeni JLRI 03 KCCM 11491P, and Rhodobacter sphaeroides JLRI 04 KCCM 11492P, And spraying the manure in powder or liquid form. Bacillus thuringiensis JLRI 02 KCCM 11490P ( Bacillus thuringiensis JLRI 02 KCCM 11490P) with odor reducing activity. Pseudomonas aeruginosa JLRI 03 KCCM 11491P ( Pseudomonas caeni JLRI 03 KCCM 11491P) Rhodobacter sphaeroides JLRI 04 KCCM 11492P, which has an odor reducing activity,