CN108707598A - A method of using filamentous fungi as the intensified anti-nitrated bacterial nitrogen removal of carrier - Google Patents
A method of using filamentous fungi as the intensified anti-nitrated bacterial nitrogen removal of carrier Download PDFInfo
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- CN108707598A CN108707598A CN201810533296.1A CN201810533296A CN108707598A CN 108707598 A CN108707598 A CN 108707598A CN 201810533296 A CN201810533296 A CN 201810533296A CN 108707598 A CN108707598 A CN 108707598A
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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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/16—Enzymes or microbial cells immobilised on or in a biological cell
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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
- 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
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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
- C12N1/20—Bacteria; Culture media therefor
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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
Abstract
The invention discloses a kind of using filamentous fungi as the method for the intensified anti-nitrated bacterial nitrogen removal of carrier, belongs to technical field of biological sewage treatment.The method of the present invention includes：It is inoculated with suitable muddy water mixed solution to cultivate in denitrification enrichment culture liquid, obtains the bacteria suspension of denitrifying bacteria；Inoculation Talaromyces flavus S1 slant mediums are inoculated in PDB culture mediums, and shaking table culture obtains ripe mycelium pellet；By denitrifying bacteria bacterial suspension inoculation to the denitrification culture solution containing ripe mycelium pellet in, with the denitrification effect of intensified anti-nitrated bacterium, and repeated collection mycelium pellet.The Talaromyces flavus S1 mycelium pellets used in the method for the present invention, denitrification effect than current existing Trichodermavirid mycelium pellets immobilization denitrifying bacteria is good, it not only has many advantages, such as that easy to operate, settling property is good and high recycling rate, the denitrification effect of also intensified anti-nitrated bacterium, foundation is provided to the practical application of denitrifying bacteria from now on, has also opened up the actual application prospect of Talaromyces flavus S1.
Description
Technical field
The present invention relates to a kind of using filamentous fungi as the method for the intensified anti-nitrated bacterial nitrogen removal of carrier, belongs at saprobia
Manage technical field.
Background technology
In recent years, the mankind unreasonably use the untreated leakage of formulation fertilizer containing nitrogen, random sewage effluent and landfill leachate,
Cause to contain a large amount of nitrate in water body.And the excess of nitrate, the health of the mankind can be threatened, such as acute siderosis red eggs
White disease and Lan Ying syndromes.Therefore, in recent years, people have been devoted to the nitrate in removal underground water.And compared to Physical,
The limitations such as the of high cost of chemical method, effect are poor, are difficult to carry out remove the azotate pollution in water body using bioanalysis, receive
The concern of people.
Nitrate removal in water body is mainly reduced by denitrification, and the microorganism for participating in denitrification is more
Sample is various.The microorganism that denitrification will usually be carried out is referred to as denitrifying microorganism, such as Proteobacteria,
Bacteroidetes, Firmicutes, Gracilibacter etc..Denitrifying bacteria utilizes under conditions of anoxic or anaerobism
Itself carbon source and additional carbon, using NO3-N and NO2-N as electron acceptor, pass through internal nitrate as electron donor
Nitrate transformation in water body is N by reductase, nitrite reductase etc.2, to complete the nitrate removal in water body,
Achieve the purpose that the removal effect and organics removal of total nitrogen.Therefore, denitrifying bacteria plays the part of in removing water body in nitrate
Important role, it is deep to be favored by everybody.
And during practice, the effect of denitrifying bacteria is directly added into water body can not obtain ideal effect
Fruit, to avoid loss of the microorganism in water body, reinforcement technique from being used in the practical application of water body.Enhancement microbiological technology,
Refer to that objective microbe is fixed in the carrier, is ensureing the original active basis of microorganism using means such as physics, chemistry
On, the concentration of microorganism is also improved, protective effect is played to microorganism, improves impact resistance.Meanwhile strengthening production recycling
On object, be conducive to the classification of product, reduce wasting phenomenon.Therefore, a kind of suitable schedule of reinforcement is found, to denitrifying bacteria
It is fixed, there is great application prospect on water body denitrification.Generally according to the fixed type of reinforcing, be divided into physical absorption,
Chemisorption and biological adsorption, common materialization sorbing material have polyurethane foam, polyvinyl alcohol, sodium alginate etc., but its
In certain degree, the activity of denitrifying microorganism is affected, affects the denitrification effect of water body.
Currently, have document report carries out denitrogenation, the mycelium pellet of this report using Trichodermavirid mycelium pellets
Training method is that a kind of secondary coccus (denitrifying bacteria) and trichoderma viride co-culture, and obtains mixing mycelium pellet to remove river dirt
Water pollutant.It is with higher removal effect, but there is mix the defects such as mycelium pellet incubation time is long, success rate is low.
Invention content
Based on the above issues, needle of the present invention puts forward a kind of highly efficient anti-nitre to the practical application of denitrifying bacteria
Change bacteria adhension method, this method is made using the mycelium pellet of filamentous fungi Talaromyces flavus S1 (blue shape Pseudomonas)
Sewage is handled in turn for the fixation support of denitrifying bacteria, is effectively improved wastewater treatment efficiency.It is existing with ball
The method that denitrogenation is carried out as fixation support using Trichodermavirid mycelium pellets compare, the present invention is using first cultivating
The mode of denitrifying bacteria is inoculated with after mycelium pellet, it greatly improves success rates prepared by mycelium pellet, effectively shorten mycelium pellet
The time of immobilization denitrifying bacteria；Meanwhile the present invention can be directed to most of denitrifying bacteria, avoid certain denitrification
Interaction between bacterium and the strain that mycelium pellet can be prepared.In short, the method for the present invention is highly efficient, it is Talaromyces
The water body processing that flavus S1 mycelium pellets are applied to denitrifying bacteria provides foundation.
The present invention prepares mycelium pellet by filamentous fungi Talaromyces flavus S1, and it is strong to become microorganism
The excellent carrier of change, and then establish a kind of Talaromyces flavus more highly efficient than Trichodermavirid mycelium pellet
S1 mycelium pellets are the denitrifying bacteria process for fixation of carrier, not only the repetition profit with Trichodermavirid mycelium pellets
It the advantages that with, strong antijamming capability, can be with the denitrification effect of intensified anti-nitrated bacterium.
The first purpose of the invention is to provide a kind of immobilization denitrifying bacteria, the immobilization denitrifying bacteria be with
Talaromyces flavus S1 mycelium pellets are obtained as fixation support.
In one embodiment, the preparation of the immobilization denitrifying bacteria includes：
(1) denitrifying bacteria bacteria suspension and Talaromyces flavus S1 mycelium pellets are obtained；
(2) denitrifying bacteria bacteria suspension and ripe Talaromyces flavus S1 mycelium pellets are inoculated into simultaneously and are waited for
In the waste water of processing, obtain using Talaromyces flavus S1 bacterium as the immobilization denitrifying bacteria of carrier.
Second object of the present invention is to provide a kind of method of intensified anti-nitrated bacterial nitrogen removal effect, the method be with
Fixation support of the Talaromyces flavus S1 mycelium pellets as denitrifying bacteria, recycles the denitrification of immobilization thin
Bacterium carries out denitrogenation processing.
In one embodiment, the method includes：
(1) denitrifying bacteria bacteria suspension and Talaromyces flavus S1 mycelium pellets are obtained；
(2) denitrifying bacteria bacteria suspension and ripe Talaromyces flavus S1 mycelium pellets are inoculated into simultaneously and are waited for
It in the waste water of processing, obtains using Talaromyces flavus S1 bacterium as the mycelium pellet of the denitrifying bacteria of carrier, culture processing
For a period of time.
In one embodiment, the preparation method of the denitrifying bacteria bacteria suspension includes：
(A) denitrifying bacteria is cultivated：The muddy water mixed solution for taking suitable sewage treatment plant's anoxic pond is inoculated into denitrification training
In nutrient solution, anaerobism shaking table culture, three times, centrifugation obtains denitrifying bacteria thalline for continuous culture；
(B) denitrifying bacteria bacteria suspension is prepared：Appropriate thalline is taken to be resuspended in aqua sterilisa, the bacterium for obtaining denitrifying bacteria is outstanding
Liquid；
In one embodiment, muddy water mixed solution need to be rushed with phosphate buffer (pH=7) in the step (A)
It washes, crosses 200 mesh sieve, remove impurity.
In one embodiment, in the step (A) activated sludge and distilled water with 1:4 volume ratios are diluted, quiet
After setting 1 hour, the mud mixture 10ml in middle level is taken, denitrification culture solution 90ml is added, is put in conical flask.
In one embodiment, the denitrification culture solution main component in the step (A) is：KNO32g/L,
C6H12O61.298g/L MgSO4·7H2O 0.2g/L, micro- 2ml/L, phosphate buffer (pH=7.5) 50ml/L,
Surplus is water.
In one embodiment, the phosphate buffer of the step (A) is by Na2HPO40.908g/L and NaH2PO4
0.25g/L is mixed.
In one embodiment, the anaerobic condition of the step (A) is to be passed through argon gas 10 minutes, to remove extra oxygen
Gas.
In one embodiment, in the step (A) denitrifying bacteria thalline be by 4000r/min, 15 minutes
Centrifugation obtains.
In one embodiment, the bacteria suspension of step (A) denitrifying bacteria is 1g/100ml.
In one embodiment, the condition of culture of the step (A) is：Rotating speed 130r/min, 28 ± 2 DEG C of temperature, training
Foster cycle time is 5 days.
In one embodiment, the preparation of the Talaromyces flavus S1 mycelium pellets includes：
(a) bacteria suspension of Talaromyces flavus S1 is prepared：The inclined-plane of Talaromyces flavus S1 is taken to train
Base is supported, is inoculated in the aqua sterilisa equipped with bead, is put in shaking table and shakes, blood count is carried out, obtains Talaromyces
The spore bacteria suspension of flavus S1；
(b) Talaromyces flavus S1 mycelium pellets are cultivated：Appropriate spore bacteria suspension is taken, PDB culture mediums are inoculated into
In, shaking table culture 3~5 days obtains ripe mycelium pellet；
In one embodiment, filamentous fungi Talaromyces flavus S1 come from Jiangnan in the step (a)
University Environment is referred to Architecture and Construction's solid waste control and recycling laboratory screening gained, concrete property《Filamentous fungi
The research of balling-up characteristics and improvement dewatering of the Talaromyces flavus S1 in sludge》.
In one embodiment, the shaking table culture condition in the step (a) is：28 ± 2 DEG C of temperature, rotating speed are
130r/min, incubation time are 5~6 days.
In one embodiment, the blood count in the step (a) is 7.8 × 107~9.3 × 107A/L.
In one embodiment, the volume ratio that the spore suspension inoculum concentration in the step (b) is 10%.
In one embodiment, the PDB culture medium main components in the step (b) are：200g/L potato decortications boil
It is ripe, 20g/L C6H12O6It is uniformly mixed, 121 DEG C, 20min high pressure sterilizations.
In one embodiment, the denitrifying bacteria bacteria suspension in the step (2) is added with 0.5-2% volume ratios,
It is 3-8g/100mL waste water with 5gTalaromyces flavus S1 mycelium pellet additive amounts.
In one embodiment, the denitrifying bacteria bacteria suspension in the step (2) is added with 1% volume ratio, with
5gTalaromyces flavus S1 mycelium pellets, while being added in the simulated wastewater of 100ml.
In one embodiment, the condition of culture in the step (2) is：28 ± 2 DEG C of temperature, rotating speed 100-
150r/min, incubation time are 1-3 days.
In one embodiment, the shaking table culture condition in the step (2) is：28 ± 2 DEG C of temperature, rotating speed are
130r/min, incubation time are 1.8 days.
Beneficial effects of the present invention：
The present invention has carried out the preparation of mycelium pellet to Talaromyces flavus S1 bacterium, using its mycelium pellet as carrier, leads to
It crosses and immobilizes denitrifying bacteria.Denitrifying bacteria utilizes the unique space structure of mycelium pellet and biocompatibility, absorption
On the surface and inside of mycelium pellet, the denitrifying bacteria Mixed Microbes ball centered on mycelium pellet is formd.It not only improves anti-nitre
The biomass for changing bacterium, also promotes impact resistance.It applies it in sewage, hence it is evident that enhance the denitrogenation energy of denitrifying bacteria
Power.
The present invention carries out the immobilization of denitrifying bacteria using Talaromyces flavus S1 as carrier, with this immobilization
The strengthened denitrification effect of denitrifying bacteria.It not only has many advantages, such as that easy to operate, settling property is good, repeatable usability is high,
Its being effectively fixed denitrifying bacteria nitrate nitrogen removal ability, enhance water body total nitrogen removal effect, so as to improve water quality
Deterioration.And the denitrifying bacteria of immobilization, recovery utilization rate is high, can reduce the pollution to environment.The present invention is not only
New application prospect is provided for the application of Talaromycesflavus S1 bacterium, is also provided more for the nitrate removal of water body
For efficient process for fixation.
Description of the drawings
Fig. 1 is the nitrate removal situation of embodiment 1；
Fig. 2 is that the total nitrogen of embodiment 1 removes situation；
Fig. 3 is the nitrate removal situation during the mycelium pellet of embodiment 1 recycles；
Fig. 4 is that the total nitrogen during the mycelium pellet of embodiment 1 recycles removes situation；
Fig. 5 is the nitrate removal situation of actual waste water；
Fig. 6 is the nitrate removal situation of actual waste water；
Fig. 7 is the nitrate removal situation that mycelium pellet is utilized in actual waste water processing cycle；
Fig. 8 is that the total nitrogen that mycelium pellet is utilized in actual waste water processing cycle removes situation.
Specific implementation mode
In order to illustrate more clearly of the present invention, the present invention is further detailed with reference to experiment case study.Ability
The technical staff in domain should understand, below the content of specific descriptions done be illustrative and be not restrictive, should not be with
This is limited the scope of the invention.
Embodiment 1：
(1) enrichment culture of denitrifying bacteria：Take the anoxic pond sludge of sewage treatment plant several.Used phosphate-buffered
Liquid (pH=7) is rinsed, and is sieved by 200 mesh, to remove extra impurity.By activated sludge and distilled water with 1:4 volume
Than being diluted.After standing 1 hour, the mud mixture 10ml in middle level is taken, is put in conical flask, denitrification culture solution is added
(Tables 1 and 2) 90ml is passed through argon gas to remove oxygen, ensures anaerobic condition.When the nitrate concentration of culture solution is reduced to centainly
After degree, mixed liquor is taken to carry out 4000r/min, centrifugation in 15 minutes obtains denitrifying bacteria.Same method repeats culture three
It is secondary, you can to obtain the higher denitrifying bacteria of relative abundance.
(2) prepared by the bacteria suspension of denitrifying bacteria：By the denitrifying bacteria deposit 1g in (1), with 100ml aqua sterilisas into
Row is resuspended, and to obtain the bacteria suspension of denitrifying bacteria, 4 DEG C of refrigerators preserve.
(3) slant medium for taking Talaromyces flavus S1 is inoculated into the aqua sterilisa equipped with bead, puts
It is shaken in shaking table, carries out blood count, obtain the spore bacteria suspension of Talaromyces flavus S1；It is inoculated into PDB cultures
In base, shaking table culture 5~6 days obtains ripe mycelium pellet；
(4) slant medium for taking Trichodermavirid is inoculated into the aqua sterilisa equipped with bead, puts
It is shaken in shaking table, carries out blood count, obtain the spore bacteria suspension of Trichoderma viride；It is inoculated into PDB culture mediums, shaking table culture
5~6 days, obtain ripe mycelium pellet；
(5) by denitrifying bacteria bacteria suspension and ripe Talaromyces flavus S1 mycelium pellets (or
Trichodermavirid mycelium pellets) while being inoculated into nitrate nitrogen a concentration of 277mg/L, COD 650mg/L, pH 7.5, temperature
For in 25 DEG C of 100ml simulated wastewaters (table 3), obtain with Talaromyces flavus S1 bacterium (or
Trichodermavirid bacterium) be carrier denitrifying bacteria mycelium pellet；
(6) it is not solid to be divided into blank group 1 (only adding culture solution group), blank group 2 (only adding culture solution and mycelium pellet) for experiment
Surely change denitrifying bacterium group (addition culture solution and denitrifying bacteria group), (addition is instead for Trichodermavirid mycelium pellet fixations group
Nitrobacteria, Trichodermavirid mycelium pellets and culture solution group) and (addition of Talaromyces flavus S1 mycelium pellets
Denitrifying bacteria, Talaromyces flavus S1 mycelium pellets and culture solution group) fixed group, surveyed every 3 hours a nitrate nitrogen and
The removal rate of total nitrogen, it is as shown in Figures 2 and 3 respectively.
(7) collection step 6) in Trichodermavirid mycelium pellet fixations group (addition denitrifying bacteria,
Trichodermavirid mycelium pellets and culture solution group) and Talaromyces flavus S1 mycelium pellets (addition denitrification it is thin
Bacterium, Talaromyces flavus S1 mycelium pellets and culture solution group) mycelium pellet in fixed group, it is carried out with phosphate buffer
It is cleaned multiple times, to remove the denitrifying bacteria in mycelium pellet, then adds the denitrifying bacteria of 1% volume ratio, renewed vaccination is to instead
It nitrifies in culture solution, with 1.8 days for a period, surveys its final nitrate nitrogen and total nitrogen concentration, to probe into the repetition profit of mycelium pellet
Use effect；
As shown in Figure 1, for the nitrate removal situation of different experiments group, ripe Trichodermavirid mycelium pellets are taken
With Talaromyces flavus S1 mycelium pellet 5g, (Tables 1 and 2) 100ml is added in denitrification culture solution, and added
The denitrifying bacteria bacteria suspension for adding 1ml analyzes the nitrate adsorption capacity of mycelium pellet.The result shows that：Itself absorption of mycelium pellet
Nitrate ability is only 24.34mg/L, and the nitrate removal amount of removal rate 8.83%, only inoculation denitrifying bacteria is
168.52mg/L, removal rate 61.20mg/L, and the denitrifying bacteria removal rate after immobilization increased, but exist
Gap.Using the Trichodermavirid mycelium pellet immobilizations of existing literature, the nitrate of 206.93mg/L, removal are only removed
Rate is 75.15%.Use the Talaromyces flavus S1 mycelium pellet immobilization denitrifying bacterias in the present invention, removal
Rate is enhanced, and is 223.40mg/L, removal rate 81.14%, compared to existing mycelium pellet, removal rate increases
5.99%.
As shown in Figure 2：Situation is removed for the total nitrogen of different experiments group.The total nitrogen removal amount of mycelium pellet itself is only
36.35mg/L, nitrogen removal rate are only 12.50%.Using the denitrifying bacteria of immobilization, total nitrogen removal effect is carried
It rises.The Talaromyces flavus S1 mycelium pellet immobilization denitrifying bacterias prepared using this experiment, the removal of total nitrogen can
Reach 225.64mg/L, removal rate 78.41%, the Trichodermavirid mycelium pellet immobilizations used than existing document
Total nitrogen removal amount 184.02mg/L, be higher by 41.62mg/L, removal rate is higher by 14.46%.
As shown in Figure 3：For the cycle of Trichodermavirid mycelium pellets and Talaromyces flavus S1 mycelium pellets
Nitrate nitrogen removal ability in utilization.The result shows that：Two kinds of mycelium pellets still keep higher fixation after 5 times recycle
Change effect.After 5 times utilize, nitrate removal rate is converted into Trichodermavirid mycelium pellets from 75.15%
74.20%, and Talaromyces flavus S1 mycelium pellets prepared by the present invention, after 5 times utilize, nitrate removal
Rate is changed into 76.68% from 81.14%.From nitrate removal rate：The repetition of Talaromyces flavus S1 mycelium pellets
The Trichodermavirid mycelium pellets of document report before utility is better than.
As shown in Figure 4：For the cycle of Trichodermavirid mycelium pellets and Talaromyces flavus S1 mycelium pellets
Total nitrogen removal ability in utilization.The result shows that：After 5 times recycle, nitrogen removal rate is still kept two kinds of mycelium pellets
In higher level.The nitrogen removal rate of Trichodermavirid mycelium pellets is converted into 62.81% from 63.95%, total nitrogen
Removal effect does not weaken.And the nitrogen removal rate of Talaromyces flavus S1 mycelium pellets is recycled by 5 times
Afterwards, it is reduced to 68.46% from 78.41%, nitrogen removal rate remains above existing document report.
1 denitrification culture solution main component of table
Trace Elements in 2 denitrification culture solution of table
Simulated wastewater main component in 3 embodiment of table
Embodiment 2：
(1) enrichment culture of denitrifying bacteria：Take the anoxic pond sludge of sewage treatment plant several.Used phosphate-buffered
Liquid (pH=7) is rinsed, and is sieved by 200 mesh, to remove extra impurity.By activated sludge and distilled water with 1:4 volume
Than being diluted.After standing 1 hour, the mud mixture 10ml in middle level is taken, is put in conical flask, denitrification culture solution is added
90ml is passed through argon gas to remove oxygen, ensures anaerobic condition.After the nitrate concentration of culture solution reduces to a certain extent, take
Mixed liquor carries out 4000r/min, and centrifugation in 15 minutes obtains denitrifying bacteria.Same method repeats culture three times, you can obtains
The higher denitrifying bacteria of relative abundance.
(2) prepared by the bacteria suspension of denitrifying bacteria：By the denitrifying bacteria deposit 1g in (1), with 100ml aqua sterilisas into
Row is resuspended, and to obtain the bacteria suspension of denitrifying bacteria, 4 DEG C of refrigerators preserve.
(3) slant medium for taking Talaromyces flavus S1 is inoculated into the aqua sterilisa equipped with bead, puts
It is shaken in shaking table, carries out blood count, obtain the spore bacteria suspension of Talaromyces flavus S1；It is inoculated into PDB cultures
In base, shaking table culture 5~6 days obtains ripe mycelium pellet；
(4) slant medium for taking Trichodermavirid is inoculated into the aqua sterilisa equipped with bead, puts
It is shaken in shaking table, carries out blood count, obtain the spore bacteria suspension of Trichoderma viride；It is inoculated into PDB culture mediums, shaking table culture
5~6 days, obtain ripe mycelium pellet；
(5) by denitrifying bacteria bacteria suspension and ripe Talaromyces flavus S1 mycelium pellets and
Trichodermavirid mycelium pellets are inoculated into nitrate nitrogen a concentration of 138mg/L, COD 693mg/L, pH 7, temperature 10 simultaneously
DEG C 100ml actual waste waters (table 4) in, obtain using Talaromyces flavus S1 bacterium as the bacterium of the denitrifying bacteria of carrier
Pompon；
(6) experiment is divided into blank group (only actual waste water group), unlockedization denitrifying bacterium group (addition actual waste water and instead
Nitrobacteria group), Trichodermavirid mycelium pellet fixations group (addition denitrifying bacteria, Trichodermavirid mycelia
Ball and actual waste water group) and Talaromyces flavus S1 mycelium pellets (addition denitrifying bacteria, Talaromyces
Flavus S1 mycelium pellets and actual waste water group) group is fixed, every the removal rate that 3 hours survey a nitrate nitrogen and total nitrogen, respectively as schemed
Shown in 5 and Fig. 6.
(7) collection step 6) in Trichodermavirid mycelium pellet fixations group (addition denitrifying bacteria,
Trichodermavirid mycelium pellets and actual waste water group) and Talaromyces flavus S1 mycelium pellets (addition denitrification it is thin
Bacterium, Talaromyces flavus S1 mycelium pellets and actual waste water group) mycelium pellet in fixed group, with phosphate buffer into
Row is cleaned multiple times, and to remove the denitrifying bacteria in mycelium pellet, then adds the denitrifying bacteria of 1% volume ratio, renewed vaccination is arrived
In denitrification culture solution, with 1.8 days for a period, its final nitrate nitrogen and total nitrogen concentration is surveyed, to probe into the repetition of mycelium pellet
Utilizing status；
Fig. 5 is nitrate removal situation of the mycelium pellet in actual waste water.The result shows that：It is thin using only 1% denitrification
Bacterium handles actual waste water, and treatment effect can only be reduced to 51.00mg/L, reduce 46.56mg/L, removal rate 47.72%.
After immobilization, nitrate removal rate increases.Use Trichodermavirid mycelium pellet immobilization groups, anti-nitre
The nitrate removal rate for changing bacterium increases to 50.48%, and uses the Talaromyces flavus S1 mycelium pellets of the present invention solid
Surely after changing denitrifying bacteria, nitrate is reduced to 20.13mg/L, reduces 77.43mg/L, and nitrate removal rate is
79.37%, the nitrate removal rate than in existing document, handling actual waste water improves 28.89%.
Fig. 6 is that total nitrogen of the mycelium pellet in actual waste water removes situation.The result shows that：Immobilization denitrifying bacteria is conducive to
The removal of total nitrogen.Denitrifying bacteria is directly added to handle actual waste water, nitrogen removal rate 47.25%.And before using
The Trichodermavirid mycelium pellets of document report immobilize denitrifying bacteria, and total nitrogen is reduced to 53.51mg/L, goes
In addition to 49.25mg/L, removal rate 47.92%.And Talaromyces flavus S1 mycelium pellets using the present invention are fixed
Change, nitrogen removal rate is greatly improved, and the total nitrogen in sewage is after the processing of 42h, from 102.76mg/L
It is reduced to 23.33mg/L, removal rate has reached 77.29%, and the Trichodermavirid mycelium pellets than existing literature are fixed,
Effect improves 29.37%.
Fig. 7 is nitrate removal situation of two kinds of mycelium pellets during recycling.The result shows that：It is recycled by 5 times
After use, the nitrate removal capacity variation of immobilization denitrifying bacteria is smaller, still has higher immobilization ability, and
The nitrate nitrogen removal rate (76.1%~79.37%) of Talaromyces flavus S1 mycelium pellets is higher than Trichodermavirid
The immobilization effect (48.12%~50.48%) of mycelium pellet.
Fig. 8 is total nitrogen removal situation of two kinds of mycelium pellets during recycling.The result shows that：Mycelium pellet is repeating
After 5 times, nitrogen removal rate is without significant change.Nitrogen removal rate after the immobilization of Trichodermavirid mycelium pellets
It is 45.15~47.92%.And pass through Talaromyces flavus S1 mycelium pellets its nitrogen removal rate (76.23 utilized for 5 times
~77.29%) it is apparently higher than the mycelium pellet nitrogen removal rate of current document report.
The main component of 4 actual sewage of table
Claims (10)
1. a kind of immobilization denitrifying bacteria, which is characterized in that the immobilization denitrifying bacteria is with Talaromyces
Flavus S1 mycelium pellets are obtained as fixation support.
2. immobilization denitrifying bacteria according to claim 1, which is characterized in that the system of the immobilization denitrifying bacteria
It is standby to include：
(1) denitrifying bacteria bacteria suspension and Talaromyces flavus S1 mycelium pellets are obtained；
(2) denitrifying bacteria bacteria suspension and ripe Talaromyces flavus S1 mycelium pellets are inoculated into simultaneously pending
Waste water in, obtain using Talaromyces flavus S1 bacterium as the immobilization denitrifying bacteria of carrier.
3. a kind of method of intensified anti-nitrated bacterial nitrogen removal effect, which is characterized in that the method is with Talaromyces
Fixation support of the flavus S1 mycelium pellets as denitrifying bacteria recycles the denitrifying bacteria of immobilization to carry out at denitrogenation
Reason.
4. according to the method described in claim 3, it is characterized in that, the method includes：
(1) denitrifying bacteria bacteria suspension and Talaromyces flavus S1 mycelium pellets are obtained；
(2) denitrifying bacteria bacteria suspension and ripe Talaromyces flavus S1 mycelium pellets are inoculated into simultaneously pending
Waste water in, obtain using Talaromyces flavus S1 bacterium as the mycelium pellet of the denitrifying bacteria of carrier, one section of culture processing
Time.
5. according to the method described in claim 4, it is characterized in that, denitrifying bacteria bacteria suspension in the step (2) with
0.5-2% volume ratios are added, and are 3-8g/100mL waste water with 5g Talaromyces flavus S1 mycelium pellet additive amounts.
6. according to the method described in claim 4, it is characterized in that, the condition of culture in the step (2) is：Temperature 28 ± 2
DEG C, rotating speed 100-150r/min, incubation time is 1-3 days.
7. according to the method described in claim 4, it is characterized in that, the preparation method of the denitrifying bacteria bacteria suspension includes：
(A) denitrifying bacteria is cultivated：The muddy water mixed solution for taking suitable sewage treatment plant's anoxic pond, is inoculated into denitrification culture solution
In, anaerobism shaking table culture, continuous culture is multiple, and centrifugation obtains denitrifying bacteria thalline；
(B) denitrifying bacteria bacteria suspension is prepared：It takes appropriate thalline to be resuspended in aqua sterilisa, obtains the bacteria suspension of denitrifying bacteria.
8. according to the method described in claim 4, it is characterized in that, the system of the Talaromyces flavus S1 mycelium pellets
It is standby to include：
(a) bacteria suspension of Talaromyces flavus S1 is prepared：The slant medium of Talaromyces flavus S1 is taken,
It is inoculated in the aqua sterilisa equipped with bead, is put in shaking table and shakes, carry out blood count, obtain Talaromyces flavus
The spore bacteria suspension of S1；
(b) Talaromyces flavus S1 mycelium pellets are cultivated：Appropriate spore bacteria suspension is taken, is inoculated into PDB culture mediums, shakes
Bed culture 3~5 days, obtains ripe mycelium pellet；
9. the method according to the description of claim 7 is characterized in that the denitrification culture solution main component in the step (A)
For：KNO32g/L, C6H12O61.298g/L MgSO4·7H2O 0.2g/L, micro- 2ml/L, phosphate buffer (pH
=7.5) 50ml/L, surplus are water.
10. according to the method described in claim 8, it is characterized in that, the spore suspension inoculum concentration in the step (b) is
10% volume ratio.
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