CN109081447A - Method for removing nitrogen and phosphorus in culture wastewater by combining chlorella, acinetobacter and pseudomonas - Google Patents
Method for removing nitrogen and phosphorus in culture wastewater by combining chlorella, acinetobacter and pseudomonas Download PDFInfo
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- CN109081447A CN109081447A CN201810974564.3A CN201810974564A CN109081447A CN 109081447 A CN109081447 A CN 109081447A CN 201810974564 A CN201810974564 A CN 201810974564A CN 109081447 A CN109081447 A CN 109081447A
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- chlorella
- culture
- acinetobacter calcoaceticus
- phosphorus
- pseudomonad
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 title claims abstract description 88
- 241000195649 Chlorella <Chlorellales> Species 0.000 title claims abstract description 69
- 239000002351 wastewater Substances 0.000 title claims abstract description 57
- 229910052757 nitrogen Inorganic materials 0.000 title claims abstract description 44
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 title claims abstract description 43
- 229910052698 phosphorus Inorganic materials 0.000 title claims abstract description 43
- 239000011574 phosphorus Substances 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims abstract description 23
- 241000589516 Pseudomonas Species 0.000 title abstract description 6
- 241000589291 Acinetobacter Species 0.000 title abstract 5
- 239000002609 medium Substances 0.000 claims description 49
- 238000009395 breeding Methods 0.000 claims description 33
- 230000001488 breeding effect Effects 0.000 claims description 33
- 241000588624 Acinetobacter calcoaceticus Species 0.000 claims description 31
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 27
- 235000015097 nutrients Nutrition 0.000 claims description 26
- 239000012530 fluid Substances 0.000 claims description 24
- 238000005119 centrifugation Methods 0.000 claims description 20
- 241000195493 Cryptophyta Species 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 18
- 239000007787 solid Substances 0.000 claims description 16
- 238000005286 illumination Methods 0.000 claims description 15
- 238000000926 separation method Methods 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 12
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 9
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 9
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 9
- 239000001110 calcium chloride Substances 0.000 claims description 9
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 9
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 9
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 9
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 9
- 239000013049 sediment Substances 0.000 claims description 9
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Inorganic materials [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 9
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 8
- 230000003014 reinforcing effect Effects 0.000 claims description 8
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 7
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 6
- LLSDKQJKOVVTOJ-UHFFFAOYSA-L calcium chloride dihydrate Chemical compound O.O.[Cl-].[Cl-].[Ca+2] LLSDKQJKOVVTOJ-UHFFFAOYSA-L 0.000 claims description 6
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 6
- 239000001963 growth medium Substances 0.000 claims description 6
- 229910052760 oxygen Inorganic materials 0.000 claims description 6
- 239000001301 oxygen Substances 0.000 claims description 6
- CDBYLPFSWZWCQE-UHFFFAOYSA-L sodium carbonate Substances [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 6
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 4
- 238000011081 inoculation Methods 0.000 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 3
- 108010010803 Gelatin Proteins 0.000 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 239000008273 gelatin Substances 0.000 claims description 3
- 229920000159 gelatin Polymers 0.000 claims description 3
- 235000019322 gelatine Nutrition 0.000 claims description 3
- 235000011852 gelatine desserts Nutrition 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 230000001376 precipitating effect Effects 0.000 claims description 3
- 238000005728 strengthening Methods 0.000 claims description 3
- 230000002195 synergetic effect Effects 0.000 abstract 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 238000005516 engineering process Methods 0.000 description 8
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 6
- 244000144972 livestock Species 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 238000012545 processing Methods 0.000 description 5
- YUWBVKYVJWNVLE-UHFFFAOYSA-N [N].[P] Chemical compound [N].[P] YUWBVKYVJWNVLE-UHFFFAOYSA-N 0.000 description 4
- 238000009360 aquaculture Methods 0.000 description 4
- 230000015556 catabolic process Effects 0.000 description 4
- 238000006731 degradation reaction Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 244000144974 aquaculture Species 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 239000003344 environmental pollutant Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 231100000719 pollutant Toxicity 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000004065 wastewater treatment Methods 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 229910052564 epsomite Inorganic materials 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000005416 organic matter Substances 0.000 description 2
- 230000008092 positive effect Effects 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000004064 recycling Methods 0.000 description 2
- 238000001226 reprecipitation Methods 0.000 description 2
- 230000003442 weekly effect Effects 0.000 description 2
- 239000004912 1,5-cyclooctadiene Substances 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- CKUAXEQHGKSLHN-UHFFFAOYSA-N [C].[N] Chemical compound [C].[N] CKUAXEQHGKSLHN-UHFFFAOYSA-N 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000001546 nitrifying effect Effects 0.000 description 1
- 230000014075 nitrogen utilization Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
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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
- C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
- C02F3/322—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae
- C02F3/325—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae as symbiotic combination of algae and bacteria
-
- 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
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
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- Hydrology & Water Resources (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Ecology (AREA)
- Botany (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for removing nitrogen and phosphorus in culture wastewater by combining chlorella, acinetobacter and pseudomonas. The invention uses chlorella to separate and culture associated acinetobacter and pseudomonas, and combines the chlorella, acinetobacter and pseudomonas to remove nitrogen and phosphorus in the culture wastewater. The chlorella, the acinetobacter and the pseudomonas are jointly used, so that the method has a synergistic effect, can obviously remove nitrogen and phosphorus in the culture wastewater, and has a good application prospect and a good market value.
Description
Technical field
The invention belongs to cultivating wastewater purification technology and environmental technology fields, it is more particularly related to a kind of bead
Algae-acinetobacter calcoaceticus-pseudomonad combined removal breeding wastewater nitrogen, phosphorus method.
Background technique
The water environment pollution problem in China is got worse at present, ammonia nitrogen, total nitrogen, organic matter and novel toxic layer of contaminants
It is not poor out.The requirement of total nitrogen prediction emission is implemented to coastal ground level and the above city in national new " ten, water ", so that
Requirement to water process improves a higher pattern.Therefore, people carry out for the improvement of the pollutions such as ammonia nitrogen, phosphorus, total nitrogen
Extensive research.The nitrogen and phosphorus pollution of breeding wastewater is more serious.Livestock culture waste water is largely discharged into local water system and causes seriously
It influences, China about generates 100,000,000 tons of aquatic bird excrement every year, be discharged daily 5,000,000 tons of livestock culture waste water or more of Guangzhou, seriously
Ground destroys water ecology balance.Especially poultry livestock culture waste water has COD high, and biodegradability is strong, the high spy of nitrogen and phosphorus content
Point；Furthermore waste water yield and aquaculture model, kind and livestock culture amount have substantial connection, the waste water of Large-scale pig farm discharge
The characteristics of amount is big and concentrates, and that there are also impact loads is big for waste water, and quantity of wastewater effluent is big when flushing, other time water very little.
The problem of returning to the field mode that traditionally uses, ecology tupe have itself can not promote.Traditional microbiological denitrogenation, dephosphorization
Technological system investment is big, operating cost is high, therefore there is an urgent need to seek a kind of low cost, efficient livestock culture wastewater treatment is new
Method.It is required based on aquaculture status and upgrading, urgently finds a kind of low cost, efficient livestock culture wastewater treatment is newly square
Method.
In general, microorganism removal nitrogen is both economical means, the neck especially cultivated in aquaculture and aquatic bird
Domain.Denitrification process includes ammoxidation process, nitrifying process, denitrification process and the continuous process of Anammox.It is several micro-
How stable organic combination is also to remove the difficult point of nitrogen to biology.The removal of phosphorus is also by microorganism to the internal super tired of phosphorus
Product removes after removing organism.On the one hand the removal of traditional biological technology and materialization technology needs to consume the energy, on the other hand need
Further to keep the activity of microorganism that could complete the efficient removal of nitrogen P elements with advanced treating.Microalgae water treatment technology is
Recycling purification techniques emerging in recent years.Microalgae is a kind of Microalgae, can largely absorb water body nitrogen P elements, and part is planted
Class can also be metabolized organic carbon source under certain conditions, to reach the removal of carbon nitrogen and phosphorus pollutants.Therefore, microalgae is applied in water
The processing of Qin aquiculture waste water, on the one hand can achieve and efficiently goes dephosphorization and nitrogen nutrition object, to avoid the eutrophy of water body
Change；On the other hand, by recycling frond, water body is completed, the dietetic products such as more common bait are converted into, to generate potential
Economic benefit.However, algal grown speed is relatively slow, and to more complex organic matters certain in breeding wastewater, organic nitrogen
Utilization ability is not strong.Therefore, it for the breeding wastewater containing large amount of organic, needs additional measure and strengthens its processing effect
Energy.
Summary of the invention
It is an object of the invention to: overcome the problems, such as that breeding wastewater nitrogen phosphorus ligands are ineffective in the prior art, provide one
The method that kind can be obviously improved nitrogen phosphorus ligands efficiency in breeding wastewater.
In order to achieve the above-mentioned object of the invention, the present invention provides a kind of chlorella-acinetobacter calcoaceticus-pseudomonad combined removals
The method of breeding wastewater nitrogen, phosphorus comprising following steps:
(1) acquisition of preliminary culture: taking 10-20L breeding wastewater, goes oil removal and large size to swim through coarse filtration, centrifugation
Biology, then filter to obtain filtrate, then collect sediment from filter membrane, is transferred to M0 fluid nutrient medium, and whole day illumination is trained at 25 DEG C
It supports 2~3 days, shaking speed 100rpm, obtains the preliminary culture of chlorella-acinetobacter calcoaceticus；
(2) separation, culture and reinforcing of chlorella: the preliminary culture of chlorella-acinetobacter calcoaceticus is taken, after dilution
5000rpm centrifugation, gained precipitating are inoculated in M0 solid medium, and separation, switching obtain chlorella algae strain；By the chlorella
Algae strain is inoculated in M1 liquid enriched medium, and whole day illumination after shaking speed 150rpm is cultivated 2~3 days at 25 DEG C, is forwarded to new
M1 liquid enriched medium culture, so repeat culture 2 weeks, the chlorella after being strengthened；
(3) acquisition of association acinetobacter calcoaceticus: the preliminary culture of chlorella-acinetobacter calcoaceticus is taken, is added after 5000rpm centrifugation
Enter M2 fluid nutrient medium, after expanding culture 3~4 days under 25 DEG C of continuation, non-illumination condition, supernatant is removed in 5000rpm centrifugation, more
Isometric fresh M1 liquid enriched medium is changed, the preliminary culture of acinetobacter calcoaceticus is obtained；Phosphorus in results of regular determination culture solution
Content takes culture solution 5000rpm to be centrifuged when tp removal rate reaches 60% or more, and gained sediment is transferred to M0 solid medium
Middle culture, separation obtain the acinetobacter calcoaceticus of chlorella association；
(4) acquisition of association pseudomonad: the chlorella after taking step (2) described reinforcing is placed in the M0 liquid of 500ml~1L
Body culture medium then takes out 5~10ml liquid, is added after 5000rpm centrifugation by cultivating 3~5 days at whole day illumination, 25 DEG C
M3 fluid nutrient medium, after continuing 25 DEG C, expanding culture 3~6 days under non-illumination condition, supernatant is removed in 5000rpm centrifugation, replaces
Isometric fresh M3 fluid nutrient medium, by the way of low dissolved oxygen culture, the content of nitrogen, phosphorus in results of regular determination culture solution,
When nitrogen removal efficiency reaches 75% or more, tp removal rate reaches 60% or more, culture solution 5000rpm is taken to be centrifuged, gained sediment
It is transferred in M3 solid medium and cultivates, separates, obtain the pseudomonad of chlorella association；
(5) pseudomonad obtained by acinetobacter calcoaceticus obtained by the chlorella, step (3) after strengthening step (2) and step (4) is mixed
It closes, being put into breeding wastewater grows it naturally 3~7 days, and the removal to nitrogen, phosphorus in breeding wastewater can be completed；
Wherein, every liter of M0 fluid nutrient medium includes following component: NaNO31500mg, K2HPO40.04mg,
MgSO4·7H2O 75mg, CaCl2·2H2O 36mg, citric acid 6mg, EDTA 1mg, Na2CO320mg；
Every liter of M0 solid medium includes following component: NaNO31500mg, K2HPO40.04mg, MgSO47H2O
75mg, CaCl2 2H2O 36mg, citric acid 6mg, EDTA 1mg, Na2CO320mg, agar powder 2wt%；
Every liter of M1 liquid enriched medium includes following component: NaNO32000mg, K2HPO40.05mg, citric acid
6mg；
Every liter of M2 fluid nutrient medium includes following component: CH3COONa 4g, Na2HPO430mg, NH4Cl 60mg,
CaCl2·2H2O 17.2mg, pH 7.0；
Every liter of M3 fluid nutrient medium includes following component: CH3COONa 0.3g, Na2HPO4 30mg, NH4Cl
100mg, CaCl2.2H2O 0.5mg, MgSO47H2O 0.2g, pH are 7.0~7.5；
Every liter of M3 solid medium includes following component: CH3COONa 0.3g, Na2HPO4 30mg, NH4Cl
60mg, CaCl2.2H2O 0.5mg, peptone 16g, gelatin freeze 18.0g, and pH is 7.0~7.5.
One kind of method as chlorella-acinetobacter calcoaceticus of the present invention-pseudomonad combined removal breeding wastewater nitrogen, phosphorus is excellent
Selecting technology scheme, in step (1), the chlorella algae strain can also pass through purchase and obtain；When purchase obtains chlorella algae strain,
Before carrying out step (1), first by the chlorella algae strain investment breeding wastewater of the purchase, subsequent step is then carried out.
One kind of method as chlorella-acinetobacter calcoaceticus of the present invention-pseudomonad combined removal breeding wastewater nitrogen, phosphorus is excellent
Selecting technology scheme, in step (5), the chlorella after the reinforcing: the acinetobacter calcoaceticus: the volume ratio of the pseudomonad is
100~200:1:1.
A kind of optimization technique side of method as chlorella of the present invention-pseudomonad combined removal breeding wastewater nitrogen, phosphorus
The volume ratio of case, mixed chlorella-acinetobacter calcoaceticus-pseudomonad inoculation volume and breeding wastewater be 1:1:500~
1000。
Compared with the existing technology, the present invention has the advantage that
Inventor has found that chlorella and pseudomonad form symbiosis shape under specific condition of culture in an experiment
State can significantly, steadily remove total nitrogen and total phosphorus in waste water.So the present invention is using separated motionless in breeding wastewater
Bacillus, pseudomonad can be used for strengthening chlorella processing breeding wastewater after domestication culture.Its principle is acinetobacter calcoaceticus, vacation
Monad often with chlorella association, wherein acinetobacter calcoaceticus is the aerobic dephosphorization bacterial of tradition, during in conjunction with chlorella,
The two can have by oxygen, the mutual-aid functions such as exchange of carbon source, nitrogen source, the macromolecule for improving heterotrophism algae conversion breeding wastewater
The ability of machine object and hardly degraded organic substance, and accelerate the ability of algae phosphorus conversion nutrient；Pseudomonad is poly- with denitrification
The function of phosphorus can complete the degradation of organic matter under aerobic condition, complete the function of denitrogenation and poly- phosphorus, the two under anoxic conditions
It is combined with chlorella, the mutual assistance system of algae border microenvironment can be formed, promotion and functionization to chlorella wastewater treatment efficiency
It is of great advantage, it has a good application prospect and market value.
Detailed description of the invention
With reference to the accompanying drawings and detailed description, chlorella-acinetobacter calcoaceticus-pseudomonad combined removal of the present invention is supported
Waste water nitrogen, the method for phosphorus and beneficial effect is grown to be described in detail.
Fig. 1 is the removal rate comparison of the ammonia nitrogen, total nitrogen and total phosphorus of four experimental groups.
Fig. 2 is the COD removal rate comparison of four experimental groups.
Fig. 3 is influence of the pseudomonad to chlorella growth.
Fig. 4 is total conversion rate of nitrogen comparison of four experimental groups.
Fig. 5 is the total phosphorus conversion ratio comparison of four experimental groups.
Specific embodiment
In order to be more clear the purpose of the present invention, technical solution and advantageous effects, with reference to embodiments, to this
Invention is further elaborated.It should be understood that embodiment described in this specification is just for the sake of this hair of explanation
It is bright, be not intended to limit the present invention, parameter, ratio of embodiment etc. can adaptation to local conditions make a choice and substance had no to result
It influences.
Culture medium used in the following embodiment are as follows:
Every liter of M0 fluid nutrient medium includes following component: NaNO31500mg, K2HPO40.04mg, MgSO4·7H2O
75mg, CaCl2·2H2O 36mg, citric acid 6mg, EDTA 1mg, Na2CO320mg；
Every liter of M0 solid medium includes following component: NaNO31500mg, K2HPO40.04mg, MgSO47H2O 75mg,
CaCl2 2H2O 36mg, citric acid 6mg, EDTA 1mg, Na2CO320mg, agar powder 2wt%；
Every liter of M1 liquid enriched medium includes following component: NaNO32000mg, K2HPO40.05mg, citric acid 6mg；
Every liter of M2 fluid nutrient medium includes following component: CH3COONa 4g, Na2HPO430mg, NH4Cl 60mg,
CaCl2·2H2O 17.2mg, pH 7.0；
Every liter of M3 fluid nutrient medium includes following component: CH3COONa 0.3g, Na2HPO4 30mg, NH4Cl 100mg,
CaCl2.2H2O 0.5mg, MgSO47H2O 0.2g, pH are 7.0~7.5；
Every liter of M3 solid medium includes following component: CH3COONa 0.3g, Na2HPO4 30mg, NH4Cl 60mg,
CaCl2.2H2O 0.5mg, peptone 16g, gelatin freeze 18.0g, and pH is 7.0~7.5.
Embodiment 1
Aquatic bird cultivates factory and obtains waste water 20L, and waste water removes oil removal and macroplankton by precipitating, coarse filtration, centrifugation,
After being aerated 30min, reprecipitation 30min；Then filtrate is obtained by filtering, sediment is collected on filter membrane；Sediment passes through nothing
Bacterium water cleans 1-2 after, is transferred to M0 fluid nutrient medium, transfers newly after cultivating 3 days under whole day illumination, 25 degrees celsius
M0 fluid nutrient medium, shaking speed 100r/min, is repeated several times.Measured weekly during culture nitrate in culture medium, total nitrogen with
The removal efficiency of total phosphorus replaces M0 fluid nutrient medium after respectively reaching 50%, 40% and 30%, until nitrate, total nitrogen and total
The removal efficiency of phosphorus obtains the preliminary culture of chlorella-acinetobacter calcoaceticus after respectively reaching 80%, 50% and 60%.Chlorella-is no
The preliminary culture of lever bacterium is divided into two parts, and a part carries out the separation of chlorella, and another part carries out the separation of acinetobacter calcoaceticus.
Chlorella separation: the preliminary culture of chlorella-acinetobacter calcoaceticus finds chlorella in low power magnifying glass after dilution
Cell, collectable sediment is inoculated in M0 solid medium after a certain amount of rear 5000 turns/min centrifugation is sucked out, and carries out scribing line purifying
Separation obtains chlorella algae strain by 3 switchings.
Chlorella algae strain obtained is connect into M1 liquid enriched medium, whole day illumination, shaking table turns under 25 degrees celsius
Fast 150r/min, after culture 2-3 days, the new M1 fluid nutrient medium after being forwarded to sterilizing continues to cultivate, and transfers 2-3 times weekly, weight
It cultivates within multiple 2 weeks, the chlorella after being strengthened.
Acinetobacter calcoaceticus separation and acquisition: the preliminary culture of chlorella-acinetobacter calcoaceticus that another part is obtained takes out 5-
M2 fluid nutrient medium is added after 5000 turns/min centrifugation in 10ml liquid, aerobic culture 3 days under 25 degrees Celsius, non-illumination condition
(dissolved oxygen is not less than 2mg/L) afterwards removes supernatant after 5000 turns/min centrifugation, replaces isometric fresh M1 culture medium,
This is the preliminary culture of pseudomonad.Then by the content of phosphorus in each period measurement culture solution, when tp removal rate reaches
60% or more, then taking precipitate after 5000 turns/min of culture solution centrifugation is taken to be transferred to the scribing line culture of M0 solid medium, it is taken turns by 2-3
Plate separation obtains the acinetobacter calcoaceticus bacterial strain of microalgae association.
Pseudomonad separation and acquisition: pseudomonad obtains from chlorella incubation.First by the chlorella after reinforcing
It is placed in the M0 fluid nutrient medium of 500ml~1L, by cultivating 3~5 days under whole day illumination, 25 degrees celsius, then takes out 5
~10ml liquid is added M3 fluid nutrient medium after 5000rpm centrifugation, expands culture 3~4 under 25 degrees Celsius, non-illumination condition
After it, remove supernatant, the isometric fresh M3 fluid nutrient medium of replacement, using low dissolved oxygen culture after 5000rpm centrifugation
Mode (slowly shakes, dissolved oxygen is less than or equal to 1mg/L) culture.The content of nitrogen, phosphorus in each period measurement culture solution, when phosphorus is gone
Except rate reaches 60% or more, nitrogen removal efficiency reaches 75% or more, then taking precipitate after liquid 5000rpm centrifugation is taken to be transferred to M3 solid
Culture medium culture obtains the pseudomonas strains of chlorella association by 2-3 wheel plate separation.
It uses:
(three's volume ratio is in use, the chlorella after reinforcing obtained, acinetobacter calcoaceticus are mixed with pseudomonad
100:1:1), it is put into aquatic bird breeding wastewater and uses that (inoculation volume ratio waste water: chlorella-acinetobacter calcoaceticus-pseudomonad is
500:1:1).The pollutant concentration of breeding wastewater is respectively as follows: total nitrogen 50-60mg/L, ammonia nitrogen 31-33mg/L, total phosphorus 30-40mg/
Between L, COD800-900mg/L, pH 6-7.
Chlorella, acinetobacter calcoaceticus, the preliminary culture of chlorella-acinetobacter calcoaceticus and chlorella-are set no simultaneously in use
Lever bacterium-four groups of pseudomonad, Fig. 1,2 show comparing four groups of experimental results, by processing in 4 days, chlorella+acinetobacter calcoaceticus
+ pseudomonad group is excellent than other three groups in COD, nitrogen, phosphorus conversion rate.In terms of removal rate, the mixing of 100:1:1 ratio makes
Used time, chlorella+acinetobacter calcoaceticus+pseudomonad group than individual chlorella group, individual acinetobacter calcoaceticus group and chlorella+
The removal efficiency of the preliminary culture group of acinetobacter calcoaceticus improves the removal rate of 20-40%, ammonia nitrogen, total nitrogen, total phosphorus removal energy
Power respectively reaches 49.01,42.89 and 43.23mg/mgVSS-1d-1.
COD removal ability improves 10% or more, and degradation rate reaches 998.23mg mgVSS-1d-1, and display addition is not
Lever bacterium, pseudomonad facilitate the transformation efficiency (Fig. 2) that chlorella improves nitrogen phosphorus.
In terms of growth rate, chlorella+acinetobacter calcoaceticus+pseudomonad group chlorella amount is 1- higher than chlorella group~
2-% or so, display acinetobacter calcoaceticus, pseudomonad have certain positive effect (Fig. 3) to the growth of chlorella.
Then setting is repeated four times culture experiment, and discovery chlorella+acinetobacter calcoaceticus+pseudomonad group total nitrogen and total phosphorus are gone
Removing solid capacity is more more stable than other three groups, shows that acinetobacter calcoaceticus and pseudomonad can stablize the nutrient discovery ability of chlorella,
Certain effect (Fig. 4,5) is played in expanding production.
Embodiment 2
From pig raising, breeding wastewater factory takes waste water 10L, after conventional exclude, removes oil removal and macroplankton,
After being aerated 25min, reprecipitation 30min obtains culture waste water and is put into waste water, after buying chlorella algae from market by M1
Enriched medium adapts to culture 1~2 week, and 5 days ammonia nitrogens, removal rate of phosphate respectively reach 60% and 70% and could use, subsequent
Step is the same as embodiment 1.
It uses:
In use, by waste water chlorella obtained, acinetobacter calcoaceticus and pseudomonad mix (three's volume ratio be 200:
1:1), it is put into aquatic bird breeding wastewater and uses (inoculation volume ratio waste water: algae 1000:1:1).The pollutant of breeding wastewater
Concentration is respectively as follows: total nitrogen 45-50mg/L, ammonia nitrogen 20-30mg/L, total phosphorus 30-40mg/L, COD800-900mg/L, pH 6-7 it
Between.
Chlorella group, acinetobacter calcoaceticus group and chlorella-acinetobacter calcoaceticus-pseudomonad group are set simultaneously in use, reflected
The degradation efficiency of degradation of organic substances microalgae association flora promotion waste water chlorella.By processing in 6 days, addition chlorella+motionless
Bacillus+pseudomonad group total nitrogen, ammonia nitrogen, total tp removal rate efficiency highest.
Than the removal ability that the removal efficiency of individual chlorella group and individual acinetobacter calcoaceticus group improves 10%, COD
Removal ability improves 13-15%, and display addition acinetobacter calcoaceticus and pseudomonad facilitate the conversion effect that chlorella improves nitrogen phosphorus
Rate.In terms of growth rate, chlorella+acinetobacter calcoaceticus+pseudomonad group algae amount is 7-10% higher than chlorella group within first four days
Left and right shows that acinetobacter calcoaceticus and pseudomonad have certain positive effect to the growth of chlorella.
According to the disclosure and teachings of the above specification, those skilled in the art in the invention can also be to above-mentioned embodiment party
Formula carries out change and modification appropriate.Therefore, the invention is not limited to the specific embodiments disclosed and described above, to this
Some modifications and changes of invention should also be as falling into the scope of the claims of the present invention.In addition, although this specification
In use some specific terms, these terms are merely for convenience of description, does not limit the present invention in any way.
Claims (4)
1. the method for a kind of chlorella-acinetobacter calcoaceticus-pseudomonad combined removal breeding wastewater nitrogen, phosphorus, which is characterized in that including
Following steps:
(1) acquisition of preliminary culture: taking 10-20L breeding wastewater, removes oil removal and macroplankton through coarse filtration, centrifugation,
It filters to obtain filtrate again, then collects sediment from filter membrane, be transferred to M0 fluid nutrient medium, whole day illumination, culture 2 at 25 DEG C~
3 days, shaking speed 100rpm, obtain the preliminary culture of chlorella-acinetobacter calcoaceticus；
(2) the preliminary culture of chlorella-acinetobacter calcoaceticus, 5000rpm after dilution the separation, culture and reinforcing of chlorella: are taken
Centrifugation, gained precipitating are inoculated in M0 solid medium, and separation, switching obtain chlorella algae strain；The chlorella algae strain is inoculated with
In M1 liquid enriched medium, whole day illumination after shaking speed 150rpm is cultivated 2~3 days at 25 DEG C, is forwarded to new M1 liquid
Enriched medium culture so repeats culture 2 weeks, the chlorella after being strengthened；
(3) acquisition of association acinetobacter calcoaceticus: the preliminary culture of chlorella-acinetobacter calcoaceticus is taken, M2 is added after 5000rpm centrifugation
Fluid nutrient medium, after continuing 25 DEG C, expanding culture 3~4 days under non-illumination condition, supernatant, replacement etc. are removed in 5000rpm centrifugation
The fresh M1 liquid enriched medium of volume, obtains the preliminary culture of acinetobacter calcoaceticus；The content of phosphorus in results of regular determination culture solution,
When tp removal rate reaches 60% or more, culture solution 5000rpm is taken to be centrifuged, gained sediment is transferred in M0 solid medium and trains
It supports, separation, obtains the acinetobacter calcoaceticus of chlorella association；
(4) acquisition of association pseudomonad: the chlorella after taking step (2) described reinforcing is placed in the M0 liquid training of 500ml~1L
Base is supported, by being cultivated 3~5 days at whole day illumination, 25 DEG C, 5~10ml liquid is then taken out, M3 liquid is added after 5000rpm centrifugation
Body culture medium, after continuing 25 DEG C, expanding culture 3~6 days under non-illumination condition, supernatant, the bodies such as replacement are removed in 5000rpm centrifugation
Long-pending fresh M3 fluid nutrient medium, by the way of low dissolved oxygen culture, the content of nitrogen, phosphorus, works as nitrogen in results of regular determination culture solution
When removal rate reaches 75% or more, tp removal rate and reaches 60% or more, culture solution 5000rpm is taken to be centrifuged, gained sediment is transferred to
It cultivates, separate in M3 solid medium, obtain the pseudomonad of chlorella association；
(5) pseudomonad obtained by acinetobacter calcoaceticus obtained by the chlorella, step (3) after strengthening step (2) and step (4) mixes,
Being put into breeding wastewater grows it naturally 3~7 days, and the removal to nitrogen, phosphorus in breeding wastewater can be completed；
Wherein, every liter of M0 fluid nutrient medium includes following component: NaNO31500mg, K2HPO40.04mg, MgSO4·
7H2O 75mg, CaCl2·2H2O 36mg, citric acid 6mg, EDTA 1mg, Na2CO320mg；
Every liter of M0 solid medium includes following component: NaNO31500mg, K2HPO40.04mg, MgSO47H2O 75mg,
CaCl2 2H2O 36mg, citric acid 6mg, EDTA 1mg, Na2CO320mg, agar powder 2wt%；
Every liter of M1 liquid enriched medium includes following component: NaNO32000mg, K2HPO40.05mg, citric acid 6mg；
Every liter of M2 fluid nutrient medium includes following component: CH3COONa 4g, Na2HPO430mg, NH4Cl 60mg,
CaCl2·2H2O 17.2mg, pH 7.0；
Every liter of M3 fluid nutrient medium includes following component: CH3COONa 0.3g, Na2HPO4 30mg, NH4Cl 100mg,
CaCl2.2H2O 0.5mg, MgSO47H2O 0.2g, pH are 7.0~7.5；
Every liter of M3 solid medium includes following component: CH3COONa 0.3g, Na2HPO4 30mg, NH4Cl 60mg,
CaCl2.2H2O 0.5mg, peptone 16g, gelatin freeze 18.0g, and pH is 7.0~7.5.
2. the method for chlorella-acinetobacter calcoaceticus according to claim 1-pseudomonad combined removal breeding wastewater nitrogen, phosphorus,
It is characterized in that, the chlorella algae strain can also be obtained by purchase in step (1)；When purchase obtains chlorella algae strain,
Before carrying out step (1), first by the chlorella algae strain investment breeding wastewater of the purchase, subsequent step is then carried out.
3. the side of chlorella-acinetobacter calcoaceticus according to claim 1 or 2-pseudomonad combined removal breeding wastewater nitrogen, phosphorus
Method, which is characterized in that in step (5), chlorella after the reinforcing: the acinetobacter calcoaceticus: the volume ratio of the pseudomonad
For 100~200:1:1.
4. the method for chlorella-acinetobacter calcoaceticus according to claim 3-pseudomonad combined removal breeding wastewater nitrogen, phosphorus,
It is characterized in that, mixed chlorella-acinetobacter calcoaceticus-pseudomonad inoculation volume and the volume ratio of breeding wastewater are 1:1:
500~1000.
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