CN102888392A - Preparation method of floating embedding bacterium agents for in-situ remediation of oil polluted water - Google Patents
Preparation method of floating embedding bacterium agents for in-situ remediation of oil polluted water Download PDFInfo
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- CN102888392A CN102888392A CN2012103822140A CN201210382214A CN102888392A CN 102888392 A CN102888392 A CN 102888392A CN 2012103822140 A CN2012103822140 A CN 2012103822140A CN 201210382214 A CN201210382214 A CN 201210382214A CN 102888392 A CN102888392 A CN 102888392A
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Abstract
The invention relates to a preparation method of floating embedding bacterium agents for in-situ remediation of oil polluted water, specifically relating to a technology for fixing oil degradation bacteria by using expanded graphite, sodium alginate and calcium chloride, absorbing and degrading to purify the petroleum polluted water. The preparation method of the floating embedding bacterium agents comprises the following steps: adding petroleum degrading bacteria liquid into sodium alginate sol containing the expanded graphite, mixing uniformly, dropping into calcium chloride solution and cross-linking overnight at 4 degrees centigrade to obtain the floating embedding bacterium agents fixing hydrocarbon degradation bacteria. The floating embedding bacterium agents prepared by the preparation method has the advantages of high mechanical strength, good floatability and mass transfer performance and high degradation efficiency; the floating embedding bacterium agents can be directly put onto the surface of the petroleum polluted water; the embedding bacterium agents can float on an oil-water interface for long term so as to ensure effective contact between the bacterium agent and the petroleum; the embedding technology increases the concentration of microorganisms, prevents the bacteria influencing by the water using environment, effectively improves the degradation efficiency, recovers and recycles conveniently and has no secondary pollution risk.
Description
Technical field
The invention belongs to water-treatment technology field, be specifically related to a kind of preparation method of the floated embedding microbial inoculum of repairing for the pollution by oil water body in-situ.
Background technology
Along with the development of oil exploration industry and the prosperity of shipping-trade, annual increasing petroleum hydrocarbon enters water body, the taking place frequently especially so that petroleum pollution becomes the focus of global concern of Accidental Oil Spill accident.Petroleum pollution can so that the dissolved oxygen of water body descends, produce stench in rapidly diffusion and form one deck oil film and hinder oxygen transfer between water body and the air of water surface, causes water quality deterioration, hydrobiological death.Polycyclic aromatic hydrocarbon compounds in the petroleum hydrocarbon has strong teratogenesis, carcinogenic effect, can enter human body from hydrobiont by food chain, finally affects human health, and the control of this type of pollution has been caused people's extensive concern with administering.
The recovery technique of petroleum pollution water body mainly comprises physics reparation, chemical reparation, biological restoration and combination recovery technique at present.The method of physics reparation adopts artificial or mechanical means that oil slick is carried out fence and absorption, but cost is high, oil removing is not thorough, the high main drawback that becomes this technology of operational requirement; Chemical repair method then is by adding chemical agent or combustion method treat surface greasy dirt, and this method easily produces secondary pollution and has certain danger; The eighties in 20th century Exxon Vadez oil tanker oil spill accident the biological restoration project, repaired at short notice oil spill, started the beginning of microorganism recovery technique in petroleum polluting water body is administered.The method does not need main equipment, has safety, economic, efficient, non-secondary pollution and simple operation and other advantages.
Although the microorganism recovery technique has obtained a series of achievement, but it uses some problems that also are faced with: at first, the oil degradation microorganism is separated to be obtained, but the impact that it in use is subjected to the factors such as competitive relation between living environment and indigenous strain is difficult to existence and application under new envrionment conditions; Next is from the pattern of microorganism panning petroleum hydrocarbon, microbiological deterioration high-carbon hydrocarbon (C 〉=12) directly contacts by cell and carries out, but because oil degradation bacteria self proportion problem, it is the sedimentation state more after throwing is added to the water, hindered it to contact with the effective of high-carbon hydrocarbon oil slick.Therefore, in order to address these problems, the use of uniting of bioremediation technology to be strengthened and other recovery technique is arranged.Chinese invention patent " a kind of biological expanded graphite preparation method for the reparation of petroleum pollution water body in-situ " (patent No. is 201010299071.8), disclose a kind of take expanded graphite as carrier, adopt the absorption fixation method, the highly effective petroleum degraded mixed bacterium of screening is fixed, the preparation biological expanded graphite, during use, biological expanded graphite is sprayed on the petroleum-polluted water surface, by the efficient adsorption Degradation of biological expanded graphite to the oil slick pollutent, realize the purification of polluted-water.Practical application shows, because the intensity of expanded graphite meterial is relatively poor, causes easily pore structure to be caved in, thereby affects the practical application performance of biological expanded graphite.In recent years, along with the development of microbial immobilized technology, research and develop novel immobilization oil microbial inoculum, be expected to realize further promoting the performance that this technology is repaired the pollution by oil water body in-situ.
Summary of the invention
The object of the invention is to overcome the deficiency that existing bioremediation technology exists, a kind of preparation method of the floated embedding microbial inoculum of repairing for the pollution by oil water body in-situ is provided.
For achieving the above object, technical scheme of the present invention is: adopt sodium alginate-calcium chloride fixation method that oil degradation bacteria is carried out embedding, realize the floating of embedding microbial inoculum take the lighter expanded graphite of proportion as inorganic additives, the expanded graphite that is about to the bacterium liquid of 1 ~ 5mL and 0.1 ~ 0.5g joins and contains in 20 ~ 50mL sodium alginate colloidal sol of 1 ~ 3%, mix, drop to dropper and to contain in 1 ~ 5% the calcium chloride solution, under 4 ℃ of conditions crosslinked 8-12 hour, clean, preserve with 0.9% NaCl solution afterwards, be fixed the floated embedding microbial inoculum of oil degradation bacteria.During use, microbial inoculum directly is added on the petroleum-polluted water surface, can by the absorption degradation effect of embedding microbial inoculum, realizes the purification of polluted-water.
Among the present invention, described oil degradation bacterial classification liquid be enlightening thatch Salmonella (
Dietzia sp.), Stenotrophomonas (
Stenotrophomonas sp.), defective shortwave Zymomonas mobilis (
Brevundimonas diminuta), pseudomonas (
Pseudomonas sp.), genus bacillus (
Bacillus sp.) and Halomonas (
Halomonas venusta) by weight mark count 0.5 ~ 1.5:0.5 ~ 1.5:0.5 ~ 1.5:1 ~ 3:1 ~ 3:2 ~ 4, constant temperature culture obtained mixed strains liquid in 4 ~ 7 days in containing the substratum that petroleum concentration is 1000mg/L.Described oil can select crude oil, diesel oil, machine oil and other different oil compositions according to processing the object needs.
Among the present invention, described sodium alginate soln is that sodium alginate is dissolved under 60 ~ 90 ℃ of conditions, stirs into uniform glue and is cooled to room temperature afterwards.
Among the present invention, described expanded graphite is that commercially available 80 ~ 200 purpose expansible black leads high temperature puffing under 600 ~ 1000 ℃ condition makes.
Principle:Take expanded graphite as inorganic additives, adopt calcium alginate embedded fixation method, the high efficient petroleum degrading bacteria that screens is carried out embedding fix, preparation floated embedding microbial inoculum.During use, the embedding microbial inoculum is added on the petroleum-polluted water surface, by the absorption of expanded graphite and the Degradation of oil degradation bacteria, realize the purification of water body.
Compare with existing petroleum pollution treatment technology, the present invention has the following advantages:
1. the present invention adopts expanded graphite and alginate calcium mixing embedding fixation method, physical strength and floatability are good, biological density is high, has effectively avoided imbedded microbe to be subjected to the impact of water body environment and indigenous strain competitive effect, has greatly improved its detergent power to the pollution by oil water body.
2. the cell surface of the oil degradation bacteria of the present invention's selection has certain hydrophobicity, is conducive to apposition growth on the hydrophobic expanded graphite, and promotes microorganism to the adhesion of oil, has improved the biodegradability to the petroleum hydrocarbon hydrophobic compound.
3. the method can be easy to use, can directly add on the water surface of petroleum pollution water body during use, the embedding microbial inoculum can swim on the oil-water interface for a long time, realization is removed the efficient adsorption degraded of oil slick, thereby realize the original position reparation of petroleum pollution water body, and effectively avoid the loss of its microorganism, and be convenient to reclaim and recycling, there is not the secondary pollution risk.
Embodiment
Following examples will the invention will be further described.
Embodiment 1
(1) preparation of oil degradation bacteria bacterium liquid
With bacterial classification enlightening thatch Salmonella, Stenotrophomonas, defective shortwave Zymomonas mobilis, pseudomonas, genus bacillus (
Bacillus sp.), Halomonas (above-mentioned bacterial classification all can obtain by commercial) by weight mark count 0.5:1:1:1.5:1.5:2, getting 2g, to be seeded in containing of 50mL former oil concentration be in the 1000mg/L minimal medium, concussion was cultivated 5 days in 30 ℃, 120 r/min constant incubators, and is stand-by.
Described minimal medium is NaCl 0.5%, (NH
4)
2SO
40.1%, MgSO
40.01%, NaNO
30.2%, KH
2PO
40.4%, K
2HPO
43H
2O 1.0%, and all the other are distilled water, and pH 7.2.Sterilising conditions is 0.1Mpa moist heat sterilization 30min.
(2) preparation of floated embedding microbial inoculum
Add respectively the water of 0.2g sodium alginate and 10mL at beaker, heating in 60 ℃ of water-baths, be stirred to the sodium alginate dissolving evenly, take out and be cooled to room temperature.The bacterium liquid and the 9mL water that add respectively the above-mentioned oil degradation bacteria of 1mL mix, and this moment, the concentration of sodium alginate was 1%.The expanded graphite that takes by weighing 0.1g adds in the beaker and mixes, and then drips CaCl
2To 1% concentration, put into 4 ℃ in refrigerator after dropwising lower crosslinked 8 hours in the solution, after the NaCl solution with 0.9% cleaned 3 times, it was stand-by to be soaked in 0.9% the NaCl solution 4 ℃ of lower preservations.
Described expanded graphite is that commercially available 80 purpose expansible black leads high temperature puffing under 700 ℃ of conditions makes.
(3) decontamination effect improving of contaminated by diesel oil
In the beaker of the 1L that fills the 500mL tap water, add 0.5g diesel oil, making diesel oil concentration is 1000mg/L, add the embedding microbial inoculum of the above-mentioned preparation of 0.2g, under 25 ℃, the condition of 150r/min, shake, through processing in 3 days, the Determination of Diesel Oil in the water did not detect, and illustrates that the embedding microbial inoculum is to being subjected to the contaminated by diesel oil water body to have good decontamination effect improving.
Embodiment 2
(1) preparation of oil degradation bacteria bacterium liquid
With enlightening thatch Salmonella, Stenotrophomonas, defective shortwave Zymomonas mobilis, pseudomonas, genus bacillus, Halomonas by weight mark count 0.5:0.5:1:1.5:2:2, getting 2g, to be seeded in containing of 50mL former oil concentration be in the 1000mg/L minimal medium, in 30 ℃, concussion was cultivated 5 days in the 120 r/min constant incubators, and is stand-by.
Described minimal medium is NaCl 0.5%, (NH
4)
2SO
40.1%, MgSO
40.01%, NaNO
30.2%, KH
2PO
40.4%, K
2HPO
43H
2O 1.0%, and all the other are distilled water, and pH 7.2.Sterilising conditions is 0.1Mpa moist heat sterilization 30min.
(2) preparation of floated embedding microbial inoculum
The water that adds 0.4g sodium alginate and 10mL in beaker heats, is stirred to the sodium alginate dissolving evenly in 80 ℃ of water-baths, take out and be cooled to room temperature.The bacterium liquid and the 9mL water that add the above-mentioned oil degradation bacteria of 1mL mix, and this moment, the concentration of sodium alginate was 2%.Take by weighing in the 0.2g expanded graphite beaker and mix, then drip CaCl
2Solution to 2% concentration was put into 4 ℃ in refrigerator lower crosslinked 10 hours after dropwising, after the NaCl solution with 0.9% cleaned 3 times, it was stand-by to be soaked in 0.9% the NaCl solution 4 ℃ of lower preservations.
Described expanded graphite is that commercially available 100 purpose expansible black leads high temperature puffing under 800 ℃ of conditions makes.
(3) decontamination effect improving of crude oil pollution
In the beaker of the 1L that fills the 500mL tap water, add a certain amount of crude oil, making former oil concentration is 800 mg/L, add the embedding microbial inoculum of the above-mentioned preparation of 0.2g, under 25 ℃, the condition of 150r/min, shake, through processing in 4 days, the crude content in the water did not detect, and illustrates that embedding microbial inoculum crude oil pollution water body has good purification repairing effect.
Claims (5)
1. preparation method who is used for the floated embedding microbial inoculum that the pollution by oil water body in-situ repairs is characterized in that concrete steps are as follows:
The expanded graphite of the bacterium liquid of 1 ~ 5mL and 0.1 ~ 0.5g joined contain in 20 ~ 50mL sodium alginate colloidal sol of 1 ~ 3%, mix, drop to and contain in 1% ~ 5% the calcium chloride solution, under 4 ℃ of conditions crosslinked 8-12 hour, NaCl solution with 0.9% cleans, and is fixed the floated embedding microbial inoculum of oil degradation bacteria; During use, the embedding microbial inoculum directly is added on the petroleum-polluted water surface, can by the absorption degradation effect of embedding microbial inoculum, realizes the purification of polluted-water.
2. the preparation method of floated embedding microbial inoculum according to claim 1 is characterized in that described sodium alginate colloidal sol is that sodium alginate is dissolved under 60 ~ 90 ℃ of conditions, stirs into uniform glue and is cooled to room temperature afterwards.
3. floated embedding bacterial preparation process according to claim 1 is characterized in that described expanded graphite is that 80 ~ 200 purpose expansible black leads high temperature puffing under 600 ~ 1000 ℃ of conditions makes.
4. floated embedding bacterial preparation process according to claim 1, it is characterized in that described oil degradation bacteria liquid be enlightening thatch Salmonella, Stenotrophomonas, shortwave Zymomonas mobilis, pseudomonas, genus bacillus and Halomonas by weight mark count the mixed strains liquid of 0.5 ~ 1.5:0.5 ~ 1.5:0.5 ~ 1.5:1 ~ 3:1 ~ 3:2 ~ 4, constant temperature culture obtained in 4 ~ 7 days in containing the substratum that petroleum concentration is 1000mg/L.
5. floated embedding bacterial preparation process according to claim 4, it is characterized in that described oil be in crude oil, diesel oil, machine oil or the different oil any.
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| CN103409338A (en) * | 2013-07-04 | 2013-11-27 | 大连施倍得生物技术有限公司 | Microbial water quality purifying agent for aquaculture pond and preparation method and strain curing method thereof |
| CN103523928A (en) * | 2013-07-16 | 2014-01-22 | 中国人民解放军海军医学研究所 | Bacterial agent for bioremediation of oil-polluted water area and preparation method |
| CN103641243A (en) * | 2013-12-03 | 2014-03-19 | 大连海洋大学 | Floating type light self-cleaning microorganism immobilization membrane for removing oil contamination polluting water and preparation method of membrane |
| CN105154367A (en) * | 2015-09-28 | 2015-12-16 | 中国石油化工股份有限公司 | Halomonas TF-1 and application thereof |
| CN105214611A (en) * | 2015-09-14 | 2016-01-06 | 同济大学 | One-step method prepares the method for porous alginate-graphene nano plural gel ball |
| CN105861365A (en) * | 2016-04-19 | 2016-08-17 | 黑龙江八农垦大学 | Pseudomonas sp. LD23 and preparation of immobilized microspheres of Pseudomonas sp. LD23 |
| CN106587358A (en) * | 2016-12-16 | 2017-04-26 | 河北大学 | Oil leakage degradation method for lakes, channels and ships |
| CN106635909A (en) * | 2016-12-29 | 2017-05-10 | 长安大学 | Crude oil degradation mixed bacterium, microbial agent and application of microbial agent |
| CN106635910A (en) * | 2016-12-29 | 2017-05-10 | 长安大学 | Brevendimonas diminuta, bacterial agent and applications of microbial agent |
| CN108996709A (en) * | 2018-08-28 | 2018-12-14 | 重庆文理学院 | A kind of round-the-clock composite photocatalyst collaboration system and preparation method thereof |
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| CN111100858A (en) * | 2019-12-23 | 2020-05-05 | 山东科技大学 | Immobilized microbial agent, offshore spilled oil treatment device carrying microbial agent and offshore spilled oil treatment method |
| CN112246228A (en) * | 2020-10-31 | 2021-01-22 | 兰州资源环境职业技术学院 | Preparation method of modified graphene material for removing heavy metal pollution in soil |
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| CN103523928A (en) * | 2013-07-16 | 2014-01-22 | 中国人民解放军海军医学研究所 | Bacterial agent for bioremediation of oil-polluted water area and preparation method |
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| CN105154367A (en) * | 2015-09-28 | 2015-12-16 | 中国石油化工股份有限公司 | Halomonas TF-1 and application thereof |
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| CN105861365A (en) * | 2016-04-19 | 2016-08-17 | 黑龙江八农垦大学 | Pseudomonas sp. LD23 and preparation of immobilized microspheres of Pseudomonas sp. LD23 |
| CN105861365B (en) * | 2016-04-19 | 2019-05-07 | 黑龙江八一农垦大学 | The preparation of one pseudomonas LD23 and its immobilized microspheres |
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Application publication date: 20130123 |