KR101861099B1 - Apparatus and method for culture of candidatus brocadia sinica oba1 using partial nitritation sequencing batch reactor supplied with ammonium oxidation bacteria(aob) granule from the connected aob granule reactor and using anaerobic ammonium oxidation reactor - Google Patents

Abstract

The present invention relates to a Candidatus Brocadia sinica OBA1 strain with a high growth speed, having an ammonia oxidation activity and separated and enriched and cultured from sludge of a bioreactor of a livestock excretion disposal plant. Moreover, to culture a large amount of Candidatus Brocadia sinica OBA1 strains with a high growth speed, separated and enriched and cultured from sludge of a bioreactor of a livestock excretion disposal plant, the present invention includes an ANAMMOX reactor stably achieving partial nitrification, which is necessary for the culture of ANAMMOA microorganisms, through an SBR reactor linked with an AOB granule generator, and operated through a completely mixed fluidized bed attachment process by attaching the Candidatus Brocadia sinica OBA1 strains to a fluidized bed carrier to prevent the strains from being lost.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a Candidatus Brocadia sinica OBA1 culture apparatus and an incubation method using an anaerobic ammonium oxidation reaction tank and a batch type partial nitrification reactor in which an ammonium oxidized bacterial granulation reactor is connected to the ammonium oxidizing bacteria granulation reactor. BACTERIA (AOB) GRANULE FROM THE CONNECTED AOB GRANULE REACTOR AND USING ANAEROBIC AMMONIUM OXIDATION REACTOR}

The present invention relates to a partial nitrite sequencing batch reactor (hereinafter referred to as 'SBR reaction tank') in which a granule production tank of ammonium oxidation bacteria (hereinafter referred to as 'AOB') is connected to an anaerobic ammonium oxidation (ANAMMOX, ANAEROBIC AMMONIUM OXIDATION) reaction tank (hereinafter referred to as 'ANAMMOX reaction tank').

More specifically, in the mass cultivation of Candidatus Brocadia sinica OBA1 strain, which has a high growth rate, by separation and enrichment culture from a biological reactor sludge of a livestock manure treatment plant, the SBR reaction vessel and the AOB granulation reactor AOB granules can be efficiently formed.

In addition, the formed AOB granules are re-introduced into the SBR reaction tank to stably attain the partial nitrification required for culturing the ANAMMOX microorganism and adhere to the fluidized bed support so that the Candidatus Brocadia sinica OBA1 strain is not lost, It is an invention using an ANAMMOX reactor operated.

High concentrations of organic wastewater such as livestock manure, excess sludge and food wastewater contain high concentrations of solids, organic matter and nitrogen. Efforts are being made to produce methane, a useful energy source, by utilizing the organic matter present in such wastewater.

In order to recover a large amount of methane, the organic matter present in the wastewater is maximally anaerobicized to maximize methane production using acid fermentation and methane fermentation microorganisms in the anaerobic state. However, the effluent wastewater is largely removed from the organic matter, Nitrogen remains as it is, so it has very low carbonitriding ratio (C / N ratio, hereinafter referred to as 'C / N ratio').

In case of livestock manure, the concentration is generally 30,000 ~ 50,000mg / L, so the treatment is usually carried out using the anaerobic digestion tank (BOD, Biochemical Oxygen Demand). After the anaerobic treatment, the wastewater containing high concentration of nitrogen is generated, and the ammonia nitrogen concentration is as high as 1,500 ~ 3,000 mg / L.

In addition, livestock manure is sprayed on agricultural land after being anaerobically treated and used as fertilizer. It is a main pollutant to pollute underground water due to continuous farmland inflow or to cause eutrophication when it is released during rainfall and worsens water quality in the water system.

Conventionally, a biological method is used for the treatment of nitrogen wastewater. However, in order to treat wastewater having a low C / N ratio, a large amount of oxygen must be supplied do.

In addition, since an external carbon source is required for denitrification, many economic advantages in resource recovery are reduced.

In order to remove the nitrogen in the wastewater biologically, several steps of biochemical reaction must be performed.

First, ammonia nitrogen in a reduced state should be oxidized to oxidized nitrogen by supplying air as shown in Reaction Scheme 1 below.

<Reaction Scheme 1>

^{_{NH 4 + + 1.5O 2 → NO}} 2 - + 2H + + H 2 O

NO ₂ ^- + 0.5O ₂ → NO ₃ ^-

NH ₄ ⁺ + 2O ₂ → NO ₃ ^- + 2H ⁺ + H ₂ O

As can be seen from the above equation, 3.43 mg of oxygen is required to oxidize 1 mg of ammoniacal nitrogen to nitrite nitrogen, and 1.14 mg of oxygen is further consumed to oxidize 1 mg of nitrite nitrogen to nitrate nitrogen, resulting in 1 mg of ammonia 4.57 mg of oxygen is needed to oxidize nitrogen to nitrate nitrogen.

The oxidized nitrate nitrogen is discharged as nitrogen gas into the atmosphere through the reaction as shown in the following Reaction Scheme 2 to finally remove nitrogen.

<Reaction Scheme 2>

NO ₃ ^- + 1.08 CH ₃ OH + 0.24H ₂ CO _{3 -} > 0.06C ₅ H ₇ O ₂ N + 0.47N ₂ + 1.68H ₂ O + HCO ₃ ^-

As can be seen from the above reaction formula (2), in order to denitrate 1 mg of nitrate nitrogen, a chemical oxygen demand (COD, hereinafter referred to as "COD") of about 4 to 5 mg is required. COD / NO ₃ -N ratio should be 4 to 5 or more.

Especially, wastewater generated from anaerobic digestion tank of high concentration organic wastewater such as livestock manure and food wastewater has an extremely low C / N ratio of less than 1, which necessitates introduction of external carbon sources such as methanol or acetic acid, resulting in an increase in operating costs .

The method developed to solve this problem is to oxidize only a part of the ammonia nitrogen to the nitrite (NO ₂ ^- ) stage, and then to remove the nitrogen through the denitrification using the remaining ammonia nitrogen as the electron donor Process is used.

This method has the advantage of saving 60% of oxygen and 100% of organic matter than the conventional method.

The single-shot nitrogen removal process is performed by ANAMMOX, which uses ANAMMOX microorganisms to convert ammonia nitrogen into nitrogen gas using nitrite nitrogen as an electron acceptor under anaerobic conditions. >.

<Reaction Scheme 3>

_{^{1.0NH 4 + + 1.32NO 2 - +}} 0.066HCO 3 - + 0.13H +

→ 1.02N ₂ + 0.26NO ₃ ^- + 0.066CH ₂ O _{0 . 5} N _{0 .15} + 2.03H ₂ O

As shown in the above Reaction Scheme 3, treatment with ANAMMOX requires nitrite nitrogen as an electron acceptor. Therefore, a partial nitrification process for converting a part (about 56.5%) of the ammonia nitrogen in the wastewater into nitrite nitrogen and converting it into nitrite nitrogen must be combined at the front stage.

The most important thing to introduce a single-shot nitrogen removal process is to achieve stable partial nitrification.

The wastewater generated from the anaerobic digestion tank for high concentration organic wastewater treatment such as livestock manure and food wastewater has a high ammonia nitrogen concentration of 1,500 ~ 3,000 mg / L and a high temperature of 25 ℃ or higher, have.

However, since nitrite nitrogen is rapidly converted to nitrate nitrogen by nitrite oxidizing bacteria (NOB), it is necessary to dominate AOB to achieve stable nitrification.

In order to ignite AOB to the right of NOB, various methods are used, but the method of keeping the solid retention time (SRT) at 1 day or less is used, which is a doubling time of NOB, Is required for more than one day, the SRT is operated for less than one day, and the AOB can be held in the system and the NOB can be washed out, thereby making it possible to dominate the AOB.

Since the ANAMMOX microorganism that performs the short-axis nitrogen removal reaction has a very slow growth rate and a long incubation time of about 11 days, it is very important to apply a reactor that does not wash out microorganisms in order to secure a stable microorganism.

Therefore, it is not easy to maintain a different SRT in the case of a single reactor in which AOB microorganisms and ANAMMOX microorganisms are grown simultaneously in one reactor to treat wastewater.

In this case, optimal operation conditions suitable for each microorganism can be established, but the number of reaction vessels is increased and a separate precipitation facility for separating microorganisms and treated water is used The number of unit processes is increased, and accordingly, the required site is increased, thereby increasing the operational management factor.

In order to solve this problem, there is a method of using granules having a high reaction rate and excellent sedimentation property. However, the conditions for forming granules and the conditions for treating contaminants are different. In the AOB and ANAMMOX reaction vessels, It is technically difficult to attain the object of forming granules at the same time.

In order to solve such a problem, methods of generating microorganisms and granules externally using a separate reaction tank and supplying them to the reaction tank are used.

Korean Patent Laid-Open Publication No. 10-2002-0072360 (hereinafter referred to as "Prior Art 1") relates to an apparatus and a treatment method for treating a high concentration nitrogen wastewater using partial nitrite oxidation and anaerobic ammonia oxidation, wherein Nitrosomonas sp. It is a method of treating high concentration nitrogen wastewater by using bacteria and ammonia oxidizing bacteria.

However, since the prior art 1 invention is difficult to be dominated by only bacteria belonging to the genus Nitrosomonas, it is required to install a plurality of processes in order to continuously flow into the SBR reaction tank in addition to slowing the reaction due to the outflow of bacteria during the inflow or outflow of wastewater. Thereby increasing the economic burden.

U.S. Published Patent Application No. 2015-0336827 (hereinafter referred to as "Prior Art 2") relates to a biofilm carrier, a treatment system and a wastewater treatment method, and a method for treating contaminated water using a mobile biofilm.

However, when the biofilm is damaged, it takes a long time to recover to the normal state, and when the microorganism grows excessively, the effluent treatment efficiency is lowered.

  Japanese Patent Publication No. 4691938 (hereinafter, referred to as 'Prior Art 3') relates to a method and apparatus for treating a nitrogen-containing liquid, wherein the nitrification reaction is carried out using ammonia-oxidizing bacterium, And a method for treating a liquid.

However, in the above-mentioned prior art document 3, since the pH is adjusted by injecting alkali into the partial nitrification process, the partial nitrification reaction hardly occurs.

Korean Patent Publication No. 10-0586535 (hereinafter referred to as "Prior Art 4") relates to a system and a method for advanced treatment of wastewater using a nitrifying bacteria granulating reactor and a method of intermittently introducing nitrified bacteria granules to denitrify wastewater to be.

However, the above-mentioned prior art document 4 can supply nitrification bacteria to the main reaction tank to enhance nitrification and nitrification. However, due to the constitution of the process, wastewater is introduced and aerobic state is passed first, so organic matter is removed and there is no organic matter flowing into the denitrification tank In order to denitrify, an external carbon source must be injected.

In addition, since methanol and acetic acid are used as an external carbon source, they are economically disadvantageous in that they must be purchased.

In addition, since the growth rate of the heterotrophic microorganisms removing the organic matter including the denitrifying microorganisms is several times faster than the growth rate of the AOB microorganisms, it is practically impossible to cause the AOB to ignite using the sludge containing such a large amount of microorganisms.

That is, when the heterogeneous microorganisms are already burned, sludge is introduced into the granular reactor to produce granules, which takes a long time to produce the granules in which the nitrification and nitrifying microorganisms generate the right-burned granules.

The ANAMMOX process, which is another process of the present invention, is difficult to commercialize despite its numerous studies and proposals, and is not generally available.

The reason for this is that the growth rate of ANAMMOX microorganisms used in the ANAMMOX process is slow and the cell yield is low.

The ANAMMOX microorganisms exist in the form of Candidatus Brocadia, Candidatus Kuenenia, Candidatus Scalindua, Candidatus Anammoxoglobus and Candidatus Jettenia in the wastewater treatment process and the nitrogen circulation system of nature.

In a typical reaction process, the growth rate of ANAMMOX microorganisms is very slow, about 11 to 15 days.

Thus, it takes a long startup time for the nitrogen removal reaction by ANAMMOX microorganisms to proceed to a certain level.

In particular, it is known that the ANAMMOX microorganisms are subjected to a nitrogen removal reaction (hereinafter referred to as &quot; reaction &quot; , Additional administration of ANAMMOX microorganisms lasting more than one year is required.

Therefore, in the nitrogen removal process using the existing ANAMMOX microorganisms, it is essential to secure the technology capable of stably and mass-culturing the ANAMMOX microorganisms in a short period of time.

In addition, stable mass culture of ANAMMOX microorganisms is greatly influenced by reactor type, influent water quality and stability of process conditions.

Thus, in the large-scale cultivation of Candidatus Brocadia sinica OBA1 strain, which is rapidly separated and concentrated from the bioreactor sludge of the livestock manure treatment plant, the SBR reactor for partial nitrite oxidation is divided into three sections and selectively sludge Stable partial nitrification required for culturing the ANAMMOX microorganism is achieved through an SBR reaction tank in which an AOB granulation unit consisting of an AOB production tank and a storage tank for introducing the AOB granulation product into the AOB granule production tank for producing high purity AOB granules is obtained and the Candidatus Brocadia sinica OBA1 strain And the culture system of Candidatus Brocadia sinica OBA1 containing the ANAMMOX reaction tank, which is attached to the fluid phase carrier so as not to be lost, is operated as a fully mixed type fluidized bed process.

<Prior Art Literature>

(Prior Art 1) Korean Patent Publication No. 10-2002-0072360

(Prior Art 2) U.S. Published Patent Application No. 2015-0336827

(Prior Art 3) Japanese Patent Publication No. 4691938

(Prior Art 4) Korean Patent Registration No. 10-0586535

(Prior Art 5) <Startup of reactors for anoxic ammonium oxidation: experiences from the first full-scale anammox reactor in Rotterdam> (Water Research Vol. 41, pp. 4149 to 4163)

The present invention recognizes the problems of the prior art, and it has been found that, in the mass culture of Candidatus Brocadia sinica OBA1 strain, which is rapidly separated and concentrated from the bioreactor sludge of the livestock manure treatment plant, granule formation is easy and sludge is selectively screened An easy SBR reactor 110; An AOB granule generating tank 121; And an AOB granule storage tank (122); And the ANAMMOX reaction tank 130, the stable partial nitrification necessary for culturing the ANAMMOX microorganism is achieved through the SBR reaction tank connected with the AOB granulation tank, and the stable partial nitrification necessary for culturing the ANAMMOX microorganism is attained by attaching to the fluidized bed support so that the Candidatus Brocadia sinica OBA1 strain is not lost The present invention provides a culture apparatus and a culture method of Candidatus Brocadia sinica OBA1 using an ANAMMOX reactor operated by a mixed fluidized bed deposition process.

AOB granules are introduced into the AOB granulation basin 121 to produce AOB granules, and the generated AOB granules are introduced into the SBR reaction basin 110 to achieve partial nitrification stably. .

In the present invention, the partially nitrified water in the SBR reaction tank 110 and the AOB granulation tank 121 is introduced into the ANAMMOX reaction tank 130, which is operated as a complete mixed-bed deposition process in the downstream stage, so that the Candidatus Brocadia sinica OBA1 strain So that the strain is adhered to the fluid phase carrier so as not to be lost.

The present invention relates to a method for recovering Candida brucadia sinica OBA1 strains grown in granular form in an ANAMMOX reaction tank 130 using solid-liquid separation means, The purpose is to replace the fluid phase carrier at 15 vol% to recover the propagated strain.

The present invention provides a method for producing a granular microorganism which is separated from a fluid phase carrier through an ANAMMOX granulation tank 140 included in an ANAMMOX reaction tank 130. When the microorganism is grown and cultured in the ANAMMOX reaction tank 130, Or to recover strains grown in the form of granules.

The objective of the present invention is to easily treat a high concentration of nitrogen by securing a large amount of Candidaatus Brocadia sinica OBA1, which has a significantly longer doubling time than that of conventional anaerobic ammonia oxidizing bacteria.

In order to achieve the above object, the present invention provides a Candidatus Brocadia sinica OBA1 strain having an ammonia oxidation activity and a high growth rate.

The strain of the present invention is deposited under the deposit number KCTC18620P and separated and concentrated from the biological reactor sludge of a livestock manure treatment plant.

The present invention relates to a method for mass production of a Candidatus Brocadia sinica OBA1 strain, which is separated and concentrated from a bioreactor sludge of a livestock manure treatment plant and has a rapid growth rate, wherein a partial nitrite oxidation with a molar ratio of ammonia nitrogen to nitrite nitrogen of 1: , The system operates in a continuous batch cycle with a supernatant discharge amount of 30%, a poor sedimentation sludge discharge amount of 30%, and a residual amount of 40%, and an air lift type air supply device is operated AOB granules with good sedimentation are kept at a lower level by keeping the sedimentation time short, and the sludge having poor sedimentability is discharged together with the treated water to form a partial nitrite sequencing batch reactor (SBR) tank 110 ); Nitrosomonas SPP. Or Nitrosospira SPP. (AOB) granules, which are spherical in shape and have a size of 200 to 400 mu m, and an AOB granulation tank 121 and an AOB granulation tank The AOB granulation unit 120 includes a granulation tank 122 for storing the AOB granules and supplying the stored AOB granules when the efficiency of the SBR tank is lowered, the smooth flow of the fluidized bed support having the Candidatus Brocadia sinica OBA1 strain A stirrer is included; An anaerobic ammonium oxidation (ANAMMOX) anaerobic ammonium oxidation reactor (130) characterized by a fully mixed fluidized bed deposition process including a discharge device for preventing carrier and bacteria from leaking out; And the like.

The SBR reaction tank 110 and the AOB granulation unit 120 of the present invention are operated as a batch type and stably achieve partial nitrification required for culturing ANAMMOX microorganisms.

The SBR reaction tank 110 of the present invention is characterized in that it includes an air supply device of an air lift type which is divided into three sections and selectively discharges sludge in the middle portion and is advantageous in AOB granule retention.

The ANAMMOX reaction tank 130 according to the present invention includes a stirrer for smoothly flowing the strain-adhered fluidized-bed carrier, wherein the ANAMMOX reaction tank 130 adheres to the fluidized-bed carrier to prevent Candidatus Brocadia sinica OBA1 from being lost; And a discharging device for preventing the carrier and the bacteria from flowing out.

The present invention relates to a method for producing Candidatus Brocadia sinica OBA1, which is grown in a granular form in an ANAMMOX reaction tank 130, using solid-liquid separation means for recovering Candida species, The fluidized bed carrier is recovered by replacing the fluidized bed carrier.

The present invention is characterized in that an ANAMMOX granulation tank (140) is further included in the ANAMMOX reaction tank (130).

The ANAMMOX granulating tank 140 of the present invention uses a water lift method to mix granular materials produced in the ANAMMOX reaction tank 130 into the ANAMMOX reaction tank 130, , The microorganism is supplied or the strain grown in the form of granules is recovered when the growth and culture of the microorganism deteriorate.

(S310) washing the bioreactor sludge in a method for separating and cultivating Candidaatus Brocadia sinica OBA1 from a bioreactor sludge of a livestock manure treatment plant of the present invention; Transferring the washed sludge to an incubator (S320); And adjusting the ammonia nitrogen and the nitrite nitrogen (S330).

The step of transferring washed sludge of the present invention to an incubator (S320) is characterized in that the temperature of the incubator is 28 to 38 ° C.

The Candidatus Brocadia sinica OBA1 strain of the present invention is characterized in that it is recovered in an SBR reaction tank connected to the AOB granule production tank and a Candidatus Brocadia sinica OBA1 culture device using the ANAMMOX reaction tank.

The Candidatus Brocadia sinica OBA1 culture apparatus and culture method using the SBR reaction tank and the ANAMMOX reaction tank combined with the AOB granule production tank according to the present invention can form an easy condition for the growth of each microorganism and the bacteria can be cultured more rapidly.

In the present invention, the AOB granulated sludge is selectively introduced into the AOB granulation tank 121 to produce high purity AOB granules, and the NO ₂ ^- is supplied faster than NOB to supply Candidatus Brocadia sinica OBA 1 There is an effect that can be cultured.

In the present invention, an air-lift type air supply device is included in the SBR reaction tank 110 to selectively introduce the granules into the AOB granulation tank 121, The volume of the AOB granulation tank 121 is reduced, and efficient and selective granulation can be achieved.

The present invention relates to a method for producing a granular microorganism produced in a granular form in an ANAMMOX granulation tank (140) by removing a microorganism released from a fluidized bed support in an ANAMMOX reaction tank (130) Or to recover the strain grown in the form of granules, thereby stably maintaining the culture of the microorganism.

The present invention can easily treat a high concentration of nitrogen by securing a large amount of Candidaatus Brocadia sinica OBA1 which has a faster growth time than a known anaerobic ammonia oxidizing bacteria.

1 shows a process using a Candidatus Brocadia sinica OBA1 culture apparatus according to the present invention.
2 shows an SBR reaction tank and an operation method of an AOB granulation tank according to the present invention.
FIG. 3 shows a sequential operation method of the SBR reaction vessel and the AOB granulation reactor according to the present invention.
FIG. 4 is a process flow chart of a Candidatus Brocadia sinica OBA1 culture apparatus including an ANAMMOX granulation tank in the process according to the present invention.
FIG. 5 is a graph illustrating the change in size of AOB granules according to operation according to the operation of the AOB granule generation tank.
6 is a graph showing the amount of AOB granules produced per ammonia removal amount under an inlet nitrogen loading condition of the AOB granule generating tank.
FIG. 7 is a microscopic view of the AOB granules in the SBR reactor sorted by size.
Fig. 8 shows Candida brucadia sinica OBA1 strain suspension according to the present invention.
FIG. 9 shows the isolation and enrichment culture of Candida strains of Candida strains OBA1 having ammonia oxidation activity in a biological reactor sludge at a livestock manure treatment plant.
FIG. 10 shows the nitrogen removal rate of the concentrated culture apparatus derived during an operation period of about 200 days or more.
FIG. 11 is a graph showing the results of evaluating growth times of the strains of Candidatus Brocadia sinica OBA1 isolated and enriched in livestock manure and a known anaerobic ammonium oxidizing bacteria.
<Description of Symbols>
110: SBR reaction tank; 111: Air lift type air supply device
120: AOB granule generator; 121: AOB granulation tank; 122: AOB granule storage tank
130: ANAMMOX Reactor
140: ANAMMOX granulation tank
S211: Step to be reacted
S212: step to be precipitated
S213: Step of draining the supernatant
S214: Step of discharging sludge
S221: Step where the supernatant is discharged
S222: Step of discharging sludge
S223: Step in which the SBR reactor is stopped
S224: Step of entering wastewater
S225, S226, S227: Step of partial nitrification and AOB granulation
S228: Step precipitated in the AOB granulation tank
S229: step of precipitating in SBR reactor
S310: washing the bioreactor sludge
S320: transferring the washed sludge to an incubator
S330: Adjusting ammonia nitrogen and nitrite nitrogen

Hereinafter, the present invention will be described in detail with reference to examples. The present invention is not limited to the embodiments disclosed herein, but may be embodied in other forms. The embodiments disclosed herein are provided so that those skilled in the art can fully understand the spirit of the present invention. Therefore, it should be understood that the present invention is not limited by the following embodiments, and all conversions, equivalents, or alternatives included in the technical idea and technical scope of the present invention are included.

The present invention is capable of various modifications and various embodiments, and particular embodiments are illustrated and described in detail in the drawings. The sizes of the elements or the relative sizes between the elements in the figures may be exaggerated somewhat for a clear understanding of the invention. Further, the shape of the elements shown in the drawings may be somewhat modified by variations in the manufacturing process or the like. Accordingly, the embodiments disclosed herein should not be construed as limited to the shapes shown in the drawings unless specifically stated, and should be understood to include some modifications.

On the contrary, the various embodiments of the present invention can be combined with any other embodiments as long as there is no clear counterpoint. Any feature that is specifically or advantageously indicated as being advantageous may be combined with any other feature or feature that is indicated as being preferred or advantageous.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

Example 1. Candidatus Brocadia sinica OBA1 culture apparatus using SBR reaction tank and ANAMMOX reaction tank connected to AOB granule producing unit>

1 shows a process using a Candidatus Brocadia sinica OBA1 culture apparatus according to the present invention.

Candidatus Brocadia sinica OBA1 includes a SBR reaction tank 110 and an ANAMMOX reaction tank 130, to which the AOB granulation unit 120 is connected in sequence, but is not limited thereto.

Partial nitrification of the wastewater is performed through the SBR reaction tank 110.

In addition, the SBR reaction tank 110 is provided with a pump for injecting wastewater containing high concentration of ammonia, a discharge device for discharging the supernatant, a sludge with poor sedimentability, and a respective motor valve for discharging AOB granules, A lift type air supply device 111 and an automatic operation control panel, but is not limited thereto.

The SBR reaction tank 110 was formed into a cylindrical shape having an effective volume of 60 liters with a diameter of 35 cm and an effective water depth of 65 cm. After the sedimentation, the volume of the sludge was set to 30%, the settable sludge discharge amount 30%, and the residual amount 40% / Day, but is not limited thereto.

In the present invention, 1 circulation means that the circulation is 6 hours in total, the granulation inlet 10 minutes, the wastewater inlet 10 minutes, the reaction 300 minutes, the sedimentation 20 minutes, the supernatant discharge 10 minutes, and the sludge discharge 10 minutes are operated in the AOB granulation tank 121 .

Further, the SBR reaction tank 110 may be divided into three sections, and only the sludge in the middle portion may be introduced into the AOB granulation tank 121, but the present invention is not limited thereto.

The AOB granule generator 120 may include an AOB granule generator 121 and an AOB granular reservoir 122, but is not limited thereto.

The AOB granulation tank 121 is a cylindrical air lift type reactor having a diameter of 20 cm and an effective water depth of 65 cm and an effective volume of 20 L. A motorized valve for discharging the treated water after settling is installed at a mid- The electric valve for discharging may be installed at the bottom of the generation tank, but is not limited thereto.

In addition, the AOB granulation tank 121 performs partial nitrification of the incoming wastewater and simultaneously produces AOB granules.

Further, the produced AOB granules can be recovered and stored in the AOB granular storage tank 122. The AOB granules stored in the AOB granular storage tank 122 can be supplied when the efficiency of the SBR reaction tank 110 is lowered.

The excess sludge containing AOB may be inoculated into the AOB granulation tank 121 and the wastewater containing ammonia nitrogen may be injected into the SBR reaction tank 110 and the AOB granulation tank 121.

The operation sequence of the AOB granule generation tank 121 may be repeated in the order of the ammonia wastewater injection step, the nitrification and AOB granule formation step, the precipitation step, the microorganisms having poor sedimentation, and the discharge step of the treated water.

The wastewater flowing into the SBR reaction tank 110 and the AOB granulation tank 121 may be an anaerobic digestion tank supernatant of the sewage treatment plant and the ammonia nitrogen concentration of the influent wastewater may be 500-1,000 mg / Do not.

The time required for wastewater inflow, aeration, settling and discharge may be varied according to the concentration of ammonia nitrogen in the feed and may be 3 to 24 hours per cycle, but is not limited thereto.

The AOB granules having good sedimentation property were kept at the bottom while the sedimentation time of the SBR reaction tank 110 was kept short. Sludge having poor sedimentation property was introduced into the AOB granulation tank 121 to reproduce the highly pure AOB granules. Sludge with poor quality can be discharged with treated water to control SRT.

The AOB granules with good sedimentation were allowed to settle down at the bottom of the production tank by keeping the sedimentation time of the AOB granulation tank 121 short, and the suspended microorganisms that did not form granules or settled at a low sedimentation rate were allowed to be washed out at the discharge stage.

When this operation is repeated, AOB granules having excellent sedimentation are formed, and NOB may be washed out, thereby selectively acting AOB as a dominant species. However, the present invention is not limited thereto.

The ANAMMOX reaction tank 130 may include, but is not limited to, a complete mixed-bed deposition process by continuously culturing a Candidatus Brocadia sinica OBA1 strain and adhering to a fluidized-bed carrier so as not to be lost.

The fluidized bed carrier having a specific gravity of 0.94 to 0.96 and a specific surface area of 500 to 800 m ² / m ³ may be charged in an amount of 40 to 50% by volume of the ANAMMOX reactor 130, It does not.

The ANAMMOX reaction tank 130 was inoculated with the culture medium containing Candida strains of Candida strains OBA1 isolated and concentrated from the bioreactor sludge of a livestock manure treatment plant.

The concentrated culture means a culture method in which when the microorganism to be separated exists in a small amount, the growth medium is selectively dominated by selectively culturing the microorganism to be separated by controlling the selection medium or sorting conditions.

The Candidatus Brocadia sinica OBA1 strain of the present invention was deposited with the Korean Collection for Type Cultures of the Korea Biotechnology Research Institute on September 26, 2017 as Deposit No. KCTC18620P as shown in FIG.

In addition, the Candidatus Brocadia sinica OBA1 strain of the present invention has a much faster doubling time than conventional ANAMMOX microorganisms, and thus it is possible to obtain a large amount of microorganisms, thereby easily treating a high concentration of nitrogen.

Furthermore, the Candida species Brocadia sinica OBA1 can rapidly treat high concentration nitrogen by oxidizing NH ₄ -N with N ₂ gas using NO ₂ -N as an electron acceptor in water under anaerobic conditions.

The fully mixed fluidized bed deposition process may include, but is not limited to, an agitator for smooth flow of the fluid phase carrier, and a discharge device for preventing the carrier and bacteria from flowing out.

<Example 2> Operation of SBR reaction tank and AOB granulation tank>

2 shows an SBR reaction tank and an operation method of an AOB granulation tank according to the present invention.

The SBR reaction tank 110 and the AOB granulation tank 121 may be operated in a batch mode, but are not limited thereto.

1) reacting (S211)

In the SBR reaction vessel 110, the partial nitrification reaction takes place using AOB granules. In the AOB granulation vessel 121, the AOB granule formation reaction occurs using sludge having poor sedimentability flowing in the SBR reaction vessel.

In addition, the present invention is not limited to the air-lift type air supply device 111 that supplies oxygen for partial nitrification in the SBR reaction tank 110 and helps maintain the granules.

2) step S212,

The precipitation starts in the SBR reaction vessel 110 and the AOB granulation vessel 121 after the reaction has been completed.

The SBR reaction tank 110 separates the supernatant and the sludge through sedimentation. At this time, the granules having good sedimentation property are located in the lower part of the SBR reaction tank 110, and the sludge having poor sedimentation property is located in the upper part, The principle that the large granules are located at the bottom is applied because the sedimentation rate is proportional to the provision of solids size.

The AOB granulation tank 121 is separated into a supernatant and a granule through precipitation.

3) the step of discharging the supernatant (S213)

The supernatant is discharged from the SBR reaction tank 110 and the AOB granulation tank 120 after the precipitation.

The AOB granules are discharged to the AOB granulation storage tank 122 and stored in the AOB granulation storage tank 122. The AOB granules are discharged into the AOB granulation tank 122, do.

4) Sludge is discharged (S214)

The sludge placed in the upper part of the granules is introduced into the AOB granulation tank 121 in the SBR reaction tank 110 from which the supernatant liquid is discharged.

In the AOB granulation tank 121 into which the sludge has been introduced, the AOB granule formation reaction occurs as in the 1) reaction step (S211) of Example 2, and is repeatedly operated.

<Example 3> Sequential operation method of SBR reaction vessel and AOB granulation reaction vessel over time>

FIG. 3 shows a sequential operation method of the SBR reaction vessel and the AOB granulation reactor according to the present invention.

1) step S221 in which the supernatant is discharged;

In the SBR reaction tank 110, the supernatant is discharged. In the AOB granulation tank 121, the granules are discharged to the AOB granular storage tank 122 and stored.

2) the step of discharging the sludge (S222)

Sludge having poor sedimentation property in the SBR reaction tank 110 is discharged to the AOB granulation tank 121.

3) the SBR reaction tank is stopped (S223)

(Raw water) containing ammonia nitrogen is introduced into the AOB granulation tank 121 into which the sludge is introduced. At this time, the SBR reaction vessel 110 is stopped.

4) In step S224,

The wastewater and the AOB granules stored in the AOB granular storage tank 122 are introduced into the SBR reaction tank 110.

The granules of the AOB granular storage tank 122 are discharged to the SBR reaction tank 110.

5) Partial nitrification and AOB granulation are formed (S225, S226, S227)

In the SBR reaction tank 110, the partial nitrification reaction proceeds due to the introduced wastewater and the AOB granules.

In the AOB granulation tank 121, the partial nitrification reaction of the introduced wastewater proceeds, and at the same time, the AOB granules are produced using sludge having poor sedimentability introduced from the SBR reaction tank 110.

Furthermore, it may be performed several times depending on the amount of wastewater and sludge, but is not limited thereto.

7) the step of precipitating in the AOB granulation tank (S228)

Precipitation for solid-liquid separation proceeds in the AOB granulation tank 121 in which the formation reaction has been completed, and the supernatant is divided into a supernatant and the granules are divided into granules.

In the SBR reaction tank 110, the partial nitrification reaction proceeds continuously.

8) the step of precipitating in the SBR reactor (S229)

In the SBR reaction tank 110 in which the partial nitrification reaction has been completed, sedimentation for solid-liquid separation proceeds, a supernatant is divided into a supernatant liquid, a sludge in the middle, and a granule in the lower part.

In addition, the AOB granulation tank 121 discharges the supernatant portion except the granules.

At this time, the suspended microorganisms having NOB and activity lowering, granules not forming, or lower sedimentation rate are discharged by washout.

In addition, AOB, which has a relatively high density and a relatively strong structure at a relatively low growth rate, is discharged from the reactor as a washout due to the influence of high free ammonia (FA, Free Ammonia) in the reactor, And is precipitated and maintained in the reactor even at a rapid precipitation time.

The process returns to the step S221 where the first step of the supernatant is released, and proceeds sequentially.

Example 4: Candidatus Brocadia sinica OBA1 incubator with ANAMMOX granulation baths>

FIG. 4 is a process flow chart of a Candidatus Brocadia sinica OBA1 culture apparatus including an ANAMMOX granulation tank in the process according to the present invention.

The ANAMMOX granulation tank 140 is further included in the Candidatus Brocadia sinica OBA1 culture apparatus including the SMB reaction tank 110 and the ANAMMOX reaction tank 130 sequentially connected to the AOB granulation unit 120 of the Example 1 But is not limited thereto.

The ANAMMOX granulation tank 140 may be a water lift system, but is not limited thereto. The ANAMMOX granulation tank 140 may be a water lift system for mixing the microorganisms separated from the fluidized bed support in the ANAMMOX reaction tank 130 into granules.

The microorganism may be any one or more of microorganisms selected from the group consisting of Candida, Broccia sinica OBA1, Candidatus Brocadia, Candidatus Kuenenia, Candidatus Scalindua, Candidatus Anammoxoglobus, and Candidatus Jettenia, and Candidatus Brocadia sinica OBA1 may be preferably used.

The Candidatus Brocadia sinica OBA1 has a faster growth time than the conventional ANAMMOX microorganisms, and thus it is possible to obtain a large amount of microorganisms, thereby easily treating a high concentration of nitrogen.

If the growth and culture of the strain are decreased in the ANAMMOX reaction tank 130, the produced microorganism can be supplied or the strain grown in granule form can be recovered.

In addition, in the method for recovering a strain, in order to recover Candidatus Brocadia sinica OBA1, which is grown in granular form in the ANAMMOX reaction tank 130, using solid-liquid separation means or recovering the cultured bacteria adhered to a fluidized- The method may further include a step of replacing the fluidized bed medium with 5 to 15% by volume of the reaction tank to recover the propagated strain.

&Lt; Example 5: AOB granule production method >

The AOB granule production reactor used in the present invention used a known technique.

The AOB granulation reactor comprises a reactor consisting of a stainless steel circular tube containing an inner circular tube to form spherical granules with an internal hydraulic shear force, a pump for injecting wastewater containing ammonia, But is not limited to, a continuous batch reactor consisting of a motorized valve, a motorized valve for the discharge of granules, a blower for air supply, a chemical pump and an automatic operation control panel.

The AOB granule production reactor has an effective volume of 2 m ³ with a diameter of 0.86 m and an effective water depth of 3.45 m. A motorized valve for discharging the treated water after settling has a middle height of the reactor and a motorized valve for discharging granules, But is not limited thereto.

The amount of air can be adjusted by controlling the flow rate in the range of 0.01 to 0.2 m / s by using a regulating valve, and the operation can be performed by adjusting the flow rate in the range of 0.05 to 0.15 m / s. However, the present invention is not limited thereto.

In addition, the area was set so that the flow rate of the inner circular tube of the reactor and the outer circular tube air were the same, and the area of the upper part of the reactor where the hydraulic shear force for granule generation was generated was reduced to 1/4 .

In addition, it is preferable that the height of the effective water depth of the reactor is at least 1: 3 ~ 4.

The surplus sludge containing AOB was inoculated into the reactor and the operation was started by injecting wastewater containing high concentration of ammonia into the reactor.

The operation sequence of the reactor was repeated in the order of the ammonia wastewater injection step, the nitrification and granulation step, the precipitation step, the microorganism having poor sedimentation, and the discharge step of the treated water.

The inlet wastewater was used for the anaerobic digestion of the primary sludge of the sewage treatment plant and the secondary sludge of the sludge, and the ammonia concentration of the influent wastewater can be operated up to 100 to 2,500 mg / L, preferably 500 to 2,000 mg / However, the present invention is not limited thereto.

The time required for wastewater inflow, aeration, settling, and discharge may be varied depending on the concentration of the incoming ammonia nitrogen, and may be 3 to 24 hours per cycle, but is not limited thereto.

If the time of the precipitation step during the operation of the AOB granulation reactor is kept short, the AOB granules having good sedimentation sufficiently fall down at the lower end of the reactor, but the suspended microorganisms that do not form AOB granules or settle at a low rate are washed out at the discharge stage. When this operation is repeated, the AOB granules having excellent sedimentation properties are formed, and the pressure is selectively applied so that the dominant species can be formed. However, the present invention is not limited thereto.

In the present invention, the AOB granules can be selectively accumulated while maintaining the sedimentation rate of the continuous batch reactor at 10 to 60 m / h, but the present invention is not limited thereto.

Example 6: Isolation and enrichment of Candidatus Brocadia sinica OBA1 strain Culture method>

The strains of Candidatus Brocadia sinica OBA1 used in Examples 1 and 4 were separated and concentrated in a sludge of a bioreactor at a livestock manure treatment plant.

1) washing the bioreactor sludge (S310)

The anaerobic ammonium oxidizing microorganisms were evaluated by the ANAMMOX activity test and the real-time polymerase chain reaction (qPCR) through batch experiments on bioreactor sludges from 10 domestic animal manure treatment plants. The target sludge for the isolation and enrichment culture of the strain having ammonia oxidation activity was selected.

The sludge having a high activity against the selected anaerobic ammonium oxidation reaction according to the present invention may be washed with an inorganic salt medium 1 to 5 times, preferably 2 to 4 times, but is not limited thereto.

The reason for washing the sludge with the inorganic salt medium is that if the washing is performed less than one time, it may contaminate other strains other than the ANAMMOX microorganisms, and if the washing is done more than 5 times, the ANAMMOX microorganisms may be washed and the culture may not be possible.

In addition, it is best to wash the sludge in an appropriate number of times in order to cultivate only ANAMMOX microorganisms without contamination.

2) transferring the washed sludge to an incubator (S320)

The washed sludge can be transferred to an upflow anaerobic reaction incubator using a carrier and sludge diluent to carry out continuous culture.

In addition, the upflow anaerobic reaction incubator may be 120-180 mL, preferably 130-170 mL, but is not limited thereto.

The incubator may be incubated in a constant temperature incubator at a temperature of 28 to 38 ° C, preferably 30 to 36 ° C, but is not limited thereto.

If the temperature of the incubator is lower than 28 ° C or higher than 38 ° C, the culture is inactivated into an environment in which the strain hardly grows, and thus it is difficult to cultivate.

Also, when the temperature of the incubator is 30 to 36 ° C., the growth of the strain becomes active and the growth of the strain becomes most active.

In addition, the artificial inorganic synthetic wastewater containing only 10 to 50 mg / L of nitrite nitrogen (NO ₂ -N) within about 6 hours of hydraulic retention time (HRT) may be used for the initial 1 to 5 days, but not limited thereto .

In addition, unnecessary microorganisms can perform a wash out operation.

3) Adjusting ammonia nitrogen and nitrite nitrogen (S330)

Thereafter, the HRT can be adjusted within the range of 11 to 13 hours, and the operation can be performed for about 30 days in the range of 10 to 25 + 5 mg / L of ammonia nitrogen and nitrite nitrogen, respectively.

In addition, it can be operated for 1 to 3 days depending on the culture conditions.

Ammonia nitrogen and nitrite nitrogen in the treated water are simultaneously removed at a similar rate. At this time, if the anaerobic ammonium oxidation reaction in which nitrate nitrogen is increased is confirmed, the ascending operation of the concentration load can be performed stepwise by shortening the influent concentration rise and residence time.

In the present invention, the total duration of the concentrated culture may be 90 to 210 days, preferably 100 to 200 days, but is not limited thereto.

EXPERIMENTAL EXAMPLE 1 Identification of AOB Granule Production Characteristics of AOB Granule Producer [

AOB granulation and partial nitrification experiments were carried out in reflux water using the AOB granulation reactor of Example 5 above.

The average ammonia nitrogen of the primary sludge anaerobic digestion liquor used in the experiments was 500 mg / L, and the pH and temperature of the reactor were maintained at 7.3 ~ 7.5 and 28 ± 2 ℃, respectively.

In addition, the oxygen required for partial nitrification was injected by an air diffuser inside the reaction tank, and the dissolved oxygen was kept at about 2 mg / L or less.

The influent water was introduced into the reactor for 5 minutes, followed by aeration and partial nitrification for 140 to 155 minutes. After 15 to 30 minutes of precipitation, the treated water was discharged for 5 minutes, resulting in 1 cycle.

In addition, 1 cycle was operated with 8 cycles per day in 3 hour cycle.

The treated water discharged 1 m ³ of the total volume of the reactor, and the total residence time was maintained at 6 hours. The precipitation time was shortened stepwise according to the formation and precipitation degree of AOB granules, Respectively.

FIG. 5 is a graph illustrating the change in size of AOB granules according to operation according to the operation of the AOB granule generation tank.

Referring to FIG. 5, the size of granules was observed under a microscope by the period in the reactor.

The sizes of the microorganisms of the activated sludge were about 10 to 50 mu m, but granules of about 100 mu m were formed from 20 days after the operation, and granules were grown up to 800 to 1,200 mu m in 60 days of operation.

In addition, it can be seen that the granules after 60 days in the reactor of Example 5 are formed into spherical and elliptical shapes.

6 is a graph showing the amount of AOB granules produced per ammonia removal amount under an inlet nitrogen loading condition of the AOB granule generating tank.

Based on FIG. 6, most of the organic materials are removed from the anaerobic digestion tank and most of the substrates available to the microorganisms are composed of ammonia nitrogen.

As a result, AOB granule production is 0.13 ~ 0.18 Kg per 1 Kg of influent ammonia nitrogen, and 0.16 Kg is produced on average.

Table 1 below shows the types and distribution of microorganisms in AOB granules through analysis of the nucleotide sequence and amount of microorganisms using Pyrosequencing method during the operation period of the AOB granule reactor.

<Table 1>

Based on Table 1, Nitrosomonas SPP. Belonging to AOB increased from about 1% at the beginning of operation to about 23% after 60 days of operation. Nitrosospira SPP. Increased from about 0.2% at the beginning of operation to about 4% after 60 days of operation Respectively.

On the other hand, Nitrobacter SPP. Belonging to NOB decreased to about 0.5% after 60 days of operation from about 3% at the beginning of operation.

These results are due to the effect of high free ammonia (FA) in the AOB granule reactor.

However, although the growth rate is relatively slow, the AOB having a large density of a strong and firm structure forms granules, so that the AOB granules are retained because they precipitate without being washed out in the reactor even at a rapid precipitation time.

EXPERIMENTAL EXAMPLE 2. Confirmation of AOB Granule Characteristics in SBR Reactor Connected with AOB Granule Production Tank [

The AOB granule production tank 121 and the SBR tank 110 of Example 1 were used to conduct AOB granule production experiments by the methods of Examples 2 and 3.

In addition, the AOB granules produced in Experimental Example 1 were operated in the SBR reaction tank 110.

The high concentration nitrogen wastewater used in the experiment was anaerobic digestion liquor combined with livestock manure and food, and the average ammonia nitrogen in wastewater was 1,500 mg / L.

In the SBR reaction tank 110, AOB granules were introduced into the AOB granulation tank 121 for 10 minutes, inflow water was introduced for 10 minutes, and aeration and partial nitrification were performed for 300 minutes. Thereafter, the sedimentation was carried out for 20 minutes, the treated water for 10 minutes and the sludge was discharged into the AOB granulation tank 121 for 10 minutes to be circulated 1 time.

The sludge was introduced from the SBR reaction tank 110 for 10 minutes, the influent was introduced for 10 minutes, and the AOB granulation tank 121 was subjected to aeration and partial nitrification for 300 minutes. Thereafter, the sedimentation was carried out for 20 minutes, the treated water for 10 minutes and the AOB granules were discharged to the SBR reaction tank 110 for 10 minutes to be circulated 1 time.

Also, the circulation of the SBR reaction vessel 110 and the AOB granulation vessel 121 was operated in four cycles per day in a 6 hour cycle, and 1/2 of the effective volume of each reaction vessel was discharged, and the overall residence time was maintained at 12 hours And the operation was carried out.

FIG. 7 is a microscopic view of the AOB granules in the SBR reactor sorted by size.

Table 2 below shows the distribution by size of AOB granules.

<Table 2>

7 and Table 2, the AOB granules in the SBR reaction tank 110 were regenerated together with the poorly precipitated granules in the AOB granulation tank 121, As shown in Fig.

Also, it can be seen that most of the AOB granules have a size of about 200 to 400 mu m and are maintained in the reactor.

Table 3 below shows the types and distribution of microorganisms in the AOB granules through analysis of the nucleotide sequence and amount of the microorganisms using the pyrosequencing method during the operation period of the SBR reaction tank 110.

<Table 3>

Based on Table 3, when granules having poor sedimentability were regenerated in the AOB granule production tank 121, NOBs and heterotrophic microorganisms with reduced activity were discharged by washout and compared with Table 1 , And Nitrosomonas SPP. Belonging to AOB were increased more than 2 times.

In addition, the distribution of AOB in the nitrification tank of the existing activated slush process is about 2 to 5% of the total microorganisms, whereas the concentration of AOB according to the present invention is more than 10 times that of the existing activated sludge, 2 times more than the method. AOB generation time was three times faster than the conventional granule generation method.

<Experimental Example 3: Isolation and enrichment culture of Candidatus Brocadia sinica OBA1>

In the present invention, the bioreactor sludge (S, C group) of a livestock manure treatment plant having high activity against the selected anaerobic ammonium oxidation reaction is washed with an inorganic salt medium 2 to 4 times, And then continuously cultured in an anaerobic reaction incubator.

The incubator used was an artificial inorganic synthetic wastewater containing only 10 to 50 mg / L of nitrite nitrogen at a temperature of 32 ° C. and a hydraulic retention time of 6 hours at an initial temperature of 1 day.

In addition, unwanted microorganisms were run with washout.

Thereafter, the residence time was adjusted within the range of 11 to 13 hours, and the operation was performed for about 30 days in the range of 10 to 25 ± 5 mg / L of ammonia nitrogen and nitrite nitrogen, respectively.

In addition, when the ammonia nitrogen and nitrite nitrogen in the treated water are removed at a similar rate and the anaerobic ammonium oxidation reaction in which the nitrate nitrogen is increased is confirmed, the influent concentration rise and the residence time are shortened, Respectively.

During the incubation period, the color change of the biomass inside the reactor was observed and the activity of the microorganism was measured by the change of inorganic nitrogen concentration in influent and effluent.

In addition, pH changes of influent and effluent were evaluated by measuring anaerobic ammonium oxidizing microorganism activity in a short time in addition to reduction of inorganic nitrogen content in the medium.

FIG. 9 shows the isolation and enrichment culturing of Candida strains from Candida strains. After about 100 days, Candida strains of Candida strains were isolated from Broccia sinica OBA1.

In addition, from about 150 days, the cultivation of the concentrate proceeded, and the cultivation of the concentrate was successful for about 200 days.

Figure 10 shows the nitrogen removal rate of the concentrated culture apparatus derived over an operating period of about 200 days or more.

As a result, the nitrogen removal rate (NRR) was 1.2 kgN / m ³ / d at a rate of 1.2 kgN / m ³ / d And the rate of nitrogen removal was more than 2 times higher than that of the study using microorganisms found in the domestic market.

EXPERIMENTAL EXAMPLE 4. Estimation of Growth Times of Doubles of Candidatus Brocadia sinica OBA1 Strains [

Two batch type reactors were continuously inoculated with culture medium containing Candidatus Brocadia sinica OBA1 and ANAMMOX microorganisms isolated and concentrated in livestock manure, respectively. Respectively.

The pH in the reaction tank was adjusted to 6.5 to 7.5 using 0.05 N NaOH solution in each reaction tank, and the temperature in the reaction tank was set at 35 ° C.

The culture solution injected into the reaction tank is basically composed of 0.25 g / L KHCO ₃ , 0.174 g / LK ₂ HPO ₄ , 0.047 mg / L MgCl ₂ and 0.055 mg / L CaCl ₂ , Add 1 mL of trace salt (II) solution.

Trace salts (Ⅰ) solution was _{10g / L Na 2 EDTA · 2H} 2 O and 5g / L FeSO ₄ · include 7H ₂ O, and trace salts (Ⅱ) solution was _{15g / L Na 2 EDTA · 2H} 2 O, 0.43 _{g / L ZnSO 4 · 7H 2} O, 0.24g / L CoCl 2 · 6H 2 O, 0.99g / L MnCl 2 · 4H 2 O, 0.25g / L CuSO 4 · 5H 2 O, 0.22g / L Na 2 MoO ₄ · 2H ₂ O, 0.19 g / L NiCl ₂ · 6H ₂ O, 0.21 g / L Na ₂ SeO ₄ · 10H ₂ O and 0.014 g / LH ₃ BO ₃ .

During the incubation, (NH ₄ ) SO ₄ and NaNO ₂ were fed into the culture medium so as to be a nitrogen inflow volume load of 0.81 kg-N / m ³ -day, respectively, as the sources of ammonium and nitrite. At this time, the molar ratio of ammonia to nitrous acid was in the range of 1: 1 to 1.2.

FIG. 11 shows the relative abundance of Candidatus Brocadia sinica OBA1 isolated and enriched in livestock manure and the known ANAMMOX microorganism according to the influent load in a batch type reactor.

The concentration of Candidatus Brocadia sinica OBA1 isolated and enriched in livestock manure according to the present invention showed a density difference of more than 20% after 15 days of experiment.

As a result of calculating the doubling time using this density difference, it was found that the doubling time of Candidatus Brocadia sinica OBA1 strain was 6 to 7 days.

In addition, it can be confirmed that the doubling time of the Candidatus Brocadia sinica OBA1 strain of the present invention is about twice as fast as that of the known ANAMMOX microorganism having a doubling time of about 13 to 15 days.

EXPERIMENTAL EXAMPLE 5. Evaluation of Culture Efficiency of Candidatus Brocadia sinica OBA1 Using AOB Granule-Based SBR Reactor and ANAMMOX Process>

The treatment efficiency of the high concentration nitrogen wastewater according to the culture efficiency of Candidatus Brocadia sinica OBA1 strain was evaluated by using the SBR reaction vessel 110 and the ANAMMOX reaction vessel 130 connected to the AOB granule production unit 120 of Example 1 above.

The SBR reaction tank 110 was formed into a cylindrical shape having an effective volume of 60 L with a diameter of 35 cm and an effective water depth of 65 cm. After the sedimentation, the tank was continuously circulated for 4 cycles / day with 30% of the supernatant, 30% of the poor sedimentation sludge, .

1 circulation is a total of 6 hours, which means that the AOB granulation is operated in 10 minutes, 10 minutes in the AOB granulation, 10 minutes in the effluent, 300 minutes in the effluent, 20 minutes in the sediment, 10 minutes in the supernatant and 10 minutes in the sludge discharge.

The AOB granulation tank 121 is a cylindrical air lift type reactor having a diameter of 20 cm and an effective water depth of 65 cm and an effective volume of 20 L and was operated in a continuous cycle of 4 cycles / day in conjunction with the SBR reaction tank 110.

The ANAMMOX reaction tank 130 was formed into a cylindrical shape having an effective volume of 25 L with a diameter of 25 cm and an effective water depth of 50 cm. A stirrer was installed in the reaction tank for the fluidized bed carrier and the complete mixing, .

The wastewater used for the treatment efficiency evaluation was the effluent from the resource recovery process where the methane recovery was performed through the anaerobic digestion of the livestock manure that was introduced into the livestock manure processing facility at Y city and the food.

The pH of the effluent of the combined anaerobic digestion process was 8.0, TCOD _Mn was 2,000 mg / L, TN was 2,000 mg / L and NH ₄ -N was 1,700 mg / L. The COD component in wastewater is mostly refractory, and it is difficult to treat it without external carbon source in general nitrification / denitrification process.

In the above evaluation process, the SBR reaction tank 110 and the ANAMMOX reaction tank 130 were subjected to rough runoff analysis to evaluate the culture efficiency of Candidatus Brocadia sinica OBA1 in the culture apparatus.

Table 4 below shows the average values of the steady state during the experiment.

<Table 4>

Based on Table 4, about 44.4% of total nitrogen was converted to nitrite nitrogen in SBR reactor effluent, and about 3% was converted to nitrate nitrogen.

It can also be seen that the molar ratio of ammonia nitrogen and nitrite nitrogen is 1: 1.05 through partial nitrification.

The consumption ratio of ammonia nitrogen and nitrite nitrogen in the ANAMMOX reactor effluent was 1: 1.2, and about 10% nitrate nitrogen was additionally produced.

As a result of the evaluation of the treatment efficiency in connection with the SBR reaction vessel 110 and the ANAMMOX reaction vessel 130 using the AOB granules as a whole, it was found that 78.1% of the total nitrogen was removed without an external carbon source.

Based on the amount of nitrogen removed through the nitrogen concentration flowing into and out of the ANAMMOX reactor, the time of doubling of the bacteria in the culture apparatus was estimated. The time of doubling of the bacteria was about 7-8 days, It was confirmed that Candidatus Brocadia sinica OBA1 strain isolated and concentrated in livestock manure can be rapidly cultured in the culture apparatus of Example 1.

Furthermore, if the partial nitrification of the highly concentrated nitrogen wastewater is sufficiently completed, the oxygen consumption rate can be reduced by up to 60% in comparison with the existing nitrification-denitration process, and the result that the organic matter required for denitrification can be reduced by 100% can be obtained.

The present invention relates to a culture apparatus and a culture method of Candidatus Brocadia sinica OBA1 using a batch SBR reaction tank and an ANAMMOX reaction tank in which AOB granule production tank is connected, and a large amount of Candidatus Brocadia sinica OBA1 strain having a significantly longer growth time than conventional ANAMMOX microorganisms Which is an industrially applicable invention capable of easily treating a high concentration of nitrogen.

Claims (9)

delete delete delete In a culturing apparatus of Candidatus Brocadia sinica OBA1 strain having a deposit number KCTC18620P,
For the partial nitrite oxidation with a molar ratio of ammonia nitrogen to nitrite nitrogen of 1: 1.05, 4 vol% of supernatant, 30% of settling sludge discharge, 30% of residual sludge and 40% ) / Day, and an air-lift type air supply device is included to keep the settling time short so that the AOB granules with good sedimentation are located at the bottom and the sludge with poor sedimentation property is treated with the treated water A partial nitrite sequencing batch reactor (SBR) reaction vessel 110,
Nitrosomonas SPP. Or Nitrosospira SPP. (AOB) granules having a spherical shape and having a size of 200 to 400 mu m are included in the AOB granulation tank 121 and the AOB granulation tank 121,
An AOB granulation unit 120 for storing the AOB granules produced in the AOB granule production tank and a granule storage tank 122 for storing the stored AOB granules when the efficiency of the SBR reaction tank deteriorates, an AOB granulation unit 120 having Candidatus Brocadia sinica OBA1 An agitator is included for smooth flow of the fluid phase carrier; An anaerobic ammonium oxidation (ANAMMOX) reactor 130, characterized by a fully mixed fluidized bed deposition process comprising an outlet for preventing the carrier and bacteria from leaking out,
Wherein the strain having ammonia oxidizing activity and having the deposit number KCTC18620P is separated and concentrated in a sludge of a livestock manure treatment plant for 90 to 210 days and has a doubling time of 6 days. The method of claim 4,
5 to 15% by volume of the reaction tank is used for recovering the bacteria cultured by adhering to the fluidized-bed carrier or by using solid-liquid separation means for recovering Candidatus Brocadia sinica OBA1 present in granular form in the ANAMMOX reaction tank 130, Characterized in that it further comprises the step of replacing the fluid phase carrier with the culture medium and recovering the proliferated strains of Candidatus Brocadia sinica OBA1. The method of claim 4,
The anaerobic ammonium oxidation reaction tank 130 further includes an ANAMMOX granulation tank 140,
The ANAMMOX granulating tank 140 uses a water lift method to mix the microorganisms separated from the fluidized bed carrier of the anaerobic ammonium oxidation reaction tank 130 in the granular form and the anaerobic ammonium oxidation reaction tank 130 Wherein the strain is cultivated when the strain grows and culture is decreased, or the strain grown in granule form is recovered. delete For the partial nitrite oxidation with a molar ratio of ammonia nitrogen to nitrite nitrogen of 1: 1.05, 4 vol% of supernatant, 30% of settling sludge discharge, 30% of residual sludge and 40% ) / Day, and an air-lift type air supply device is included to keep the settling time short so that the AOB granules with good sedimentation are located at the bottom and the sludge with poor sedimentation property is treated with the treated water A partial nitrite sequencing batch reactor (SBR) reaction vessel 110,
Nitrosomonas SPP. Or Nitrosospira SPP. (AOB) granules having a spherical shape and having a size of 200 to 400 mu m are included in the AOB granulation tank 121 and the AOB granulation tank 121,
An AOB granulation unit 120 for storing AOB granules produced in the AOB granule production tank and a granule storage tank 122 for supplying stored AOB granules when the efficiency of the SBR reaction tank is lowered, Candidatus Brocadia sinica OBA1 having accession number KCTC18620P An agitator is included for smooth flow of the strained carrier; An anaerobic ammonium oxidation (ANAMMOX) reactor 130, characterized by a fully mixed fluidized bed deposition process comprising an outlet for preventing the carrier and bacteria from leaking out,
Ammonia-oxidizing activity, and the deposit No. KCTC18620P is obtained from Candidatus Brocadia, which is recovered in a Candidatus Brocadia sinica OBA1 culture apparatus, which is separated and concentrated from sludge of a livestock manure treatment plant for 90 to 210 days, sinica OBA1 strain. A method for recovering a strain according to claim 8,
In order to recover the Candidatus Brocadia sinica OBA1 strain, which is present in the granular form in the ANAMMOX reaction tank 130, and which is present in the deposit number KCTC18620P, or using a solid-liquid separating means, or to deposit the bacteria on the fluidized- To 15 vol.%, To recover the propagated strain,
Wherein the strain having an ammonia oxidizing activity and having a deposit number of KCTC18620P is separated and concentrated in a sludge of a livestock manure treatment plant bioreactor for 90 to 210 days and a doubling time is 6 days.

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