CN101386822B - Special effect phosphate accumulating organisms and waste water processing method using thereof - Google Patents
Special effect phosphate accumulating organisms and waste water processing method using thereof Download PDFInfo
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- CN101386822B CN101386822B CN2007100769606A CN200710076960A CN101386822B CN 101386822 B CN101386822 B CN 101386822B CN 2007100769606 A CN2007100769606 A CN 2007100769606A CN 200710076960 A CN200710076960 A CN 200710076960A CN 101386822 B CN101386822 B CN 101386822B
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Abstract
The invention discloses a specific phosphorus-accumulating bacterium. The strain is a phosphorus-accumulating bacterial strain HJP07, wherein the preserving number of the bacterial strain is CGMCC No. 2164 and the preserving date of the bacterial strain is September 13th, 2007. The phosphorus-accumulating bacterial strain is separated from sludge, cultured in an enriched medium and separated in a separation culture medium, wherein the pH value of the phosphorus-accumulating bacterium at a temperature of between 25 and 35 DEG C is between 6.5 and 8.0. By adopting the phosphorus-accumulating bacterium to treat waste water, the cost is low, the efficiency is stable and no secondary pollution produces, thereby effectively treating phosphor-containing waste water and protecting the environment.
Description
[technical field]
The present invention relates to biological technical field, especially about a special effect phosphate accumulating organisms and handle the method for waste water.
[background technology]
Phosphorus is the main factor of body eutrophication, therefore removes the phosphorus in the water body particularly important to preventing body eutrophication.Make a general survey of domestic sewage work, dephosphorization technique is the difficult problem of puzzlement sewage work operation always.Traditional materialization dephosphorization technique needs a large amount of medicaments, has the running cost height, the shortcoming that sludge yield is big.Although existing municipal sewage plant technology mostly comprises the denitrogenation dephosphorizing link, through the design improvement of sewage biological treatment system or the change of operation scheme polyP bacteria colony can be got the mastery in the substrate competition of treatment system, to guarantee phosphor-removing effect.Yet these technologies exist the startup of dephosphorization system and recover problems such as slow, dephosphorization efficiency by using instability in the actual motion, and organic content is lower in waste water, or phosphorus content is when surpassing 10mg/L, and water outlet is difficult to satisfy the emission standard of phosphorus.
[summary of the invention]
Technical problem to be solved by this invention is, overcomes the deficiency of prior art, provides that a kind of cost is low, dephosphorization efficiency by using stable, the special effect phosphate accumulating bacterium of non-secondary pollution and handle the method for waste water.
The technical solution adopted for the present invention to solve the technical problems is: a special effect phosphate accumulating organisms, this bacterial strain are HJP07, Pseudomonas fluorescens (Pseuodomonas fluorescens), and preserving number is CGMCC № 2164, preservation date is on 09 13rd, 2007.
Described polyP bacteria bacterial strain separates from mud and obtains, and this polyP bacteria is at 25 ℃～35 ℃, and pH value 6.5～8.0 is cultivated in the enrichment medium, separates in the isolation medium.
Described polyP bacteria bacterial strain separates from bed-silt and obtains, and it is 30 ℃ that this polyP bacteria bacterial strain suits in temperature, and the pH value is 7.2～7.4, cultivates in the enrichment medium, separates in the isolation medium.
Described enrichment culture based component is: anhydrous sodium acetate 5.0g, MAGNESIUM SULPHATE HEPTAHYDRATE 99.5 0.5g, calcium chloride 0.2g, ammonium sulfate 2.0g, potassium primary phosphate 2～20mg, micro-1ml, zero(ppm) water are to 1000ml, and the pH value is 7.2～7.4.
Described separation and Culture based component is: Carnis Bovis seu Bubali cream 3g, peptone 10g, sodium-chlor 5g, potassium primary phosphate 15mg, agar 15～20g, zero(ppm) water are to 1000ml, and the pH value is 7.2～7.4.
Described polyP bacteria bacterial strain separates from bed-silt and obtains, and it is 30 ℃ that this polyP bacteria bacterial strain suits in temperature, and shaking culture is 3 days on the shaking table of 150r/min; In described enrichment medium, cultivate, after substratum becomes muddiness, increase the content of potassium primary phosphate in the substratum; When temperature is 30 ℃, in isolation medium, separate.
Described polyP bacteria bacterial strain belongs to Rhodopseudomonas (Pseudomonas sp.), and principal character is: colonial morphology is circular, white, smooth, moistening, low protruding, opaque, diameter 2mm; The bacterial strain individuality is shaft-like, and wide is 0.7～0.8 μ m, long 2.0～2.8 μ m.
The physiological and biochemical property of described polyP bacteria bacterial strain shows as: Gram-negative, flagellum number＞1; Oxidase positive, denitrification is positive, and methyl red, V-P feminine gender are utilized glucose, fructose, produce oxydase, and edwardsiella hoshinae does not produce H
2S does not need growth factor.
A kind of method of utilizing a described special effect phosphate accumulating organisms to handle phosphorus-containing wastewater is characterized in that: comprise the steps:
A) cultivation of polyP bacteria bacterial strain;
B) separation of polyP bacteria bacterial strain;
C) polyP bacteria bacterial strain and sewage reaction dephosphorization;
D) reclaim the polyP bacteria bacterial strain, recycle
The invention has the beneficial effects as follows, handle waste water through polyP bacteria, cost is low, stabilised efficiency, non-secondary pollution, thereby has handled phosphorus-containing wastewater effectively, the protection environment.
[description of drawings]
Fig. 1 is the influence curve figure of differing temps to the HJP07 bacterial strain;
Fig. 2 is the influence curve figures of different pH to the HJP07 bacterial strain;
Fig. 3 is that the HJP07 bacterial strain is to river sewage dephosphorization usefulness pilot scale with put into practice engineering experiment diagram as a result.
[embodiment]
Use at present the biological process dephosphorization, have advantages such as less investment, operation are simple, non-secondary pollution.It is the excess phosphorus absorbing phenomenon that utilizes active sludge; Be that the phosphorus amount that mikrobe absorbs surpasses the needed phosphorus amount of mikrobe normal growth; The mikrobe of the excessive Absorption of Phosphorus of this type ability is polyP bacteria (Phosphorus Accumulating Organisms; PAOs), polyP bacteria plays a decisive role to biological phosphate-eliminating.
A kind of special effect phosphate accumulating bacterium of the present invention belongs to Rhodopseudomonas (Pseudomonas sp.), is Pseudomonas fluorescens, called after HJP07.Be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) on 09 13rd, 2007, preserving number is CGMCC № 2164.
HJP07 bacterial strain of the present invention is taken from Shenzhen's Buji bed-silt, obtains a strain and gathers the strong bacterial strain of phosphorus ability through 6 enrichment culture, 5 separation and purification.Its colony characteristics is: colonial morphology is circular, white, smooth, moistening, low protruding, opaque, diameter 2mm; The bacterial strain individuality is shaft-like, and wide is 0.7～0.8 μ m, long 2.0～2.8 μ m.Its physiological and biochemical property shows as: Gram-negative, flagellum number＞1; Oxidase positive, denitrification is positive, and methyl red, V-P feminine gender are utilized glucose, fructose, produce oxydase, and edwardsiella hoshinae does not produce H
2S does not need growth factor.Can find out that from Fig. 1, Fig. 2 HJP07 polyP bacteria optimum growing condition is: the pH value is 6.5～8.0,25～35 ℃ of temperature.
The technology contents that the present invention relates to also comprises: the enrichment culture based formulas of screening HJP07 bacterial strain is that the enrichment culture based component is: anhydrous sodium acetate 5.0g, MgSO
4.7H
2O 0.5g, CaCl
20.2g, (NH
4)
2SO
42.0g, KH
2PO
42～20mg, micro-1ml, zero(ppm) water are to 1000ml, and the pH value is 7.2～7.4, KH during enrichment culture
2PO
4Content increase progressively successively; The culture medium prescription that is used for pure strains separation is: Carnis Bovis seu Bubali cream 3g, peptone 10g, NaCl 5g, KH
2PO
415mg, zero(ppm) water are to 1000ml, and the pH value is 7.2～7.4.When being applied to engineering throwing bacterium, inoculation HJP07 bacterial strain is to enrichment medium enlarged culturing (KH
2PO
4(cell concentration is to add in right amount after 108～109CFU/L) 10mg/L) to be cultured to logarithmic phase.
Embodiment 1: efficient polyP bacteria is selected good strains in the field for seed and is educated and gather the phosphorus rate and measure
(1) collected specimens
Collection in worksite Shenzhen Buji bed-silt sample is smashed with glass strain vibration behind the adding distil water in the aseptic triangular flask of 500ml.
(2) bacterial strain enrichment, separation
Extracting sample solution 10ml is to the triangular flask that 150ml enrichment medium (sterilizing) is housed, and shaking culture is 3 days on 30 ℃, the shaking table of 150r/min.Enrichment culture based component: anhydrous sodium acetate 5.0g, MgSO
4.7H
2O 0.5g, CaCl
20.2g, (NH
4)
2SO
42.0g, KH
2PO
42～20mg, micro-1ml, zero(ppm) water is to 1000ml; The pH value is 7.2～7.4, after substratum becomes muddiness, shifts 8ml again to new enrichment medium; Shaking culture is 3 days under the similarity condition, repeats so 4 times again, and each transfer amount is respectively: 5ml, 3ml, 1ml, 0.5ml.Increase KH in the substratum successively
2PO
4Content be: 2mg, 5mg, 8mg, 10mg, 15mg, 20mg.
After the bacteria suspension of getting 1m1 is diluted to 10000 times, get the dull and stereotyped coating of dilution bacterium liquid 0.2ml, cultivated 2～3 days in 30 ℃, promptly have bacterium colony to occur.Select the different bacterium colony of those forms to separate, repeat to separate more than five times obtaining the pure bacterial strain of many strains according to the purebred isolating ordinary method plate streaking of mikrobe.The separation and Culture based component is: Carnis Bovis seu Bubali cream 3g, peptone 10g, NaCl 5g, KH
2PO
415mg, agar 15～20g, zero(ppm) water are to 1000ml, and the pH value is 7.2～7.4.
(3) gather the mensuration of phosphorus rate
With the multiple bacterial strain behind the purifying; Access contains in the enrichment culture liquid of different concns phosphorus (5,10,15,20mg/L); Shaking culture 48h, 60h through the centrifugal 20min of 4000r/min, get supernatant and measure total phosphorus with molybdenum-antimony anti-spectrophotometric method then under 30 ℃, 150r/min; And with the total phosphorus value contrast that gathers the phosphorus nutrient solution that does not connect bacterium, calculate and gather the phosphorus rate.Gather phosphorus rate=(A
TP-B
TP)/A
TP* 100%; A wherein
TP: do not connect the total phosphorus of the enrichment culture liquid of bacterium, B
TP: the total phosphorus after the enrichment culture liquid that connects bacterium is handled.Obtain a strain and gather the strongest bacterial strain of phosphorus ability, be bacterial strain HJP07 of the present invention, gather phosphorus rate such as table 1 under the different gradient phosphorus concentrations.
Table 1 HJP07 bacterial strain gather the phosphorus rate
| Phosphorus concentration | 5mg/L | 10mg/L | ?15mg/L | 20mg/L |
| 48h 60h | 93.1％ 97.2％ | 81.5％ 88.7％ | ?68.3％ ?73.8％ | 48.7％ 52.2％ |
Gather the test of phosphorus rate and show that when KH2PO4 concentration was 10mg/L, 48h, 60h gathered the phosphorus rate and be respectively 81.5%, 88.7%.
Embodiment 2:HJP07 bacterial strain is to river sewage dephosphorization usefulness pilot experiment
Experimental technique adopts microbial film-active sludge composite treatment process, and sewage is taken from Buji river, Shenzhen sewage, and total phosphorous 1～6mg/L, seed sludge take from municipal sewage plant's sbr reactor device, the aerobic operation of system.With HJP07 inoculation (phosphorus concentration 10mg/L) in the enrichment medium of 300mL triangular flask, on 30 ℃, 150r/min shaking table, cultivate, using colony counting method to survey its cell concentration to the later stage of logarithmic phase is 10
8During CFU/L, add certain volume bacterium liquid to reactor drum, making its cell concentration is 5 * 10
6CFU/L, earlier to the aeration 48h of system, reactor drum moves according to aerobic mode then, measures its water outlet total phosphorous.Test finds that the HJP07 bacterial strain has good reinforced phosphor-removing effect; The domestication time is also shorter; Dephosphorizing rate has reached more than 82.7% in the time of can knowing 3d by Fig. 3, and system's phosphor-removing effect is very stable always behind the 7d, and the water inlet phosphorus concentration is below 5mg/L; The water outlet phosphorus concentration is less than 0.2mg/L, and the clearance of total phosphorus is more than 93%.And common Sewage treatment systems is not adding under any microbial inoculum, behind 20d to clearance＞44% (Liu Yanan, 2005) of phosphorus.
Embodiment 3:HJP07 bacterial strain in Shenzhen the waste water control of Buji river put into practice the effect of engineering
Buji river matter evolution factory adopting process: strengthen and flood microbial film-active sludge (SBF-AS) compound bio treatment process (EHYBFAS technology, the patent No.: 200620017991.5).After the method laboratory culture of HJP07 bacterial strain according to embodiment 2, get 5% inoculum size to 5000L sterilization culture tank fermentation culture, be furnished with the whisking appliance of adjustable speed and the microporous aeration device of adjustable aeration rate in the culture tank, being cultured to cell concentration is 10
8～10
9CFU/L.System adds a certain amount of bacterium liquid and adds to the startup with accelerating system of the aerobic section of SBF-AS reaction tank when starting, system's run duration polyP bacteria concentration in reaction tank is lower than 10
4During CFU/L, replenish and add an amount of HJP07 bacterium liquid.The result shows that shorten dramatically the start time of dephosphorization system; System stable operation behind the 14d; The total phosphorus index request reaches " urban wastewater treatment firm pollutant emission standard " (GB18918-2002) one-level A standard, and the clearance of phosphorus is stabilized in (like table 2, Fig. 3) more than 86.2%.
Table 2 HJP07 bacterial strain is applied to the removal effect of Shenzhen Buji river matter purification plant to phosphorus
| Date | Water temperature (℃) | The pH value | Water inlet TP concentration (mg/L) | Water outlet TP concentration (mg/L) | The clearance of TP (%) |
| 9/8 9/10 9/12 9/15 9/17 9/21 9/27 10/2 10/8 10/13 10/20 | 31.4 30.5 26.8 29.8 30.7 28.7 24.4 29.5 26.7 28.4 27.6 | 7.0 7.2 7.2 7.3 7.1 7.4 7.3 7.5 7.4 7.4 7.3 | 1.17627 4.11802 2.47802 2.008452 3.220982 4.565788 3.529626 5.888548 4.631926 2.40528 4.521696 | 0.71502 1.80152 0.98152 0.222726 0.840014 0.619554 0.487278 0.6416 0.5416 0.20068 0.26206 | 39.2 56.3 60.4 88.9 73.9 86.4 86.2 89.1 88.3 91.7 94.3 |
Above content is to combine concrete preferred implementation to the further explain that the present invention did, and can not assert that practical implementation of the present invention is confined to these explanations.For the those of ordinary skill of technical field under the present invention, under the prerequisite that does not break away from the present invention's design, can also make some simple deduction or replace, all should be regarded as belonging to protection scope of the present invention.
Claims (2)
1. a special effect phosphate accumulating organisms, it is characterized in that: this bacterial strain is Pseudomonas fluorescens (Pseudomonas fluorescens.) HJP07, and its preserving number is CGMCC № 2164.
2. a method of utilizing the described special effect phosphate accumulating organisms of claim 1 to handle phosphorus-containing wastewater is characterized in that: comprise the steps:
A) cultivation of polyP bacteria bacterial strain;
B) separation of polyP bacteria bacterial strain;
C) polyP bacteria bacterial strain and sewage reaction dephosphorization;
D) reclaim the polyP bacteria bacterial strain, recycle.
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| CN101386822B true CN101386822B (en) | 2012-07-04 |
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| CN101935632B (en) * | 2010-06-29 | 2012-02-01 | 东华大学 | Strain for degrading wool scale layer |
| CN103540546A (en) * | 2013-10-12 | 2014-01-29 | 西南大学 | High-efficiency phosphorus-accumulating bacterium Pseudomonas sp. GPA1 |
| CN103482773B (en) * | 2013-10-15 | 2015-01-21 | 江苏商达水务有限公司 | Dephosphorization reagent for rural domestic sewage and application thereof |
| CN103952365B (en) * | 2014-05-19 | 2016-02-10 | 安徽大学 | Phosphorus-accumulating bacteria with higher phosphorus removal effect at low temperature and screening method and application thereof |
| CN105254120A (en) * | 2015-10-16 | 2016-01-20 | 上海纳米技术及应用国家工程研究中心有限公司 | Advanced treatment method of phosphate sewage |
| CN113652364B (en) * | 2021-06-11 | 2024-03-01 | 安徽农业大学 | Phosphorus indicating bacteria and application thereof in detecting phosphorus content in water body |
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| CN1807585A (en) * | 2005-12-23 | 2006-07-26 | 南京农业大学 | Highly effective phosphorus removal bacteria and its produced bacteria formulation |
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