KR20060122207A - Method and apparatus for decomposing sludge using alkalophilic strain - Google Patents

Abstract

A method and an apparatus for decomposing the sludge by using the alkalophilic strain are provided to enhance the removing efficiency of the sludge. An alkalophilic strain is derived from sludge, barnyard manure and soil and includes Bacillus cereus or Bacillus megaterium. The alkali condition of the sludge has pH of 8-11 at 20-40 degrees in centigrade. The alkalophilic strain is fixed and used at a carrier. The carrier is manufactured by one or more material selected from a group consisting of sodium alginate, calcium alginate, carrageenan, gelatin, chitosan, polyacrylamide jel and carboxymethyl cellulose. The sludge is sewage, polluted water, excrement waste water, stock raising waster water, industrial waste water, high-concentrated organic waste water or indegradable waste water.

Description

Sludge treatment method and apparatus therefor for solubilizing sludge using alkalophi strain {Method and apparatus for decomposing sludge using alkalophilic strain}

1 is a photograph showing the activity of protease as a clear zone by culturing the isolated alkaline strain (E-18, C-10) according to the present invention in a non-fat solid medium,

Figure 2 is a graph showing the quantitative measurement of the activity of the protease enzyme of the isolated alkaline strain according to the present invention,

Figure 3 is a graph showing the measurement of sludge degradation activity (S-COD) of the isolated alkaline strains according to the present invention,

4 is an analysis result showing a fatty acid profile included in the E-18 strain of the present invention as a microorganism identification system,

5 shows the 16s-rRNA nucleotide sequence of the E-18 strain according to the present invention.

6 is an analysis result showing a fatty acid profile included in the C-10 strain of the present invention as a microorganism identification system,

Figure 7 shows the 16s-rRNA nucleotide sequence of the C-10 strain according to the present invention,

Figure 8 is a graph showing the growth according to the skim milk concentration and pH of the E-18 and C-10 strain of the present invention,

9 is a graph showing the growth according to the culture temperature of the E-18 and C-10 strain of the present invention and the amount of dissolved oxygen in the culture medium,

10 is a graph showing the growth curve of Bacillus cereus of the present invention,

11 is a graph showing the measured value of S-COD generated after inoculating the Bacillus cereus of the present invention in sludge,

12 is a graph showing the fatty acid profile of the resulting degradation products incubated for 12 hours in the experimental group and control group inoculated with the alkaline alkali in the sludge,

13 is a graph showing the dry weight of microorganisms in the sludge of an aeration tank cultured using sludge solubilized by a strain as a carbon source;

FIG. 14 is an electrophoretic photograph of Bacillus cereus and sludge treated according to the present invention, extracting chromosomal DNA from the processed product, and processing by RFLP method.

15 is an electrophoretic photograph showing the effect of sludge treatment according to the present invention and microbial diversity in the sludge through RFLP,

16 is a graph showing the viability of Bacillus cereus water fixed with sodium alginate,

17 is a schematic view showing a sludge treatment apparatus according to a preferred embodiment of the present invention.

The present invention relates to a sludge treatment method for solubilizing sludge using alkalescent strains.

In Korea, due to the lack of awareness of environmental conservation in the course of rapid economic development, the pollution of the air as well as the water quality has reached a very serious level. In particular, domestic sewage, agricultural and livestock wastewater, and industrial wastewater are contaminating factors such as lakes, public waters such as inner bays and inland seas, and water quality of urban small and medium rivers.

In addition, as interest in environmental problems increased due to the improvement of living standards, the demand for improvement of environmental quality increased, and the infrastructure was gradually expanded to prevent water pollution of major rivers. Among these basic facilities, sewage treatment plants such as wastewater and household sewage are the most urgent projects, and large cities have almost completed sewage treatment plants, and recently, the targets have been expanded to small and medium cities.

However, as the sewage treatment plant is operated, a large amount of sludge is incidentally generated. Most of these sludges are simply disposed of landfills. Recently, the sludge has been attempted to landfill due to lack of landfills, but there are many problems in treatment due to the strengthening of various regulations. Particularly, in the case of simple landfill, sewage sludge contains about 80% of water, which causes secondary pollution in transportation and transportation, and shows difficulty in compacting the ground when bringing in landfill. Avoiding the situation. Most of them are organic wastes that cause secondary environmental pollution such as odors, leachate, and pests in landfills due to decomposition, and the NIMBY phenomenon for waste disposal makes it more difficult to dispose of sewage sludge.

As a method for solving this problem, many methods for reducing the amount of sludge for each sludge treatment process, such as wastewater / wastewater, have been studied.

This can bring about various side effects as the effect of sludge reduction. It is easy to manage, which reduces transportation and disposal expenses, and facilitates disposal. This is because there is less secondary pollution and the landfill can take up less landfills, which can extend the life of the landfill.

Among them, activated sludge method (Active sludge method) is BOD (biological oxygen) by aeration and agitation in an aeration tank together with a bacterial group that decomposes wastewater under aerobic conditions. The required amount) and the colloidal or dissolved substance in the waste water precipitates or is adsorbed on the activated sludge to purify the water.In this case, the activated sludge is purified and then separated by precipitation. After dehydration, concentration, etc., it is discarded or returned to the main treatment process. In general, the raw sludge is reacted at 20 to 30 days and 30 to 37 in an aeration tank. The macromolecules are hydrolyzed by the bacterial group to lower their molecular weight, and lower fatty acids (organic acids), alcohols and inorganic salts are produced as intermediate products. This process is also called a hydrogen fermenter because it generates a liquefaction process, sporadic effect tablets, and a large amount of hydrogen, and the pH is reduced to about 5-6. In addition, intermediate products such as organic acids and alcohols produced in sporadic effect tablets are decomposed into methane, carbon dioxide and ammonia by methane in the next step, which is also referred to as methane fermentation and gasification. That is, most of the generated carbon dioxide and methane diffuses, but ammonia and alkalinity are dissolved into ammonium nitrite and the methane fermentation tank rises to a pH of 7.4. For this reason, methane fermentation is sometimes called alkaline fermenter.

In addition, an inorganic flocculant, which is a chemical agent, is used to forcibly change the basicity of the suspended solids including acidified organic matter in the wastewater and to cause the suspended solids to aggregate and precipitate. It is used to alkalinize the conditions of sludge or wastewater containing sludge.

However, until now, most strains that degrade or remove sludge have been limited to decomposing alkalized sludge because most of them are acidophilic.

Accordingly, the present inventors have been trying to find a method that can be used in the sludge treatment method using a conventional microorganism to increase the decomposition or removal efficiency of the sludge of various wastewater / waste water by decomposing the sludge under alkaline conditions, various sludge, compost and soil By selecting and identifying mesophilic alkalophilic strains, the present invention was completed.

Accordingly, the present invention provides a sludge treatment method for solubilizing sludge by using a alkaline strain in a sludge treatment method using microorganisms.

In addition, the present invention is to provide a sludge treatment method that can improve the treatment water quality as well as sludge control efficiency in connection with the solubilized sludge sewage water treatment process.

In order to achieve the above object, the present invention provides a sludge treatment method for solubilizing sludge by using a alkaline strain in the sludge treatment method using a microorganism.

In addition, in order to achieve another object, the present invention is a sludge treatment apparatus using a microorganism, the sludge treatment apparatus further comprises a solubilization tank which can solubilize the waste water or sludge containing the alkalescent strain. to provide.

Hereinafter, the present invention will be described in detail.

At this time, if there is no other definition in the technical terms and scientific terms used, it has a meaning commonly understood by those of ordinary skill in the art.

In addition, repeated description of the same technical configuration and operation as in the prior art will be omitted.

The present invention can be applied to the conventional activated sludge method (active sludge method) as it is, and the medium temperature alkaline alkali strains, which are optimized for the sludge solubilization, are selected from sludge, compost or soil, so that they can be solubilized (decomposed) by microorganisms even in alkaline sludge. By identifying the solubilization efficiency of the sludge as high as possible.

This is because in the early stage of development, it was intended to target high temperature aerobic bacteria with excellent sludge removal efficiency, but when applied to the actual industry, it is more preferable that the mesophilic strain is more preferred than the high temperature strain which should consider the cost of energy to maintain the high temperature.

In this case, sludge in the present invention refers to sludge derived from sewage, sewage, manure wastewater, animal wastewater, industrial wastewater, high concentration organic wastewater or hardly degradable wastewater.

In addition, since the strain used in the present invention is a strain identified and selected from sludge, compost, and soil, it can be used to treat sludge such as various kinds of sewage / wastewater and sewage.

In addition, the present invention can be used for sludge dewatered and caked.

In particular, in the present invention, it is preferable to use Bacillus cereus or Bacillus megaterium having excellent sludge resolution and efficiency.

In addition, the solubilization efficiency of the sludge can be used not only for the aeration tank of the conventional activated sludge method, but also for the sludge under alkaline conditions by the use of an alkaline agent such as alkalinized specific industrial wastewater or flocculant. It is much superior to this conventional activated sludge method.

At this time, in the present invention, the alkaline conditions are preferably pH 8-11 at 20-40 ° C, more preferably 30 ° C, pH 10. This is because the protein resolution of the alkalescent strain of the present invention is lowered in the range exceeding the temperature and pH range.

And, the alkaline alkali strain used in the present invention is preferably used to treat the sludge fixed to the carrier.

In general, the immobilization of cells can improve economics by reducing raw materials, labor saving, and facility scale due to increased cell load per unit volume, which can be used in a wide range of biological industries such as antibiotics, coffee, soy, ethanol production, and industrial wastewater treatment. In the present invention, both an adsorption method and an entrapment trapped in the material can be used, and a comprehensive method for fixing a large amount of cells can be used. do.

In addition, the non-reactive carrier is a cell alginate, sodium alginate in the present invention, in consideration of the chemical properties, because heat or chemical treatment may impair the cell viability due to the stability of the cell in the cell fixation process (Na-alginate), carrageenan (carrageenan), gelatin (gelatin), chitosan (chitosan), polyacrylamide gel (polyacrylamide gel) and made of at least one material selected from the group consisting of carboxymethyl cellulose (carboxymethyl cellulose) Preference is given to using porous carriers.

In particular, the present invention can be easily obtained in large quantities, the immobilization process is cheap, simple, excellent mechanical strength for a long period of time and large pore size (porous size), the activity of the strain is excellent, non-toxic natural polymer sodium alginate or alginic acid It is more preferable to use a comprehensive method using calcium-alginate.

In addition, the sludge treatment apparatus according to the present invention can be used by applying a solubilization tank capable of solubilizing wastewater or sludge in which the alkalescent strain contains sludge or a wastewater treatment apparatus using a conventional microorganism or a treatment apparatus of the separated sludge.

Specifically, the sludge treatment apparatus according to the present invention, as shown in Figure 17, in the wastewater treatment apparatus using a conventional microorganism, the biological treatment tank (1) for biotreating the organic wastewater introduced and the sludge and the biological treatment material A solubilization tank 3 capable of solubilizing a part of the sludge separated from the precipitation tank to an alkaline bacterium is used in a sedimentation tank 2 separating the treatment liquid. At this time, in the solubilization tank (3), the conveying means (10) is connected to the bioprocessing tank (1) so as to convey the soluble cargo to the biological device. That is, the treated material in the biological treatment tank 2 is separated into sludge and treated water in the settling tank 2, and a part of the sludge is returned to the biological treatment apparatus 1, and the remaining part is returned to the solubilization tank 3. By repeating these cycles, the amount of sludge contained in the treated water discharged to the outside is minimized, thereby improving the water quality environment. At this time, by installing a solid-liquid separator such as a sedimentation device, flotation device, centrifuge device, membrane separation device in the sedimentation tank to facilitate the separation of sludge and treated water and install a concentration device between the settling tank and the solubilization tank (3). By concentrating the sludge, it is preferable to form a nutrient condition suitable for the growth of the alkaline alkali strain so as to exhibit a high solubilization rate.

In another specific example, the present invention can be applied to an apparatus for directly treating a sludge cake separated by filtering sludge only through wastewater and sewage treatment plants or by dredging sludge.

However, the sludge treatment apparatus of the present invention is not limited to the above-described apparatus, and if the apparatus is capable of treating sludge or wastewater using microorganisms, a solubilization tank for partitioning with other strains and using an alkaline alkali strain may be separately installed. Can be used.

Hereinafter, the present invention will be described in more detail with reference to specific examples. However, the present invention is not limited to the following examples, and it will be apparent to those skilled in the art that various changes or modifications can be made within the spirit and scope of the present invention.

EXAMPLE

In this embodiment, to increase the sludge solubilization efficiency using microorganisms, isolates excellent strains of protease activity in the natural environment, such as compost, soil, sludge, and the like, the concentration of the substrate, culture temperature, culture The purpose of this study was to determine the environmental factors affecting the growth of isolated strains such as pH and dissolved oxygen in culture.

Then, the isolated strain was added to the sludge and then cultured to determine the sludge solubilization effect by the added strain.

In addition, the microbial community in the sludge was confirmed by the Restriction Fragment Length Polymorphism (RFLP) method, the effect of the added microorganism on the microbial community in the sludge, the culture factors on the growth of the isolated effective strain, and the microorganisms prepared by immobilizing the microorganisms. It was intended to confirm the stabilization of the formulation.

(1) Securing strains for sludge solubilization

① Strain Screening

In order to secure an effective strain for sludge decomposition, candidate strains having various properties were separated in various environments. Samples such as compost, sludge, soil, etc. from high temperature aerobic, high temperature alkaline alkali, and medium temperature alkaline alkaline strains were used as a medium for nutrient broth (Nutrient Broth) as a medium. The mixture was separated through enrichment culture in alkaline and pH 10 conditions.

In this example, a total of 310 candidate strains were obtained from compost, sludge, and soil, and among the strains having various characteristics, the strains having the properties of mesophilic alkalis were targeted.

At this time, a list of the strains isolated under each condition is shown in Table 1 below.

② Selection of effective strain for solubilization of sludge

Generally, cell wall components of sewage sludge are mostly composed of protein, carbohydrate, and lipid, so microorganisms with excellent secretion ability of enzymes that can decompose these substances are excellent in sludge decomposition. It can be said.

Thus, in this embodiment, among the above-described material decomposition ability, in particular, protein (protein) degradation ability has the greatest effect on sludge degradation, protease enzymes are selected from 310 candidate strains selected primarily to select strains with excellent sludge degradation performance. Protease activity was investigated.

First, in order to select strains having excellent protease activity, 310 types of effective strains which form a clear zone are formed by culturing at 30 ° C for 24 hours using a solid medium of skim milk. (See FIG. 1). As shown in Table 2 below, after a certain time, a strain having a diameter of at least 1.2 cm in diameter of the formed clear zone was selected to have excellent protease activity of 38 species among 310 candidate strains selected first. Mid / high strains were selected.

In particular, quantitatively measured their protease activity (Lowry assay) of 11 strains excellent in protease activity among the isolated medium temperature alkaline bacteria (see Fig. 2), these about 1.0% Sludge digestion activity of each strain was confirmed by inoculating each of the sludge. The microorganisms were treated with sludge and incubated at 30 ° C. for 20 hours, and the amount of S-COD produced was compared with the control group. The E-18 and C-10 strains, the experimental group in which S-COD production increased by about two times or more compared to the amount of S-COD produced in the sludge without adding microorganisms, were confirmed as effective strains (see FIG. 3).

③ Profile analysis of separated effective strains

(1) Strain Identification [16r-rRNA sequencing and FA analysis (Fatty Acid analysis)]

Selected E-18 strains and C-10 strains were identified using 16s-rRNA and fatty acid microorganism identification system.

In general, molecular biology methods can be applied to identify strains, and among them, 16s-rRNA is used as an analysis target, which is a region in which 16s-rRNA genes contain information for classifying bacteria into species levels. Changes in the nucleotide sequence are useful for determining the relationship between the microorganisms, and because of the relatively large accumulated data, the information in the rRNA database can be used to classify and identify specific bacteria, and PCR primers at various taxa levels. And DNA probe design is possible.

First, the fatty acid profile of the isolated E-18 strains was found in most fatty acids of ISO-15: 0 (33.04%), ISO-13: 0 (10.66%), ISO-17: 0 (7.08%) has 74.7% similarity with Bacillus cereus (see FIG. 4), and the identified 16s-rRNA nucleotide sequence (see SEQ ID NO: 1) was analyzed by Genbank Blast. The -18 strain showed 100% homology with Bacillus cereus , and as a result, the E-18 strain was Bacillus cereus (hereinafter referred to as B. cereus ) (see FIG. 5).

In the case of isolated C-10 strains, the fatty acid profile shows that most of the fatty acids are ISO-15: 0 (45.10%) and ANTEISO-15: 0 (32.50%) with Bacillus megaterium and 78.8%. Similarity (see Fig. 6), 16s-rRNA sequence (see SEQ ID NO: 2) shows that 99.9% similarity to Bacillus megaterium and bacillus megaterium (hereinafter referred to as B. megaterium ) .) (See FIG. 7).

(2) Culture conditions on the growth of the strain

Two strains isolated [E-18; Bacillus cereus , C-10; Bacillus megaterium ] to determine the activity of the strain according to the environmental conditions by varying the concentration of skim milk (skim milk), culture temperature, pH, oxygen concentration in the culture medium.

The amount of skim milk was 2.5, 5.0, 7.5, 10.0 and 20.0 g / L incubated for 24 hours at 30 ℃, pH 10 conditions.

In the case of B. cereus strains, the growth of the strain increased with increasing amount of skim milk in the culture medium, which was about 1.5 times higher in the medium containing 20 g / L skim milk than the growth in the medium containing 2.5 g / L skim milk. It showed increased growth capacity. On the other hand, in the case of B. megaterium strains, the overall growth pattern was lower than that of B. cereus strains. As the amount of skim milk in the culture increased to 7.5 g / L, the growth pattern increased but the growth of the skim milk medium was inhibited. I could confirm the pattern. The pH in the culture was 7, 8, 9, 10 and 11, respectively, and cultured for 24 hours under conditions of 30 ° C and 5 g / L skim milk. As a result, the B. cereus strain showed the optimum growth pattern at pH 10, and it was confirmed that the growth was inhibited at higher pH11 culture conditions. In the case of B. megaterium strains, growth was most excellent at pH 8 and growth was clearly inhibited as the pH was increased more than that. At pH 10 and above, growth was reduced by 50% or more compared with the pH 8 condition. The optimum growth pH of the strain was confirmed to be neutral pH of about 8 (see Figure 8).

And, it was incubated for 24 hours under the condition of 5g / L skim milk, pH 10 at the culture temperature of 20 ℃, 30 ℃, 40 ℃. As a result of confirming each activity, both strains showed excellent activity at the low temperature 20 ℃ or high temperature 40 ℃ compared to the high temperature 40 ℃ confirmed that these two strains are mesophilic bacteria. 50, 100, 200, and 300 ml of the culture solution were put into 500 ml flasks, respectively, to change the amount of dissolved oxygen that may be present in the culture solution, and for 24 hours under skim milk 5g / L, 30 ° C, and pH 10. Culture was confirmed the effect of dissolved oxygen on the growth of microorganisms. It can be confirmed that B. megaterium strain is an aerobic bacterium that requires relatively high dissolved oxygen compared to B. cereus strain, and medium containing very low dissolved oxygen in B. cereus strain grows in medium containing high dissolved oxygen. It showed similar growth as in, rather it was confirmed that high dissolved oxygen inhibits the growth of the strain (see Fig. 9).

As a result, when protease activity was confirmed in solid medium of skim milk in alkaline environment at pH 10 for the same time, B. cereus strain showed higher protease activity than B. megaterium strain and confirmed growth pattern. It was confirmed that the growth pattern of B. cereus strain was superior to B. megaterium strain under the culture condition of pH 10. Therefore, the culture conditions of B. cereus strain was confirmed that the aerobic condition is not too high at a temperature of 30 ℃, pH10 in a medium containing more than 10g / L skim milk. On the other hand, B. megaterium strains were mesophilic bacteria that require high dissolved oxygen, and showed optimum growth patterns at neutral pH of about 8, and aerobic bacteria were required for high dissolved oxygen. Therefore, B. cereus strains with excellent growth and protease activity under alkaline conditions were determined as the final effective strain, and pH 10, 30 ℃, 0.5 vvm (aeration volume / medium volume / minute) according to the flask test results. , Fermentation was performed under the condition of 300 rpm. As a result, it was confirmed that the B. cereus strain reached the stationary phase after 15 hours of incubation when 10 g / L skim milk was used as a medium under alkaline conditions of pH 10 (see FIG. 10).

2. Effective strain effect and sludge property analysis

(1) Effective strain effect

In order to check the sludge resolution of B. cereus , the selected effective strain under the condition of maintaining the environment of mesophilic arc alkali, the control was maintained at 30 ℃, 250rpm, 0.5 vvm, and pH 10 using a 5L jar fermentor. The sludge was incubated for 24 hours by inoculating the sludge with B. cereus , an effective strain.

Inoculation of sludge with effective strain was inoculated after washing with sterile distilled water twice to completely remove the media components in the seed. There was no significant difference between the control group and the experimental group, but the S-COD concentration increased when the microorganisms were inoculated by about 30%. The degree of sludge solubilization peaked around 12 hours of incubation. This was confirmed that the solubilization of the sludge increased due to the addition of the alkaline alkali strain in alkaline conditions maintained at pH 10 (see Fig. 11).

(2) Characterization of decomposition products

In order to compare the characteristics of the sludge decomposition products produced in the alkaline conditions and the sludge degradation products generated by the addition of microorganisms, the B. cereus strain was added to the sludge 2L using a 5L fermenter and the cases were not.

After 12 hours of incubation, the culture medium was harvested and only the supernatant was obtained. The sludge degradation products were measured by gas chromatography (GC) to compare the composition and ratio of fatty acids in order to analyze lipids in the culture.

As shown in FIG. 12, when comparing the results of the left GC graph (A), which is a control without adding microorganisms, to the GC graph (B) on the right, in which microorganisms affected sludge solubilization, sludge was decomposed by the microorganisms. It can be seen that the types of the produced products are more diverse than those of the right control group. In addition, the fatty acid having a long carbon number which was not produced in the decomposition products of the control group was identified in 18, 20, 22 minutes, and the composition ratio was also found in the case of a short carbon number fatty acid observed in the 5, 7, 10, 11, 12 minutes. It was confirmed that solubilization of sludge using microorganisms was much more diverse than that of the control group. Thus, when solubilizing the sludge using a microorganism, the resulting degradation product is a polymer that was not degraded in the control group without the action of microorganisms can be further broken down to low molecules by the secretion enzyme of the microorganisms can be confirmed that the fatty acids of various compositions.

In addition, to determine the efficiency of the sludge decomposition products of the control group and the experimental group having different compositions and ratios as described above, when used as a carbon source of activated sludge during wastewater treatment, the aeration tank sludge was incubated at 30 ° C. for 20 hours. In addition, the same amount of S-COD of the degradation product was adjusted to exclude the effect of the amount of S-COD.

That is, the sludge solubilization product B produced by the action of the microorganism with a S-COD concentration of 5,600 ppm is diluted with A (4,000 ppm), which is equal to the S-COD concentration of the control group (4,000 ppm), and the microorganism is used as a control group. Activated sludge was incubated using the product (C, S-COD 4,000 ppm) produced by solubilizing the sludge without addition. After incubation at the same time, each sample was collected (harvest) and freeze-dried, and the dry weight was measured to determine the effect of the carbon source under each condition on the growth of activated sludge. As can be seen from the analysis results of the above solubilization products, when the microorganism is added to the sludge and the solubilization of the sludge occurs due to exocrine enzymes such as proteolytic enzymes, the polymer secreted from the sludge is actively decomposed into low molecules. When using A and B generated as a carbon source, it can be seen that the growth of activated sludge increased 1.3 and 1.6 times than that in C (see FIG. 13). This increases the efficiency of sludge decomposition by adding microorganisms to the sludge, and microbial enzymes act on the by-products of dead cells, resulting in better decomposition of polymer by-products into low molecules. You'll find it easier to use.

As a result, when the sludge was decomposed by the addition of B. cereus , an alkalescent strain, the solubilization effect was about 30% or more compared to the case without using the strain. In addition, the solubilized decomposition product produced at this time was found to have a variety of organic composition compared to the product without the effect of microorganisms, and when used as a carbon source was confirmed a positive (positive) effect on the growth of aeration tank sludge.

(3) Sludge cluster analysis using molecular ecological techniques

In this example, one of the approaches for understanding the microbial community without using the culture method was to use the molecular biological method.

Among them, using a conventional RFLP analysis method, nucleic acid is extracted from field samples, amplified by PCR, and then subjected to restriction enzymes and electrophoresed to identify various band patterns on the gel. The total sum of these bands was intended to confirm the species diversity of the samples.

First, the inoculated B. cereus strains were inoculated with raw sludge having a pH of about 7, sludge adjusted to pH 10, and sludge having a pH of 10, and then incubated for 6 hours and 12 hours, respectively. When the effective strain was treated in the sludge together with the strain characteristics, changes in the microbial community in the sludge were examined (see FIG. 14A).

In addition, the chromosomal DNA of B. cereus and the experimental sludge, which are effective strains, were extracted using a genomic DNA kit (QIAGEN Co.) and used as template DNA for PCR. It was. In addition, 16S rDNA of microorganisms in the target strain and sludge was subjected to PCR using 27F 5 '(AGA GTT TGA TCM TGG CTC AG) and 785R 5' (ACT ACC RGG GTA TCT AAT CC) as a universal primer. Amplified (see FIG. 14B).

At this time, lanes 1 and 17 in Figure 14 (A) 1kb ladder (ladder), lanes 2 to 4 is pH 7, lanes 5 to 7 is pH 10, incubated for 0 hours; Lanes 8 to 10 were incubated at pH 10 for 12 hours; Lanes 11 to 13 were sludge + B. cereus, incubated for 6 hours; And lanes 14 to 16 show sludge + B. Cereus, 12 h incubation, in FIG. 14 (B) lane 1 was pH7; Lane 2 was incubated at pH 10, 0 hours; Lane 3 was incubated at pH 10 for 12 hours; Lane 4 was sludge + B. cereus, incubated for 6 hours; Lane 5 was sludge + B. Cereus, incubated for 12 hours; And lane 6 used the 1kb ladder as a sample.

As a result, a specific portion of 16s-rRNA was amplified by PCR, and the amplified DNA fragment was treated with HaeIII restriction enzyme to cut the DNA fragment cut into a DNA fragment cut using a 3.5% Nusieve: agarose (3: 1) gel. Confirmation was confirmed the diversity of microorganisms in the sludge.

As a result of the RFLP analysis, the chromosomal DNA extracted from the raw sludge and the sludge under pH alkaline condition was treated with a restriction enzyme of HaeIII to confirm the DNA fragment pattern. As shown in Fig. 16, in both pictures (A, B), as shown in lanes 1 and 2, the DNA bands found in the initial sludge are blurry but smeared so many DNA fragments are attracted. The numbers indicate indirectly the diversity of microbial communities present. The DNA band pattern observed in the sludge after 12 hours of incubation without treatment of the strain was found to be less than 10 DNA fragments compared to the original sludge or the DNA band pattern immediately after adjusting to pH 10 (lane 3). B. On the other hand, if a band of DNA was added to decompose the sludge cereus was found that unique DNA band was observed in the more simplified to check the tendency, B. cereus strains compared to lane 3 not treated with the control group of microorganisms . In particular, as shown more clearly the microorganism and the DNA band thickness after 12 hours incubation compared to the thickness of the DNA bands visible sludge to the rear at 6 hours of incubation with the right picture illustrates the significantly thick, making B. cereus strain it is effective for sludge decomposition And the dominant species in the group.

On the other hand, the pattern of DNA fragments after a culture by treating a B. cereus strain in sludge were to be simple compared to the control group, especially B. cereus strain 6 hours to inoculated with B. cereus strains to DNA fragment patterns of the sludge itself, but 12 When comparing the DNA fragment patterns (lanes 4 and 5) after time incubation, DNA bands appearing specifically in B. cereus strains that were not observed in the control group appeared, and the thickness of the band became thicker as the incubation time passed. Losing tendency could be confirmed.

At this time, lane 1 of (A) in Figure 15, pH 7, lane 2 is pH 10, incubated for 0 hours; Lane 3 was incubated at pH 10 for 12 hours; Lane 4 was sludge + B. Cereus, incubated for 12 hours; Lane 5 is B. cereus; Lane 6 represents a 1 kb ladder, lane 1 in (B) had pH 7, lane 2 had pH 10, incubated for 0 hours; Lane 3 was incubated at pH 10 for 12 hours; Lane 4 was sludge + B. cereus, incubated for 6 hours; Lane 5 was sludge + B. Cereus, incubated for 12 hours; Lane 6 is B. cereus ; Lane 7 is 100bp ladder; And lane 8 shows a sample of 1 kb ladder.

As a result, when B. cereus is added to the sludge and cultured, the strain effectively decomposes other sludge microorganisms by an enzyme such as protease secreted by the B. cereus strain under alkaline conditions, thereby increasing the amount of S-COD, The sludge and other microorganisms are decomposed at the same time as the ingestion of some of its decomposition products in alkaline conditions, it is believed that the colonies in the sludge is further simplified and added strain is expected to predominate in this community.

(4) Strain immobilization using sodium alginate

In this example, B. cereus , an effective strain, was cultured in skim milk for 18 hours, concentrated by centrifugation, washed once with sterile distilled water, mixed with 2.4 and 3.4% sodium alginate solution, and the final sodium alginate solution was 2.0, B. cereus strain was immobilized to 3.0%. Alginate beads (Alginate beads) was added to the sterilized syringe of a mixture of the strain and sodium alginate drop in 2% CaCl 2 solution to prepare a sodium alginate beads having a diameter of 3 mm. And, after stabilization at room temperature for 2 hours for stabilization, the bead and the unfixed B. cereus strain as a control were cultured in distilled water to confirm the viability of the B. cereus strain. After collecting and diluting a certain amount of each sample, inoculated in Nutrient agar (NA) solid medium, and incubated for 18 hours at 30 ° C. The survival rate is shown in comparison to the number of viable cells.

As shown in FIG. 16, the viability of the strain was reduced to about 40% after one day of cultivation when the B. cereus strain was not fixed and shaken in distilled water without additional nutrients at pH 10 and 30 ° C. After 2 days, the survival rate of the strain was confirmed to be less than about 20%. On the other hand, when the beads fixed in sodium alginate were cultured under the same conditions as the control strain, the viability of the strain was maintained at about 90% or more after 3 days of culture, and the amount of sodium alginate was 3.4%. If the 2.4% rather than the survival rate of the strain can be confirmed to be maintained a little higher. In other words, at 3 days of cultivation of the strain, the strain fixed with 2.4% alginic acid showed 97% survival rate and the strain survival rate with 3.4% alginic acid showed 92% strain survival rate.

Therefore, when fixing the strain to sodium alginate, it was confirmed that the survival rate of the strain is maintained at 90% or more.

As described above, according to the present invention, in the activated sludge method, by using a medium temperature alkalescent strain, not only can the sludge removal efficiency be improved, but also the sludge which has not been treated conventionally can be treated.

Therefore, when the present invention enters the same industry, it is easy to apply the conventional treatment method, thereby reducing the processing cost and reducing environmental problems and complaints.

In addition, the present invention is economical solubilization of sludge by using microorganisms, such as ultrasonic treatment, ozone treatment, microwave treatment, electron beam irradiation, thermal treatment, solubilization of the sludge has a high cost of treatment, odor generation, secondary environment, etc. Problems such as contamination can be solved. In addition, unlike conventional physicochemical solubilization methods, solubilization products are converted into organic biodegradable organic compounds (RBDCOD) by the solubilization method using metabolic activity of microorganisms, so that solubilized sludge is reintroduced in connection with the wastewater treatment process. As the utilization of carbon as a municipal carbon source is very high, not only the sludge reduction efficiency but also the treated water quality can be improved.

<110> Hanwha Engineering and Construction Corp. <120> Method and apparatus for decomposing sludge using alkalophilic          strain <160> 2 <170> KopatentIn 1.71 <210> 1 <211> 1372 <212> DNA <213> E-18 <220> <221> rRNA (222) (1) .. (1372) <223> E-18 16s-rRNA <400> 1 agagtttgat cctggctcag gatgaacgct ggcggcgtgc ctaatacatg caagtcgagc 60 gaatggatta agagcttgct cttatgaagt tagcggcgga cgggtgagta acacgtgggt 120 aacctgccca taagactggg ataactccgg gaaaccgggg ctaataccgg ataacatttt 180 gaaccgcatg gttcgaaatt gaaaggcggc ttcggctgtc acttatggat ggacccgcgt 240 cgcattagct agttggtgag gtaacggctc accaaggcaa cgatgcgtag ccgacctgag 300 agggtgatcg gccacactgg gactgagaca cggcccagac tcctacggga ggcagcagta 360 gggaatcttc cgcaatggac gaaagtctga cggagcaacg ccgcgtgagt gatgaaggct 420 ttcgggtcgt aaaactctgt tgttagggaa gaacaagtgc tagttgaata agctggcacc 480 ttgacggtac ctaaccagaa agccacggct aactacgtgc cagcagccgc ggtaatacgt 540 aggtggcaag cgttatccgg aattattggg cgtaaagcgc gcgcaggtgg tttcttaagt 600 ctgatgtgaa agcccacggc tcaaccgtgg agggtcattg gaaactggga gacttgagtg 660 cagaagagga aagtggaatt ccatgtgtag cggtgaaatg cgtagagata tggaggaaca 720 ccagtggcga aggcgacttt ctggtctgta actgacactg aggcgcgaaa gcgtggggag 780 caaacaggat tagataccct ggtagtccac gccgtaaacg atgagtgcta agtgttagag 840 ggtttccgcc ctttagtgct gaagttaacg cattaagcac tccgcctggg gagtacggcc 900 gcaaggctga aactcaaagg aattgacggg ggcccgcaca agcggtggag catgtggttt 960 aattcgaagc aacgcgaaga accttaccag gtcttgacat cctctgaaaa ccctagagat 1020 agggcttctc cttcgggagc agagtgacag gtggtgcatg gttgtcgtca gctcgtgtcg 1080 tgagatgttg ggttaagtcc cgcaacgagc gcaacccttg atcttagttg ccatcattaa 1140 gttgggcact ctaaggtgac tgccggtgac aaaccggagg aaggtgggga tgacgtcaaa 1200 tcatcatgcc ccttatgacc tgggctacac acgtgctaca atggacggta caaagagctg 1260 caagaccgcg aggtggagct aatctcataa aaccgttctc agttcggatt gtaggctgca 1320 actcgcctac atgaagctgg aatcgctagt aatcgcggat cagcatgccg cg 1372 <210> 2 <211> 1372 <212> DNA <213> C-10 <220> <221> rRNA (222) (1) .. (1372) <223> C-10 16s-rRNA <400> 2 agagtttgat cctggctcag gatgaacgct ggcggcgtgc ctaatacatg caagtcgagc 60 gaactgatta gaagcttgct tctatgacgt tagcggcgga cgggtgagta acacgtgggc 120 aacctgcctg taagactggg ataacttcgg gaaaccgaag ctaataccgg ataggatctt 180 ctccttcatg ggagatgatt gaaagatggt ttcggctatc acttacagat gggcccgcgg 240 tgcattagct agttggtgag gtaacggctc accaaggcaa cgatgcatag ccgacctgag 300 agggtgatcg gccacactgg gactgagaca cggcccagac tcctacggga ggcagcagta 360 gggaatcttc cgcaatggac gaaagtctga cggagcaacg ccgcgtgagt gatgaaggct 420 ttcgggtcgt aaaactctgt tgttagggaa gaacaagtac aagagtaact gcttgtacct 480 tgacggtacc taaccagaaa gccacggcta actacgtgcc agcagccgcg gtaatacgta 540 ggtggcaagc gttatccgga attattgggc gtaaagcgcg cgcaggcggt ttcttaagtc 600 tgatgtgaaa gcccacggct caaccgtgga gggtcattgg aaactgggga acttgagtgc 660 agaagagaaa agcggaattc cacgtgtagc ggtgaaatgc gtagagatgt ggaggaacac 720 cagtggcgaa ggcggctttt tggtctgtaa ctgacgctga ggcgcgaaag cgtggggagc 780 aaacaggatt agataccctg gtagtccacg ccgtaaacga tgagtgctaa gtgttagagg 840 gtttccgccc tttagtgctg cagctaacgc attaagcact ccgcctgggg agtacggtcg 900 caagactgaa actcaaagga attgacgggg gcccgcacaa gcggtggagc atgtggttta 960 attcgaagca acgcgaagaa ccttaccagg tcttgacatc ctctgacaac tctagagata 1020 gagcgttccc cttcggggga cagagtgaca ggtggtgcat ggttgtcgtc agctcgtgtc 1080 gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt gatcttagtt gccagcattt 1140 agttgggcac tctaaggtga ctgccggtga caaaccggag gaaggtgggg atgacgtcaa 1200 atcatcatgc cccttatgac ctgggctaca cacgtgctac aatggatggt acaaagggct 1260 gcaagaccgc gaggtcaagc caatcccata aaaccattct cagttcggat tgtaggctgc 1320 aactcgccta catgaagctg gaatcgctag taatcgcgga tcagcatgcc gc 1372  

Claims (9)

In the sludge treatment method using microorganisms, Sludge treatment method that solubilizes sludge using alkalescent strain. The method of claim 1, The alkalescent strain is sludge, compost, sludge treatment method characterized in that the soil-derived alkalescent strain. The method of claim 2, The alkalescent strain is Bacillus cereus ( Bacillus cereus ) or Bacillus megaterium ( Bacillus megaterium ) characterized in that the sludge treatment method. The method of claim 1, A sludge treatment method, which is used for sludge under alkaline conditions. The method of claim 4, wherein Sludge treatment method characterized in that the alkali conditions are pH 8-11 at 20-40 degreeC. The method of claim 1, Sludge treatment method characterized by using a fixed alkaline strain on a carrier. The method of claim 6, The carrier is sludge treatment method characterized in that the carrier is made of at least one material selected from the group consisting of sodium alginate, calcium alginate, carrageenan, gelatin, chitosan, polyacrylamide gel and carboxymethyl cellulose. The method according to any one of claims 1 to 7, The sludge is sludge treatment method characterized in that the sludge derived from sewage, sewage, manure wastewater, livestock wastewater, industrial wastewater, high concentration organic wastewater or hardly degradable wastewater. In the sludge treatment apparatus using microorganisms, And a sludge solubilization tank for solubilizing the sludge generated during the sewage water treatment process by a alkaline strain and a conveying device for re-injecting the solubilized sludge into the waste water treatment process.

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