CN106698676A - Enrichment method and application of methane oxidative coupling perchlorate reducing florae - Google Patents
Enrichment method and application of methane oxidative coupling perchlorate reducing florae Download PDFInfo
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- CN106698676A CN106698676A CN201611246493.2A CN201611246493A CN106698676A CN 106698676 A CN106698676 A CN 106698676A CN 201611246493 A CN201611246493 A CN 201611246493A CN 106698676 A CN106698676 A CN 106698676A
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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
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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
- C02F3/341—Consortia of bacteria
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/12—Halogens or halogen-containing compounds
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
- C02F2101/163—Nitrates
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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
- C02F2203/00—Apparatus and plants for the biological treatment of water, waste water or sewage
- C02F2203/004—Apparatus and plants for the biological treatment of water, waste water or sewage comprising a selector reactor for promoting floc-forming or other bacteria
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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
- C02F2305/00—Use of specific compounds during water treatment
- C02F2305/06—Nutrients for stimulating the growth of microorganisms
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- Hydrology & Water Resources (AREA)
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- Water Supply & Treatment (AREA)
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- Separation Using Semi-Permeable Membranes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to the technical field of biodegradation treatment, and aims at providing an enrichment method and application of methane oxidative coupling perchlorate reducing florae. The enrichment method of the methane oxidative coupling perchlorate reducing florae comprises the following steps: preparing an inorganic culture medium; inoculating ANMO-D florae in a hollow fiber membrane bioreactor; introducing the inorganic culture medium added with nitrite into a water inlet of the reactor; operating until NO2- is completely reduced to finish inoculation of initial florae; and introducing the inorganic culture medium added with ClO4- into the water inlet of the reactor and after continuously operating for 10 days, enriching florae capable of degrading perchlorate in the reactor. The florae disclosed by the invention have high-efficiency perchlorate degradation efficiency, and further can degrade perchlorate and nitrate in wastewater, and the degradation rates of perchlorate and nitrate can be close to 100%. A microbial bioreactor is used for removing perchlorate in water, so that the enrichment method and application of methane oxidative coupling perchlorate reducing florae have the advantages of low cost, long validity period, stable degradation effect and the like and are suitable for treating large quantities of low perchlorate contaminated wastewater for a long time.
Description
Technical field
The invention belongs to biodegradable processing technology field, and in particular to a kind of methane oxidation couples perchlorate reduction bacterium
The application for being enriched with and its being carried out in membrane bioreactor biodegradable treatment of group.
Background technology
With the fast-developing and industrial prosperity of China's economy, perchlorate is extensive as a kind of strong oxidizer
Apply in such as rocket fuel, fireworks, during ammunition etc. is produced and generated, so as to also bring follow-up pollution problem.Typical case
Underground water perchlorate content be 100 μ g/L, but in some cases, its content can reach 20mg/L or more.And it is high
Chloranion because there are close structure and properties with iodide ion, thus when people drink perchlorate content drink higher
When using water, perchlorate can substitute iodide ion and be had influence on so as to have influence on the generation of thyroid hormone necessary to human body
The normal development of human body.
And it is current, the perchlorate reduction carried out in body of groundwater using microorganism is with certain development prospect.
Using the vegetative activity of microorganism, using the energy source needed for perchlorate as its production, while using methane as carbon source,
Carbon dioxide, water and chlorion are formed in the presence of enzyme.Following reaction is carried out in vivo：
CH4+ClO4 -=HCO3 -+Cl-+H2O ΔG0’=-792KJmol-1CH4
Because its final metabolite is pollution-free, and the cost chemical treatment method that compares is more cheap, therefore such as
It is highly significant that what reaches efficient microorganism perchlorate degraded.
Meanwhile, nitrate anion is a kind of oxide anion, in underground water generally and perchlorate together.In the feelings of high concentration
Under condition, nitrate anion is possible to suppress the reduction of perchlorate, so the efficiently reduction simultaneously of research perchlorate and nitrate anion
Have very important significance.
In the prior art, membrane bioreactor is there is no at present can carry out the efficient reduction of perchlorate.Main chlorine high
Hydrochlorate biodegrading process by charcoal absorption, physical method and the chemical reduction method such as ion-exchange.
The content of the invention
The invention solves the problems that technology contents be to overcome the deficiencies in the prior art, there is provided a kind of perchlorate that can degrade
The enrichment method of flora and application.
To solve technical problem, solution of the invention is：
A kind of enrichment method of the flora of the perchlorate that can degrade is provided, is comprised the steps：
(1) inorganic medium is prepared
By 1mL acidity trace element solution, 1mL alkalescence trace element solution, 1mg CaCl2、0.3g NaHCO3、5mg
MgSO4·7H2O、0.2g KH2PO4With 0.4g Na2HPO4·12H2O is added into 1 liter of deionized water, and pH is adjusted to 7 with HCl,
It is made into culture medium；
The acid trace element solution refers to：Every liter of solution HCl containing 100mM, 2.085g FeSO4·7H2O、68mg
ZnSO4·7H2O、14mg H3BO3、120mg CoCl2·6H2O、500mg MnCl2·4H2O、320mg CuSO4、95mg
NiCl2·6H2O, balance of water；
The alkaline trace element solution refers to：Every liter of solution NaOH containing 10mM, 67mg SeO2、50mgNa2WO4·
2H2O、242mg Na2MoO4·2H2O, balance of water；
(2) initial inoculation source enrichment
10mL ANMO-D floras are inoculated with hollow-fiber membrane bioreactor；Containing by quality in the ANMO-D floras
Following strains that accounting is calculated：β-mycetozoan 2.88%, α-mycetozoan 4.29%, γ-mycetozoan 3.27%, acidfast bacilli
0.73%th, green bacterium 1.11%, surplus is the miscellaneous bacteria of non-enrichment object；
It is 2mg/L, Ran Houtong to nitrogen content during nitrite to culture medium is added in the inorganic medium obtained by step (1)
Pump is crossed to be introduced in the water inlet of hollow-fiber membrane bioreactor；Once, the amount of supply is 5L every time, is made hollow for supply in every two days
Nitrite concentration in the water inlet of fiber membrane biological reaction device remains constant；Continuous supply stops introducing culture medium after 40 days, directly
To NO2 -It is completely reduced, obtains being vaccinated with the hollow-fiber membrane bioreactor of initial flora；
(3) stage of reaction
ClO is added in the inorganic medium obtained by step (1)4 -Until its concentration is 5.5mg/L, inoculation is then introduced to
Run in the hollow-fiber membrane bioreactor of initial flora and continuously；Control temperature for 29 DEG C in whole process, enter water speed
Rate is 0.5mL/min, and half retention time is 130min, and methane partial pressure is 15psi (1.03bar)；After operation 10 days, the hollow fibre
The flora of the perchlorate that can degrade is enriched in dimension membrane bioreactor.
It is using being enriched energy invention further provides the application of the flora in perchlorate of degrading of foregoing acquisition
The hollow-fiber membrane bioreactor of the flora of degraded perchlorate, carries out the treatment containing perchlorate sewage；Specifically include：
(1) inorganic medium is prepared by claim 1 methods described, and is aerated to removal oxygen with nitrogen；
(2) by gained inorganic medium be introduced to it is described be enriched it is in the hollow-fiber membrane bioreactor of flora and continuous
Operation；The sewage containing perchlorate, and ClO are introduced at water inlet4 -Concentration in total amount is introduced into is 5.5mg/；In whole process
It is 29 DEG C to control temperature, and water inlet speed is 0.5mL/min, and half retention time is 130min, and methane partial pressure is 15psi
(1.03bar)。
It is using being enriched and can degrade present invention also offers the application of the flora in perchlorate of degrading of foregoing acquisition
The hollow-fiber membrane bioreactor of the flora of perchlorate, carries out the treatment of the sewage containing perchlorate and nitrate；Specifically
Including：
(1) inorganic medium is prepared by claim 1 methods described, and is aerated to removal oxygen with nitrogen；
(2) by gained inorganic medium be introduced to it is described be enriched it is in the hollow-fiber membrane bioreactor of flora and continuous
Operation；The sewage containing perchlorate and nitrate, and ClO are introduced at water inlet4 -Concentration in total amount is introduced into is 1mg/L,
NO3 -Concentration in total amount is introduced into is 4mg/L；It is 29 DEG C that temperature is controlled in whole process, and water inlet speed is 0.5mL/min, half
Retention time is 130min, and methane partial pressure is 15psi (1.03bar).
Compared with prior art, its advantage is the present invention：
1st, the flora in the present invention has the efficiency of efficient degraded perchlorate, can in half retention time of 130min
With by the ClO of 5.5mg/L4 -It is degradable.
2nd, the reactor in the present invention can degrade the perchlorate and nitrate in sewage simultaneously, while the degraded of the two
Rate can be reached close to 100%.
3rd, prior art is compared, the perchlorate that present invention microorganism reactor is gone in water removal has low cost, effectively
Phase is long, the advantages of degradation effect stabilization, is adapted to process for a long time the waste water of a large amount of low perchlorate contaminateds.
Brief description of the drawings
Reactor carries out perchlorate sewage degradation data figure in Fig. 1 present invention；
Reactor carries out perchlorate sewage degradation efficiency figure in Fig. 2 present invention；
Reactor carries out perchlorate and nitrate sewage mixed degradation perchlorate datagram in Fig. 3 present invention；
Reactor carries out perchlorate and nitrate sewage mixed degradation nitrate datagram in Fig. 4 present invention；
Reactor carries out perchlorate and nitrate sewage mixed degradation efficiency chart in Fig. 5 present invention.
Specific embodiment
In the present invention, as initial inoculation source is ANMO-D floras.The acquisition modes of ANMO-D floras are：Collection sewage
The activated sludge (the applicant is gathered from the lattice sewage treatment plant of Hangzhou seven) for the treatment of plant, using nitrate as electron acceptor, with first
Alkane is then seeded into methane MBfR reactors until stable, you can obtain as electron donor enrichment culture about half a year
The ANMO-D floras of strain stable content.Contain the following strains calculated by quality accounting in ANMO-D floras：β-mycetozoan
2.88%th, α-mycetozoan 4.29%, γ-mycetozoan 3.27%, acidfast bacilli 0.73%, green bacterium 1.11%, surplus are that non-enrichment is right
The miscellaneous bacteria of elephant.
Applicant promises to undertake：The ANMO-D floras are provided to the public from from the patent application 20 years, for realizing,
Using technical scheme of the present invention.
In the present invention, the reactor for being used is hollow-fiber membrane bioreactor, can voluntarily be built in the usual way.Should
Reactor is two-tube hollow-fiber membrane bioreactor, and cumulative volume is 65mL, each 32 hydrophobic micropore polyethylene in each pipe
Film (external diameter is 280 μm, and internal diameter is 180 μm, and micro-pore diameter is 0.1-0.15 μm), the film is by Japanese Mitsubishi company systems
Make, model MHF-200TL).Reactor configuration can refer to MBfR reactors, specific visible bibliography (Zhao et.al,
2011,Interactions between perchlorate and nitrate reductions in the biofilm
of a hydrogen-based membrane biofilm reactor))。
Embodiment 1：
In the present invention, the enrichment method of the flora of the perchlorate that can degrade comprises the steps：
(1) inorganic medium is prepared
By 1mL acidity trace element solution, 1mL alkalescence trace element solution, 1mg CaCl2、0.3g NaHCO3、5mg
MgSO4·7H2O、0.2g KH2PO4With 0.4g Na2HPO4·12H2O is added into 1 liter of deionized water, and pH is adjusted to 7 with HCl,
It is made into culture medium；
The acid trace element solution refers to：Every liter of solution HCl containing 100mM, 2.085g FeSO4·7H2O、68mg
ZnSO4·7H2O、14mg H3BO3、120mg CoCl2·6H2O、500mg MnCl2·4H2O、320mg CuSO4、95mg
NiCl2·6H2O, balance of water；
The alkaline trace element solution refers to：Every liter of solution NaOH containing 10mM, 67mg SeO2、50mg Na2WO4·
2H2O、242mg Na2MoO4·2H2O, balance of water；
(2) initial inoculation source enrichment
10mL ANMO-D floras are inoculated with hollow-fiber membrane bioreactor；Containing by quality in the ANMO-D floras
Following strains that accounting is calculated：β-mycetozoan 2.88%, α-mycetozoan 4.29%, γ-mycetozoan 3.27%, acidfast bacilli
0.73%th, green bacterium 1.11%, surplus is the miscellaneous bacteria of non-enrichment object；
To in the inorganic medium obtained by step (1) add nitrite to culture medium in nitrogen content be 2mg/L (i.e.
6.57mg/L nitrite), then it is introduced in the water inlet of hollow-fiber membrane bioreactor by pump；Supply one in every two days
Secondary, the amount of supply is 5L every time, and the nitrite concentration in making hollow-fiber membrane bioreactor intake remains constant；It is continuous to supply
To stopping introducing culture medium after 40 days, until NO2 -It is completely reduced, obtains being vaccinated with the doughnut membrane bioreaction of initial flora
Answer device；
(3) stage of reaction
ClO is added in the inorganic medium obtained by step (1)4 -Until its concentration is 5.5mg/L, inoculation is then introduced to
Run in the hollow-fiber membrane bioreactor of initial flora and continuously；Control temperature for 29 DEG C in whole process, enter water speed
Rate is 0.5mL/min, and half retention time is 130min, and methane partial pressure is 15psi (1.03bar)；After operation 10 days, the hollow fibre
The flora of the perchlorate that can degrade is enriched in dimension membrane bioreactor.
(4) Bacterial community identification
Structure of community to enrichment flora in step (3) carries out sequencing identification, and main bacteria seed includes in flora：β-mycetozoan
(Betaproteobacteria), α-mycetozoan (Alphaproteobacteria), γ-mycetozoan
(Gammaproteobacteria), acidfast bacilli (Acidobacteria) and green bacterium (Chlorobia), respective ratio is：β-
(remaining is for mycetozoan 41.15%, α-mycetozoan 27.08%, γ-mycetozoan 11.12%, acidfast bacilli 7.38%, green bacterium 2.40%
The miscellaneous bacteria of all kinds of non-lipid collection objects).
Embodiment 2：
The present invention can be further used for the perchlorate in degraded sewage.
Can be carried out containing perchlorate sewage by using the hollow-fiber membrane bioreactor for being enriched the flora
Treatment；Specifically include：
(1) inorganic medium is prepared by preceding method, and is aerated to removal oxygen with nitrogen；
(2) gained inorganic medium is introduced to be enriched in the hollow-fiber membrane bioreactor of the flora and continuous
Operation；The sewage containing perchlorate, and ClO are introduced at water inlet4 -Concentration in total amount is introduced into is 5.5mg/；In whole process
It is 29 DEG C to control temperature, and water inlet speed is 0.5mL/min, and half retention time is 130min, and methane partial pressure is 15psi
(1.03bar)。
Through inspection, in half retention time of 130min, can be by the ClO of 5.5mg/L in sewage4 -It is degradable.
Embodiment 3：
The present invention can be additionally used in the perchlorate and nitrate in degraded sewage simultaneously.
The treatment containing perchlorate sewage is carried out by using the hollow-fiber membrane bioreactor for being enriched the flora；
Specifically include：
(1) inorganic medium is prepared by claim 1 methods described, and is aerated to removal oxygen with nitrogen；
(2) gained inorganic medium is introduced to be enriched in the hollow-fiber membrane bioreactor of the flora and continuous
Operation；The sewage containing perchlorate and nitrate, and ClO are introduced at water inlet4 -Concentration in total amount is introduced into is 1mg/L,
NO3 -Concentration in total amount is introduced into is 4mg/L；It is 29 DEG C that temperature is controlled in whole process, and water inlet speed is 0.5mL/min, half
Retention time is 130min, and methane partial pressure is 15psi (1.03bar).
Through inspection, in half retention time of 130min, can be by the ClO of 1mg/L in sewage4 -With the NO of 4mg/L3 -It is complete
Degradable.
Claims (3)
1. a kind of methane oxidation couples the enrichment method of perchlorate reduction's flora, it is characterised in that comprise the steps：
(1) inorganic medium is prepared
By 1mL acidity trace element solution, 1mL alkalescence trace element solution, 1mg CaCl2、0.3g NaHCO3、5mg
MgSO4·7H2O、0.2g KH2PO4With 0.4g Na2HPO4·12H2O is added into 1 liter of deionized water, and pH is adjusted to 7 with HCl,
It is made into culture medium；
The acid trace element solution refers to：Every liter of solution HCl containing 100mM, 2.085g FeSO4·7H2O、68mg
ZnSO4·7H2O、14mg H3BO3、120mg CoCl2·6H2O、500mg MnCl2·4H2O、320mg CuSO4、95mg
NiCl2·6H2O, balance of water；
The alkaline trace element solution refers to：Every liter of solution NaOH containing 10mM, 67mg SeO2、50mg Na2WO4·2H2O、
242mg Na2MoO4·2H2O, balance of water；
(2) initial inoculation source enrichment
10mL ANMO-D floras are inoculated with hollow-fiber membrane bioreactor；Containing by quality accounting in the ANMO-D floras
The following strains for calculating：It is β-mycetozoan 2.88%, α-mycetozoan 4.29%, γ-mycetozoan 3.27%, acidfast bacilli 0.73%, green
Bacterium 1.11%, surplus is the miscellaneous bacteria of non-enrichment object；
It is 2mg/L to nitrogen content during nitrite to culture medium is added in the inorganic medium obtained by step (1), then by pump
It is introduced in the water inlet of hollow-fiber membrane bioreactor；Once, the amount of supply is 5L every time, makes doughnut for supply in every two days
Nitrite concentration in membrane bioreactor water inlet remains constant；Continuous supply stops introducing culture medium after 40 days, until NO2 -
It is completely reduced, obtains being vaccinated with the hollow-fiber membrane bioreactor of initial flora；
(3) stage of reaction
ClO is added in the inorganic medium obtained by step (1)4 -Until its concentration is 5.5mg/L, then it is introduced to be vaccinated with just
Run in the hollow-fiber membrane bioreactor of beginning flora and continuously；Temperature is controlled in whole process for 29 DEG C, water inlet speed is
0.5mL/min, half retention time is 130min, and methane partial pressure is 15psi；After operation 10 days, the doughnut membrane biological reaction
The flora of the perchlorate that can degrade is enriched in device.
2. application of the flora for being obtained in claim 1 methods described in perchlorate of degrading, it is characterised in that be using richness
Collect the hollow-fiber membrane bioreactor of the flora of the perchlorate that can degrade, carry out the treatment containing perchlorate sewage；Specifically
Including：
(1) inorganic medium is prepared by claim 1 methods described, and is aerated to removal oxygen with nitrogen；
(2) gained inorganic medium is introduced to described be enriched in the hollow-fiber membrane bioreactor of flora and continuously transport
OK；The sewage containing perchlorate, and ClO are introduced at water inlet4 -Concentration in total amount is introduced into is 5.5mg/；Controlled in whole process
Temperature processed is 29 DEG C, and water inlet speed is 0.5mL/min, and half retention time is 130min, and methane partial pressure is 15psi.
3. application of the flora for being obtained in claim 1 methods described in perchlorate of degrading, it is characterised in that be using richness
Collect the hollow-fiber membrane bioreactor of the flora of the perchlorate that can degrade, carry out the sewage containing perchlorate and nitrate
Treatment；Specifically include：
(1) inorganic medium is prepared by claim 1 methods described, and is aerated to removal oxygen with nitrogen；
(2) gained inorganic medium is introduced to described be enriched in the hollow-fiber membrane bioreactor of flora and continuously transport
OK；The sewage containing perchlorate and nitrate, and ClO are introduced at water inlet4 -Concentration in total amount is introduced into is 1mg/L, NO3 -
Concentration in total amount is introduced into is 4mg/L；It is 29 DEG C that temperature is controlled in whole process, and water inlet speed is 0.5mL/min, and half retains
Time is 130min, and methane partial pressure is 15psi.
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| CN201611246493.2A CN106698676B (en) | 2016-12-29 | 2016-12-29 | Enrichment method and application of methane oxidation coupling perchlorate reducing flora |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101786763A (en) * | 2010-01-26 | 2010-07-28 | 华东理工大学 | Batch-type groundwater denitrifying method |
| CN102061281A (en) * | 2010-11-30 | 2011-05-18 | 哈尔滨工业大学 | Method for enriching and culturing anaerobic ammonia oxidizing bacteria by using membrane bioreactor |
| CN103275868A (en) * | 2013-05-22 | 2013-09-04 | 北京工业大学 | Device and method for enriching denitrification-dependent anaerobic methane oxidation bacteria based on intermittent membrane bioreactor |
| CN103723893A (en) * | 2014-01-10 | 2014-04-16 | 中国科学院生态环境研究中心 | Method for removing nitrate nitrogen from water |
| CN105271513A (en) * | 2015-11-02 | 2016-01-27 | 湖南大学 | Methane substrate biomembrane reactor and process for removing perchlorate from drinking water |
-
2016
- 2016-12-29 CN CN201611246493.2A patent/CN106698676B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101786763A (en) * | 2010-01-26 | 2010-07-28 | 华东理工大学 | Batch-type groundwater denitrifying method |
| CN102061281A (en) * | 2010-11-30 | 2011-05-18 | 哈尔滨工业大学 | Method for enriching and culturing anaerobic ammonia oxidizing bacteria by using membrane bioreactor |
| CN103275868A (en) * | 2013-05-22 | 2013-09-04 | 北京工业大学 | Device and method for enriching denitrification-dependent anaerobic methane oxidation bacteria based on intermittent membrane bioreactor |
| CN103723893A (en) * | 2014-01-10 | 2014-04-16 | 中国科学院生态环境研究中心 | Method for removing nitrate nitrogen from water |
| CN105271513A (en) * | 2015-11-02 | 2016-01-27 | 湖南大学 | Methane substrate biomembrane reactor and process for removing perchlorate from drinking water |
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