CN102978195A - Preparation method for embedding floating bacteria agent of nutritive salt and expandable graphite and microorganism together - Google Patents
Preparation method for embedding floating bacteria agent of nutritive salt and expandable graphite and microorganism together Download PDFInfo
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- CN102978195A CN102978195A CN2012104646644A CN201210464664A CN102978195A CN 102978195 A CN102978195 A CN 102978195A CN 2012104646644 A CN2012104646644 A CN 2012104646644A CN 201210464664 A CN201210464664 A CN 201210464664A CN 102978195 A CN102978195 A CN 102978195A
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Abstract
The invention relates to a preparation method for an embedding floating bacteria agent of nutritive salt, expandable graphite and microorganism together. The preparation method for the embedding floating bacteria agent of the nutritive salt, the expandable graphite and the microorganism together comprises the steps of using a sodium alginate solution as an embedding frame, adding a nutritive salt solution, the expandable graphite and petroleum degrading bacteria to the sodium alginate solution, mixing evenly, dripping into a calcium chloride solution to embed, crosslinking for the night under the condition of 4 DEG C, and obtaining an embedding bacteria agent capable of strengthening bioremediation effect. The bacteria agent prepared by the preparation method for the embedding floating bacteria agent of the nutritive salt, the expandable graphite and the microorganism together is suitable for in-situ remediation of marine petroleum pollution, and has the advantages of being long in floating time, good in mass transfer performance, high in petroleum degrading efficiency, and the like. The method that the nutritive salt and bacteria solutions are embedded together is adopted by the embedding bacteria agent, defects that nutrition of marine environment is poor and additionally the nutritive salt is diluted quickly are overcome, and floating of the bacteria agent is achieved by addition of the expandable graphite. When the embedded bacteria agent is used, the embedded bacteria agent can be directly added to the surface of a polluted water body, effective contact with petroleum pollutant is ensured, biodegradation efficiency is effectively improved, and recycling is facilitated, and risk of a secondary pollution does not exist.
Description
Technical field
The invention belongs to water-treatment technology field, be specifically related to the preparation method that a kind of nutritive salt, expanded graphite and microorganism for the purification of pollution by oil water body in-situ is total to embedding floated microbial inoculum.
Background technology
Fast development along with industry, so that market raises year by year to the demand of oil and derived product thereof, and the prosperity of the development of oil exploration industry and shipping-trade causes annual increasing petroleum hydrocarbon to enter water body, and the water body petroleum-type is polluted has become the focus that environmental area is paid close attention to.
At present, the recovery technique of oil pollution mainly comprises physics reparation, chemical reparation, biological restoration and combination recovery technique.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; Bioremediation technology and the emphasis that will become following marine oil spill recovery technique development around the combination technique of biological restoration with its safety, characteristics economic, efficient, non-secondary pollution.
Although the microorganism recovery technique is with the obvious advantage, also exist some problems: at first, the oil degradation microorganism in use is subjected to the impact of the factors such as competitive relation between living environment and indigenous strain, is difficult to existence and application under new envrionment conditions; Secondly, 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 afterwards in dropping into water, hindered it to contact with the effective of high-carbon hydrocarbon oil slick; The 3rd, most of petroleum-polluted Water phosphorus nutrition element is deficient, such as the ocean, do not utilize microorganism growth, and entering open water body environment, the nutritive salt that adds can be diluted at short notice, low by the microorganism utilization ratio, and the deficiency that causes secondary pollution is arranged.Therefore in order to address these problems, remain bioremediation technology is improved and strengthens.
Summary of the invention
The object of the invention is to overcome the deficiency that prior art exists, with the microorganism recovery technique together the embedded immobilization technology combine, provide a kind of and pollute the floating microbial inoculum of common embedding that original position is repaired for marine oil spill.
For achieving the above object, technical scheme of the present invention is: adopt sodium alginate-calcium chloride fixation method to nutritive salt, oil degradation bacteria carries out common embedding, realize the floating of embedding microbial inoculum take the lighter expanded graphite of proportion as inorganic additives, be about to the nutrient salt solution of 1 ~ 5ml, the oil degradation bacteria of 1 ~ 5ml and 0.1 ~ 0.3g expanded graphite join and contain in 1% ~ 3% the sodium alginate colloidal sol, mix, drop to dropper and to contain in 1% ~ 5% the calcium chloride solution, under 4 ℃ of conditions crosslinked 8-12 hour, clean with 0.9% NaCl solution afterwards, preserve, obtain nutritive salt, expanded graphite and microorganism be the floating microbial inoculum of embedding altogether.During use, this 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 nutrient salt solution main component is nitrogen and phosphorus, and wherein nitrogenous source can be selected NaNO
3, NH
4Cl, NH
4NO
3Or in the urea etc. more than one; KH can be selected in the phosphorus source
2PO
4, K
2HPO
4, NaH
2PO
4Or Na
2HPO
4Deng in more than one.Nitrogen content in the embedding system is that 20 ~ 40mg/L, phosphorus content are 5 ~ 10mg/L.
Among the present invention, described oil degradation bacterial classification liquid be alkane eating bacteria (
Alcanivorax sp.), Alcaligenes (
Alcal igenes sp.), the Ge Dengshi mattress (
Gordonia sp.), enlightening thatch Salmonella (
Dietzia 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 expanded graphite is that commercially available 50 ~ 200 purpose expansible black leads high temperature puffing 10 ~ 30s under 600 ~ 1000 ℃ condition makes.
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.
Principle:Take expanded graphite as inorganic additives, adopt calcium alginate embedded fixation method, nutrient salt solution and highly effective petroleum degraded composite bacteria to be carried out embedding fix, preparation is the floating microbial inoculum of embedding altogether.During use, to be total to the floating microbial inoculum of embedding adds on the petroleum pollution water surface, the interpolation of expanded graphite is so that microbial inoculum floats on oil-water interface, realize that oil degradation bacteria contacts with the effective of petroleum pollution, the nutritive salt of embedding microbial inoculum inside provides microbial growth and metabolism required nutrition, strengthen biodegradable efficient, realized the purification of water body.
Compare with existing petroleum pollution treatment technology, the present invention has the following advantages:
1. the present invention adopts calcium alginate embedded fixation method, and physical strength is good, biological density is high, has effectively avoided imbedded microbe on the impact of polluted-water environment and the impact of indigenous strain competitive effect.
2. to select expanded graphite be the embedding additive in the present invention, and expanded graphite proportion is lighter, can reduce the proportion of embedding microbial inoculum and realize that it is floating.The embedding microbial inoculum of floated can carry out efficient solution with petroleum hydrocarbon and remove, thereby realizes the fast degradation of pollutent, and expanded graphite has good bioaffinity simultaneously, and its distinctive mesopore and macroporous structure are suitable for the Growth and reproduction of oil degradation bacteria.
3. the present invention adopts the altogether mode of embedding of nutritive salt and oil degradation bacteria, so that the residing microenvironment of imbedded microbe is more suitable for the growth of himself and the metabolism of petroleum hydrocarbon, overcome ocean water body nutrition scarcity so that the inefficient shortcoming of microbiological deterioration realizes the efficient removal of petroleum pollution.
4. the method is easy to use, directly adds during use on the petroleum pollution water surface, and the embedding microbial inoculum can swim on the oil-water interface for a long time altogether, realization is to the original position reparation of petroleum pollution water body, and effectively avoided the microorganism loss, be convenient to reclaim, do not had the secondary pollution risk.
Embodiment
Embodiment 1
(1) preparation of oil degradation bacteria bacterium liquid
With bacterial classification alkane eating bacteria, Alcaligenes, Ge Dengshi mattress, enlightening thatch Salmonella, genus bacillus and 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.Add respectively that 1mL nutritive salt is molten, the bacterium liquid of the above-mentioned oil degradation bacteria of 1mL and 8mL water mixes.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 800 ℃ 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 1500mg/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 alkane eating bacteria, Alcaligenes, Ge Dengshi mattress, enlightening thatch Salmonella, genus bacillus and Halomonas (above-mentioned bacterial classification all can obtain by commercial) 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 ° of C, 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.Adding 2mL nutritive salt bacterium liquid and 7mL water molten, the above-mentioned oil degradation bacteria of 1mL mixes.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 700 ℃ 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 1000 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 the embedding microbial inoculum has good purification repairing effect to the crude oil pollution water body.
Claims (5)
1. a nutritive salt, expanded graphite and the microorganism preparation method of the floating microbial inoculum of embedding altogether is characterized in that concrete steps are as follows:
The petroleum hydrocarbon degradation bacterium of the nutrient salt solution of 1 ~ 5mL, 1 ~ 5mL and 0.1 ~ 0.3g expanded graphite joined contain in 1% ~ 3% the sodium alginate colloidal sol, 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, obtain nutritive salt, expanded graphite and send out " microorganism is the floating microbial inoculum of embedding altogether; During use, the floating microbial inoculum of this embedding 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. a kind of nutritive salt according to claim 1, expanded graphite and microorganism are total to the preparation method of the floating microbial inoculum of embedding, it is characterized in that described nutrient salt solution main component is nitrogen and phosphorus, and wherein: nitrogenous source is NaNO
3, NH
4Cl, NH
4NO
3Or in the urea more than one; The phosphorus source is KH
2PO
4, K
2HPO
4, NaH
2PO
4Or Na
2HPO
4In more than one; Nitrogen content in the floating microbial inoculum of gained embedding is that 20 ~ 40mg/L, phosphorus content are 5 ~ 10mg/L.
3. a kind of nutritive salt according to claim 1, expanded graphite and microorganism be the preparation method of the floating microbial inoculum of embedding altogether, described petroleum hydrocarbon degradation bacterium is by alkane eating bacteria, Alcaligenes, the Ge Dengshi mattress, enlightening thatch Salmonella, genus bacillus and Halomonas form, alkane eating bacteria, Alcaligenes, the Ge Dengshi mattress, enlightening thatch Salmonella, genus bacillus and Halomonas by weight mark are counted (0.5 ~ 1.5): (0.5 ~ 1.5): (0.5 ~ 1.5): (1 ~ 3): (1 ~ 3): (2 ~ 4), constant temperature culture obtained petroleum hydrocarbon degradation bacterium in 4 ~ 7 days in containing the substratum that petroleum concentration is 1000mg/L, and described oil adopts crude oil, diesel oil, in machine oil or other the different oil compositions any.
4. a kind of nutritive salt according to claim 1, expanded graphite and microorganism are total to the preparation method of the floating microbial inoculum of embedding, it is characterized in that described sodium alginate colloidal sol is that sodium alginate is dissolved under 60 ~ 90 ℃ of conditions, stir into uniform glue and be cooled to room temperature afterwards.
5. a kind of nutritive salt according to claim 1, expanded graphite and microorganism are total to the preparation method of the floating microbial inoculum of embedding, it is characterized in that described expanded graphite is that 80 ~ 200 purpose expansible black leads high temperature puffing under 600 ~ 1000 ℃ of conditions makes.
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
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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 |
| CN103695412A (en) * | 2013-12-28 | 2014-04-02 | 天津北洋百川生物技术有限公司 | Embedding microbial agent for in-situ purification of water body and preparation method thereof |
| CN105087540A (en) * | 2015-09-25 | 2015-11-25 | 辽宁海澳科技有限公司 | Microbe embedding repairing agent as well as preparation method and application thereof |
| CN105152341A (en) * | 2015-07-11 | 2015-12-16 | 北京化工大学 | Self-floating type ecological floating bed capable of being used for water body self-purification |
| CN106630180A (en) * | 2016-11-10 | 2017-05-10 | 中国海洋石油总公司 | Preparation method of slow release nutritional agent for increasing biochemical treatment effect of chemical industry production wastewater |
| CN106699322A (en) * | 2015-09-25 | 2017-05-24 | 北京菲迪威尔科技有限公司 | Nitrogen-supplementing embedded growth-promoting agent, and preparation method and application thereof |
| CN110577949A (en) * | 2019-10-10 | 2019-12-17 | 浙江海洋大学 | Oil spill repairing agent, preparation method thereof and oil spill in-situ repairing method |
| CN110628754A (en) * | 2019-10-10 | 2019-12-31 | 浙江海洋大学 | Microbial foam type carrier, preparation method thereof and in-situ treatment method for petroleum pollution on water surface |
| CN110980960A (en) * | 2019-11-26 | 2020-04-10 | 浙江永续环境工程有限公司 | Oil-removing microbial inoculum and sludge treatment technology using same |
| 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 |
| CN112960776A (en) * | 2020-12-17 | 2021-06-15 | 云南中贸环境节能科技投资股份有限公司 | Microbial carrier particle for sewage treatment, preparation method and application |
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Cited By (15)
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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 |
| CN103695412A (en) * | 2013-12-28 | 2014-04-02 | 天津北洋百川生物技术有限公司 | Embedding microbial agent for in-situ purification of water body and preparation method thereof |
| CN103695412B (en) * | 2013-12-28 | 2016-05-04 | 天津北洋百川生物技术有限公司 | A kind of imbedded microbe microbial inoculum purifying for water body in-situ and preparation method thereof |
| CN105152341A (en) * | 2015-07-11 | 2015-12-16 | 北京化工大学 | Self-floating type ecological floating bed capable of being used for water body self-purification |
| CN106699322A (en) * | 2015-09-25 | 2017-05-24 | 北京菲迪威尔科技有限公司 | Nitrogen-supplementing embedded growth-promoting agent, and preparation method and application thereof |
| CN105087540A (en) * | 2015-09-25 | 2015-11-25 | 辽宁海澳科技有限公司 | Microbe embedding repairing agent as well as preparation method and application thereof |
| CN106630180A (en) * | 2016-11-10 | 2017-05-10 | 中国海洋石油总公司 | Preparation method of slow release nutritional agent for increasing biochemical treatment effect of chemical industry production wastewater |
| CN106630180B (en) * | 2016-11-10 | 2019-05-28 | 中国海洋石油集团有限公司 | Improve the preparation method of the slow-release nutrient of chemical production wastewater Biochemical Treatment |
| CN110577949A (en) * | 2019-10-10 | 2019-12-17 | 浙江海洋大学 | Oil spill repairing agent, preparation method thereof and oil spill in-situ repairing method |
| CN110628754A (en) * | 2019-10-10 | 2019-12-31 | 浙江海洋大学 | Microbial foam type carrier, preparation method thereof and in-situ treatment method for petroleum pollution on water surface |
| CN110980960A (en) * | 2019-11-26 | 2020-04-10 | 浙江永续环境工程有限公司 | Oil-removing microbial inoculum and sludge treatment technology using same |
| 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 |
| CN111100858B (en) * | 2019-12-23 | 2023-08-15 | 山东科技大学 | Immobilized microbial agent, offshore spilled oil treatment device carrying immobilized microbial agent and offshore spilled oil treatment method |
| CN112960776A (en) * | 2020-12-17 | 2021-06-15 | 云南中贸环境节能科技投资股份有限公司 | Microbial carrier particle for sewage treatment, preparation method and application |
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Application publication date: 20130320 |