CN101168736B - Introduced bacterium microorganism immobilization method used for repairing soil and special-purpose device for the same - Google Patents
Introduced bacterium microorganism immobilization method used for repairing soil and special-purpose device for the same Download PDFInfo
- Publication number
- CN101168736B CN101168736B CN2006101340699A CN200610134069A CN101168736B CN 101168736 B CN101168736 B CN 101168736B CN 2006101340699 A CN2006101340699 A CN 2006101340699A CN 200610134069 A CN200610134069 A CN 200610134069A CN 101168736 B CN101168736 B CN 101168736B
- Authority
- CN
- China
- Prior art keywords
- carrier
- bacterium
- immobilization
- particle
- shutoff
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Abstract
The invention relates to allotopia repair of polycyclic aromatic hydrocarbon contaminated soil, in particular to an introduced bacteria micro organism immobilization method used for soil repair, namely, the embedment to introduced bacteria is realized through a microbe immobilization granulator, particularly bacteria liquid in the logarithmic growth phase is immobilized and embedded, namely, the bacteria liquid with 20 to 30 percent volume taking carrier solution as base is added in the carrier solution, after being mixed uniformly, cross linking is performed by entering in cross linking solution through a fine pore assistant granulator, a granule is produced, and then the granule is added in solidifying liquid to perform strengthening, washing is performed with sterilized water, the cultivation and the multiplication of plain bumper is performed, so as to obtain the immobilized granule. The adoption of the method of the invention can be used for the immobilization of the single plantof the culture and the common immobilization of a plurality of plants, and the quantity and the activation of thallus inside the carrier are guaranteed. The carrier adopted by the invention has nontoxicity to organism, the price is cheap, the source is wide, the micro organism embedded by the carrier has strong resistance power to external toxicity, the bacteria quantity is increased, and the degradation capacity to pollutant is also greatly improved.
Description
Technical field
The present invention relates to the dystopy reparation of polycyclic aromatic hydrocarbon pollution, a kind of specifically introduction bacterium microorganism immobilization method and isolated plant that is used for soil remediation, promptly realize the particle that obtains being introduced the dystopy reparation that bacterium is used for polycyclic aromatic hydrocarbon pollution to introducing the embedding of bacterium by microbial immobilized pellet fabrication device.
Background technology
Along with industrialized continuous expansion, the pollution grows worse of toxic and harmful substance has become the main Environmental Problems that countries in the world face in soil, water body and the air, seeks a kind of efficient, economical, lasting and the little improvement method of environmental perturbation is become the focus of present concern.Because soil pollution has characteristics such as tangible disguise, latent, non-reversibility, its environmental risk of bringing is on the rise.During particularly organic polluted soil is repaired polycyclic aromatic hydrocarbons (PAHs) be considered to a kind of carcinogenic, cause mutagen, can directly jeopardize human health by food chain.Biological restoration is promptly utilized hazardous and noxious substances in the microbiological deterioration environment, and decontamination person reduces the restorative procedure of its concentration.
In general, immobilization method of microorganism has absorption method, entrapping method, and crosslinking and chemical covalent method etc. are several.Immobilized microorganism have free bacterium incomparable advantage, particularly will screen have some specific function efficient degrading bacteria after immobilization, not only improved the density of thalline, strengthened the tolerance of bacterial classification simultaneously to poisonous substance, and can be according to concrete processing requirements, adopt different process for fixation, and treatment effect is strengthened greatly.The mechanism of immobilized microorganism technique is that microorganism is fixed on the carrier, makes its high-density intensive and keep its biological function, can also breed under suitable condition, to satisfy the requirement of treatment process.The improvement that immobilization technology is used for hardly degraded organic substance has its particular performances, and promptly immobilized cell all has obvious enhancing to the ability to bear and the degradation capability of toxic substance.
The prospect of immobilized microorganism technique in environmental engineering is immeasurable.
Summary of the invention
The object of the present invention is to provide a kind of a kind of specifically introduction bacterium microorganism immobilization method and isolated plant that is used for soil remediation, promptly realize the particle that obtains being introduced the dystopy reparation that bacterium is used for polycyclic aromatic hydrocarbon pollution to introducing the embedding of bacterium by microbial immobilized pellet fabrication device.
For achieving the above object, the technical solution adopted in the present invention is: the bacterium liquid of logarithmic phase is capable immobilization embedded, and soon be that the bacterium liquid of 20-30% volume of benchmark joins in the carrier soln, fully behind the mixing, at 1.05-1.2*10 with the carrier soln
5Pa enters in the crosslinked fluid by the pore of 2-3.5mm down, and stagnates 22-25 hour in crosslinked fluid, makes the small-particle that diameter is 3-4mm, reinforced 22-25 hour adding in the solid solution behind the particle filtration, sterile water wash is carried out shaking table and is cultivated propagation, the particle of being fixed; Described carrier soln is 10-15% polyvinyl alcohol, 0.3-0.6% sodium alginate and 3-6% gac by weight percentage, and surplus is a sterilized water.
Described sterile water wash was soaked 22-25 hour.The microorganism that contains logarithmic phase in the described bacterium liquid is 10
5-10
7Individual/ml.Described crosslinked fluid is 5-8% boric acid by weight percentage, 1-3% CaCl
2With 1-3% FeCl
3, surplus is a water.
Described solid solution is the metabisulfite solution of mass concentration 8%-15%.Described bacterial classification: convex pseudomonas (Pseudomonas convexa chester), crooked pseudomonas (Pseudomonasgenicula tachester), one or more in the pseudomonas-1.130 (Pseudomonas sp.).
The isolated plant of process for fixation is: an end is provided with the wound packages structure of shutoff, is provided with piston at the other end of wound packages structure, offers pore in shutoff, and the outer side of piston is provided with handle.
Be connected with Glass tubing in the pore in the described shutoff, the length of Glass tubing is increased to the center to outer rim gradually by shutoff.
When immobilization microorganism particles was used to degrade the mud contamination thing, the water and soil mass ratio in the mud was 3-4:1, and the inoculum size of immobilization particle is for by weight percentage: 4-8%, and degradation process is carried out under 28-30 ℃.
Principle: realize adopting the adding pressure type pellet fabrication device in the immobilization embedded process first to introducing the embedding of bacterium by microbial immobilized pellet fabrication device, the use of adding pressure type pellet fabrication device can improve the multiple technologies index of immobilization particle greatly.The particle that obtains is introduced the dystopy reparation that bacterium is used for polycyclic aromatic hydrocarbon pollution.
The present invention has following advantage:
1. the present invention is by the use of adding pressure type pellet fabrication device, and the immobilization particle of preparation is even, has significantly reduced the phenomenon of particle hangover.Guarantee the uniform distribution of granule interior hole simultaneously, improved the particulate mass-transfer performance.Particle is difficult for broken.
2. the individual plant that adopts method of the present invention to can be used for bacterial classification is fixed and being total to of multi-strain bacteria fixed.Can guarantee the quantity and the activity of the thalline of carrier inside.
3. cost is low, and is effective.Adopt carrier of the present invention be PVA to biological nontoxic, simultaneously cheap, wide material sources, through the microorganism of this carrier embedding, toxic to external world tolerance strengthens, the bacterium number increases, the degradation capability to pollutent has also improved much simultaneously.
Description of drawings
Fig. 1 introduces the growth curve of bacterium for the present invention.
Fig. 2 is the use figure of the special-purpose adding pressure type pellet fabrication device of the present invention.
Fig. 3 is one of fractionation synoptic diagram of the special-purpose adding pressure type pellet fabrication device of the present invention.
Fig. 4 is two of the fractionation synoptic diagram of the special-purpose adding pressure type pellet fabrication device of the present invention.
Fig. 5 is the upward view of the special-purpose adding pressure type pellet fabrication device of the present invention.
Label is: 1. the wound packages structure of shutoff, 2. piston, 3. handle, 4. pore, 5. Glass tubing.
Embodiment
Culturing micro-organisms: (B61) with convex pseudomonas (Pseudomonas convexa chester), crooked pseudomonas (Pseudomonas genicula tachester) (B67) and pseudomonas-1.130 (Pseudomonas sp.) (MB) three strains mix in twos and three strains mixing, by the inclined-plane insert contain pollutent pyrene and benzo (a) pyrene concentration be respectively 10 μ g/ml in seed culture medium, be placed on shaking table and cultivate 24h, switching goes into to contain the pollutent pyrene and benzo (a) pyrene concentration is 30 μ g/ml successively afterwards, 50 μ g/ml, in the liquid nutrient medium of 70 μ g/ml and 100 μ g/ml, be placed on and cultivate 20-25h on the shaking table.To tame good bacterial strain at last and transfer and preserve on the slant medium that contains a certain amount of pyrene and benzo (a) pyrene pollutent, standby.
Microorganism immobilization embedded: will preserve standby convex pseudomonas (Pseudomonas convexachester) (B61), crooked pseudomonas (Pseudomonas genicula tachester) (B67) and pseudomonas-1.130 (Pseudomonas sp.) (MB) three strains mix in twos and three strain mixing microorganismss are inserted in the seed culture medium by the inclined-plane, put into constant temperature culture on 120-130r/min, 30 ℃ the shaking table, the microbial growth curve is measured.Referring to Fig. 1, with microorganism culturing 20-30h, to guarantee the high reactivity of microorganism, then that the bacterium liquid of logarithmic phase is capable immobilization embedded, the microorganism that contains logarithmic phase in the bacterium liquid is 10 according to measurement result
5Individual/ml, soon be that the bacterium liquid of 20% volumetric concentration of benchmark joins in the carrier soln with the carrier soln, fully behind the mixing, put into the adding pressure type pellet fabrication device of the pore of 2mm and stagnate 24 hours with 1.05*10 with crosslinked fluid
5Pa makes the small-particle that diameter is 2-3.5mm, reinforces 24 hours adding in the solid solution behind the particle filtration, soaks 24 hours with sterilized water again, then cultivates propagation by shaking table, and the particle of being fixed is standby.
Described carrier soln is 10% polyvinyl alcohol, 0.3% sodium alginate and 3% gac by weight percentage, and surplus is a sterilized water; Seed culture medium and proliferated culture medium are by weight percentage: grape 2%, peptone 2%, yeast extract paste 1%, pH nature; Described crosslinked fluid is 5% boric acid by weight percentage, 1% CaCl
2With 1% FeCl
3Described solid solution is 8% metabisulfite solution.
5% immobilization particle is by weight percentage gone up shaking table after inserting the mud substratum, wherein: the concentration of mud internal contamination thing polycyclic aromatic hydrocarbon pyrene and benzo (a) pyrene records B61 for 100 μ g/ml degradation processes respectively after carrying out 96 hours under 120r/min29 ℃, B67, the degradation rate of my polycyclic aromatic hydrocarbon pyrene and benzo (a) pyrene is respectively 48.89% and 35.46%, 56.04% and 38.03%, 45.03% and 30.97%.
The proportioning of mud substratum is: grape 2%, and peptone 2%, yeast extract paste 1%, the pH nature, soil ratio is 3:1.
The isolated plant of described process for fixation is: an end is provided with the wound packages structure 1 of shutoff, is provided with piston 2 at the other end of wound packages structure, offers pore 4 in shutoff, and the outer side of piston 2 is provided with handle 3.
Be connected with Glass tubing 5 in the pore 4 in the described shutoff, the length of Glass tubing 5 is increased to the center to outer rim gradually by shutoff.
Difference from Example 1 is:
With bacterium B61, B67, this three strain of MB mixes in twos and three strains mix, and cultivates and then gets with the carrier soln bacterium liquid of 25% volumetric concentration that is benchmark respectively.Stagnate 22 hours with 1.1*10 in the adding pressure type pellet fabrication device of the pore of 2mm with in the crosslinked fluid
5Pa makes the small-particle that diameter is 3-4mm, reinforces 22 hours adding in the solid solution behind the particle filtration, soaks 22 hours with sterilized water again, then cultivates propagation by shaking table, and the particle of being fixed is standby.Making particle grain size is 2.5mm.
Carrier soln is by weight percentage: 12% polyvinyl alcohol, and 0.4% sodium alginate, 4% gac, surplus is a sterilized water;
Described crosslinked fluid is by weight percentage: 8% boric acid, 2% CaCl
2, 2% FeCl
3
4% immobilization particle is by weight percentage gone up shaking table after inserting the mud substratum, water and soil mass ratio in the mud is 3:1, degradation process records B61 after carrying out 96 hours under 28 ℃, B67, MB is respectively 50.12% and 34.09% to the degradation rate of pyrene and benzo (a) pyrene, 50.37% and 36.16%, 42.24% and 32.64%.
Difference from Example 1 is:
With bacterium B61, B67, this three strain of MB mixes in twos and three strains mix, and cultivates and then gets with the carrier soln bacterium liquid of 30% volumetric concentration that is benchmark respectively.2.5mm the adding pressure type pellet fabrication device of pore in crosslinked fluid in stagnate 23 hours with 1.15*10
5Pa makes the small-particle that diameter is 3mm, reinforces 23 hours adding in the solid solution behind the particle filtration, soaks 23 hours with sterilized water again, then cultivates propagation by shaking table, and the particle of being fixed is standby.
Carrier soln is by weight percentage: 13% polyvinyl alcohol, and 0.5% sodium alginate, 5% gac, surplus is a sterilized water; Described crosslinked fluid is by weight percentage: 8% boric acid, 2% CaCl
2, 2% FeCl
3
8% immobilization particle is by weight percentage gone up shaking table after inserting the mud substratum, and the water and soil mass ratio in the mud is 3:1, and degradation process records B61 after carrying out 96 hours under 30 ℃, B67, MB to the degradation rate of pyrene and benzo (a) pyrene referring to table 1.
Table 1
Difference from Example 1 is:
The pellet fabrication device of packing into is made the small-particle that particle diameter is 3.5mm.
5% immobilization particle is by weight percentage gone up shaking table after inserting the mud substratum, record degradation rate referring to table 2 after 96 hours.
Table 2
Claims (5)
1. introduction bacterium microorganism immobilization method that is used for soil remediation, it is characterized in that: the bacterium liquid of logarithmic phase is capable immobilization embedded, the bacterium liquid that soon with the carrier soln will be the 20-30% volume of benchmark joins in the carrier soln, fully behind the mixing, in (1.05-1.2) * 10
5Pore by 2-3.5mm under the Pa enters in the crosslinked fluid, and stagnates 22-25 hour in crosslinked fluid, makes the small-particle that diameter is 3-4mm, reinforced 22-25 hour adding in the solid solution behind the particle filtration, sterile water wash is carried out shaking table and is cultivated propagation, the particle of being fixed; Described carrier soln is the polyvinyl alcohol of 10-15%, the sodium alginate of 0.3-0.6% and the gac of 3-6% by weight percentage, and surplus is a sterilized water;
The microorganism that contains logarithmic phase in the described bacterium liquid is 10
5-10
7Individual/ml;
Described crosslinked fluid is the boric acid of 5-8% by weight percentage, the CaCl of 1-3%
2FeCl with 1-3%
3, surplus is a water.
2. by the described introduction bacterium microorganism immobilization method that is used for soil remediation of claim 1, it is characterized in that: described sterile water wash is meant sterile water wash immersion 22-25 hour.
3. by the described introduction bacterium microorganism immobilization method that is used for soil remediation of claim 1, it is characterized in that: described solid solution is the metabisulfite solution of mass concentration 8%-15%.
4. by the described introduction bacterium microorganism immobilization method that is used for soil remediation of claim 1, it is characterized in that: described bacterial classification: convex pseudomonas, one or more in the crooked pseudomonas.
5. the isolated plant of the described process for fixation of claim 1, it is characterized in that: it is the wound packages structure (1) that an end is provided with shutoff, the other end in the wound packages structure is provided with piston (2), offers pore (4) in shutoff, and the outer side of piston (2) is provided with handle (3);
Be connected with Glass tubing (5) in the pore in the described shutoff (4), the length of Glass tubing (5) is increased to the center gradually by the outer rim of shutoff.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2006101340699A CN101168736B (en) | 2006-10-27 | 2006-10-27 | Introduced bacterium microorganism immobilization method used for repairing soil and special-purpose device for the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2006101340699A CN101168736B (en) | 2006-10-27 | 2006-10-27 | Introduced bacterium microorganism immobilization method used for repairing soil and special-purpose device for the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101168736A CN101168736A (en) | 2008-04-30 |
| CN101168736B true CN101168736B (en) | 2011-08-03 |
Family
ID=39389502
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2006101340699A Expired - Fee Related CN101168736B (en) | 2006-10-27 | 2006-10-27 | Introduced bacterium microorganism immobilization method used for repairing soil and special-purpose device for the same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101168736B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109365498A (en) * | 2018-12-20 | 2019-02-22 | 北京市环境保护科学研究院 | A kind of formula and soil treatment device reducing P in soil AHs biological effectiveness |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102114492A (en) * | 2010-12-06 | 2011-07-06 | 沈阳工业大学 | Novel method for degrading pollutants through immobilized microorganism carrier |
| CN102199430B (en) * | 2010-12-27 | 2014-02-26 | 北京师范大学 | Immobilized microsphere for remedying calcium carbonate-containing petroleum polluted soil as well as preparation method and application |
| CN102505011A (en) * | 2011-12-08 | 2012-06-20 | 沈阳工业大学 | Microbial immobilized embedding method for degrading pollutants and dedicated device therefor |
| CN103374560B (en) * | 2012-04-19 | 2016-01-20 | 中国石油化工集团公司 | A kind of preparation method of polyvinyl alcohol immobilized microorganism gel |
| CN103923904A (en) * | 2013-01-16 | 2014-07-16 | 浙江海洋学院 | Preparation method for immobilized microbe oil-spill repairing agent |
| CN104208841A (en) * | 2014-08-25 | 2014-12-17 | 徐州工程学院 | Method for degrading organophosphorus pesticide in soil by immobilized engineering bacteria |
| CN106623413B (en) | 2017-02-14 | 2017-12-05 | 温铁军 | A kind of devices and methods therefor for being used to extract the microorganism for completing heavy-metal contaminated soil reparation |
| CN110153177B (en) * | 2019-03-08 | 2020-07-31 | 中国科学院广州地球化学研究所 | Method for repairing polycyclic aromatic hydrocarbon polluted soil by using fungi |
| CN111470615B (en) * | 2020-05-06 | 2022-03-29 | 北京工业大学 | Preparation and application of sulfate-reducing-reinforced composite bacteria embedded bioactive filler |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1740317A (en) * | 2004-08-27 | 2006-03-01 | 中国科学院沈阳应用生态研究所 | A kind of method for preparing immobilized particles that is used for the surface water reparation |
-
2006
- 2006-10-27 CN CN2006101340699A patent/CN101168736B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1740317A (en) * | 2004-08-27 | 2006-03-01 | 中国科学院沈阳应用生态研究所 | A kind of method for preparing immobilized particles that is used for the surface water reparation |
Non-Patent Citations (4)
| Title |
|---|
| 徐新阳等.污染地表水修复用微球菌的化学包埋法.东北大学学报(自然科学版)27 10.2006,27(10),p1146-1148. |
| 徐新阳等.污染地表水修复用微球菌的化学包埋法.东北大学学报(自然科学版)27 10.2006,27(10),p1146-1148. * |
| 蔡军.凝聚法固定产丙烯腈水合酶菌株的研究.化工科技市场 3.2005,(3),P40. |
| 蔡军.凝聚法固定产丙烯腈水合酶菌株的研究.化工科技市场 3.2005,(3),P40. * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109365498A (en) * | 2018-12-20 | 2019-02-22 | 北京市环境保护科学研究院 | A kind of formula and soil treatment device reducing P in soil AHs biological effectiveness |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101168736A (en) | 2008-04-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101168736B (en) | Introduced bacterium microorganism immobilization method used for repairing soil and special-purpose device for the same | |
| Cruz et al. | Biological deterioration of alginate beads containing immobilized microalgae and bacteria during tertiary wastewater treatment | |
| De-Bashan et al. | Immobilized microalgae for removing pollutants: review of practical aspects | |
| CN102031230B (en) | Bacillus amyloliquefacien and preparation method of polluted water reparation agent by using same | |
| Germaine et al. | Ecopiling: a combined phytoremediation and passive biopiling system for remediating hydrocarbon impacted soils at field scale | |
| CN105836900B (en) | A kind of ecological purified water system | |
| CN111254137A (en) | Immobilized carbon algae material and preparation method and application thereof | |
| CN110078329A (en) | A kind of sediment in-situ is biological reinforced with benthic environment ecological restoring method | |
| CN108624582A (en) | Microorganism formulation for soil remediation | |
| US8586342B2 (en) | Artificial biomembrane and the method for manufacturing the same | |
| CN102505011A (en) | Microbial immobilized embedding method for degrading pollutants and dedicated device therefor | |
| KR101543134B1 (en) | Method for producing thalli of lichens, method for restoring the degraded ecology by them, and compositions therefor | |
| CN103045579A (en) | Microbial remediation curing adsorbing bacterial preparation applicable to marine environment petroleum pollution as well as preparation method and application of same | |
| CN102424806A (en) | DDT (dichlorodiphenyltrichloroethane)-degrading pseudomonad strain | |
| CN102676432A (en) | Efficient water body denitrification pseudomonas stutzeri DB-33 and culture method thereof | |
| CN106399176A (en) | Paenibacillus and its application in water body purification | |
| CN112514737A (en) | Method for restoring water environment pollution by using mycorrhizal plant and dominant microorganisms | |
| Abe et al. | Removal of inorganic nitrogen sources from water by the algal biofilm of the aerial microalga Trentepohlia aurea | |
| CN103013885A (en) | Benzene composite degrading microbe and immobilized benzene composite microbial inoculum, and preparation method thereof | |
| CN108423838B (en) | Microbial preparation for ecological safety water system and preparation method thereof | |
| CN102060389A (en) | Composite microecological preparation capable of degrading nitrites in water body rapidly | |
| CN116515665A (en) | Pseudomonas capable of degrading microcystin, immobilized microbial agent and application | |
| Li et al. | Fabrication of immobilized algal-bacterial beads and its application in synthetic mariculture tail water treatment | |
| CN206089209U (en) | Ecological water purification system | |
| CN104694527A (en) | Method for strengthening sanitary sewage treatment capacity of constructed wetland through fixed type bacillus with polymeric sponges as carrier |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110803 Termination date: 20121027 |