JP2014121692A - Method for treating organic effluent by using activated sludge - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method capable, by quantitatively and qualitatively analyzing the variation of a microbial flora included within an activated sludge of an organic effluent treatment system, of accurately grasping operative states for each treatment system.SOLUTION: The provided method for treating an organic effluent includes: in a treatment system of an organic effluent by using activated sludge, detection means for visualizing and detecting information on genes derived from a microbial flora within the activated sludge of the treatment system; and monitoring means for comparing the gene information obtained by the detection means with data provided by visualizing, detecting, and recording sets of information on genes derived from the microbial flora within the activated sludge and obtained when the treatment system is operated in an activation mode and in a standard mode, respectively so as to determine the operative state of the treatment system. The method for treating an organic effluent also includes control means for executing a control for recovering a stable operation of the treatment system on the basis of the results of the monitoring.

Description

  The present invention relates to a method for treating organic wastewater using activated sludge.

  Organic wastewater treatment technology using activated sludge is widely used to treat a wide variety of organic wastewater such as manure, washing water, domestic wastewater, and factory wastewater. However, since the main treatment is a microflora composed of a wide variety of microorganisms, it is necessary to accurately grasp changes in the microflora and manage the operating state of the treatment system so that stable activity can be maintained. However, in the past, since there was no means for quantitatively or qualitatively grasping the change in the microflora in the activated sludge during operation, operation management was left to the experience and intuition of skilled workers. Therefore, a lot of knowledge and experience are required for stable operation of the treatment system.

  In recent years, in an organic wastewater treatment system using activated sludge, a method for grasping the microflora contained in the activated sludge during operation has been reported. For example, in a biological sewage treatment system for nitrogen-containing wastewater, a method for measuring the nitrification ability in a biological nitrification reaction system by conducting microbial analysis of activated sludge by T-RFLP has been reported (Patent Document 1). ). This is to determine the nitrification ability of microorganisms in the nitrification reaction system based on the fragment size obtained by T-RFLP analysis using a specific restriction enzyme. Furthermore, it has been reported that a reproducible nitrification system can be constructed by introducing microbial information by fragment size analysis into a control system using this (Patent Document 2). In addition, a method has been reported that comprehensively grasps the structure of the microbial flora of methanogenic bacteria in sludge using primers capable of comprehensively detecting genes derived from methanogenic bacteria (Patent Document 3).

  However, the techniques described in Patent Documents 1 and 2 relate to an analysis of a specific ability called nitrification ability. On the other hand, organic wastewater such as human waste contains a wide variety of organic substances and nitrogen compounds, and the biological activity of a wide variety of microbiota contained in sludge is used to treat this. Therefore, the techniques of the cited documents 1 and 2 cannot comprehensively grasp the operating state of the organic wastewater treatment system. Moreover, although the structure of the microflora contained in activated sludge differs for every individual processing system, and comprises a complicated microbial ecosystem, the technique of patent document 1 and 2 does not respond | correspond to an individual processing system. Moreover, only the relationship between the fragment size by specific restriction enzyme treatment and the nitrification ability was found, and the fragment size fluctuated when other restriction enzymes were used. Therefore, it has not been sufficient for quantitatively or qualitatively grasping the change of the diverse microflora contained in the activated sludge. The technique of Patent Document 3 is also intended only for methanogens, and this technique does not quantitatively or qualitatively grasp changes in the diverse microflora, but operates individual treatment systems. The situation could not be accurately grasped.

JP 2006-254747 A JP 2006-255548 A JP 2003-56081 A

  There was still a need for the construction of technology that can quantitatively and qualitatively analyze changes in the microflora contained in the activated sludge of organic wastewater treatment systems and accurately grasp the operating state for each treatment system.

  Therefore, as a result of intensive studies to solve the above problems, the present inventors have made a visual detection of the genetic information derived from the microbiota in the activated sludge of the organic wastewater treatment system using activated sludge, and the detection means, Data obtained by detecting and recording the genetic information obtained by the detecting means by visualizing the genetic information derived from the microbiota in the activated sludge when the operation of the treatment system is in the activated state and when in the standard state, respectively. It was found that the transition of the microbiota can be grasped accurately and quickly by determining the operating state of the treatment system as compared with the above. Furthermore, based on the results, it was found that by controlling so that the activated state can be maintained, the microflora can be optimized, and the processing efficiency can be improved and the processing cost can be reduced. The invention has been completed.

That is, the invention shown in the following [1] to [6] is provided.
[1] In an organic wastewater treatment system using activated sludge, detection means for visualizing and detecting genetic information derived from microbial flora in the activated sludge of the treatment system, and gene information obtained by the detection means , When the operation of the treatment system is in an activated state and when the treatment system is in a standard state, the genetic information derived from the microbiota in the activated sludge is visualized, compared with data recorded and detected, respectively. An organic wastewater treatment method having a monitoring means for determining a state.
[2] The organic waste water is human waste and / or septic tank sludge.
[3] The gene information is a band pattern obtained by electrophoresis of DNA derived from the microbiota.
[4] The band pattern is obtained by subjecting the microbiota-derived DNA to denaturant concentration gradient gel electrophoresis.
[5] Control means for performing control for recovery of the activated state when the monitoring means determines that the operation state of the processing system is the standard state or a state approaching the standard state.
[6] The control means is supplying one or more of humus, humus extract, humic acid, fulvic acid, cinnabar and aragonite to the activated sludge.

  According to the configuration of [1] above, changes in the microflora in the activated sludge can be visually analyzed and monitored as genetic information. This allows visual confirmation of changes in the microflora in activated sludge, comparison of specific microbial species such as dominant microbial species, and the like. Therefore, the change of the microflora in activated sludge can be confirmed simply and rapidly, and the operation state of the organic waste water treatment system using activated sludge can be monitored in real time. Based on the monitoring result, the microflora of the treatment system can be optimized, and the treatment efficiency can be improved and the treatment cost can be reduced. Furthermore, according to the said structure, the genetic information of the microflora at the time of driving | running | working in an activation state and a standard state is acquired and recorded for every separate processing system. And since this gene information and the gene information of the microflora of the activated sludge in operation are compared, the operation state of the treatment system can be accurately grasped.

  According to the configuration of [2] above, it is possible to contribute to stable operation of a human waste treatment facility using a complex microbiota ecosystem such as nitrogen removal by nitrification and denitrification.

  According to the configuration of [3] above, the genetic information of the microflora in the activated sludge can be visualized and acquired as a band pattern. Thereby, the structure of a microflora can be confirmed simply and reliably. In the band pattern, one band corresponds to one type of microorganism, and the intensity of the band reflects the amount of the microbial species, so that the dominance of the species can be determined. Therefore, by observing the band pattern over time, it is possible to visually analyze changes in the microbial flora, comparison of specific microbial species such as dominant microbial species, and the like. In addition, since the comparison with the genetic information of the microflora when the operation is in the activated state and the standard state can also be performed by comparing the band pattern, the operation state of the treatment system can be confirmed easily and quickly, and the treatment system Can be optimized.

  According to the configuration of [4] above, gene information derived from the microflora can be obtained by a simple operation because the denaturant concentration gradient gel electrophoresis (DGGE), which is a general-purpose technique, is used.

  According to the configuration of [5], it is possible to provide a control means for controlling the processing system based on the determination result of the monitoring means. Thereby, since it can control, monitoring the change of microflora in real time, further optimization of the microflora of the said processing system, improvement of processing efficiency, and reduction of processing cost are realizable.

  According to the configuration of [6] above, the treatment system can be controlled by supplying one or more of humus, humus extract, humic acid, fulvic acid, cinnabar, and meteorite. . Thus, the treatment system is suitably controlled through optimization of the microflora, improvement of sludge sedimentation, suppression of malodor generation, and the like, and further improvement in treatment efficiency and reduction in treatment cost can be realized.

FIG. 1 is a schematic diagram of a schematic configuration of a human waste treatment facility using a high-load membrane separation system to which the method of the present invention is applied. Example 1 FIG. 2 is a graph showing the results of confirming the solid-liquid separability of activated sludge in a human waste treatment facility by a high-load membrane separation method to which the method of the present invention is applied, and a photograph of the activated sludge. (Example 2) FIG. 3 is an electrophoresis pattern showing the result of analyzing the microflora in the activated sludge of the human waste treatment facility by the high load membrane separation method to which the method of the present invention is applied, by DGGE. (Example 3)

  The organic wastewater treatment method of the present invention comprises a detecting means for visualizing and detecting microbiota-derived gene information in the activated sludge of the treatment system in the treatment system of organic wastewater using activated sludge, and the detection A database in which the genetic information obtained by the means is visualized, detected and recorded, respectively, from the microbiota-derived genetic information in the activated sludge when the operation of the treatment system is in an activated state and in a standard state; It has a monitoring means for determining the operating state of the processing system by comparison. By comprising in this way, the change of the microflora in activated sludge can be visually analyzed and monitored as gene information. This allows visual confirmation of changes in the microflora in activated sludge, comparison of specific microbial species such as dominant microbial species, and the like. Therefore, the change of the microflora in activated sludge can be confirmed simply and rapidly, and the operation state of the treatment system can be monitored in real time. Based on the monitoring result, the microflora of the treatment system can be optimized, for example, the generation of malodor and sludge generation can be reduced, and the treatment efficiency can be improved and the treatment cost can be reduced. Furthermore, the method of the present invention acquires and records the genetic information of the microflora when the operation is in the activated state and the standard state for each individual processing system. And since this gene information and the gene information of the microflora of the activated sludge during driving | operation are compared, the driving | running state can be grasped | ascertained correctly.

  The activated sludge method in organic wastewater treatment means that sludge is artificially added to organic wastewater, and organic matter in the wastewater is decomposed and purified by the biodegradation action of microorganisms that are constituents of sludge. By removing by solid-liquid separation, clean water is obtained. In addition, the organic waste water used as the object of the present invention is waste water containing organic matter, and examples thereof include human waste, septic tank sludge, domestic waste water, sewage, and organic industrial waste water.

  The basic configuration of the treatment system based on the activated sludge method includes a biological treatment tank and a solid-liquid separation tank. First, the outflow of the waste water downstream is quantitatively and qualitatively adjusted by a flow rate adjusting means configured as a flow rate adjusting tank or the like. Subsequently, the wastewater is received in a biological treatment tank and mixed with sludge. In the biological treatment tank, the organic matter contained in the wastewater is decomposed by the biodegradation action of the microorganisms in the sludge. Here, after maintaining a sufficient contact time, the mixture of treated water and sludge is guided to a solid-liquid separation tank. Sludge is solid-liquid separated in the solid-liquid separation tank, and the treated water is discharged to the outside through treatment such as disinfection as required. The sludge is returned to the biological treatment tank as return sludge and again used for organic matter decomposition work. On the other hand, an amount of sludge corresponding to the grown microorganisms is collected as excess sludge and is appropriately treated.

  Activated sludge is mainly composed of bacteria and fungi, forms a complex microbial ecosystem that contains a wide variety of microorganisms, including protists, and contributes to the decomposition of organic matter contained in organic wastewater. Continue to grow. And activated sludge can be acquired from soil, compost, sludge, etc. by accumulation culture or acclimatization culture.

  In the treatment system, a reactor filled with one or more of humus, humus extract, humic acid, fulvic acid, cinnabar, aragonite and the like can be installed. A part of the sludge is guided to the reactor, and the sludge is reformed by the humus or the like and supplied to the biological treatment tank. By continuously adding the humus and the like to the activated sludge, the microflora constituting the activated sludge is changed. Thus, the decay of sludge can be suppressed, the concentration and sedimentation of the sludge in the treatment system can be improved, the malodor can be controlled, and the amount of sludge generated can be reduced. Here, humus refers to polyphenols, quinones, and amino compounds produced by the decomposition of animal and plant bodies and microorganism bodies in the soil by biological communities, which are catalysts such as enzymes, microorganism oxidases, inorganic ions, and clay minerals. It is a substance that has been polycondensed by the action, and has changed to a dark amorphous colloidal polymer compound unique to soil, and a biological remains component in the process of decomposition, which can preferentially maintain soil-based microorganisms, and coagulate sludge and odor Has a component removing action. That is, the soil fungus group can be induced by installing the reactor.

  In the biological treatment tank, the organic matter contained in the organic waste water is decomposed by the biological activity of microorganisms in the sludge. The biological treatment tank has stirring means for mixing wastewater and sludge, oxygen supply means such as an aeration device for adjusting activated sludge to an aerobic atmosphere, air shut-off means for adjusting to anaerobic conditions, etc. Consists of including. When human waste is treated, human waste contains a large amount of nitrogen-containing compounds such as ammonia nitrogen and organic nitrogen in addition to BOD. The biological treatment tank is configured so that the nitrification / denitrification reaction using the microorganisms of the above progresses. For example, a nitrification tank and a denitrification tank configured as a deep reaction tank can be provided. The nitrification tank is configured to oxidize ammonia nitrogen by nitrifying bacteria and nitrite bacteria in an aerobic atmosphere to generate nitric acid and oxidatively decompose BOD. The denitrification tank is nitrified tank in an anaerobic atmosphere. The nitric acid produced in step 1 can be reduced by denitrifying microorganisms, and means for circulating the nitric acid as a nitrifying solution is provided between the nitrification tank and the denitrification. Thereby, total nitrogen, BOD, COD, etc. in human waste can be removed.

  In the solid-liquid separation tank, the treated water biodegraded in the preceding biological treatment tank is separated from the activated sludge. Even if the solid-liquid separation tank is configured so that the mixed liquid of sludge and treated water in the biological treatment tank is allowed to stand, and sludge is allowed to settle as a flock for solid-liquid separation. You may comprise so that solid-liquid separation may be carried out by filtering the mixture of treated water. As the microfiltration membrane, for example, a hollow fiber membrane or a flat membrane having fine pores with a pore diameter of 0.1 to 0.4 μm can be used, and tens to hundreds (several hundreds to thousands) of membranes are combined. It is preferable to configure as a membrane unit. In addition to being installed as a solid-liquid separation tank, the filtration membrane may be installed in a biological treatment tank, and there is no particular limitation on the immersion membrane method, the external membrane separation method, and the like.

  The present invention is obtained by collecting sludge from the biological treatment tank of the treatment system configured as described above, and visualizing and detecting gene information derived from microorganisms constituting the sludge. Prior to the analysis of gene information, it is preferable to apply after performing known nucleic acid extraction or the like for the purpose of revealing DNA. For example, after separating microorganisms from sludge by a method such as centrifugation, treatment with a surfactant such as SDS, enzyme treatment such as lysozyme and proteinase, heat treatment, ultrasonic treatment, treatment with chaotropic salt, etc. It can be exemplified that DNA is isolated by destroying microbial cells and the like by combination.

  The detection means for visualizing and detecting genetic information is not particularly limited as long as it can visualize and detect genetic information of DNA derived from the microbiota. For example, it can be performed by visualizing the state of the microflora as a band pattern using an electrophoresis apparatus. Since the electrophoresis result changes depending on the structure of the microflora constituting the activated sludge, the structure of the microbiota can be confirmed by the band pattern. In the band pattern, one band corresponds to one type of microorganism, and the intensity of the band reflects the amount of the microbial species, so that the dominance of the species can be determined. Therefore, by observing the band pattern over time, it is possible to visually analyze changes in the microbial flora, comparison of specific microbial species such as dominant microbial species, and the like. In addition, since it is possible to visually compare the genetic information of the microflora when the operation is in the activated state and the standard state, the operation state of the treatment system can be confirmed easily and quickly, and the treatment system can be optimized. Can be achieved.

  Examples of detection means for visualizing and detecting specific genetic information include, for example, denaturing gradient gel electrophoresis (DGGE), temperature gradient gel electrophoresis (TGGE), A terminal-fluorescence modified restriction enzyme fragment polymorphism (T-RFLP) can be used. DGGE is an electrophoresis method that separates DNA based on the difference in stability of double-stranded DNA against denaturing agents. Since the stability of the double-stranded DNA to the denaturant depends on the base sequence, the double-stranded DNA can be separated according to the difference in the base sequence even if the molecular weight and charge are the same. TGGE is an electrophoresis method that separates DNA based on the difference in stability of double-stranded DNA with respect to thermal stability, and utilizes the thermal denaturation process of double-stranded DNA that accompanies an increase in temperature. T-RFLP uses a restriction enzyme that has the property of cleaving a specific base sequence site of DNA. By using an appropriate restriction enzyme, DNA of different lengths can be obtained based on the difference in base sequence. Are to be separated.

  When genetic analysis of microbial flora in sludge is performed using DGGE, for example, DNA extracted from sludge is amplified using a nucleic acid amplification technique such as PCR. One of the primers used for amplification is designed to contain a sequence rich in GC bases (GC clamp). Thereby, a GC clamp is added to the terminal portion on one side of the amplified fragment. Then, the amplified fragment is electrophoresed in a gel having a denaturant concentration gradient. The concentration gradient may be parallel or perpendicular to the direction of travel, but is preferably electrophoresed in a denaturant concentration gradient gel prepared such that the concentration of the denaturant increases in the direction of electrophoresis. Perform electrophoresis. The denaturing agent is not particularly limited as long as a part of the double-stranded DNA can be dissociated, but urea, formamide, or a combination thereof can be preferably used, and polyacrylamide gel can be preferably used as the gel. . Part of the hydrogen bond of the double-stranded DNA dissociates at a certain position on the gel, while the GC clamp part has a strong binding force and maintains the double-stranded state, so the part is partially dissociated. . When a part of the gel dissociates, the speed of migration in the gel decreases due to an increase in resistance. Since the concentration of the denaturing agent that is partially dissociated differs depending on the base sequence of the double-stranded DNA, the double-stranded DNA can be separated based on the base sequence according to the mobility. Specifically, since the binding between double-stranded DNAs is stronger than the binding between ATs, the mobility on the gel varies depending on the GC content of double-stranded DNA, that is, the difference in base sequence. DNA from each microorganism can be isolated. Reagents, gel preparation procedures, sample application amounts, electrophoresis, staining, and detection can be performed according to conventional methods.

  Primers can be designed as universal primers that target genes commonly held by a wide range of microorganisms such as bacteria and fungi, or designed to target at a specific microorganism group, eg, genus and species level You can also. In particular, it is preferable to target ribosomal RNA genes such as 16S rRNA gene and 18S rRNA gene. The ribosomal RNA gene has a conserved region that is universally conserved among all microorganisms and a variable region that includes a species-specific mutated sequence. Therefore, the target region can be set according to the purpose, for example, when it is desired to detect a wide range of microbial communities or to focus on a specific microbial species.

  Once the amplification target region has been determined, a primer is designed to include a sequence complementary to that region in order to amplify the region. Primer design can also be designed using, for example, primer design support software. And a primer can be prepared based on a conventional method. For example, it can be chemically synthesized by solid phase synthesis used in the phosphoramidite method or the like, and can be synthesized automatically by a commercially available automatic nucleic acid synthesizer. The synthesized primer is purified by a known method such as HPLC as necessary. Commercially available primers may be selected and used according to the purpose. As such a primer, an oligonucleotide composed of 10 or more, preferably 15 or more, more preferably about 20 to 50 bases is designed with a desired amplification region sandwiched based on the base sequence of the target nucleic acid molecule. Illustrated. As described above, a GC clamp sequence is added to one of the primers for DGGE.

  Here, the term “complementary” means that the primer and the target nucleic acid molecule can specifically bind according to the base pairing rule to form a stable duplex structure. Here, not only perfect complementarity, but only a few nucleobases fit along the base-pairing rules as long as the primer and target nucleic acid molecule are sufficient to form a stable duplex structure with each other Even partial complementarity is acceptable. The number of bases must be long enough to specifically recognize the target nucleic acid molecule. However, if the length is too long, a nonspecific reaction is induced, which is not preferable. Therefore, the appropriate length is determined depending on many factors such as target nucleic acid sequence information such as GC content and amplification reaction conditions such as reaction temperature and salt concentration in the reaction solution.

  Here, when the operation of the treatment system is in an activated state, the microflora in the activated sludge is in a dominant state of useful microorganism species, and BOD, nitrogen compounds, etc. are effectively removed by biological treatment. It means being in a state. For example, it can be determined whether or not it is in an activated state by confirming the solid-liquid separation property of sludge, and when the concentration of sludge after treatment improves and settles well, It can be determined that the driving is in an activated state. Solid-liquid separation can be confirmed by determining the activated sludge sedimentation rate such as SV60 and SV30, and sludge capacity index such as SVI. It is also possible to determine whether or not the activated flocs are in the activated state by observing the size of the flocs of the sludge. When the formation of large flocs is confirmed, the cohesiveness and concentration of the sludge are improved. It can be determined that it is in an activated state. It is also possible to determine whether or not the activated sludge is in an activated state by confirming the amount of surplus sludge generated, the generation of malodors or the transparency of the treated water. In this case, the amount of surplus sludge generated is reduced. It is judged to be in an activated state by suppressing the generation of malodor and improving the transparency of treated water.

  The standard state means that the useful microorganisms of the microflora in the activated sludge are attenuated compared to the activated state. In the same manner as the determination of the activation state, it can be determined whether or not it is in the standard state. For example, it is possible to determine whether or not it is in the standard state by confirming the solid-liquid separation property of sludge, the size of floc, the amount of excess sludge generated, the generation of malodor, the transparency of treated water, and the like. Therefore, it is determined that the product is in the standard state due to a decrease in solid-liquid separation property, a decrease in floc-forming ability, an increase in the amount of surplus sludge, generation of malodor, a decrease in transparency of treated water, and the like.

  The gene information when in the activated state and the gene information when in the standard state are acquired for each individual processing system. Each treatment system has a different microflora configuration depending on the treatment object, treatment conditions, activated sludge used, and the like. Accordingly, the composition of the sludge microflora in the activated state and the standard state is different for each individual processing system, and thus the genetic information is also different for each individual processing system. Therefore, by acquiring genetic information for each individual processing system, it is possible to control an appropriate operating state for each processing system.

  By comparing the genetic information of the microflora collected from the sludge during the operation of the treatment system with the genetic information of the microflora of the sludge when in the activated state and the standard state, the operation of the treatment system is in the activated state The operating state of the processing system can be monitored in real time, such as whether it is in the standard state or in which state it is approaching. In order to facilitate search and comparison, data obtained by recording the acquired gene information may be managed as a database or the like.

  Furthermore, the organic wastewater treatment method of the present invention includes a control means for controlling the treatment system based on the determination result of the monitoring means. When it is determined that the operating state is the standard state or a state approaching the standard state, control for recovery to the activated state is performed. For example, control can be performed by adding one or more of humus, humus extract, humic acid, fulvic acid, cinnabar, aragonite and the like. Since it can be controlled while confirming changes in the microflora in real time, the timing and amount of addition of the humus and the like can be optimized. Also, control to maintain the sludge microbial flora when activated, such as adjustment of the amount of returned sludge, addition of useful microorganisms, optimization of treatment conditions such as aerobic atmosphere and anaerobic atmosphere It can be performed. Thereby, further optimization of the microflora of a processing system, improvement of processing efficiency, and reduction of processing cost can be realized.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, these Examples do not limit this invention. Here, as an example, an embodiment in which the method of the present invention is applied to a human waste treatment facility using a high-load membrane separation method is shown.

Example 1
In this embodiment, a human waste treatment facility using a high-load membrane separation system to which the method of the present invention is applied will be described. FIG. 1 is a schematic diagram of a schematic configuration of a human waste treatment facility using a high-load membrane separation method. A denitrification tank 1, a nitrification tank 2, a membrane raw water tank 3, 4, a membrane unit tank 5, a mixing tank 6, a coagulation tank 7, a coagulation sedimentation tank 8, a return sludge tank 9, a sludge conditioning tank 10, and a sludge storage tank 11. . The denitrification tank 1, the nitrification tank 2, and the membrane raw water tanks 3 and 4 serve as a biological treatment system in which biodegradation treatment with activated sludge is performed.

  Human waste and / or septic tank sludge to be treated flows into the biological treatment system while the inflow is adjusted in quality and quantity by a flow rate control tank or the like. First, it flows into the denitrification tank 1, and in the denitrification tank 1, the denitrifying bacteria reduce nitric acid produced in the nitrification tank 2 to nitrogen in an anaerobic atmosphere using human waste and septic tank sludge as an electron donor. And in the nitrification tank 2, ammonia in human waste and / or septic tank sludge is converted into nitric acid by nitric acid bacteria in an aerobic atmosphere. And it is comprised so that nitric acid may circulate as a nitrification liquid between the nitrification tank 2 and the denitrification tank 1, and decomposes organic matter (BOD) in human waste and / or septic tank sludge to carbon dioxide. The denitrification tank 1 and the nitrification tank 2 are configured as a deep reaction tank. In the membrane raw water tank 3 provided downstream of the denitrification tank 1 and the nitrification tank 2, if there is residual ammonia that cannot be treated in the nitrification tank 2, it is converted into nitric acid by nitrifying bacteria in an aerobic atmosphere. In the membrane raw water tank 4, if there is residual nitric acid that cannot be treated in the denitrification tank 1, it is reduced to nitrogen in an anaerobic atmosphere. At that time, methanol or the like is added as an electron donor. In this way, in the denitrification tank 1, the nitrification tank 2, and the membrane raw water tanks 3 and 4, organic substances and nitrogen compounds contained in human waste are biologically decomposed. Specifically, in the denitrification tank 1 and the nitrification tank 2, 90% of BOD and nitrogen compounds are decomposed and in the membrane raw water tanks 3 and 4, the remaining 10% is decomposed and removed. And the treated water decomposed | disassembled is sent to the membrane unit tank 5 with sludge. In the membrane unit tank 5, the treated water and the sludge are solid-liquid separated by a flat membrane, and the nitrogen produced in the previous stage is degassed.

  Then, the membrane permeate separated in the membrane unit tank 5 is sent to the mixing tank 6. In the mixing tank 6, ferric sulfate is added in a strong stirring atmosphere to neutralize and condense the surface charge of the colloidal turbid material charged and dispersed negatively. Subsequently, in the coagulation tank 7 installed downstream, the polymer coagulant is added in a weakly stirred atmosphere to neutralize the surface charge against the positively charged fine condensed sludge generated in the mixing tank 6. A coarse floc is formed by agglomerating fine particle sludge by the cross-linking adsorption action. In the coagulation sedimentation tank 8, COD and phosphorus that could not be biologically processed are coagulated by the coagulant, and the coagulated sludge is removed from the membrane permeate (treated water). And about the supernatant after coagulation sludge, COD, chromaticity, etc. which could not be coagulated and settled in the previous stage are removed by activated carbon. Subsequently, the treated water is disinfected and discharged into a river outside the facility.

  The concentrated sludge that has been solid-liquid separated in the membrane unit tank 5 is sent to the return sludge tank 9. Therefore, a part of the concentrated sludge is returned to the denitrification tank 1 in order to maintain a sludge concentration at which the biodegradation process can proceed efficiently in the previous biological treatment system. On the other hand, of the sludge grown in the biological treatment system in the previous stage, the amount of sludge exceeding the sludge necessary for maintaining the above concentration is regarded as surplus sludge and sent to the sludge refining tank 10. In the sludge refining tank 10, the volume is reduced by aeration of excess sludge. That is, the sludge is self-decomposed by aerobic digestion. Subsequently, the downstream sludge storage tank 11 is a pump tank for introducing excess sludge into a subsequent dehydrator. Subsequently, the excess sludge can be dehydrated with a dehydrator, then dried and reused as organic fertilizer or the like.

  To the human waste treatment facility of Example 1, humus and meteorite were added as inducers on November 2, 2011 to prioritize useful microorganisms. On March 15, 2012, a reactor filled with humus and meteorite was installed in the membrane unit tank. Humus and meteorites were continuously added from the reactor into the treatment system, thus preserving useful microorganisms.

Example 2 Improvement of Solid-Liquid Separation In this example, changes in sludge settling during the induction period after adding humus and meteorite as inducers to the human waste treatment facility of Example 1 were observed.

<Procedure>
The sludge in the membrane raw water tank 4 of the urine treatment facility of Example 1 was collected, and the change in sludge sedimentation was observed.
1,000 mL of the above sludge was added to a 1 L glass graduated cylinder, the value at the sludge interface after 60 minutes was read, and the value divided by 10 was SV60. The sludge concentration during the observation period was approximately 10,000 mg / L.

<Result>
The results are shown in FIG. The graph in FIG. 2 shows the change over time in the SV60 value of sludge, and the photographs are enlarged photographs (× 40) of sludge at each date and time. Immediately after the introduction of the inducer on November 2, 2011, the sludge floc was small, the SV60 value was high, and the sedimentation was not good. The SV60 value and form of the sludge on November 24 of the same year were similar to those immediately after the introduction of the inducer, and the sedimentation was not good. About one month after the introduction of the inducer, the SV60 value decreased and the sedimentation property of sludge improved, and the SV60 value reached about 70% around December 8 of the same year. Observing the sludge on December 26 of the same year, a large floc was confirmed. After that, around February 1, 2012, the SV60 value returned to the same level as immediately after the introduction of the inducer. After the reactor installation on March 15 of the same year, the sludge settling started to improve again, and after that, the sludge continued to maintain good settling, with the SV60 value falling below 50%. When the sludge on May 17 of the same year was observed about two months after the installation of the reactor, a large floc was confirmed.

Example 3 Analysis of microflora of activated sludge In this example, changes in the microflora of sludge during the induction period after adding corrosion and meteorite as inducers to the human waste treatment facility of Example 1 were analyzed. PCR-DGGE was used as an analysis means, and the band pattern of microorganisms (bacteria) contained in the sludge was analyzed. In the band pattern obtained by PCR-DGGE, one band corresponds to one type of microorganism, the intensity of the band indicates species dominance, and the band pattern indicates the composition of the microflora in the sample. Yes. The methods and conditions used in this experiment, the products used, etc. are shown below.

<1> DNA extraction <1> -1. DNA extraction using UltraClean (registered trademark) Microbial DNA Isolation Kit <1> -1-1. Reagents used
DNA extraction kit (Ultra Clean (registered trademark) Microbial DNA Isolation Kit from MO BIO)

<1> -1-2. Equipment used (a) Centrifuge Centrifuge with centrifugal force output of 10000 xg or more and cooling capacity (b) Vortex mixer

<1> -1-3. Procedure (a) 1.8 mL of sludge collected from the membrane raw water tank 4 of the human waste treatment facility of Example 1 was added to a 2 mL microtube (in the DNA extraction kit), and centrifuged at room temperature for 30 seconds at 10,000 × g. After discarding the supernatant, it was centrifuged again at 10000 × g for 30 seconds at room temperature to obtain a precipitate. When discarding the supernatant, Pipetman was used.
(B) 300 μL of MicroBeadSolution (MBS) was added to the precipitate, gently vortexed and stirred. Subsequently, the entire amount was transferred to the MicroBeadTube.
(C) 50 μL of reagent MD1 was added to MicroBeadTube. When precipitation was observed in reagent MD1, it was added after dissolving the precipitate at 60 ° C.
(D) Vortex the MicroBeadTube at maximum power for 10 minutes.
(E) It was confirmed whether MicroBeadTube was compatible with a centrifuge and centrifuged at 10,000 × g for 30 seconds at room temperature to obtain a supernatant. In addition, since MicroBeadTube may be damaged, consideration was given to avoiding centrifugal force of 10,000 xg or more.
(F) The supernatant was transferred to a 2 mL microtube (about 300 to 350 μL).
(G) 100 μL of the reagent MD2 was added to the microtube, vortexed for 5 seconds, and then cultured at 4 ° C. for 5 minutes in a heat block.
(H) Centrifuged at 10000 × g for 1 minute at room temperature.
(I) The entire amount of the supernatant was transferred to a 2 mL microtube while avoiding the precipitate (about 450 μL)
).
(J) 900 μL of reagent MD3 was added to the microtube and vortexed for 5 seconds. The reagent MD3 was shaken and mixed before use.
(K) 700 μL of the sample of the step (j) was dispensed into a SpinFilter, centrifuged at 10000 × g for 30 seconds at room temperature, and the flowing-down solution after centrifugation was discarded. The remaining sample was dispensed into SpinFilter and centrifuged in the same manner, and the falling liquid was discarded.
(L) Further, 300 μL of the reagent MD4 was added, and centrifuged at 10,000 × g for 30 seconds at room temperature, and the falling liquid after centrifugation was discarded.
(M) SpinFilter was set in a new 2 mL microtube while paying attention to liquid spillage.
(N) 50 μL of the reagent MD5 was added to the filter center of the SpinFilter, and centrifuged at 10,000 × g for 30 seconds at room temperature to allow the DNA extract to flow down. The filter was discarded.
(O) The obtained DNA extract was stored at −30 ° C. until use.

<2> PCR
<2> -1. Reagents used (a) PCR mix (GENE GENE RED PCR mix)
(B) BSA (20 mg / mL) (TaKaRa Bovine Serum Albumin solution)
BSA was added to prevent PCR inhibition by corrosive components contained in the sludge.
(C) As the template DNA, the DNA obtained in the section “DNA extraction” in <1> above was used.
(D) Sterilized distilled water

<2> -2. Primer (a) For bacteria
F984GC (10 μM) and R1378 (10 μM) were used. This primer was designed to target the bacterial 16S rRNA gene V6-8 variable region. The primer sequence information is summarized in Table 1.

<2> -3. Equipment used Thermal cycler (ABI GeneAmp (registered trademark) PCR system 9700)

<2> -4. Procedure Gloves were worn throughout the process.
(A) Reagents were added to a 0.2 mL tube as shown in Table 2. When the amplification efficiency of DNA is poor, the amount of Template DNA added can be adjusted as appropriate. The negative control was prepared by adding the same volume of sterile distilled water without adding Template DNA.
(B) The thermal cycler was started and a 0.2 mL tube was set on the heat block.
(C) The thermal cycler was set to the following reaction cycle, and PCR was performed. The reaction cycle is as follows.
Bacteria: 94 ℃-2 minutes → [94 ℃ -15 seconds, 55 ℃ -30 seconds, 68 ℃ -30 seconds] x 34

<3> DGGE
<3> -1. Equipment used (a) DCode system (manufactured by BIORAD)
(B) Power supply

<3> -2. Procedure (a) Gel preparation conditions (a) -1. The gel concentration was set to 6%, and the denaturant concentration gradient was set from 50 to 70%.
(A) -2. The solidification time was set at room temperature for 3 hours.

(B) Electrophoresis and staining conditions (b) -1. The amount applied to the gel was 5 μL.
(B) -2. The electrophoresis voltage was set to 50 V, the electrophoresis temperature was set to 58 ° C., and the electrophoresis time was set to 18 hours.
(B) -3. After electrophoresis, it was stained with SYBR GOLD (20 minutes) and then washed for 10 minutes.
(B) -4. As a marker, DGGE Marker III (manufactured by Nippon Gene) was used.
(B) -5. Microbial flora was analyzed in the same manner for sludge collected from human waste treatment facilities implemented by the soil microorganism activation method (ASB) (trademark registration pending) conducted elsewhere. Here, the soil microorganism activation method (ASB) is a method in which the treatment by the biological activity of the soil microorganism group is added to the standard activated sludge method, and the soil fungus group is induced and cultured in a reactor installed in the treatment system. This is a method for decomposing and purifying organic matter.

<4> Results The results are shown in FIG. Lane 1 is November 1, 2011, Lane 2 is November 5, 2011, Lane 3 is December 8, 2012, Lane 4 is February 1, 2012, Lane 5 is February 21, 2012, and Lane 6 is On March 28, the lane 7 shows the results of electrophoresis of the soil microorganism activation method (ASB) facility, and M shows the molecular weight marker electrophoresis. The band patterns on November 1, 2011 and November 5, 2011 immediately after the induction were similar (lanes 1 and 2), and the dominant microbial species were also the same (circles in lanes 1 and 2). This microbial species is unique to the human waste treatment facility of this experiment, and the microbial species could not be confirmed in the above-mentioned soil microorganism activation method (ASB) facility (lane 7). However, the band pattern on December 8 of the same year confirmed that the microbial flora changed immediately after induction (comparison between lane 3 and lanes 1 and 2), and the dominant microorganisms also changed (circled in lane 3). . And this microbial species was confirmed also in the above-mentioned soil microorganisms activation method (ASB) facility (circled lane 7, the lower part). The band pattern confirmed on February 1, 2012 and February 21, 2012 was similar to that immediately after induction (comparison between lanes 4 and 5 and lanes 1 and 2), confirming that the microbiota was restored. did it. In addition, microorganisms specific to the human waste treatment facility of this experiment confirmed immediately after the induction were also confirmed (circles in lanes 4 and 5). Subsequently, from the band pattern on March 28 of the same year two weeks after the installation of the reactor on March 15 of the same year, it was found that the microbiota changed and the dominant microorganisms also changed (lane 6 and lane 1). ~ 5 comparison). This dominant microbial species was also confirmed in the above-mentioned soil microorganism activation method (ASB) facility (circled in lane 7, ordinary method). The dominant microbial species confirmed on December 8, 2011 and March 28, 2012, which were inhabited in trace amounts in the original activated sludge, were dominated by the addition of humus it is conceivable that. Since it has been confirmed in the soil microorganism activation method (ASB) facility described above, it may be a useful microbial species in the process of human waste treatment, and such useful knowledge is used to visualize genetic information by the method of the present invention. As a result, it could be confirmed easily and quickly.

  Next, when the result of Example 3 and the result of Example 2 were considered, based on the visualized genetic information analyzed by the method of the present invention, the correlation between the change in the microflora and the sedimentation property of the sludge was obtained. I was able to clarify. For example, in Example 3, the sludge of December 8, 2011, which was confirmed to have changed from the microflora immediately after induction, showed good sedimentation properties as shown in Example 2. Similarly, the sludge on March 28, 2012, which was confirmed to have changed from the microbiota immediately after induction, showed good sedimentation properties as shown in Example 2. Moreover, the sedimentation properties of the sludge on November 1, 2011, November 5, 2011, February 1, 2012 and February 21, in which the microflora was judged to be similar in Example 3, were similar. Therefore, it was found that the state of the sludge and the dominant microbial species can be accurately grasped by tracking the change in the microbial flora in the sludge.

  In this way, the operation state of the human waste treatment facility can be accurately determined by the method of the present invention. Specifically, December 8, 2011 (lane 3) and March 28, 2012 (lane 6) are in the activated state, and other time points (lanes 1-2, 4-5) are in the standard state. It can be judged that there is. In order to support this, Table 3 shows the results of evaluation of the driving state using various indicators. In Table 3, SV60 (%) of effect 1 was calculated based on the description in Example 2. The surplus sludge generation amount of effect 2 is the surplus sludge generation amount per 1 liter of human waste and septic tank sludge that was put into this facility, and was calculated by dry weight. Effects 3 and 4 indicate the amounts of antifoaming agent and sodium hypochlorite used, respectively.

  From the results in Table 3, when it is determined that DGGE is in the activated state, the solid-liquid separation property of sludge is good, the amount of surplus sludge and bubbles generated and the odor are reduced, and in a good operating state. I can understand. Therefore, it was supported by this result that the operation state of the human waste treatment facility can be accurately determined by the method of the present invention.

  The method of the present invention can be used in all technical fields where a method for treating organic wastewater such as human waste treatment facilities is required.

DESCRIPTION OF SYMBOLS 1 Denitrification tank 2 Nitrification tank 3 Membrane raw water tank 4 Membrane raw water tank 5 Membrane unit tank 6 Mixing tank 7 Coagulation tank 8 Coagulation sedimentation tank 9 Return sludge tank 10 Sludge refining tank 11 Sludge storage tank

Claims (6)

  In the organic wastewater treatment system using activated sludge, the detection means for visualizing and detecting the gene information derived from the microflora in the activated sludge of the treatment system, and the genetic information obtained by the detection means, the treatment The genetic information derived from the microbiota in the activated sludge in the activated sludge when the system is in the activated state and when in the standard state is respectively visualized, detected, and compared with the recorded data to determine the operational state of the treatment system An organic wastewater treatment method having a monitoring means.   The method for treating organic wastewater according to claim 1, wherein the organic wastewater is human waste and / or septic tank sludge.   The organic wastewater treatment method according to claim 1 or 2, wherein the genetic information is a band pattern obtained by electrophoresis of DNA derived from the microbiota.   The organic wastewater treatment method according to claim 3, wherein the band pattern is obtained by subjecting the DNA derived from the microbiota to a denaturing gradient gel electrophoresis method.   The control unit according to any one of claims 1 to 4, further comprising a control unit that performs control for recovery of the activated state when the monitoring unit determines that the operation state of the processing system is the standard state or a state approaching the standard state. The method for treating organic waste water according to one item.   The method for treating organic waste water according to claim 5, wherein the control means is supply of one or more of humus, humus extract, humic acid, fulvic acid, cinnabar, and aragonite to the activated sludge. .