WO2010147345A2 - Bacillus subtilis gu1 and use thereof - Google Patents

Abstract

The present invention relates to novel microorganism Bacillus subtilis GU01 separated from bamboo smoke distillates, and to a use thereof. The microorganism of the present invention can be grown in a culture medium containing 0.1 to 50 weight % of pyroligneous liquor, and therefore, can be used as an ingredient of feed additives containing a pyroligneous liquor, and further can be used as a microorganism for a bio-filter which sprays a pyroligneous liquor for removing malodorous substances.

Description

Bacillus subtilis ＧＵ１ and use

The present invention relates to a microorganism isolated from bamboo vinegar and the use of the microorganism, the microorganism of the present invention is used in conjunction with the vinegar solution to bring the effect of reducing the mortality rate of livestock, shorten the shipping period, increase the history of history, reduce the smell of meat and improve meat quality It can be used as a feed additive and used as a microorganism for biofilters to absorb, adsorb, and decompose complex odorous gases such as ammonia, hydrogen sulfide, hydrocarbons, lower fatty acids and VOCs.

Wood vinegar contains about 100 kinds of trace components such as organic acids, alcohols, neutral components and basic components, and main components include organic acids such as acetic acid and functional substances such as alcohols and polyphenols. It is used.

With the recent increase in consumer interest in hygiene and safe livestock products due to food safety accidents caused by new harmful substances, the reality is that now more emphasis is placed on taste and safety than on sheep. In addition, food safety management standards are being strengthened internationally. Securing safety is also the most important task for overseas export of domestic livestock products. As the response strategy for the full opening of livestock products is urgently needed, the high-level branding of ducks, broilers, and pork, which Koreans prefer as a health food, is increasing.

 Meanwhile, ammonia, sulfur compounds, fatty acids, amines, hydrocarbons, VOCs, etc. generated from odorous sources such as livestock manure treatment plant, food storage and treatment plant, sewage treatment plant, by-product fertilizer plant, waste storage and treatment plant, sewage relay pump plant, and village sewage treatment plant. Adsorption deodorization method, chemical liquid cleaning method and biofilter method are mainly used to collect and remove air pollution (odor) substances.

 Adsorption deodorization method passes odorous substances through adsorbents such as activated carbon and removes them by adsorbing gaseous air pollution (odor) substances into fine pores, and adsorbents filled with air pollution (odor) substances are frequently used to maintain high removal efficiency. It has to be recycled or exchanged and has low processing efficiency.

 The chemical liquid cleaning method is a method of removing the odorous substances to be deodorized by selecting and using a chemical that removes the odorous substances to be deodorized by spraying finely diluted circulating water in the reactor to make contact with air pollution (odor) substances. Although there is an advantage in height, the use of chemicals not only causes problems of worker health hygiene and polluting the surrounding environment, but also has disadvantages such as increased water pollution due to the treatment of waste deodorizer.

 The biofilter method is environmentally friendly and high in deodorization efficiency to remove odor gas by mechanisms such as absorption, adsorption and decomposition of microorganisms, but in case of high concentration of odor gas generated in large quantities, it requires excessive installation site to secure sufficient contact time. Since the initial investment cost is high and the ammonia gas in the atmosphere is absorbed and treated in the circulating water, there is a disadvantage of greatly increasing the total nitrogen content in the water treatment process of the circulating water waste liquid.

 In addition, the method of spraying deodorant and disinfectant with high pressure sprayer is mainly used for the improvement of livestock environment such as odor gas removal and disinfection of livestock house (pig, duck, chicken, etc.), but liquid deodorant or disinfectant is diluted with water and sprayed. When the stress is applied to the livestock and the effect is greatly effected, but the duration is short, because it has to be sprayed periodically, there is a drawback of manpower consumption and drug consumption.

 The deodorization efficiency of antibiotics to improve the safety of food and functional microorganisms for improving meat quality and existing deodorization methods are low, and to overcome the problems of handling and environmental pollution, the efficiency of deodorizers and the efficiency of deodorizing devices The need for development is increasing.

The present invention is used together with the wood vinegar solution, which is difficult to achieve by the wood vinegar alone, the effect as a feed additive and the odorous substance removal effect to improve the weight gain, carcass rate, meat color and physicochemical properties and physical properties, the content of unsaturated fatty acids, etc. In the present invention, new strains having a synergistic effect are isolated and identified, and the new strains are used in a biofilter apparatus using a feed additive containing wood vinegar and wood vinegar.

The present invention provides Bacillus subtilis GU1 [Accession No. KCCM 10890P], which is isolated from bamboo vinegar and is capable of growing in a medium containing 0.1 to 50% by weight of wood vinegar.

The present invention provides a culture in which Bacillus subtilis GU1 [Accession No. KCCM 10890P] strain is fermented in a medium containing 0.5 to 10% by weight of wood vinegar.

In the culture of the present invention, the medium is characterized in that the solid medium.

The present invention provides a feed additive comprising a culture fermented Bacillus subtilis GU1 [Accession No. KCCM 10890P] strain in a medium containing 0.5 to 10% by weight of wood vinegar.

The present invention provides a biofilter comprising a blower for introducing a malodorous substance into the biofilter device, a humidifier for supplying moisture into the biofilter device, and a carrier having microorganisms attached thereto, wherein the humidifier is a wood-choice solution composition. Bacillus subtilis GU1 [Accession number KCCM, characterized in that the sprayed washing herbaceous scrubber, wherein the microorganisms attached to the carrier are separated from the bamboo vinegar solution and grow in a medium containing 0.5 to 50% by weight of bamboo vinegar solution. 10890P] to provide a biofilter device for removing odorous substances.

In the biofilter device for removing odorous substances of the present invention, the microorganism is Nitrosomonas sp., Bacillus sp., Pseudomonas sp., Nitrosomonas sp. .) Strains, nitrobacter sp. ( Nitrobacter sp.) Strains, genus Bacillus (Thiobacill us sp.) Is characterized in that any one or more strains are further included.

In the bio-filter device for removing the odor substance of the present invention, the ablution vinegar disk rabeo the derived Bacillus from wood vinegar with wood vinegar composition subtilis (Bacillus subtilis) GU1 [Accession No. KCCM 10890P] strain and Bacillus subtilis (Bacillus subtilis ) GU2 [Accession No. KCCM 10891P] is characterized in that any one or more strains selected from among the strains are sprayed together.

Bacillus subtilis GU1 of the present invention is mixed with wood vinegar or fermented in a medium containing wood vinegar solution as a feed additive that can not be achieved conventional animal vinegar mortality mortality reduction, shorten the shipping period, increase the history of history, reduce the smell of meat and meat quality It brings the effect of improvement.

In addition, the biofilter device of the present invention is excellent in removing odorous substances due to Bacillus subtilis GU1 [Accession No. KCCM 10890P] strain isolated from bamboo vinegar solution, and particularly, a scrubber for spraying wood vinegar solution with a humidifier. By using it can significantly reduce the concentration of foreign matter and ammonia-odorous odor substances introduced into the biofilter.

1 is a schematic view of a biofilter apparatus using a cleaning throat (chochu) vinegar scrubber as one of the embodiments of the present invention.

2 is a schematic view of a biofilter apparatus to which a chemical cleaning apparatus or a metal catalyst deodorizing apparatus, which is one of embodiments of the present invention, is added.

Figure 3 is a graph showing the number of bacteria after treatment of the strain of the present invention at each temperature for 10 minutes.

[Code Description of Main Part of Drawing]

10: biofilter unit 11, 21, 31: spray nozzle

12, 22, 32: inlet 13, 23, 33: outlet

14: carrier 15: nutrient medium supply tank

16: Alkaline supply tank 17: Acid supply tank

18: microbial incubator 19: blower

20: humidifier [washing wood vinegar scrubber] 24, 34: filling material layer

25, 35: reservoir 26, 36: circulation pump

27: wood vinegar 30: chemical cleaning device

37: chemical

The strain of the present invention was isolated and identified in bamboo vinegar solution of pH 2.6 produced in Damyang, Jeollanam-do, and the isolated strain was identified as Bacillus subtilis . The strain was named Bacillus subtilis GU1 and was deposited internationally in accordance with the Budapest Treaty under the deposit number KCCM 10890P on November 21, 2007.

The strain of the present invention has the effect of reducing livestock mortality that can not be achieved alone, in the form of a culture solution fermented in the medium containing or mixed with the wood vinegar solution, shorten the shipping period, increase the history of history, reduce the smell of stalk and improve the meat quality Bring.

The medium for the culture of the strain of the present invention, the wood vinegar used as an active ingredient of feed additives or odorant removal agent is conifers, deciduous trees, trees, shrubs, evergreens, deciduous trees, evergreen conifers, deciduous conifers, evergreen softwoods, deciduous softwoods Cooling and condensing liquids or essential oils or vapors produced by one or more processes selected from pyrolysis, bath, heating, distillation, extraction, and weeding of any one or more selected from leaves, stems, roots, fruits and flowers of vegetation; It can be prepared through one or more processing selected from, separation, ripening, filtration, evaporation, and distillation. For example, the wood vinegar of the present invention is one or two selected from conifers, deciduous trees, arbors, shrubs, evergreens, deciduous trees, evergreen conifers, deciduous conifers, evergreen hardwoods, deciduous hardwoods, and leaves, stems, roots, fruits, and flowers of vegetation. Mixing the above to direct or indirect pyrolysis at 150 to 350 ℃, repeated heating in hot water, warmed to 80 to 150 ℃ in atmospheric pressure or reduced pressure in a cauldron, extraction by solvent extraction or supercritical fluid extraction method, Or steaming and cooking in a cauldron, collecting and cooling liquid condensate or vapor generated during the pyrolysis, bath, warming, extraction, or steaming process, and cooling down and condensing the plant essential oil components contained in the cooled or condensed wood vinegar solution. The oil and water are separated for recovery, the foreign matter and tar contained in the wood vinegar are filtered and the wood vinegar is Aged at room temperature or warmed for stabilization, and is prepared by separating the foreign matter and tar contained in the wood vinegar. Particularly, the process is carried out in a cauldron at a temperature not pyrolyzed and carbonized, and heated at 80 to 150 ° C. under atmospheric pressure or reduced pressure, or repeated heating and heating, followed by extraction, heating, distillation, or steaming by solvent extraction or supercritical fluid extraction. The wood vinegar composition prepared by capturing a liquid substance or steam or essential oil which occurs in the process of cooling, condensation and separation is preferable because of its low tar content. The wood vinegar of the present invention can be used by mixing two or more kinds of wood vinegar with different raw materials or manufacturing methods.

Cultures fermented by the strain of the present invention in a medium containing wood vinegar significantly increases the nutritional or immunological characteristics of wood vinegar and at the same time increases the odor removal efficiency. The medium may be liquid or solid, and when used as a feed additive, it is preferable to use a solid medium because it may be easily mixed uniformly with a solid feed, and the solid medium may be a nutrient source. The preferred raw material for the solid medium may be a by-product of plant processing including cereals, soybeans, yeast or wood, which are used as raw materials for feed, and various raw materials such as skimmed steel, soybean meal, sawdust, etc. may be used in combination as necessary. Can be.

The content of wood vinegar is 0.5 to 10% by weight, more preferably 2 to 10% by weight, most preferably 2 to 5% by weight, based on the weight of the medium added to the medium to be inoculated with the strain of the present invention. In addition, if the culture of the present invention is preferably incubated at 15 ~ 45 ℃, preferably 20 ~ 42 ℃, more preferably 30 ~ 40 ℃, the incubation time is 12 hours or more, preferably 24 hours or more, more Preferably 36 hours or more, even more preferably 48 hours, the maximum incubation time is not particularly limited, but even if culture for 48 hours or more because the strain amount of the present invention is not increased within 60 hours.

In addition, even if the strain of the present invention is mixed with wood vinegar and formulated without fermentation, and used as a feed additive or odor removing agent, post-fermentation proceeds after the formulation is achieved to achieve the effect as a feed additive of the present invention can do.

Hereinafter, a biofilter device for removing contaminants of the present invention will be described with reference to the drawings illustrated in FIGS. 1 and 2. The biofilter device 10 includes a blower 19 for introducing malodorous substances into the biofilter device, a humidifier 20 for supplying moisture into the biofilter device, and a carrier 14 to which microorganisms are attached. Any conventional biofilter device may be used.

In the biofilter device of the present invention, the humidifying apparatus 20 may be a cleaning wood-vinegar scrubber sprayed with a wood-choice solution composition. In the biofilter body, the wood vinegar composition reacts with ammonia gas, hydrogen sulfide, and methyl mercaptan, which are odor pollutants, and is neutralized and adsorbed with ammonium acetate salt, hydrogen sulfide ion, methyl mercaptan ion, etc., and is discharged to the outside together with sludge as shown in the following reaction formula. Wastewater discharged from the exchange of scrubber circulating water does not affect the increase of nitrogen compound content in the sewage water treatment process, which is the biggest problem in the existing biofilter process. Deviation is not a big problem. In addition, even if a high concentration of malodorous gas is introduced, since 80 to 90% is removed from the cleansing wood-vinegar scrubber sprayed with the wood-vinegar composition, the low-density residual gas is introduced into the biofilter device, so the deodorizing efficiency is high.

The removal of ammonia gas from the atmosphere is carried out by dissolving acetic acid (CH ₃ COOH), the main component of the wood vinegar composition, into acetate ions (CH ₃ COO ⁻ ) and hydrogen ions (H ⁺ ) as shown in Scheme 1 below. In addition, ammonia (NH ₃ ) gas among the malodorous substances flowing into the wood vinegar scrubber is hydrolyzed in an aqueous solution as in Scheme 2 and dissociated into ammonium ions (NH ₄ ⁺ ) and hydroxide ions (OH ⁻ ). Therefore, the ammonia gas introduced into the wood vinegar scrubber body increases the pH of the wood vinegar aqueous solution due to the increase of ammonium ions and hydroxyl ions produced by the hydrolysis reaction in the wood vinegar aqueous solution. The removal of the ammonia gas from the odorous substance is determined to remove the odor as ammonium acetate (CH ₃ COONH ₄ ) dissociated in the aqueous solution of the wood vinegar to the acid ions and acid-base reaction as in Scheme 3. The expected reaction scheme of the wood vinegar composition is as follows.

<Scheme 1>

^{_{CH 3 COOH → CH 3 COO -}} + H +

<Scheme 2>

_{NH 3 + H 2 O → NH} 4 + + OH -

<Scheme 3>

_{^{CH 3 COO - + NH 4 +}} → CH 3 COONH 4

The removal of hydrogen sulfide gas from the sulfur compounds in the air is odor due to the formation of fixed non-volatile salts of acetic acid (CH ₃ COOH) and HS ^- by reaction with acetic acid ion and acid base generated by dissociation of acetic acid in aqueous solution of wood vinegar. In the case of methyl mercaptan (CH ₃ SH) gas, a reaction mechanism is expected in which odor is removed as fixed nonvolatile salts of acetic acid (CH ₃ COOH) and CH ₃ S ⁻ are generated as in Scheme 5. The expected reaction scheme of the wood vinegar composition is as follows.

<Scheme 4>

^{_{H 2 S + CH 3 COO -}} → CH 3 COOH + HS -

Scheme 5

^{_{CH 3 SH + CH 3 COO -}} → CH 3 COOH + CH 3 S -

Devices for satisfying the conditions necessary for the growth of microorganisms inside the biofilter device may be further added. Those that may be added to the biofilter device include a nutrient medium supply tank 15, an alkali supply tank 16 or an acid supply tank 17 for adjusting pH conditions to a range suitable for microbial growth, and also a temperature for microbial growth. And a thermostat equipped with a heater or a cooler outside the biofilter for adjusting to a suitable range. The heater or cooler may be installed in the warmer or cooler inside the biofilter, or may be installed in such a manner as to surround the outside of the biofilter device like a jacket. In addition, if necessary, air or an oxygen supply device may be further provided to increase the oxygen concentration in the biofilter.

In the present invention, as the carrier 14 to which the microorganism is attached, any organic material such as pit, bark, compost, or organic carrier such as porous foamed polymer may be used as well as inorganic carrier such as porous ceramic. Preferably, a porous ceramic carrier having high microbial activity per unit weight, easy maintenance and high water content is used.

The microorganism attached to the carrier of the biofilter device of the present invention is Bacillus subtilis GU1 [Accession No. KCCM 10890P] isolated from bamboo liquor, and the strain is excellent in removing odorous substances, and bamboo vinegar solution (usually pH 2.6). Because it can grow inside and outside), even if the internal pH of the biofilter device drops sharply, the microorganisms are not affected by pH and do not interfere with the treatment of odorous substances. In particular, Bacillus subtilis GU1, which is highly resistant to acidic acid, has a low pH because organic acids are continuously produced during fermentation in addition to carbon dioxide and water as odorous substances are decomposed by microorganisms fixed on the carrier. It is advantageous properties as a microorganism for biofilters.

A microorganism attached to the carrier of the present invention wherein the Bacillus subtilis in other nitro consumption eggplant is used to remove malodorous substances with GU1 strain (Nitrosomonas sp.) Strain, Bacillus (Bacillus sp.) Strain, Pseudomonas species (Pseudomonas sp.) strain, Nitrosomonas sp., Nitrobacter sp., Thiobacillus sp. It is preferable in that the biofilter device can be stably operated without the need.

In particular, in the present invention, using Bacillus subtilis GU2 [Accession No. KCCM 10891P] strain isolated from bamboo shoot liquid by the present inventors together with Bacillus subtilis GU1 is to be used by Bacillus subtilis GU1 alone. It is more advantageous in that the removal efficiency is increased in adsorbing and decomposing odorous substances such as ammonia and sulfur compounds.

Both Bacillus subtilis GU1 and GU2 strains were isolated from the bamboo vinegar solution at pH 2.6 produced in Damyang, Jeollanam-do, and were reported to Korea Gene Bank on November 21, 2007, under the deposit numbers KCCM 10890P and KCCM 10891P, respectively. It has been deposited internationally under the Treaty.

The biofilter device of the present invention further comprises a microbial incubator 18 and a microbial supply device for transferring the microorganisms cultured in the microbial incubator to the biofilter device, the microorganisms being supplied from the outside and injected into the carrier of the biofilter. It is advantageous in lowering the cost. The microorganism incubator 18 preferably treats the outside of the incubator with a heat insulator to maintain a constant temperature in the incubator in order to adjust the growth conditions of the microorganisms, or more preferably, is provided with a heater. The incubator may be equipped with a cooler, but most of the microorganisms for removing odorous substances are mesophilic bacteria, and the use frequency is low since domestic summer temperatures rarely exceed 37 ℃. The microorganisms cultured at a predetermined concentration or a predetermined number of bacteria in the microorganism incubator are supplied into the biofilter device through a microorganism supply device consisting of a transfer pump and a transfer tube. Preferably, it is preferable to supply through the spray nozzle 11 so as to be evenly attached to the carrier, it is preferable to supply to a position spaced a certain distance from the acid or alkali supply device. The microorganism is supplied to the biofilter device may be used either in a continuous or intermittent manner of supplying a certain amount or a method of supplying a variable by a pre-programmed formula. In addition, the microbial incubator 18 is preferably added with a supply device for supplying air or oxygen.

When the microorganism is discharged from the microbial incubator, it is preferable that the nutritional medium supply device 15 is connected to the microbial incubator 18 so that the nutritional medium can be supplied to supplement the microorganisms.

In the biofilter apparatus of the present invention, the humidifier 20 is preferably a cleaning wood-choice scrubber to which the wood-choice composition is injected, in that it can simultaneously perform the removal of humidification and odorous substances, especially ammonia-based substances. Any type of clean scrubber can be used without limitation, but the demister must be removed from the outlet through which the treated air is discharged from the clean scrubber so that water can be supplied to the biofilter device.

In one embodiment of the cleaning wood vinegar scrubber of the present invention is provided with a blower 19 in the inlet 22, the cleaning scrubber provided with a wood vinegar composition spray nozzle 21 in the upper portion of the main body to the wood vinegar composition supply unit Can be used. It is preferable to form a swirl flow to increase the time for which odorous substances forced into the main body through the blower stay in the main body, and forced inflow of pollutants by the blower increases the effect of contact with the deodorant of the present invention. It also increases efficiency and prevents odors from escaping. One or more spray nozzles are installed in the upper region of the main body for the widespread spraying of the wood vinegar composition. The number of spray nozzles to be installed is determined by the pressure of the blower, the rate of rise of odorous substances, and the desired treatment efficiency. The spray nozzles are installed upwards or downwards and it would be economically advantageous for the number and spacing of these spray nozzles not to overlap each other with the total volume of the spray space and the sprayed up or down regions. In addition, the lower portion of the main body may be provided with a reservoir 25 for storing the falling wood vinegar composition, a circulation pump 26 may be provided to recycle the wood vinegar composition stored in the reservoir to the spray nozzle. In addition, a filler layer 24 such as polling having a large surface area of various forms may be provided to increase the contact efficiency of the vinegar liquor in the form of droplets sprayed by the spray nozzle inside the main body and odorous substances.

In the wood vinegar of the present invention or the wood vinegar mixed with two or more kinds of wood vinegar, or a mixture or the above-mentioned wood vinegar, plant pyrolysis composition, metal catalyst, photocatalyst, plant essential oil, plant extract, oxidizing agent, reducing agent, phytonutrient, organic acid, fermenting agent, flavoring agent, adsorbent , Microorganisms, microbial agents, clay solution, ocher solution, mud flats, surfactants or fragrances may be mixed (hereinafter, wood-vinegar composition) to be used. Mixing ratio may be used to 0.01 to 90% by weight based on the total weight of wood vinegar. In addition, 0.01 ~ 10% by weight of deodorant, deodorant, disinfectant, fungicide, etc. may be added as necessary in a range that does not inhibit the growth or proliferation of the Bacillus subtilis GU1 strain of the present invention.

In addition, to increase the reaction rate of wood vinegar and odorous substances, to control the amount of evaporation or to promote evaporation, wood vinegar is mixed with a solvent, such as water of 2 to 1000 times the weight of wood vinegar solution or alcohol of 1 to 4 carbon atoms, or separately evaporated Accelerators may be added.

Bacillus subtilis GU1 strain, Bacillus subtilis GU2 strain, or mixtures thereof may be added to the wood vinegar to increase the odor gas removal efficiency which is treated in the cleaned wood vinegar scrubber before entering the biofilter device of the present invention. The concentration is 1.00E + 02 ~ 1.00E + 08 cfu / ml, preferably 1.00E + 03 ~ 1.00E + 08 cfu / ml, more preferably 1.00E + 04 ~ 1.00E + It is preferably added at a concentration of 08 cfu / ml.

In the biofilter device of the present invention, one or more deodorizers may be used in series at the front end or the rear end, and a separate chemical cleaning device, a filter deodorizer, a membrane vaporizer, an ozone deodorizer, a biofilter deodorizer, and a metal catalyst deodorizer may be used. Adsorption deodorizer, plasma deodorizer, incineration deodorizer and single or two or more types can be connected and used in series. Preferably, the addition of the acid gas or VOCs chemical liquid cleaning device 30 or the metal catalyst (photocatalyst) deodorizer to the front stage of the biofilter device or the scrubber wood scrubber attached to the biofilter device is the microorganisms in the acid gas. It is not preferable to the impact caused by the impact, and to increase the efficiency of removing the odorous substances of the biofilter device, it is desirable in terms of economic and operational.

Hereinafter, the present invention will be described in detail by Examples and Experimental Examples. However, the following Examples and Experimental Examples are merely illustrative of the present invention, but the content of the present invention is not limited thereto.

Example 1 Isolation, Identification and Characterization of Strains

(1) separation

In order to separate useful acid-resistant strains from bamboo vinegar, bamboo vinegar with pH 2.6 produced in Damyang, Jeollanam-do, which had been aged for more than 6 months, was used. In order to isolate the microorganisms present in the bamboo vinegar solution was plated on LB agar medium, MRS agar medium, PDA agar medium and mannitol eggyolk polymyxine (MEP) medium. After the first selection of microorganism-grown PDA agar medium and strains grown in MEP medium, the strains were grown at pH 2.6, and the final strain was selected with excellent amylase and protease activity. The final selected strains were identified using an optical microscope and Gram staining was performed. The strain was identified as a strain capable of growing in a solid medium and difficult to grow other microorganisms at low pH conditions of bamboo vinegar solution.

(2) Molecular Biology Identification

DNA extracted from the strain and 16s rDNA analysis showed the highest homology with Bacillus subtilis HDYM-11 strain and identified as Bacillus subtilis (1511 matches among 1515 sequences, 99% phase). Homosexual). The sequence of 16s rDNA obtained is shown in SEQ ID NO: 1. Another strain was identified as Bacillus subtilis with 99% homology with 1512 sequences among Bacillus subtilis and 1517 sequences, and the sequence of 16s rDNA obtained was shown in SEQ ID NO: 2.

(3) heat resistance

To be used as a feed additive, it must be able to survive under high temperature and high pressure conditions such as general pelleting (80 ° C.) or extrusion (120 ° C.). In general, Bacillus microorganisms produce a variety of useful substances, including enzymes, non-toxic antibacterial substances, pesticides, etc., and is also a gram-positive bacteria, characterized by the formation of spores at high temperatures.

In order to test the heat resistance of the Bacillus subtilis GU01 strain of the present invention, when the absorbance (Opical Density, 600nm) value is 1.0, the number of bacteria after heat treatment for 10 minutes at each temperature was measured and shown in FIG. As shown in FIG. 3, the viability was excellent even when treated at 80 ° C., far exceeding 60 ° C., which is a common killing temperature of lactic acid bacteria used as a useful microorganism of feed additives. In addition, the Bacillus subtilis GU02 strain also did not differ from before the heat treatment up to 45 ℃ bacteria number 1.00E + 08 cfu / ml, and maintained at more than 1.00E + 08 cfu / ml at 60 ℃.

(4) Change of microbial bacteria in bamboo vinegar

Bamboo vinegar is a strong acidic condition of pH 2.6, after diluting the bamboo vinegar in distilled water at a constant ratio, the growth change according to the incubation time of the strain (GU01) of the present invention at each concentration is measured by the change in absorbance at 600 nm is shown in Table 1 In addition, the change in the number of microbial microorganisms with incubation time is shown in Table 2. Bamboo liquor was diluted 10, 20, 30, 40 and 50% by weight in distilled water, respectively, and the initial microbial inoculation concentration was 1.00E + 05 cfu / ml and incubated at 37 ° C.

Table 1

time	10%	20%	30%	40%	50%
0	0.239	0.254	0.280	0.288	0.319
24	0.335	0.378	0.423	0.456	0.375
48	0.375	0.385	0.477	0.523	0.516

TABLE 2

time	10%	20%	30%	40%	50%
0	4.00E + 05	8.00E + 05	6.00E + 05	2.00E + 05	1.40E + 06
24	1.00E + 07	1.20E + 07	7.00E + 06	6.00E + 06	3.00E + 07
48	1.03E + 07	1.04E + 07	1.04E + 07	1.07E + 07	1.07E + 07

The growth (absorbance of 600 nm) of the strain of the present invention was increased as the incubation time was increased, increased to 40% by weight of bamboo vinegar concentration, but no longer increased from 50% by weight. In addition, the number of bacteria of the strain of the present invention was increased up to 24 hours, even if incubated for 48 hours, the change in the number of bacteria was insignificant, and the change in the number of other bacteria in the bamboo shoot concentration was also insignificant.

Therefore, the strain of the present invention was able to grow in a medium containing more than 0.1% by weight of bamboo vinegar, especially in the medium added up to 50% bamboo vinegar was determined that the growth will not be significantly affected.

Experimental Example 1: Effect on broiler productivity

In order to confirm the effect of the bamboo vinegar solution contained in the feed additive of the embodiment of the present invention on the broiler productivity, the degreasing steel whose water content was adjusted to 40% by weight was added to the group (Comparative Example 1) and degreasing steel instead of the feed additive. Addition of 2% by weight of bamboo vinegar solution and a feed additive (Comparative Example 2) in which the water content was adjusted to 40% by weight were used by adding 0.2% by weight to the feed for general broilers.

175 broiler chickens of the same size were stocked in the barns of Comparative Examples 3 and 4 and Example 3, respectively. After breeding broilers for 35 days, the number of shoots and growth rate were measured, and 10 broilers were randomly selected from each experimental group to analyze the start weight, end weight, weight gain and broiler chemical characteristics.

TABLE 3

Item	Comparative Example 1	Comparative Example 2
Test	175	175
Starting weight (g)	44.82 ± 0.15	44.86 ± 0.10
End weight (g)	2079.29 ± 1.67	2055.71 ± 9.18
Breeding days	35	35
Weight gain (g)	2034.46 ± 1.79	2010.86 ± 9.17
Feed Intake (g)	3453.99 ± 1.78	3416.86 ± 1.81

Comparing Comparative Examples 1 and 2, only the addition of bamboo vinegar to the feed did not show any difference in the addition of bamboo vinegar and the weight gain and feed intake, but rather worse, the addition of bamboo vinegar was positive for the weight gain and feed intake of broilers. It was judged not to affect.

In order to confirm the effect of the feed additive of the present invention strain and the bamboo vinegar solid fermentation, 2% by weight of bamboo vinegar solution in the skimmed steel, the strain dilution 2 weight diluted in distilled water at a concentration of 1.00E + 05 cfu / ml strain of the present invention % Was added and the water content was adjusted to 40% by weight, followed by fermentation at 37 to 48 hours to prepare a solid feed additive (Example 1).

The effect of the present invention was confirmed in comparison with the group 1 (Comparative Example 1) in which the degreasing steel, in which the moisture content was adjusted to 40% by weight, was added instead of the feed additive. The test animal barn was 638 m ² (11m X 8m), respectively, and 0.2 wt% of the feeds of Example 1 and Comparative Example 1 were added to the general broiler feed. 14,000 broiler chicks of the same size were stocked in stalls of Comparative Example 1 and Example 1, respectively. After breeding broilers for 35 days, the number of shoots and growth rate were measured, and 100 broilers were randomly selected from each experimental group to analyze the start weight, end weight, weight gain, and chemical characteristics of broilers.

Table 4

Item	Comparative Example 1	Example 1
Test	14,000	14,000
Starting weight (g)	42.0	42
End weight (g)	1,789	1,870
Breeding days	35	35
Daily gain (g / d)	49.91	52.23
Feed Intake (kg)	2.68	3.24
Dead shooter	426	419
Growth rate (%)	96.96	97.01

As a result of the breeding test, the weight gain per day was 52.23g in Example 1 and 49.91g in Comparative Example 1, which was 2.32g higher than in Comparative Example 1, and Example 1 was 97.09% and Comparative Example 1 in the growth rate. 96.96%, which is 0.13% higher (Table 4). By feeding the feed additive of the present invention to broiler chickens, it is thought that the reduction of mortality and the increase in the daily weight gain are caused by the increase of immunity by the microorganisms and the death by disease is reduced. In addition, it is believed that the synergistic effect of the mixed strain of the present invention and bamboo vinegar increased the digestibility of broilers and inhibited the growth of intestinal harmful bacteria.

Experimental Example 2 Effects on Meat Quality, Meat Color, General Components and Fatty Acid Composition of Broilers

In Example 1 and Comparative Example 1 in Experimental Example 1 after the 35 days of broiler meat was collected and compared to the effects on meat quality, meat color, general components and fatty acid composition are shown in Tables 5 to 8, respectively.

The heating loss (%) is a sample (250 ± 50g) formed into a certain shape, then put in a polyethylene bag and placed in a water bath at 80 ° C to be completely immersed in water, heated for 40 minutes, and cooled in water for 20 minutes. . After the water of the cooled sample was removed, the weight loss was calculated. Shear force was measured by using a salter (Warner Bratzler Shear, USA) as the shear force measurement sample, which was removed from the surface portion of the sample from which the heating loss was measured.

The sample was placed in a filter tube (VIDAS tube, BIOMERIEUX, France) for the measurement of water holding capacity, placed in an 80 ° C water-bath, heated for 20 minutes, and allowed to cool for 10 minutes. The cooled tube was centrifuged at 2000 ° C. for 10 minutes at 10 ° C., and the free moisture and the water holding capacity were calculated by the formula.

Free moisture = [(difference in weight before and after centrifugation) / sample weight] X fat coefficient (1-fat content) X 100

Water-retaining power (%) = [(whole water-free water) / total water] X100

Table 5

Analysis item	Comparative Example 1	Example 1
Heating loss	22.48	22.36
Shear force (kg / 0.5inch 2 )	1.81	1.21
Conservative	60.53	61.18
pH	5.96	5.88

It was found that Example 1 was significantly lower than that of Comparative Example 1 in shearing force, and the meat was lightly cut well, and in Example 1, it was confirmed that the water holding capacity was excellent.

Table 6

Analysis item	Comparative Example 1	Example 1
L (brightness)	45.66	51.79
a (red)	3.53	4.77
b (yellow)	4.46	4.25

In flesh color, Example 1 had a higher brightness and a lower yellowness than Comparative Example 1, indicating that it had a bright flesh color, and the redness was lower.

TABLE 7

Analysis item	Comparative Example 1	Example 1
moisture	75.73	75.26
Crude fat	0.17	0.49
Crude protein	22.95	23.13
View minutes	1.16	1.12

In Example 1, the content of crude ash was lower than that of Comparative Example 1, and it was found that crude fat and crude protein were higher to provide better meat quality.

Fatty acid content was measured using gas chromatography (model name: Varian 3600).

Table 8

fatty acid	Comparative Example 1	Example 1
Myristic acid (C14: 0)	0.89	0.83
Palmitic acid (C16: 0)	25.06	23.55
Stearic acid (C18: 0)	6.82	6.41
Palmitoleicacid (C16: ln7)	5.32	5.65
Oleic acid (C18: ln9)	44.76	45.76
Vaccenic acid (C18: ln7)	0.01	0.02
Linoleic acid (C18: 2n6)	15.52	15.91
v-Linoleic acid (C18: 3n6)	0.16	0.19
Linolenic acid (C18: 3n3)	0.66	0.68
Eicosenoic acid (C20: ln9)	0.50	0.55
Arachidonic acid (C20: 4n6)	0.30	0.44
Eicosapentaenoicacid (EPA) (C20: 5n3)	0.00	0.00
Docosatetraenoic acid (C22: 4n6)	0.00	0.00
Docosahexaenoic acid (DHA) (C22: 6n3)	0.00	0.00
Total	100.00	100.00
saturated fatty acid	32.76	30.79
Unsaturated fatty acid	67.24	69.21

Also in the fatty acid composition, Example 1 was found to provide a broiler of good quality with a lower content of saturated fatty acids and a higher content of unsaturated fatty acids than Comparative Example 1.

Experimental Example 3: Effect on Duck Productivity

In order to confirm the effect of the feed additive in the solid fermentation of the strain of the present invention and bamboo vinegar, 2% by weight of bamboo vinegar solution in the skim steel, 2% by weight of the dilution of the strain diluted in distilled water at a concentration of 1.00E + 05 cfu / ml strain of the present invention, lactose 0.2 wt% strain diluted in distilled water at a concentration of Bacillus ashidophilus 1.00E + 05 cfu / ml, 0.2 wt% strain diluted in distilled water at a concentration of 1.00E + 05 cfu / ml Lactobacillus plantarum, 0.5 wt% By weight, bamboo charcoal powder 0.1% by weight was added and the water content was adjusted to 40% by weight and then fermented at 37 to 48 hours to prepare a solid feed additive (Example 2). As a control, a group in which degreasing steel having a water content of 40% by weight was added instead of a feed additive (Comparative Example 1) was used.

The scale of the test animal house was 400 m ² (10m X 40m), respectively, and Example 2 and Comparative Example 4 were used by adding 0.2% by weight of the general duck breeding feed to the general duck breeding feed. 10,000 ducks of the same size were stocked in stalls of Example 2 and Comparative Example 1, respectively. After 43 days of duckling, the number of dead and growth rates were measured, and 100 groups of ducks were randomly selected from each experimental group to analyze the initiation weight, end weight, weight gain and feed efficiency.

Table 9

Item	Comparative Example 1	Example 2
Test	10,000	10,000
Starting weight (g)	48	48
End weight (g)	3.190	3.280
Breeding days	43	43
Daily weight gain (g)	74.19	76.27
Feed Intake (kg)	65.709	65.922
Average weight (g)	3.190	3.280
Dead shooter	141	118
Feed requirement (g)	2.10	2.03
Growth rate (%)	96.96	97.09

In Example 2, the weight gain, feed efficiency and growth rate per day were improved compared to Comparative Example 1. In Comparative Example 1, the daily weight gain was 74.19 g, whereas Example 2 was 76.27 g, and Example 2 was higher, and the growth rate was 98.82% in Example 2, 98.57% in Comparative Example 1, and 0.25% in Comparative Example 1. High.

Experimental Example 4: Effect on productivity of swine

60 pigs with an average body weight of 79.66 ± 0.12 kg were divided into three groups with the same average body weight, and the water content was adjusted to 40% by weight to replace the feed additive (Comparative Example 1). It was grown in the same conditions for 4 weeks by dividing into the group added by weight, 0.1 wt% of the feed additive of Example 2 to the feed for general pigs, and 0.2 wt% added to the feed additive of Example 2 to the feed for general pigs. While increasing the daily weight gain, daily feed intake and feed efficiency are shown in Table 10.

Table 10

division	Comparative Example 1	Example 2	Example 2	SE
Amount	0.2 wt%	0.1 wt%	0.2 wt%	-
Daily weight gain (g)	772 b	825 a	821 a	13.3
Feed Intake (g)	2,356	2,480	2,413	65.3
Feed efficiency (%)	32.8	33.3	34.0	0.009

Compared to Comparative Example 1, in Example 2, the addition amount was 0.1 wt% or 0.2 wt%, and the weight gain per day was significantly increased, the feed intake was increased, and the feed efficiency was also confirmed.

On the other hand, the feed additive of Example 2 of the present invention in the pig farm in Jeongeup replaces the feed additive including the general lactic acid bacteria and added 0.2% by weight to the feed for general pigs, as a result, compared to the feed additive containing the common lactic acid bacteria Feeding date of pigs was reduced by 9 days and mortality decreased from 7 ~ 8% to less than 3%.

Experimental Example 5: Removal of odorous substances by direct mixing

In order to confirm the odor removing effect of the strain of the present invention, Example GU1 strain inoculated 1.00E + 05 cfu / ml of the GU1 strain of the present invention in 100 ml of water, 1.00E of the GU1 strain of the present invention in 100 ml of bamboo vinegar solution Example 4 inoculated with +05 cfu / ml, GU2 strain of the present invention in 100 ml of water Example 5 inoculated with 1.00E + 05 cfu / ml, GU1 strain and GU2 strain in 100 ml of water, respectively Example 6 was prepared inoculated with 5.00E + 04 cfu / ml. As a control, a group to which only 100 ml of water was added (Comparative Example 2) and a group to which only 100 ml of bamboo vinegar solution was added (Comparative Example 3) were used for the experiment. The bamboo vinegar used in this experiment was produced in Damyang, Jeonnam, and indirectly heated bamboo cut to 30cm at 250 ~ 300 ℃, and the sap vapor contained in smoke of stack was collected at 80 ~ 150 ℃ for cooling and The condensed crude bamboo vinegar was aged for at least 6 months and filtered once with a filter having a pore size of 5.0 μm.

A test method for removing odorous substances using a PVC column used at the University of Iowa was used (www.nationalhogfarmer.com). 4 ft. Pig manure on a PVC column 15 inches long. The strain dilution of Examples 3 to 6, Comparative Example 2 is added to the distilled water, bamboo paste solution of Comparative Example 3 0.2% by weight of pig manure after measuring the change in the content of odorous substances (unit ppm) as shown in Table 11 It was.

Table 11

division	hr	ammonia	Hydrogen sulfide
Comparative Example 2	0	18	6.0
	12	10	5.5
	24	12	5.5
	48	14	5.5
Comparative Example 3	0	15	6.5
	12	4.5	5.0
	24	7.5	4.5
	48	8.0	5.0
Example 3	0	16	6.0
	12	6.5	3.5
	24	3.0	3.0
	48	3.0	2.0
Example 4	0	16	6.0
	12	4.5	3.0
	24	3.5	2.5
	48	2.5	2.0
Example 5	0	16	6.0
	12	9.0	4.0
	24	5.0	3.0
	48	3.5	2.5
Example 6	0	16	6.0
	12	5.0	3.0
	24	3.0	2.0
	48	2.5	1.5

Compared with Comparative Example 2, Comparative Example 3, in which only bamboo vinegar solution was added, decreased to 4.5 ppm from the initial 15 ppm to 12 hours, but increased again after 24 hours. On the other hand, in Examples 3 to 6 of the present invention was found to decrease from 4.5ppm to 9.0ppm from the initial 16ppm to 12 hours and lowered by the metabolism of the strain over time, compared to the initial odorous substance concentration is reduced The ratio was constantly kept low. In particular, in Examples 3, 5 and 6, the ammonia-reducing effect at 12 hours was slightly higher than that of Comparative Example 3 containing only bamboo liquor solution. In contrast, the concentration of ammonia was increased in 2, but in Examples 3, 5, and 6, it was confirmed that the concentration was significantly lower.

The reduction effect of hydrogen sulfide was compared with Comparative Example 2 in Comparative Example 3 added only bamboo shoot liquid ammonia reduction effect from the initial 6.5ppm to 12ppm to 5.5ppm and then maintained for 48 hours, in Examples 3 to 6 the initial 6.0ppm The elongation decreased with time and decreased to 1.5 ~ 2.5ppm at 48 hours.

Experimental Example 6: Effect of Removing Odor Substances in a Biofilter Device

In the biofilter apparatus of the present invention, 200 g of portant cement, 260 g of silica sand powder, 20 g of lime component, 40 g of alumina cement, and 80 g of peat material were strongly stirred for about 3 minutes with 290 g of water mixed with 20 g of a surfactant, and the prepared mixture was stirred. After casting to a mold of the size of 100mm x 100mm x l00mm, the carrier was pre-cured for 6 hours at 50 ℃ and hydrothermally cured for 8 hours at 180 ℃ in an oatclave. The dry specific gravity of the carrier was found to be about 0.33, and the pH of the solution was measured by mixing 10 g of the prepared carrier with 100 g of water and the pH of the solution was about 7.7.

The microorganisms of Table 12 were fixed to the carriers, respectively, to obtain biofilter devices of Examples 7 to 9 and Comparative Example 4.

Table 12

division	microbe
Example 7	Bacillus subtilis GU1
Example 8	Bacillus subtilis GU1, Nitrosomonas europaea strain, Nitrobacter agilis strain, Nitrosomonas sp.
Example 9	Bacillus subtilis GU1, Bacillus subtilis GU2, Pseudomonas sp., Thiobacillus sp., Bacillus sp.
Comparative Example 4	Activated Sludge Collected from Sewage Treatment Plant

5 ml of the effective volume filled with the carrier was inoculated with 500 ml of the culture medium (absorbance of 0.5) of each microorganism cultured in a general nutrient medium. Examples 8 and 9 were inoculated in 500 ml total aliquots of each microorganism. However, in Comparative Example 4, the carrier was supported on an activated sludge of 5 g / L of MLSS and aerated for 24 hours, followed by filling into a biofilter of 5 l of effective volume.

The biofilter filled with the microorganism-attached carrier passes through the odor gas generated in the livestock manure public treatment plant collection tank (standard 18,000W × 14,000L × 5,000H, effective capacity: 1,008 m ³ ) for 10 weeks by 10 L / min. To stabilize the microorganisms.

After stabilizing the microorganisms of the biofilter, the concentration of the tower stay in the biofilter was adjusted to 5 seconds, and the concentration of the malodorous gas generated in the livestock manure public treatment plant collection tank was measured at the inlet and the outlet, respectively. Malodorous items are ammonia (NH ₃ ), hydrogen sulfide (H ₂ S), methylmerethane (CH ₃ SH), and complex odor, and the measurement method is shown in Table 13.

Table 13

division	How to measure	Analysis method
ammonia	The detective law	UV / VIS
Hydrogen sulfide	Tedlar Bag	GC / FPD
Methylmerethane	Tedlar Bag	GC / FPD
Compound odor	Tedlar Bag	Air dilution sensory method

Table 14 shows the results of removing pollutants by measuring the inflow and outflow concentrations on each of the malodorous substances on the 1st, 31st and 91st days.

Table 14

division		Example 7			Example 8			Example 9			Comparative Example 4
		1 day	31 days	91 days	1 day	31 days	91 days	1 day	31 days	91 days	1 day	31 days	91 days
Ammonia (ppm)	inflow	23	24	24	25	25	26	26	25	26	25	26	25
	outflow	1.5	1.0	ND	1.5	0.5	ND	1.0	1.0	ND	4.0	2.0	1.5
Hydrogen sulfide (ppm)	inflow	3.5	3.0	3.0	3.5	4.0	4.0	3.0	3.5	4.0	3.5	3.0	3.0
	outflow	1.0	0.5	0.2	0.5	0.2	0.2	0.5	ND	ND	2.0	1.0	0.6
Methylmerethane (ppm)	inflow	1.5	1.0	1.3	1.5	1.0	1.0	1.0	1.5	1.0	1.0	1.0	1.0
	outflow	1.0	0.3	0.2	0.5	0.2	ND	0.3	ND	ND	0.5	0.5	0.3
Compound Odor (Dilution Drainage)	inflow	5000	6000	6000	6000	6000	7000	6000	5000	7000	6000	6000	6000
	outflow	Less than 1000	500 or less	300 or less	Less than 1000	500 or less	300 or less	Less than 1000	300 or less	300 or less	Less than 2000	500 or less	300 or less

Experimental Example 7: Removal of odorous substances in a pilot biofilter device

In order to construct the pilot biofilter device of the present invention, a biofilter device, a cleaning scrubber used as a humidifier of the biofilter device, and an acid gas removal chemical cleaning device installed in front of the cleaning scrubber are prepared on a pilot scale, respectively. Connected.

The biofilter device of Example 10 is 1,800W × 1,400L × 2,600H (6,550 L), and the carrier with the microorganism of Example 6 attached therein is filled with an effective volume of 1,400 L in a single layer, Without adjusting the pH adjusting agent such as medium, acid, or alkali, the internal temperature of the biofilter device was adjusted to be maintained at 15 ° C or higher, and the air column residence time was 16 seconds and the air velocity (SV) was 0.16 m / s. The water was circulated in the following scrubber as a humidifier.

The size of the main body of the cleaning scrubber is 1,600W × 1,000L × 1,600H (2,500L), and a blower (capacity 25 m ³ / min) is installed at the bottom of the main body so that the residence time is 6 seconds. (Capacity 1,000 L) was made to continuously spray water into two spiral nozzles by a circulating water supply pump (capacity 50 L / min).

The biofilter apparatus of Example 11 uses the apparatus of Example 10, but uses bamboo vinegar dilution water diluted with 0.5% by weight of bamboo vinegar in place of water in a scrubber.

The biofilter device of Example 12 was inoculated at the concentration of 1.00E + 05 cfu / ml of Bacillus subtilis GU1 strain by using the device of Example 10, but diluted with 0.5% by weight of bamboo vinegar diluted with bamboo vinegar instead of water of the scrubber. One GU1- bamboo liquor dilution water was used.

The biofilter device of Example 13 is an acid gas removal chemical cleaning device in front of the bamboo vinegar cleaning scrubber of the biofilter device of Example 11, the chemical cleaning device is 1,600W × 1,000L × 1,600H (2,500 L), and a blower (capacity 25 m ³ / min) was installed at the bottom of the main body so that the residence time was 6 seconds, and the bottom of the main body was supplied with a circulating water supply pump (capacity 50 L / min) ) To continuously spray into two spiral nozzles.

Table 15 shows the removal efficiency of pollutants by measuring the inflow and outflow concentrations for each odorous substance at 1st, 31st and 91st day.

Table 15

division		Example 10			Example 11			Example 12			Example 13
		1 day	31 days	91 days	1 day	31 days	91 days	1 day	31 days	91 days	1 day	31 days	91 days
Ammonia (ppm)	inflow	45	45	50	35	40	40	35	40	35	32	36	37
	outflow	0.5	0.3	ND	0.5	ND	ND	0.2	ND	ND	0.3	ND	ND
Hydrogen sulfide (ppm)	inflow	3.5	3.0	3.0	5.0	6.0	6.0	6.5	6.0	5.5	5.5	6.5	5.0
	outflow	0.3	0.2	ND	0.2	ND	ND	0.2	ND	ND	ND	ND	ND
Methylmerethane (ppm)	inflow	1.5	1.0	1.3	1.5	2.0	1.5	2.0	1.0	1.5	1.5	2.0	1.5
	outflow	0.3	ND	ND	0.2	ND	ND	0.2	ND	ND	ND	ND	ND
Compound Odor (Dilution Drainage)	inflow	5000	6000	6000	4000	5000	5000	5000	5000	5000	5000	6000	5000
	outflow	500 or less	500 or less	300 or less	500 or less	300 or less	300 or less	500 or less	300 or less	300 or less	500 or less	300 or less	300 or less

Table 15 shows the deodorization efficiency test using the pilot plant. The deodorization efficiency was higher than the test results using the laboratory scale test apparatus shown in Table 14. In the case of Example 13, the concentration of hydrogen sulfide and methyl mercaptan at the outlet was significantly lower than that of Comparative Example 9, which was determined by the addition of an acid gas removal chemical cleaning device in front of the bamboo vinegar cleaning scrubber of the biofilter device of Example 11. do. In addition, in the case of Example 12, the concentration of ammonia, hydrogen sulfide, methylmercaptan, and complex odor at the outlet was lower than that of Example 11. The Bacillus subtilis GU1 strain in the bamboo vinegar-cleaning scrubber circulating water of the biofilter device of Example 11 It is judged to be the result of using GU1- bamboo vinegar dilution water.

Claims (7)

1. Bacillus subtilis GU1 [Accession No. KCCM 10890P], characterized in that it is isolated from bamboo vinegar and growable in a medium containing 0.1 to 50% by weight of wood vinegar.
2. Bacillus subtilis ( Bacillus subtilis ) GU1 [Accession No. KCCM 10890P] A culture in which the strain was fermented in a medium containing 0.5 to 10% by weight of wood vinegar.
3. The culture according to claim 2, wherein the medium is a solid medium.
4. Bacillus subtilis ( Bacillus subtilis ) GU1 [Accession No. KCCM 10890P] feed additive comprising a culture fermented in a medium containing 0.5 to 10% by weight of wood vinegar.
5. In the biofilter apparatus comprising a blower for introducing a malodorous substance into the biofilter device, a humidifier for supplying moisture into the biofilter device, and a carrier to which microorganisms are attached, the humidifier comprises: Bacillus subtilis GU1 [Accession No. KCCM 10890P], which is a formal wood vinegar scrubber, and the microorganisms attached to the carrier are separated from the bamboo vinegar solution and are capable of growing in a medium containing 0.5 to 50% by weight of bamboo vinegar solution. Biofilter device for removing odorous substances, characterized in that.
6. The method of claim 5, wherein the microorganism is Nitrosomonas sp., Bacillus sp., Pseudomonas sp., Nitrosomonas sp., Nitro Bacterial genus ( Nitrobacter sp.) Strain, Genus Bacillus (Thiobacill us sp.) Strain of any one or more biofilter device for removing the odorous substance, characterized in that it further comprises.
7. The method according to claim 5, wherein in the washed wood-vinegar scrubber Bacillus subtilis GU1 [Accession No. KCCM 10890P] strain and Bacillus subtilis GU2 [Accession No. KCCM 10891P] Biofilter device for removing odorous substances, characterized in that any one or more strains selected from among the strains are sprayed together.

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