CN107858295A - One plant is applied to high ammonia nitrogen landfill leachate processing bacterium and its application - Google Patents
One plant is applied to high ammonia nitrogen landfill leachate processing bacterium and its application Download PDFInfo
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- CN107858295A CN107858295A CN201711220474.7A CN201711220474A CN107858295A CN 107858295 A CN107858295 A CN 107858295A CN 201711220474 A CN201711220474 A CN 201711220474A CN 107858295 A CN107858295 A CN 107858295A
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- percolate
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/77—Fusarium
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/347—Use of yeasts or fungi
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
Abstract
The invention discloses one plant to be applied to high ammonia nitrogen landfill leachate processing bacterium and its application.It is related to one plant of method with COD in the Fusariumverticillioides for reducing COD of waste leachate ability, and application this bacterium removal percolate.Inoculation 30 DEG C of shaken cultivations 2 days, is inoculated in garbage leachate wastewater with 2% inoculum concentration and handled, as a result show that COD removal effects are obvious to improveing in martin substratum.The bacterium has broad application prospects in percolate sewage treatment project.
Description
Technical field
The present invention relates to the screening and its application of percolate degraded new strains, more particularly to one plant has reduction rubbish
COD method in the Fusariumverticillioides of percolate COD abilities, and application this bacterium removal percolate.
Background technology
As economy rapid development and urban population quantity increase, also rapid growth, control rubbish are dirty for municipal refuse yield
Dye has turned into China's great environmental problem urgently to be resolved hurrily.Landfill method is still the main method of garbage disposal at present, but the party
The problem of method maximum is to form substantial amounts of percolate.After rubbish enters landfill yard, due to aerobic inside junk-heap body or
Anaerobic fermentation, natural sleet wash away leaching, and the sewage of the formation such as immersion of earth's surface and interflow subsurface drainage, i.e. garbage filter
Liquid, it is that a kind of ammonia-nitrogen content is high, organism kinds are more and the complicated, sewage of water quality and quantity change greatly, wherein containing a large amount of carcinogenic
Material and noxious material, if cannot handle well, underground water and surrounding environment can be caused seriously to pollute, garbage filter
The processing of liquid is problem generally acknowledged in the world.
The processing method of percolate mainly has physico-chemical process and the major class of bioanalysis two.Bioremediation is with its running cost
With the features such as low, treatment effeciency is high, turn into current most important refuse leachate treatment technology.Biological treatment be divided into it is aerobic and
Anaerobic Treatment.Aerobic treatment technology is using microorganism metabolic activity vigorous under aerobic condition, in the condition that oxygen is sufficient
Under, the organic matter of microorganism easily biological-degradable using in sewage is as metabolic raw material, and degraded is dirty while synthesizing living matter
Contaminate thing.The technology biological domestication adaptation time is short, simple to operate, technology maturation, to COD in waste waterCr、BOD5, ammonia nitrogen etc. it is organic
Thing removal effect is good, and can remove the metals such as iron removaling, manganese simultaneously.But microorganism is by factor shadows such as toxic metals, ambient temperature fluctuations
Sound is larger, and energy consumption is larger, and sludge yield is more.Anaerobic Methods in Treating is the dirt in the presence of anaerobism and amphimicrobe
Complicated organic matter in water is degraded to simple organic matter CH4、CO2、H2The end-products such as O.The advantages of technology is that technique is steady
Fixed, can bear larger impact load, saving energy, surplus sludge volume few, simple to operate, emission-free pollution, investment and running cost
With cheap.But operation requires strict, bacterium easy in inactivation under aerobic conditions.The general COD for working as percolate is higher than 50000mg/L
When, handled using Anaerobic cultural methods；When COD is less than 5000mg/L, handled using aerobic process；COD 5000mg/L~
During 50000mg/L, handled using Aerobic-anaerobic integrated processes.Although aerobic technology and anaerobic technology respectively have advantage and disadvantage, the two
Use in conjunction can also learn from other's strong points to offset one's weaknesses, but due to the unique component characteristic of percolate, there is stronger suppression to microbial activity
It effect, can often make largely to leak beneficial to thalline mass extinction, harmful substance, cause the waste water after processing not up to standard, cause biology
Method processing is very limited.Bioaugnentation technology can make up these shortcomings of conventional bioanalysis, by adding thing into system
The dominant bacteria with special role first tamed, with enhancement microbiological to the anti-of a certain specific environment or specific pollutant
Should, and then strengthen microorganism to the adaptability of former hostile environment, the treatment effect to organic matter is improved, so as to more effectively remove
Object noxious material, improve internal system Sludge Property, strengthening system stability.The source of dominant bacteria have nature screening and
Technique for gene engineering.Liu Qin etc. screens the dominant microflora for isolating high adaptability and high degradation capability from oxidation pond sludge,
It is added in anaerobic mud bed reactor, wastewater treatment performance can be strengthened significantly, COD clearances is maintained at 90% or so.
Qiu Zhong equalitys filter out the dominant strain of 5 plants of COD of waste leachate of effectively degrading, and research shows that hybrid bacterial strain compares single bacterial strain
Good degrading effect.Percolate water quality is complicated, is influenceed by component of refuse, landfill age, land-fill method, deals with difficulty pole
Greatly, existing dominant bacteria far can not meet actual demand at present.Therefore, screening amplification is applied to processing percolate
Dominant bacteria is particularly important.
The content of the invention
An object of the present invention is the provision of one plant of dominant strain for being applied to the processing of high ammonia nitrogen landfill leachate
Fusarium chlamydosporum TJPU02。
The bacterial strain screening of the present invention is from the soil sample of Tianjin Beichen District twoport Sanitary Landfill, in potato grape
Filamentous growth is shown as on sugared flat board, the conidium of yellow can be produced.It is sequenced by ITS and is compared in ncbi database
To finding that the bacterial strain is fusarium chlamydosporum (Fusarium chlamydosporum), Fusarium is named as
chlamydosporum TJPU02.Colonial morphology is as shown in Figure of description 1, and qualification result is as shown in Figure of description 2, figure
Middle Query is bacterial strain to be identified.
The second object of the present invention is to remove percolate using Fusarium chlamydosporum TJPU02
Middle COD.
To realize this purpose, the present invention takes following technical scheme：100mL garbage filters are accurately added into triangular flask
Liquid, taken out after sterilizing 30min under the conditions of 121 DEG C and be cooled to room temperature.Martin fluid nutrient medium mycelium pellet is taken out, use is sterile
Water cleans 2~3 times.20 sizes of access are homogeneous in each triangular flask and eugonic mycelium pellet, are put into after bottleneck is sealed
Shaking table, rotating speed 200rpm, 30 DEG C of temperature.Appropriate percolate water determination COD value is taken per 24h.
Brief description of the drawings
Fig. 1 is colonial morphology figure.
Fig. 2 is the genealogical classification result carried out by NCBI blast functions to bacterial strain.
Fig. 3 is that Fusarium chlamydosporum TJPU02 remove COD figures in percolate.
Specific embodiment
With reference to instantiation, the present invention will be further described.Bacterial strain used is Fusarium in embodiment
Chlamydosporum TJPU02, different culture mediums and solution formula are as follows：
(1) screening and culturing medium：Agar 1g, percolate 100mL, pH 7.0.
(2) martin substratum is improved：Glucose 10g, peptone 5g, KH2PO41g, MgSO4·7H2O 0.5g, NaCl
20g, distilled water 1000ml, pH are natural.
(3) garbage filter liquid culture medium：(NH4)2SO40.4g, glucose 0.5g, NaH2PO40.1g, pH 6.5-7.5, ooze
2 times of filtrate wastewater dilution, 100mL.
Embodiment 1Fusarium chlamydosporum TJPU02 screening
Soil sample of the experiment material from Tianjin Beichen District twoport Sanitary Landfill.Soil sampling is matched somebody with somebody with distilled water
Into bacterium solution, take 0.1ml to be coated on screening and culturing medium flat board, cultivated in biochemical cultivation case, the bacterium colony that picking can mushroom out
Line purifying 2-3 times is repeated, obtains purifying bacterial strain.
Embodiment 2Fusarium chlamydosporum TJPU02 identification
(1) Morphological Identification
By mycelium inoculation on PDA culture medium flat board, yellow, radial filamentous can be formed after being cultivated 2 days at 28 DEG C,
Mycelia covers rapidly whole planar surface.
(2) Molecular Identification
The spore of the aseptic inoculation bacterium and mycelia are into potato dextrose broth, 28 DEG C of shaken cultivations 24 hours.
Mycelium is obtained by filtering method.The genome of mycelia is extracted using the fast of TIANGEN Biotech (Beijing) Co., Ltd.
Prompt type plant genome DNA extraction system (Cat：DP321), expand the primer of ITS sequence for universal primer ITS1 and
ITS4.PCR, using PrimeSTAR HS (Premix) hi-fi systems (Cat of Takara companies：R040).Amplification system is：
PrimeSTAR HS (Premix, 2X) 25ul, ITS1 0.5ul, ITS4 0.5ul, template DNA 1ul, deionized water 23ul.Expand
Increasing condition is：98 DEG C of 10s, 60 DEG C of 5s, 72 DEG C of 1min, 30 circulations.PCR primer is purified and sent after reclaiming through gel electrophoresis
Hua Da Gene science limited company is sequenced.Taxonomic identification is carried out by ITS sequencings and constructing system tree, qualification result is as said
Shown in bright book accompanying drawing 2.
Embodiment 3Fusarium chlamydosporum TJPU02 handle the application of percolate
Fusarium chlamydosporum TJPU02 inoculations are to martin substratum, 30 DEG C of shaken cultivations 1 day,
It is inoculated in 0.5% inoculum concentration in garbage leachate wastewater, 30 DEG C of shaken cultivations, maintenance rotating speed is 200rpm；Daily centrifuging and taking
Supernatant, with titration measurement COD changes, after 7 days, the COD clearances of percolate have reached 80%, such as Figure of description 3
It is shown.
Claims (5)
1. one plant is applied to high ammonia nitrogen landfill leachate processing bacterium, it is characterised in that the Strain Designation is Fusarium
chlamydosporum TJPU02。
2. the screening technique of bacterial strain described in claim 1, comprises the following steps：
The soil sample of Tianjin Beichen District twoport Sanitary Landfill is taken, bacterium solution is made into distilled water, takes 0.1ml to be coated on
On screening and culturing medium flat board, cultivated in biochemical cultivation case, the bacterium colony that picking can mushroom out repeats line purifying 2-3 times, obtains
To purifying bacterial strain；Wherein screening and culturing medium：Agar 1g, percolate 100mL, pH7.0；Improve martin substratum：Grape
Sugared 10g, peptone 5g, KH2PO41g, MgSO4·7H2O 0.5g, NaCl 20g, distilled water 1000ml, pH are natural.
3. utilizing Fusarium chlamydosporum TJPU02, carry out landfill leachate treatment and remove COD, including following step
Suddenly：
By inoculation into martin substratum, 30 DEG C of shaken cultivations 2 days, percolate is inoculated in 2% inoculum concentration
In sewage, 30 DEG C of shaken cultivations 2 days, rotating speed maintains 200rpm, centrifuging and taking supernatant, uses KCr2O7Titration measuring COD changes.
4. the method for bacterial strain processing percolate according to claim 3, it is characterised in that inoculation in the step
Measure as 2%.
5. the method for bacterial strain processing percolate according to claim 3, it is characterised in that sewage disposal in the step
Temperature is 30 DEG C.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711220474.7A CN107858295A (en) | 2017-11-27 | 2017-11-27 | One plant is applied to high ammonia nitrogen landfill leachate processing bacterium and its application |
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| CN201711220474.7A CN107858295A (en) | 2017-11-27 | 2017-11-27 | One plant is applied to high ammonia nitrogen landfill leachate processing bacterium and its application |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102992539A (en) * | 2012-10-31 | 2013-03-27 | 杭州创达环境工程有限公司 | Method for treating high-concentration ammonia-nitrogen landfill leachate |
| CN106701602A (en) * | 2017-03-01 | 2017-05-24 | 青岛秾贝尔生物工程有限公司 | Strain of fusarium chlamydosporum and application thereof |
-
2017
- 2017-11-27 CN CN201711220474.7A patent/CN107858295A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102992539A (en) * | 2012-10-31 | 2013-03-27 | 杭州创达环境工程有限公司 | Method for treating high-concentration ammonia-nitrogen landfill leachate |
| CN106701602A (en) * | 2017-03-01 | 2017-05-24 | 青岛秾贝尔生物工程有限公司 | Strain of fusarium chlamydosporum and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| MARC E.SAVARD 等: "Chlamydosporoi, a new metabolite from Fusarium chlamydosporum", 《MYCOPATHOLOGIA》 * |
| 贺晓凌 等: "垃圾渗滤液处理菌株的筛选及处理工艺条件的优化", 《天津工业大学学报》 * |
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Application publication date: 20180330 |