CN102676433B - Pseudomonas capable of removing nitrogen and phosphorus synchronously at low temperature and application thereof - Google Patents
Pseudomonas capable of removing nitrogen and phosphorus synchronously at low temperature and application thereof Download PDFInfo
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- CN102676433B CN102676433B CN201210158468.4A CN201210158468A CN102676433B CN 102676433 B CN102676433 B CN 102676433B CN 201210158468 A CN201210158468 A CN 201210158468A CN 102676433 B CN102676433 B CN 102676433B
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Abstract
The invention discloses pseudomonas capable of removing nitrogen and phosphorus synchronously at low temperature and application thereof. The Pseudomonas sp. Strain HA11 provided by the invention is preserved in China General Microbiological Culture Collection Center (CGMCC) on September 15, 2010, and the preservation number is CGMCC No. 4169. The strain is suitable to be grown at low temperature, especially at the temperature of 15 DEG C, and simultaneously have the capabilities of heterotrophic nitrification, aerobic denitrification and aerobic phosphorus uptake. Under aerobic condition at low temperature, the strain can be used for synchronous nitrification and denitrification so as to effectively remove total nitrogen in waste water and remove phosphate simultaneously, so that simultaneous nitrogen and phosphorus removal at a next aerobic stage at low temperature is effectively realized, and the pseudomonas strain has wide application prospect.
Description
Technical field
The present invention relates to biological carbon and phosphorous removal field, particularly a kind of pseudomonad strain for low-temperature biological denitrogenation dephosphorizing and application thereof.Under cold condition, this bacterial strain has the function of heterotrophic nitrification-aerobic denitrification, and can be in the process of synchronous nitration denitrification denitrogenation, remove simultaneously phosphoric acid salt in sewage-, thereby reach the object of Simultaneous Denitrification dephosphorization.
Background technology
The sanitary sewage quantity discharged that mankind's daily life produces is large, contains the nutritive substances such as organism and nitrogen, phosphorus many, has become the important source of pollution of water body.China is at present because the body eutrophication problem that nitrogen and phosphorus pollution causes is particularly outstanding.The monitoring result of environment department shows, in the large water system of China seven, section over half water quality is polluted at present, the water body in the whole nation 1/3 is unsuitable for Fish Survival, 1/4 water body is unsuitable for irrigating, 90% urban waters is seriously polluted, 50% water source, cities and towns does not meet drinking water standard, and 40% water source can not be drunk, and principal pollutant are ammonia nitrogen, COD
mn, BOD, volatile phenol and petroleum substance.For effectively control and pollution control of water, realize qualified discharge, country has proposed the emission standard of the pollutents such as strict organism, nitrogen, phosphorus in < < urban wastewater treatment firm pollutant emission standard > > (GB18918-2002).
Municipal sewage plant mostly adopts biological treatment stable, with low cost at present, wherein has the A of denitrogenation dephosphorizing function
2o, inversion A
2the technique such as O, SBR can be removed the nutritive elements such as organism in water and nitrogen, phosphorus conventionally more effectively, realizes qualified discharge.But in cold winter, temperature at discharging condensate can reach 10 ℃ even lower, severe inhibition microbic activity in biological treatment system, pollutant removal speed is obviously reduced.Wherein, the restraining effect that the nitrobacteria of autotrophy is subject to low temperature is more remarkable, under cold condition growth extremely slow, very easily from treatment system, run off, make that low temperature is nitrated cannot carry out; The also disadvantageous effect that causes to anoxic denitrification of the obvious rising of oxygen in water concentration under cold condition; Under cold condition, mud increasess slowly, sludge age obviously extends, and this also makes to rely on excess sludge discharge and realizes the biological phosphate-eliminating efficiency that phosphorus removes and obviously reduce.Therefore, under cold condition, carry out biological carbon and phosphorous removal and become town domestic sewage treatment plant and realize a great problem of qualified discharge.
In engineering, usually adopt and reduce sludge loading, increase sludge reflux amount, increase dwell time of sewage and to part structures, take the measures such as insulation or intensification to guarantee effluent quality.But these measures not only can increase construction investment and working cost, and be difficult to guarantee wastewater treatment efficiency, also often can cause the problems such as sludge bulking.
Separate, screen highly active psychrophile and contribute to fundamentally to solve low temperature dephosphorization denitrogenation problem for sanitary sewage disposal in winter.By add Autotrophic nitrification mud in biological treatment tank, can improve the removal efficiency of ammonia nitrogen, but under at low temperature, compared with high-solubility oxygen condition, denitrification is difficult to carry out, thereby the nitre nitrogen, the nitrite nitrogen that cause nitration reaction to produce cannot effectively be removed, and water outlet total nitrogen is still difficult to up to standard.The accumulation of oxidation state nitrogen discharges and has certain restraining effect the anaerobism of phosphorus, thereby phosphor-removing effect cannot be guaranteed conventionally.In addition, because Autotrophic nitrification bacterium grows extremely slowly under cold condition, easily, along with excess sludge discharge wash-out from system for handling is gone out, often need to add continuously, increased working cost.
For the problems referred to above, that the present invention filters out is cold-resistant, and have the bacterial strain of denitrogenation and phosphorus removal functional for the processing of low-temperature sewage simultaneously.Under low temperature, aerobic condition, this bacterial strain can carry out synchronous nitration and denitrification and effectively remove in sewage total nitrogen and remove phosphoric acid salt simultaneously.In addition, this bacterium is well-grown under cold condition, can in biological treatment system, hold and stay for a long time, need not add continuously, significant for a difficult problem that solves low-temperature denitrification phosphorous removal.
Summary of the invention
The object of the present invention is to provide a kind of pseudomonad strain for low-temperature biological denitrogenation dephosphorizing and application thereof.
Pseudomonas provided by the invention (Pseudomonas sp.) bacterial strain HA11 be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (be called for short CGMCC) on September 15th, 2010, preserving number is CGMCCNo.4169, depositary institution address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode is 100101.
Bacterial strain HA11 separates, screens the strain gram negative bacterium obtaining from active sludge.
Morphological specificity: bacterial strain HA11, at 15-20 ℃, cultivates after 24-36h bacterium colony smooth surface, non-pigment on nutrient agar; By being negative under the microscope after gramstaining; It is shaft-like that thalline is, single appearance.
The 16S rRNA gene sequence characteristic of this bacterial strain: its 16S rRNA has the nucleotide sequence as shown in sequence in sequence table 1, sequence length is 1417bp.
According to its morphological specificity and 16S rRNA gene order thereof, identify that this bacterial strain is pseudomonas (Pseudomonas sp.).
This bacterial strain HA11 can, at denitrogenation dephosphorizing under cold condition, in actual applications, can be placed in waste water by bacterial strain HA11, carries out biological carbon and phosphorous removal.
The temperature 10-20 ℃ of described waste water.
The dissolved oxygen of described waste water can be 2-6mg/L.
The carbon source of described waste water is for being sodium-acetate or sodium succinate.
The nitrogen of described waste water is ammonia nitrogen, nitrate nitrogen and/or nitrite nitrogen.
The phosphorus of described waste water is phosphoric acid salt.
The pH of described waste water can be 6-9.
Another object of the present invention is to provide a kind of microbiobacterial agent for low-temperature biological denitrogenation dephosphorizing, and its activeconstituents is described bacterial strain HA11.
Pseudomonas of the present invention and application thereof have following beneficial effect compared with prior art:
1. bacterial strain HA11 of the present invention is strong to the tolerance of low temperature, can Fast Growth under cold condition and remove ammonia nitrogen, effectively overcome traditional Autotrophic nitrification bacterium difficult difficult problem of staying that is difficult to effectively hold of grow in system for handling under cold condition;
2. bacterial strain HA11 can carry out aerobic denitrification when carrying out nitration reaction, has realized the thorough removal of nitrogen under a step aerobic condition, compares traditional anaerobic-aerobic denitrification process, has simplified technical process, has saved initial cost and working cost;
3. bacterial strain HA11 can effectively inhale phosphorus in denitrogenation under aerobic condition, realizes the removal of phosphorus, the different and contradiction that produces of required sludge age when also efficiently solving denitrogenation dephosphorizing and being carried out respectively by two kinds of dissimilar floras of Autotrophic nitrification bacterium and heterotrophism polyP bacteria;
4. bacterial strain HA11 of the present invention is applicable to the denitrogenation dephosphorizing processing of various waste water, has a extensive future, and has good economic benefit, environmental benefit and social benefit.
Accompanying drawing explanation
Fig. 1 is the temperature profile of bacterial strain HA11;
Fig. 2 is bacterial strain HA11 removal to phosphoric acid salt and nitrogen at 10 ℃;
Fig. 3 is bacterial strain HA11 removal to nitrogen under differing temps;
Fig. 4 is bacterial strain HA11 removal to phosphorus under differing temps.
Embodiment
The invention will be further described in conjunction with specific embodiments, but the present invention is not limited to following examples.
In following embodiment, if no special instructions, be ordinary method.
In following embodiment, described percentage composition if no special instructions, is quality percentage composition.
The temperature profile of embodiment 1. bacterial strain HA11
Under 10 ℃ of different temperature, 15 ℃, 20 ℃, 25 ℃, 30 ℃ conditions, investigate the growth performance of bacterial strain, concrete implementation step is as follows:
Bacterial strain HA11 is inoculated in in the BM substratum of 150ml, (every liter containing sodium-acetate: 2.73g, NH
4cl:0.306g, KH
2pO
4: 0.1g, MgSO
47H
2o:0.1g, FeSO
47H
2o:0.001g, pH:7.0-7.3,121 ℃ of 20min steam sterilizings), respectively at 10 ℃ of shaking tables, 15 ℃, 20 ℃, 25 ℃, 30 ℃, 160r/min carries out preculture.Until strain growth, during to the logarithmic phase later stage, the bacterium liquid of getting 10ml accesses in fresh 150ml BM substratum, and respectively at 10 ℃, 15 ℃, 20 ℃, 25 ℃, 30 ℃, 160r/min carries out shaking culture.Extract reaction solution at set intervals, measure OD
600, the growing state of sign bacterial strain.
As shown in Figure 1, bacterial strain is best 15 ℃ of growths, 10 ℃, 20 ℃, 25 ℃ growth velocitys, all has by a small margin and declines for result.In addition, bacterial strain is difficult to grow under 30 ℃ of conditions.As can be seen here, this bacterial strain is the psychrotroph that low temperature is had to well tolerable ability.Its temperature profile is obviously different from general optimum temperuture higher than 30 ℃, at 15 ℃ of following growth velocity degradations, and the difficult mesophilic bacteria of growing in the time of 10 ℃.The growth velocity of this bacterial strain under cold condition far exceedes traditional Autotrophic nitrification bacterial strain, has good application potential.
Embodiment 2. bacterial strain HA11 removal to phosphoric acid salt and nitrogen at 10 ℃
Take sodium-acetate as organic carbon source, ammonia nitrogen is nitrogenous source, and phosphoric acid salt is phosphorus source, and described bacterial strain HA11 measures the removal ability of nitrogen, phosphorus.Concrete implementation step is as follows:
Bacterial strain HA11 is inoculated in the BM substratum of 150ml, in 10 ℃ of shaking tables, 160r/min carries out preculture.Until strain growth, during to the logarithmic phase later stage, the bacterium liquid of getting 10ml accesses in fresh 150ml BM substratum, and 10 ℃, 160r/min carries out shaking culture.Extract reaction solution at set intervals, after the centrifugal 10min of 8000rpm, get supernatant liquor and measure various nitrogenous compounds and phosphate concn.
As shown in Figure 2, in whole experimentation, nitrogen, phosphorus concentration all constantly decline result.Within 24h, the clearance of ammonia nitrogen exceedes 80%, and the average degradation rate of ammonia nitrogen reaches 2.35mg-N/L/h.In addition, in whole degradation process, nitre nitrogen and nitrite nitrogen are almost without any accumulation, and in waste water, most ammonia nitrogen is converted into gaseous product by heterotrophic nitrification-aerobic denitrification effect, has realized effective removal of total nitrogen.In addition, in denitrogenation, phosphorus is also effectively removed, and concentration has reduced 11.06mg/L, and bacterial strain embodies the ability except good aerobic suction phosphorus.Therefore, this bacterial strain has effectively been realized the Fast synchronization removal of nitrogen and phosphorus under low-temperature aerobic condition.
Under 10 ℃ of different temperature, 15 ℃, 20 ℃, 25 ℃ conditions, investigate the nitrogen removal performance of bacterial strain, concrete implementation step is as follows:
Bacterial strain HA11 is inoculated in the BM substratum of 150ml, respectively at 10 ℃ of shaking tables, 15 ℃, 20 ℃, 25 ℃, 160r/min carries out preculture.Until strain growth, during to the logarithmic phase later stage, the bacterium liquid of getting 10ml accesses in fresh 150mlBM substratum, and with 10 ℃, 15 ℃, 20 ℃, 25 ℃, 160r/min carries out shaking culture respectively.Extract reaction solution at set intervals, after the centrifugal 10min of 8000rpm, get supernatant liquor and measure the concentration of various nitrogenous compounds.
As shown in Figure 3, this bacterial strain, in the temperature range of 10 ℃-25 ℃, all can effectively be removed total nitrogen to result.They are in the time of 15 ℃, and the average removal rate of total nitrogen is the highest, are 3.85mg-N/L/h.Under 10 ℃, 20 ℃ and 25 ℃ of conditions, TN removel rate also can reach respectively 2.35mg-N/L/h, 2.22mg-N/L/h and 2.16mg-N/L/h.
Under 10 ℃ of different temperature, 15 ℃, 20 ℃, 25 ℃ conditions, investigate the phosphorus removal property of bacterial strain, concrete implementation step is as follows:
Bacterial strain HA11 is inoculated in the BM substratum of 150ml, respectively at 10 ℃ of shaking tables, 15 ℃, 20 ℃, 25 ℃, 160r/min carries out preculture.Until strain growth, during to the logarithmic phase later stage, the bacterium liquid of getting 10ml accesses in fresh 150mlBM substratum, and with 10 ℃, 15 ℃, 20 ℃, 25 ℃, 160r/min carries out shaking culture respectively.Extract reaction solution at set intervals, after the centrifugal 10min of 8000rpm, get supernatant liquor and measure phosphatic concentration.
Result as shown in Figure 4, this bacterial strain in the temperature range of 10 ℃-25 ℃, all effectively dephosphorization.In the time of 10 ℃, phosphorus concentration can reduce about 11mg/L within 24h.When temperature is 25 ℃, bacterial strain dephosphorization efficiency by using slightly declines.This may be the born of the same parents' Systox effect that not only relies on bacterial strain due to the removal of phosphorus, also relevant with the outer EPS Adsorption of born of the same parents of bacterial strain secretion.And under higher temperature condition, the EPS of bacterial strain secretion reduces, also make the dephosphorization efficiency by using of bacterial strain have decline by a small margin.
Claims (1)
1. a pseudomonas (Pseudomonas sp.) HA11 carries out the application of low-temperature denitrification phosphorous removal in the aqueous solution, it is characterized in that described pseudomonas HA11, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCC No.4169, described low temperature is 10-20 ℃, the dissolved oxygen of the described aqueous solution is 2-6mg/L, nitrogen in the described aqueous solution is one or several in ammonia nitrogen, nitrate nitrogen, nitrite nitrogen, and the phosphorus in the described aqueous solution is phosphoric acid salt.
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| CN103525734B (en) * | 2013-10-17 | 2015-05-20 | 北京沃土天地生物科技有限公司 | Denitrification compound bacteria agent as well as preparation method and application thereof |
| CN103966129A (en) * | 2014-04-30 | 2014-08-06 | 深圳市格瑞斯特环保技术有限公司 | Screening cultivating method for denitrifying phosphorus removing bacterial strain and application thereof |
| CN104388342B (en) * | 2014-10-30 | 2017-06-23 | 临沂大学 | A kind of short distance nitration bacterium pseudomonad and application |
| CN105907678A (en) * | 2016-05-10 | 2016-08-31 | 东北师范大学 | Low-temperature-resisting pseudomonas fragi and application thereof |
| CN106148227B (en) * | 2016-07-05 | 2020-01-07 | 国家海洋局第一海洋研究所 | Low-temperature nitrifying pseudomonas and application thereof |
| CN110106097A (en) * | 2019-04-25 | 2019-08-09 | 黄山市益天士生物科技有限公司 | Accelerate the strain enrichment procedure of reparation eutrophication water |
| CN110407338B (en) * | 2019-08-16 | 2020-06-09 | 中国科学院上海高等研究院 | Low-temperature denitrification and dephosphorization bacteria and application thereof |
| CN116622556B (en) * | 2023-04-25 | 2024-01-05 | 海南立昇净水科技实业有限公司 | Low-temperature-resistant high-alkalinity high-efficiency denitrifying bacterium, screening method and application thereof |
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Application publication date: 20120919 Assignee: Beijing Gaia Environmental Technology Co., Ltd. Assignor: Beijing Gaiya Techn Center Co., Ltd. Contract record no.: 2016110000013 Denomination of invention: Pseudomonas capable of removing nitrogen and phosphorus synchronously at low temperature and application thereof Granted publication date: 20140423 License type: Exclusive License Record date: 20160919 |
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