CN102826661A - Treatment method of contaminated water body by promoting degrading bacteria to be fixedly planted at biological film - Google Patents
Treatment method of contaminated water body by promoting degrading bacteria to be fixedly planted at biological film Download PDFInfo
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Abstract
The invention discloses a treatment method of a contaminated water body by promoting degrading bacteria to be fixedly planted at a biological film. The method comprises the following steps of: screening bridging bacteria and degrading bacteria with wide conglutination capability, and respectively collecting bacteria bodies; preparing bacterium suspension in which OD660=1 by a liquid which contains CaCl2.2H2O and MgSO4.7H2O and is used for promoting the conglutination; and adding the mixed bacterium suspension of the bridging bacteria and the degrading bacteria into a biofilm process waste water treatment pond containing toxic and difficult-degradable organic matters, or a water area polluted by the toxic and difficult-degradable organic matters. Aiming at the phenomenon that the waster water or the contaminated water body normally contains various difficult-degradable organic matters, the bacteria which are wide in conglutination capability are adopted to be mixed with various high-efficiency degrading bacteria, so that the conglutination function among various functional bacteria can be further enhanced due to the improvement of a bacteria cultivating condition or a conglutination condition, various degrading bacteria can be promoted to be immobilized at the biological film, the loss of the degrading bacteria can be reduced, the stability can be remained, and the treatment effect of the contaminated water body can be effectively improved.
Description
Technical field
The present invention relates to a kind of sewage disposal technology, particularly a kind ofly promote degradation bacteria to be colonizated in microbial film to be used for treatment process polluted-water.
Technical background
Adding biological reinforced (bioaugmentation) technology of special efficacy degradation bacteria to biochemical treatment of wastewater, is that to remove poisonous hardly degraded organic substance in the environment effective, easy and do not have a method of secondary pollution.The different complicated organic Microbial resources of degrading for many years constantly come to light; As Chinese invention patent CN 101343616 B one one-strain high-ring polycyclic aromatic hydrocarbon degradation bacterium and application thereof are disclosed, CN 101638630 B disclose styrene-degrading bacteria MJ001 and separation thereof; Though these valuable degradation bacteria resources have been hopes of knowing clearly of biological intensified process band, but still exist degradation bacteria loss that adds or the problem that is difficult for field planting.
Aggegation is the mutual absorption of carrying out through special polysaccharide of cell surface and protein lectin molecule between two bacteriums; The common aggegation between aggegation and different genera bacterium between bacterium of the same race plays an important role in biomembranous formation, and the bacterium that exists with the microbial film form can better stay in the water treatment system.
Chinese invention patent (CN 101255403 B) disclosed " pyridine degradation bacterium strain and application thereof " has utilized pyridine degradable bacteria to have autoflocculation (aggegation) effect simultaneously, has improved the shock-resistance of system.Diana etc. reported do not have degradation capability but can with a bacillus of NP degradation bacteria generation conglutination (
BacillusSp. VA160) degradation efficiency ([J] Research in Microbiology, 2004,155 (9): 761-769) of NP have been promoted.
Aggegation altogether generally only can take place with other a few bacterium in most of bacteriums.From oral biological film, find can with multiple bacterium take place extensively altogether the bridge formation bacterium-Fusobacterium nucleatum of agglutinability (
Fusobacteriumnucleatum) back (and [J] FEMS Microbiology Letters, 2000,182 (1): 57-62), Buswell in 1997 etc. are separated to 19 strain bacteriums from the water body microbial film of artificial culture, discover micrococcus luteus (
Micrococcus luteus) can obviously aggegation altogether take place with 11 strain bacteriums wherein, played the part of the role ([J] Journal of Applied Microbiology 1997,83,477 – 484) of bridge formation bacterium; People such as Rickard were separated to 19 strain bacteriums from the limnobios film in 2002, found
Blastomonas natatoria2.1 can with bacterial strain generation specific agglutination ([J] Applied and Environmental Microbiology, 2002,68 (7): 3644-3650) of other 18 kinds; People such as Sim es discovery in 2008 derives from the tap water environment
Acinetobacter calcoacticusNot only have from agglutinability power, can also be total to aggegations, do not have with 4 generations in other 5 strain isolateds
A. calcoacticusCommon aggegation ([J] Applied and Environmental Microbiology, 2008,74 (4): 1259 – 1263) does not then take place.These have the bridge formation bacterium that extensively is total to agglutinability to belong to a plurality of Pseudomonas, and dispersive is still compared in present its distribution in evolutionary system of discovering.Before the present invention makes; In document " the common aggregation studies of bacterium in the Waste Water Treatment microbial film " ([J] Anhui agricultural sciences; 2010; 38 (11): 5752-5754) in, the bridge formation bacterium that conglutination takes place for 2 kinds of abilities and many strains bacterium is disclosed, and proof filter out have extensively altogether 2 strain bacteriums of agglutinability have many strains bacterium is fixed in biomembranous function.
Chinese invention patent (CN 1076323 C) disclosed " method of wastewater treatment "; Be employed in the sludge activation operation; Add the stripping composition that contains at least a packing material in Humus, stripping property silica, the stripping property silica precursor; Contact the blended technical scheme, can improve the aggegation adsorptive power of active sludge and the formation ability of throw out.
The effect that prior art is reached after the screening to the bridge formation bacterium is from aggegation or can only aggegation altogether takes place with a kind of degradation bacteria, thereby improves the shock-resistance of system or a kind of degradation efficiency of pollutent.The bacterium of agglutinability makes up with multiple degradation bacteria and have extensively altogether, and further improves agglutination activity through the improvement of aggegation condition, and is used for biological reinforced technology and does not also appear in the newspapers.
Summary of the invention
The objective of the invention is to overcome the deficiency that prior art exists, provide a kind of and can promote that multiple degradation bacteria be colonizated in microbial film, reduce the loss of degradation bacteria, keep stablely, effectively improve method the polluted-water regulation effect.
The technical scheme that realizes the object of the invention is a kind ofly to promote degradation bacteria to be colonizated in microbial film to be used for the treatment process to polluted-water; From biomembrance process purification tank for liquid waste or river course, the pool, lake water body, gather diffraction patterns for biomembrane samples; Screening has extensively the bridge formation bacterium of agglutinability altogether, carries out following steps again:
(1) bridge formation bacterium and degradation bacteria were cultivated in liquid medium separately 20～30 hours respectively, collected thalline after the spinning;
(2) promote that with containing agglutinative liquid is processed OD altogether
660=1 bacteria suspension, said containing promotes that agglutinative liquid is altogether: the CaCl that in concentration is the phosphoric acid buffer of 0.1～0.2 mol/L, contains 1～3 mmol
22H
2The MgSO of O and 1～3 mmol
47H
2O, pH=7.0～7.5;
(3) 1～3:1 mixes the bridge formation bacterium with the bacteria suspension of degradation bacteria by volume; After leaving standstill 3 hours; Be added in the biomembrance process purification tank for liquid waste that contains poisonous hardly degraded organic substance, or receive in the waters of poisonous hardly degraded organic substance pollution, throwing the bacterium amount is 1～10% V/V.
The liquid medium of bridge formation bacterium is in every 1L water, comprises peptone 5～10 g, yeast powder 2～5 g, (NH
4)
2NO
31.0～1.5 g, NaCl 0.1～0.5 g, MgSO
47H
2O 0.1～0.2 g; CaCl
22H
2O 0.05～0.2 g, FeCl
36H
2O 0.01～0.05 g, K
2HPO
40.5～1.0 g regulate pH=7.2～7.4.
The liquid medium of bacterium for degrading is in every 1L water, comprises peptone 0～1.0 g, glucose 0～1.0 g, poisonous hardly degraded organic substance 0.1～1 g, (NH
4)
2NO
31.0～1.5 g, NaCl 0.1～0.5g, MgSO
47H
2O 0.1～0.2 g; CaCl
22H
2O 0.05～0.2 g, FeCl
36H
2O 0.01～0.05g, K
2HPO
40.5～1 g regulates pH 7.2 ~ 7.4.
Described poisonous hardly degraded organic substance comprises benzene, toluene, phenol, oil of mirbane, aniline, chlorobenzene and pyridine.
In the waters that polluted by poisonous hardly degraded organic substance, be placed with the filler that supplies the microbial film apposition growth, described filler is fixed in box frame or the mesh bag.
Compared with prior art; Advantage of the present invention is: to the phenomenon that usually contains multiple hardly degraded organic substance in waste water or the polluted-water, employing has the bacterium that extensively is total to agglutinability and mixes with multiple efficient degrading bacteria, through microbial culture condition and the improvement of aggegation condition altogether; Further strengthen the conglutination between various function yeast; Impel multiple degradation bacteria to be fixed in microbial film, reduce the loss of degradation bacteria, keep stable degradation effect.
Description of drawings
Fig. 1 is the common aggegation rate correlation curve figure of G5 and A3 in bacteria suspension that present embodiment provides and common phosphoric acid buffer;
Fig. 2 is that the agglomeration bacteria suspension that provides of the embodiment of the invention 1 and Comparative Examples are inoculated in behind the reactor drum in the water outlet 3, the correlation curve figure of 5-dinitrobenzoic acid content;
Fig. 3 is the stereoscan photograph of the agglomeration that provides of the embodiment of the invention 2;
Fig. 4 is that the agglomeration bacteria suspension that provides of the embodiment of the invention 2 and Comparative Examples are put in the water body correlation curve figure of phenol content in the pond water outlet of back.
Embodiment
Below in conjunction with accompanying drawing and embodiment technical scheme of the present invention is further elaborated.
(1) the altogether cultivation of aggegation bacterium (bridge formation bacterium): get prior separation screening to the bridge formation bacterium of agglutinability altogether arranged extensively
Bacillus cereusG5; Separating screening method is referring to document " the common aggregation studies of bacterium in the Waste Water Treatment microbial film " ([J] Anhui agricultural sciences, 2010,38 (11): 5752-5754); Be inoculated into the bridge formation inoculum after the activated cultivation; 30 ℃, cultivated centrifugal collection thalline 20 hours for 150 rev/mins.The prescription of bridge formation inoculum is for to comprise in every 1L tap water: peptone 5 g, yeast powder 5 g, (NH
4)
2NO
31.5 g, NaCl 0.1 g, MgSO
47H
2O 0.2 g; CaCl
22H
2O 0.05 g, FeCl
36H
2O 0.01 g, K
2HPO
40.5 g regulates pH=7.2.
(2) cultivation of degradation bacteria: degradation bacteria can select to use the multiple degradation bacteria of reporting or obtaining through screening, like Comamonas testosteroni A3, in the present embodiment, get prior separation screening to have 3,5-dinitrobenzoic acid bacterium
Comamonas testosteroneA3, concrete grammar is referring to document " 3, separation and the degradation characteristic of 5-dinitrobenzoic acid degradation bacteria A3 " ([J] China Environmental Science, 2007,27 (1): 106～110).Be inoculated into after the activated cultivation in the degradation bacteria nutrient solution, A3 falls the prescription of degradation bacteria nutrient solution in every 1L tap water, to comprise: peptone 0.5 g, 3,5-dinitrobenzoic acid 0.1 g, (NH
4)
2NO
31.5 g, NaCl 0.1 g, MgSO
47H
2O 0.2 g; CaCl
22H
2O 0.05 g, FeCl
36H
2O 0.01 g, K
2HPO
41 g regulates pH 7.2.The microbial culture liquid temp is 30 ℃, cultivates centrifugal collection thalline 20 hours for 150 rev/mins.
(3) preparation of agglomeration: above-mentioned 2 kinds of thalline of centrifugal collection are resuspended in respectively contain 2 mmol CaCl
22H
2O and 2 mmol MgSO
47H
2In the phosphoric acid buffer of 0.1 mol/L of O (pH 7.0), the adjusting cell concentration is OD
660=1, press the 1:1 volume above-mentioned 2 kinds of bacteria suspensions mixed, leave standstill 3 hours after, obtain G5A3 agglomeration bacteria suspension.
Referring to accompanying drawing 1, it is the G5 in G5A3 agglomeration bacteria suspension and common phosphoric acid buffer that provides of present embodiment and the common aggegation rate correlation curve figure of A3; Can find out that by Fig. 1 in the bacteria suspension of calcic that present embodiment provides, mg ion, the common aggegation effect of G5 and A3 is apparently higher than the common phosphoric acid buffer of calcic, mg ion not.
(4) in the reactor drum that 3 useful volumes are 5L, add 4 suspension ball fillers respectively.By inoculum size is 5% (V/V), in 3 reactor drums, adds the G5A3 agglomeration bacteria suspension that present embodiment provides respectively, and comparative example A's 3 bacteria suspensions and city domestic sewage treatment plant aeration tank contain the bacteria suspension of active sludge.3 reactor drum inoculation vexed 24h that expose to the sun in back, waste water 1 L of synthetic, 28 ± 2 ℃ of reactor drum operating temperatures are changed in beginning in second day every day.The prescription of synthetic waste water is: peptone 0.2 g, 3,5-dinitrobenzoic acid 0.2 g, (NH
4)
2NO
31.5 g, NaCl 0.1 g, MgSO
47H
2O 0.2 g; CaCl
22H
2O 0.05 g, FeCl
36H
2O 0.01 g, K
2HPO
41 g, tap water 1L regulates pH 7.2 ~ 7.4.
Referring to accompanying drawing 2, the bacterial suspension inoculation that it is the G5A3 agglomeration bacteria suspension that provides of present embodiment and A3 bacteria suspension, contain active sludge behind reactor drum, 3 in the water outlet, the correlation curve figure of 5-dinitrobenzoic acid content.Can find out by Fig. 2; The reactor drum that adds G5 and A3 agglomeration than the reactor drum that only adds degradation bacteria A3 to 3; The degradation speed of 5-dinitrobenzoic acid is faster, and only inoculates sanitary sewage disposal factory reactor with active sludge to 3, and the degraded of 5-dinitrobenzoic acid is the slowest.
(1) the altogether cultivation of aggegation bacterium: by embodiment 1 technical scheme separation screening to having the bridge formation bacterium that extensively is total to agglutinability
Bacillus megateriumT1 is inoculated into the bridge formation inoculum after the activated cultivation, 30 ℃, cultivated centrifugal collection thalline 20 hours for 150 rev/mins.The prescription of bridge formation inoculum comprises: peptone 5 g, yeast powder 2 g, (NH
4)
2NO
31.5 g, NaCl 0.1 g, MgSO
47H
2O 0.2 g; CaCl
22H
2O 0.05 g, FeCl
36H
2O 0.01 g, K
2HPO
40.5 g, tap water 1L regulates pH=7.4.
(2) cultivation of degradation bacteria: get the phenol degrading bacterium
PseudomonasSp. FY-6, concrete grammar is referring to document " preliminary evaluation of phenol degrading bacterial strain FY-6 and phenol degrading The Characteristic Study " ([J] science and technology circular, 2009; 25 (4): 441-444); Be inoculated into after the activated cultivation in the degradation bacteria nutrient solution, the prescription of degradation bacteria nutrient solution is an inoculum, 30 ℃; Cultivated centrifugal collection thalline 20 hours for 150 rev/mins.The prescription of degradation bacteria nutrient solution is: peptone 0.5 g, phenol 0.2 g, (NH
4)
2NO
31.5 g, NaCl 0.1 g, MgSO
47H
2O 0.2 g; CaCl
22H
2O 0.05 g, FeCl
36H
2O 0.01 g, K
2HPO
41 g, tap water 1L regulates pH=7.4.
(3) preparation of agglomeration: above-mentioned 2 kinds of thalline of centrifugal collection are resuspended in respectively contain 2 mmol CaCl
22H
2O and 2 mmol MgSO
47H
2In the phosphoric acid buffer of 0.1 mol/L of O (pH 7.0), the adjusting cell concentration is OD660=1, presses the 1:1 volume 2 kinds of bacteria suspensions were mixed 3 hours, and the stereoscan photograph of 2 kinds of bacterium agglomeratioies is seen Fig. 3, can be found out by Fig. 3, altogether the aggegation bacterium
B. megateriumT1 (escherichia coli) and degradation bacteria P (dialister bacterium) aggegation have closely formed the agglomeration than bulk together.
(4) at 0.5 m
3The pond in introduce river and in river, add phenol, make phenol concentration reach 250mg/L, pond depth of water 0.8m; With the suspension ball filler of the diameter 8cm rectangle mesh bag of packing into, tighten the mesh bag mouth, make suspension ball filler in mesh bag, pile up 5～10 layers; Be several nylon ropes of hanging masonry on mesh bag simultaneously, make mesh bag be positioned at the position about the 20cm of underwater, with present embodiment step (3) make extensively altogether the aggegation body fluid of aggegation bacterium and degradation bacteria be added in the pond of placing biofilm packing in the ratio of 2%d; Common agglomeration is attached in settled process on the suspension ball filler of stacked in multi-layers; With the whole emptyings of the water in the pond (treating to form minute colony attached to the agglomeration of filling surface), change again and add the river (as 0 day) of phenol to 250mg/L after 2 days, change and add the river 50L of phenol to 250mg/L every day after 1 day; Whenever opened 1 water-circulating pump in the operational process at a distance from 12 hours; Each 1 hour, water temperature was 24～28 ℃, in the time of the 5th day in the water outlet of pond phenol content be 50mg/L; With the same treatment mode but the pond that does not add the pond of common agglomeration and only add degradation bacteria P is a Comparative Examples, phenol content is seen Fig. 4 in the water outlet.Can be found out that by Fig. 4 do not add in the pond of degradation bacteria, the phenol degrading situation is the poorest, behind the emptying Chi Shui, phenol concentration is higher for 241.3mg/L when running to the 3rd day; Only add in the pond of degradation bacteria, the degraded situation of phenol is slightly more better than the pond that does not add degradation bacteria, in the time of the 3rd day phenol concentration higher be 203.2mg/L, explain that most of degradation bacteria have been lost in emptying Chi Shui; In the pond of the agglomeration that adds bridge formation bacterium T1 and degradation bacteria, the degradation effect of phenol is best, and phenol content is 94.3mg/L in the water outlet in the 3rd day; Explanation is in the emptying pond during water; Because the aggegation bacterium impels degradation bacteria to be colonizated in filling surface altogether, degradation bacteria is not along with emptying pond current lose, in afterwards 24 days; Phenol content in the water outlet is kept at lower concentration all the time, explains that degradation bacteria is present in the pond always.
(1) by conventional microbiology experimental technique, have extensively the bacterium of agglutinability and degradation bacteria streak inoculation altogether to the LB test tube slant with what cultivation in advance obtained, cultivated 24 hours for 30 ℃.
(2) thalline on the picking test tube is inoculated in the triangular flask of 250 mL that 100mL LB liquid nutrient medium is housed, and 30 ℃, 150 rev/mins of shaking culture 24 hours.
(3) bacterial classification in the triangular flask is inoculated into respectively in the triangular flask of 1000 mL that 300 mL LB liquid nutrient mediums are housed by 10% inoculum size, 30 ℃, 150 rev/mins of shaking culture 24 hours.
(4) bacteria suspension of 2 kinds of thalline of collection, 6000 rev/mins centrifugal, respectively with containing 2 mmol CaCl
22H
2O and 2 mmol MgSO
47H
20.1 phosphoric acid buffer of Omol/L (pH 7.0) is processed OD
660=1 bacteria suspension, two kinds of bacteria suspensions are pressed the 1:1 mixed, leave standstill 3 hours, make the common aggegation of carrying out that extensively is total to aggegation bacterium and degradation bacteria, form agglomeration altogether.
(5) will be total to agglomeration by the technical scheme of embodiment 1 or 2 and be added in the biomembrance process purification tank for liquid waste that contains poisonous hardly degraded organic substance, or receive in the waters that poisonous hardly degraded organic substance pollutes waste water or polluted-water to be carried out biological treating.
Claims (5)
1. one kind promotes degradation bacteria to be colonizated in microbial film to be used for the treatment process to polluted-water; From biomembrance process purification tank for liquid waste or river course, the pool, lake water body, gather diffraction patterns for biomembrane samples; Screening has extensively the bridge formation bacterium of agglutinability altogether, it is characterized in that carrying out following steps again:
(1) bridge formation bacterium and degradation bacteria were cultivated in liquid medium separately 20～30 hours respectively, collected thalline after the spinning;
(2) promote that with containing agglutinative liquid is processed OD altogether
660=1 bacteria suspension, said containing promotes that agglutinative liquid is altogether: the CaCl that in concentration is the phosphoric acid buffer of 0.1～0.2 mol/L, contains 1～3 mmol
22H
2The MgSO of O and 1～3 mmol
47H
2O, pH=7.0～7.5;
(3) 1～3:1 mixes the bridge formation bacterium with the bacteria suspension of degradation bacteria by volume; After leaving standstill 3 hours; Be added in the biomembrance process purification tank for liquid waste that contains poisonous hardly degraded organic substance, or receive in the waters of poisonous hardly degraded organic substance pollution, throwing the bacterium amount is 1～10% V/V.
2. according to claim 1ly a kind ofly promote degradation bacteria to be colonizated in microbial film to be used for the treatment process to polluted-water, to it is characterized in that: the liquid medium of bridge formation bacterium comprises peptone 5～10 g, yeast powder 2～5 g, (NH in every 1L water
4)
2NO
31.0～1.5 g, NaCl 0.1～0.5 g, MgSO
47H
2O 0.1～0.2 g; CaCl
22H
2O 0.05～0.2 g, FeCl
36H
2O 0.01～0.05 g, K
2HPO
40.5～1.0 g regulate pH=7.2～7.4.
3. according to claim 1ly a kind ofly promote degradation bacteria to be colonizated in microbial film to be used for treatment process to polluted-water; It is characterized in that: the liquid medium of bacterium for degrading comprises peptone 0～1.0 g, glucose 0～1.0 g in every 1L water; Poisonous hardly degraded organic substance 0.1～1 g, (NH
4)
2NO
31.0～1.5 g, NaCl 0.1～0.5g, MgSO
47H
2O 0.1～0.2 g; CaCl
22H
2O 0.05～0.2 g, FeCl
36H
2O 0.01～0.05g, K
2HPO
40.5～1 g regulates pH 7.2 ~ 7.4.
4. according to claim 1ly a kind ofly promote degradation bacteria to be colonizated in microbial film to be used for treatment process that it is characterized in that: described poisonous hardly degraded organic substance comprises benzene, toluene, phenol, oil of mirbane, aniline, chlorobenzene and pyridine to polluted-water.
5. according to claim 1ly a kind ofly promote degradation bacteria to be colonizated in microbial film to be used for treatment process to polluted-water; It is characterized in that: in the waters that polluted by poisonous hardly degraded organic substance; Be placed with the filler that supplies the microbial film apposition growth, described filler is fixed in box frame or the mesh bag.
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| CN106186348A (en) * | 2016-07-13 | 2016-12-07 | 盐城工学院 | Improve microorganism to degrade in the liquid phase the bio-enhancer of m-dichlorobenzene efficiency and adding method thereof |
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| CN106186348A (en) * | 2016-07-13 | 2016-12-07 | 盐城工学院 | Improve microorganism to degrade in the liquid phase the bio-enhancer of m-dichlorobenzene efficiency and adding method thereof |
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Application publication date: 20121219 |