CN105254025B - A kind of method for improving oil-removing bacteria oil removing efficiency - Google Patents
A kind of method for improving oil-removing bacteria oil removing efficiency Download PDFInfo
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- CN105254025B CN105254025B CN201510632606.1A CN201510632606A CN105254025B CN 105254025 B CN105254025 B CN 105254025B CN 201510632606 A CN201510632606 A CN 201510632606A CN 105254025 B CN105254025 B CN 105254025B
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
It is an object of the invention to provide a kind of method for improving oil-removing bacteria oil removing efficiency.The externally-applied magnetic field around oil-removing bacteria, by adjusting magnetic field intensity and magnetic fields mode, oil-removing bacteria can be effectively improved to the degradation effect of oil and the resistivity of impact load, the microorganism deoiling method of process runs well is improved, belong to oil-polluted water biological treatment field.The permanent magnet for being used is circular ferrite permanent magnet, and action condition refers to magnetic field intensity between 18 35mT, magnetic action mode is continuous action.Magnetic field of the present invention is produced by commercially available permanent magnet, and required magnetic field intensity is small, it is not necessary to consume electric energy, invest low, effect good, do not change existing process flow, big equipment is not increased, be a suitable China's national situation, economically viable reinforcing oily waste water treatment technology.
Description
Technical field
The present invention relates to a kind of oil-polluted water bioremediation, specifically a kind of magnetic field-intensification oil-removing bacteria is carried
The method of oil removing efficiency high.
Background technology
Oil has influence on many aspects of social development as the main energy sources of industrial civilization.Oil is referred to as " work by people
The blood of industry ".Substantial amounts of oil and its product enter environment during the exploitation to oil, transport, processing and accumulating etc., right
It causes serious oil pollution.Oil pollution surface water, destroys the growth of aquatile, threatens water supply security；Polluted Soil
Earth, destruction soil texture hinders plant normal growth and is enriched with by crops to enter human body, endangers human health；Pollution sea
Ocean, the entrance of influence light and oxygen, destruction marine ecology balance etc., the economy and safety hazard for bringing some row serious.
Oily waste water be it is a kind of have a large capacity and a wide range and endanger serious industrial wastewater, with COD, BOD values are high, there is certain gas
Taste and colourity, inflammable, oxidizable decomposition, the characteristics of be insoluble in water.Due to bioanalysis deoiling effect stabilization, the economical and practical state of method
It is interior, it is widely used at home.NW Hebei is by oil-polluted water by sedimentation oil removal and the oil slick being filtered to remove in water
The suspended oil larger with particle, enters back into biochemical treatment unit and oil emulsion and dissolved oil is degraded, final outflow water row up to standard
Put.Dagang Oilfield is to water quality modification technology, stabilization pond biologic treating technique, suspend the oil-containings such as attachment anaerobic-aerobic treatment technology
Sewage disposal technology carries out the comparative analysis of oil field practice application, it is believed that bioremediation is economical and practical, is at organic sewage
The first-selected technique of reason.Bioanalysis oil removing is the metabolism using microorganism, is made in water in dissolving, the organic contamination of colloidal state
Material is converted into the innocuous substance of stabilization.Current handling process comparative maturity and using it is more be activated sludge process and biological filter
Chi Fa.Activated sludge process be in aeration tank by the use of flow regime activated sludge as purification microorganism carrier, by absorption,
The microorganism being concentrated on activated sludge surface carrys out decomposing organic matter.Biological filter process is in biofilter, to make microorganism attached
On filtrate, waste water is flowed through during filter material surface from top to down, organic pollution just by microorganism adsorption and decomposes destruction.
Microbial method degreasing technique it is critical only that biological inoculum and biological treatment, and the particularity according to oily waste water develops height
The biological inoculum of effect and efficient handling process are the focuses of the area research, but the treatment of existing microbial method contains waste water
It is vulnerable to the influence of environmental factor, causes oil removal efficiency to reduce.
It is an emerging in recent years biological reinforcing technology that magnetic field promotes microbial degradation organic matter, and microorganism is strong through magnetic field
After change effect, activity is improved, metabolism enhancing, while water physicochemical properties after magnetization treatment can also become
Change.Magnetic field can produce influence to the enzymatic activity of microorganism in water and metabolism, and different microbe species are in different magnetic
Different magnetic effects can be all shown under the conditions of field type, magnetic field intensity, action time etc..Magnetic technology is introduced as saprobia
Handling process research provides new approaches.Magnetic technology couples small with floor space with biologic treating technique, and energy consumption is low, be easy to behaviour
The advantages such as work, non-secondary pollution, are a kind of New Wastewater Treatment Technology arts with good development prospect.
The content of the invention
It is an object of the invention to provide a kind of degradation effect and impact load for being effectively improved oil-removing bacteria to oil
Resistivity, improve process runs well raising oil-removing bacteria oil removing efficiency method.
The object of the present invention is achieved like this：
1st, actication of culture
The bacterium colony of the culture presevation medium slant for being stored in 4 DEG C of going bail for coats activation medium flat board, 30 DEG C of cultures 24
~36h；The strain is Acinetobacter sp.B11, has been deposited in that " China Committee for Culture Collection of Microorganisms is general
Logical microorganism " center ", accession designation number is CGMCC Nc.10283, preservation date on January 6th, 2015, depositary institution address：Beijing
No. 3 Institute of Microorganism, Academia Sinica of institute of city Chaoyang District North Star West Road 1；
2nd, prepared by liquid seeds
Picking single bacterium colony is inoculated into seed liquid culture medium, 30 DEG C, 200r/min shaking table cultures on activation medium flat board
12h, OD600Bacteria suspension concentration is determined under wavelength and bacteria suspension is diluted to 0.8, it is stand-by as seed liquor；
3rd, oil-removing bacteria decomposing petroleum hydrocarbon under magnetic action
Bacterial strain seed liquor is inoculated into the degraded culture medium of oil concentration 100mg/L-2000mg/L according to 5% inoculum concentration
In, the magnetic field of additional 18-60mT intensity carries out degradation treatment.
The present invention can also include：
The composition of culture presevation culture medium is beef extract 5.0g/L, peptone 10.0g/L, NaCl 5.0g/L and agar
20g/L, regulation pH is 7.2~7.4.
The composition of seed liquid culture medium is beef extract 5.0g/L, peptone 10.0g/L and NaCl 5.0g/L, and regulation pH is
7.2~7.4.
The composition of degraded culture medium is NH4NO32.0g/L、K2HPO41.5g/L、KH2PO43.0g/L、MgSO4·7H2O
0.1g/L, anhydrous CaCl20.01g/L and Na2EDTA·2H2O 0.01g/L, regulation pH is 7.2~7.4.
It is outer around oil-removing bacteria it is an object of the invention to provide a kind of method that magnetic field-intensification improves oil-removing bacteria oil removing efficiency
Plus magnetic field, by adjusting magnetic field intensity and magnetic fields mode, oil-removing bacteria can be effectively improved to the degradation effect of oil and liquidated
The resistivity of load is hit, the microorganism deoiling method of process runs well is improved.
Provided by the present invention is a kind of method for effectively improving oil-removing bacteria oil removing efficiency.Oil-removing bacteria is one plant useless from recovering the oil
The isolated bacterial strain for being capable of efficient degradation petroleum hydrocarbon in the activated sludge of water biochemical treatment tank collection, single bacterium colony is rounded,
1~2mm of diameter, surface is smooth, moistening, and center is slightly swelled, neat in edge, in tooth yellow.Bacterial strain be in rod-short, two ends blunt circle,
Typically in single or paired, gemma is not produced.Bacterial strain is carried out into Gram's staining, observation finds that bacterial strain can be redyed by sarranine, is
Gram-negative.Bacterial strain catalase positive, the oxidase negative, it is not necessary to growth factor, indoles experiment do not develop the color, it is impossible to
Make gelatin liquefaction, methyl red test is negative.D-Glucose, D- xyloses, L-arabinose, D-ribose can not be utilized, lemon can be utilized
Lemon acid sodium, and can be by the use of potassium nitrate and ammonium sulfate as only nitrogen source used as unique carbon source.The 16S rDNA genes of the bacterial strain
Sequence has been delivered to GenBank databases, it is found that the sequence is same with Acinetobacter sp. strain genes sequence height
Source, is named as Acinetobacter sp.B11, has been deposited in that " China Committee for Culture Collection of Microorganisms is commonly micro-
Bio-Centers ", accession designation number is 10283.
The oil-removing bacteria decomposing petroleum hydrocarbon under magnetic action, magnetic field is produced by commercially available ferrite permanent-magnet iron, is used forever
Magnet is circular ferrite permanent magnet.Magnet sizeThick 10mm.Magnet and its magnetic field producing method are as shown in figure 1, institute
Magnetic field intensity is stated by adjusting permanent magnet spacing to adjust.
Taken in the 150mL conical flasks that 5mL seed liquors access culture medium of being degraded equipped with 50mL with the liquid-transfering gun after sterilizing, placed
In constant-temperature table, 100r/min, 30 DEG C of culture 7d.Using residual petroleum content in determined by ultraviolet spectrophotometry culture medium.
The present invention has following features compared with prior art：
(1) externally-applied magnetic field promotes the growth of oil-removing bacteria.Magnetic field promotes adaptability and growth of the oil-removing bacteria to new environment
Speed, when magnetic field intensity is 25mT, externally-applied magnetic field is not than plus the microorganism in magnetic field shifts to an earlier date 12 hours biomass and reaches stabilization.
(2) externally-applied magnetic field generates obvious facilitation to oil-removing bacteria oil removing efficiency.When externally-applied magnetic field, magnetic field intensity
When between 18mT-35mT can all facilitation be produced to oil-removing bacteria oil degradation efficiency, and when magnetic field intensity reaches 25mT
Microorganism brings up to 66.0% to the clearance of oil from 54.1%, is improve compared to the situation oil removal rate for being not added with magnetic field
11.9%, oil removal efficiency is effectively improved.
(3) externally-applied magnetic field can effectively improve the resistivity of oil-removing bacteria impact load.When additional 25mT magnetic fields, remove
Oily bacterium oil removal rate in the range of 100mg/L-2000mg/L all increases.As initial concentration of oil 1000mg/L, magnetic field pair
The reinforcing effect of oil-removing bacteria is the most obvious, and oil removal rate brings up to 69.4% from 61.5%, improves 7.9%, effectively enhances
The resistivity of oil-removing bacteria impact load.
(4) magnetic field of the present invention is produced by commercially available permanent magnet, and required magnetic field intensity is small, it is not necessary to consume electric energy, investment
Low, effect is good, and existing process flow is not changed, and big equipment is not increased, is a suitable China's national situation, economically viable reinforcing
Oily waste water treatment technology.
Brief description of the drawings
Fig. 1 magnet and magnetic field producing method；
Fig. 2 is influence of the magnetic field to oil removing bacteria growing；
Fig. 3 is influence of the magnetic field intensity to oil-removing bacteria oil removing efficiency；
Fig. 4 is influence of the magnetic field to oil-removing bacteria anti-shock loading.
Specific embodiment
The method for improving oil-removing bacteria oil removing efficiency of the invention specifically includes following process：
1st, strain selection
Selection is deposited in " China Committee for Culture Collection of Microorganisms's common micro-organisms center ", and accession designation number is
10283, it is named as the strain of Acinetobacter sp.B11.The oil-removing bacteria is one plant and is gathered from oil extraction waste water biochemical treatment tank
Activated sludge in the isolated bacterial strain for being capable of efficient degradation petroleum hydrocarbon.Single bacterium colony is rounded, 1~2mm of diameter, surface light
Sliding, moistening, center is slightly swelled, neat in edge, in tooth yellow.Bacterial strain is in rod-short, two ends blunt circle, typically in single or paired,
Do not produce gemma.Bacterial strain is carried out into Gram's staining, observation finds that bacterial strain can be redyed by sarranine, is Gram-negative.The bacterial strain
Catalase positive, oxidase negative, it is not necessary to growth factor, indoles experiment do not develop the color, it is impossible to make gelatin liquefaction, methyl red
Negative.D-Glucose, D- xyloses, L-arabinose, D-ribose can not be utilized, can be by the use of sodium citrate as unique carbon
Source, and can be by the use of potassium nitrate and ammonium sulfate as only nitrogen source.
2nd, culture medium is prepared
(1) culture presevation and activation medium (solid)：Beef extract 5.0g/L, peptone 10.0g/L, NaCl 5.0g/L,
Agar 20g/L, regulation pH is 7.2~7.4；
(2) seed liquid culture medium (liquid, 1L)：Beef extract 5.0g/L, peptone 10.0g/L, NaCl 5.0g/L, regulation
PH is 7.2~7.4；
(3) degraded culture medium (liquid, 0.05L)：NH4NO32.0g/L, K2HPO41.5g/L, KH2PO43.0g/L,
MgSO4·7H2O 0.1g/L, anhydrous CaCl20.01g/L, Na2EDTA·2H2O 0.01g/L, regulation pH is 7.2~7.4, one
Quantitative oil.
Above culture medium is in 121 DEG C of 20~30min of sterilizing.
3rd, actication of culture
The bacterium colony of the culture presevation medium slant for being stored in 4 DEG C of going bail for coats activation medium flat board, 30 DEG C of cultures 24
~36h.
4th, prepared by liquid seeds
Picking single bacterium colony is inoculated into seed liquid culture medium, 30 DEG C, 200r/min shaking table cultures on activation medium flat board
12h。OD600Bacteria suspension concentration is determined under wavelength and bacteria suspension is diluted to 0.8, it is stand-by as seed liquor.
5th, oil-removing bacteria decomposing petroleum hydrocarbon under magnetic action
The magnetic field is produced by commercially available ferrite permanent-magnet iron, and the permanent magnet for being used is circular ferrite permanent magnet.
Magnet sizeThick 10mm.Magnet and its magnetic field producing method are as shown in figure 1, the magnetic field intensity is by adjusting forever
Magnet spacing is adjusted.
Taken in the 150mL conical flasks that 5mL seed liquors access culture medium of being degraded equipped with 50mL with the liquid-transfering gun after sterilizing, placed
In constant-temperature table, 100r/min, 30 DEG C of culture 7d.Using residual petroleum content in determined by ultraviolet spectrophotometry culture medium.
Illustrate below and the present invention is described in more detail.
Magnetic field is produced by commercially available ferrite permanent-magnet iron, and the permanent magnet for being used is circular ferrite permanent magnet.Magnet
SizeThick 10mm.Magnet and its magnetic field producing method are as shown in Figure 1.
Magnetic field intensity is defined by filler Surface field intensity, i.e., be defined by culture dish or triangular flask bottom outer wall.Magnetic field
Generation direction be N poles vertically upward.The magnetic field intensity is adjusted by adjusting permanent magnet spacing.
Circular magnet is positioned in the stainless steel clamp of constant-temperature table, no permeable material is filled in magnet surface, by it is special this
Draw meter actual measurement adjustment Surface field intensity.
Embodiment 1：Influence of the magnetic field to oil removing bacteria growing
Go bail for be stored in 4 DEG C of medium slants Acinetobacter sp.B11 strains coat nutrient agar put down
Plate, after 30 DEG C of 24~36h of culture, picking single bacterium colony is inoculated into seed liquid culture medium, 30 DEG C, 200r/min shaking table cultures 12h.
Bacteria suspension concentration is determined under OD600 wavelength and bacteria suspension is diluted to 0.8, it is stand-by as seed liquor.Will according to 5% inoculum concentration
Bacterial strain seed liquor is inoculated into the degraded culture medium of 200mg/L petroleum concentrations.Difference additional 0,18,25, the 35, magnetic of 60mT intensity
, being compareed with the blank cultures for not connecing bacterium, 30 DEG C, shaking table culture 7 days, as a result see Fig. 2 under conditions of 100r/min.By scheming
2 experimental results understand that the microorganism under the effect of different magnetic field intensity differs greatly to the adaptability of oil-polluted water.35mT magnetic fields
Under intensity effect, there is appropriate shortening the laundering period of bacterial strain, and other magnetic field intensities have no significant effect to the bacterial strain laundering period.Work as bacterial strain
During into logarithmic phase, externally-applied magnetic field significantly promotes the speed of growth of microorganism.Wherein under the magnetic field intensity effect of 25mT,
Growth of microorganism is fastest, and exponential phase is most short, and bacterial strain biomass has maximum biology after reaching stabilization and stabilization at first
Amount.
Embodiment 2：Influence of the magnetic field intensity to oil-removing bacteria oil removing efficiency
Go bail for be stored in 4 DEG C of medium slants Acinetobacter sp.B11 strains coat nutrient agar put down
Plate, after 30 DEG C of 24~36h of culture, picking single bacterium colony is inoculated into seed liquid culture medium, 30 DEG C, 200r/min shaking table cultures 12h.
Bacteria suspension concentration is determined under OD600 wavelength and bacteria suspension is diluted to 0.8, it is stand-by as seed liquor.Will according to 5% inoculum concentration
Bacterial strain seed liquor is inoculated into the degraded culture medium of 200mg/L petroleum concentrations.Difference additional 0,18,25, the 35, magnetic of 60mT intensity
, compareed with the blank cultures for not connecing bacterium, 30 DEG C, shaking table culture 7 days under conditions of 100r/min.Microorganism is investigated not
Influence under being acted on magnetic field intensity to oil degradation efficiency.Microorganism sees Fig. 3 to the degradation effect of oil.Different magnetic field is strong
Spend and different influences are produced to microbiological petroleum degradation effect.When not having externally-applied magnetic field, microorganism is to the degradation rate of oil
54.1%, remaining oil concentration is 88.3mg/L, and when applied field strengths are improved, oil removal rate is improved therewith, and in magnetic field intensity
It is 66.0% that microorganism reaches maximum to the clearance of oil when reaching 25mT, is carried compared to the situation oil removal rate for being not added with magnetic field
It is high by 11.9%.
Embodiment 3：Influence of the magnetic field to oil-removing bacteria anti-shock loading
It is inoculated into after oil-removing bacteria is activated in fresh liquid petroleum culture medium, 25mT magnetic field intensities and is being not added with magnetic field
Under the conditions of, be placed in 30 DEG C, 100r/min shaking table cultures 7 days, initial concentration of oil is respectively 100,200,500,1000,1500,
2000、3000mg/L.Petroleum degradation rate and biomass are determined respectively, to investigate initial concentration of oil to microbiological petroleum degradation rate
Influence and magnetic field determine the optimum crude oil drop of microorganism to the reinforcing effect of oil removal rate under microorganism difference initial concentration of oil
Solution concentration.Degradation results of the microorganism under the conditions of different initial concentration of oil are shown in Fig. 4.Microorganism is to the degradation rate of oil with first
The rising of beginning oil concentration is in downward trend after first rising.As initial concentration of oil 1000mg/L, degraded of the microorganism to oil
Rate highest, the bacterial strain oil clearance for being not added with magnetic field and externally-applied magnetic field reinforcing respectively reaches 61.5% and 69.4%, and oil removal rate is carried
It is high by 7.9%.And the oil removing of magnetic field-intensification microorganism promotes highest to appear in 1500mg/L than being not added with the strains for degrading rate in magnetic field, now
The bacterial strain oil clearance for being not added with magnetic field and externally-applied magnetic field reinforcing respectively reaches 45.3% and 59.7%, magnetic field-intensification bacterial strain oil removing
Rate improves 14.4%.When initial concentration of oil is more than 1000mg/L, regardless of whether externally-applied magnetic field oil removal rate starts decline occur,
When initial concentration of oil reaches more than 2000mg/L, there is decline by a relatively large margin in oil removal rate, now regardless of whether externally-applied magnetic field
Petroleum degradation rate is not all influenceed significantly, degradation effect is poor.
Implementation of the invention is described in detail above by specific embodiment, it is therefore intended that help reader to more fully understand this hair
Bright Spirit Essence, but not as the restriction to the scope of the present invention.
Claims (5)
1. it is a kind of improve oil-removing bacteria oil removing efficiency method, it is characterized in that：
(1) actication of culture
The bacterium colony of the culture presevation medium slant for being stored in 4 DEG C of going bail for coats activation medium flat board, and 30 DEG C of cultures 24~
36h；The strain is Acinetobacter sp.B11, has been deposited in that " China Committee for Culture Collection of Microorganisms is common
Microorganism " center ", accession designation number is CGMCC Nc.10283；
(2) prepared by liquid seeds
Picking single bacterium colony is inoculated into seed liquid culture medium on activation medium flat board, 30 DEG C, 200r/min shaking table culture 12h,
OD600Bacteria suspension concentration is determined under wavelength and bacteria suspension is diluted to 0.8, it is stand-by as seed liquor；
(3) oil-removing bacteria decomposing petroleum hydrocarbon under magnetic action
Bacterial strain seed liquor is inoculated into the degraded culture medium of oil concentration 100mg/L-2000mg/L according to 5% inoculum concentration, outward
Plus the magnetic field of 18-60mT intensity carries out degradation treatment.
2. it is according to claim 1 improve oil-removing bacteria oil removing efficiency method, it is characterized in that：The group of culture presevation culture medium
As beef extract 5.0g/L, peptone 10.0g/L, NaCl 5.0g/L and agar 20g/L, regulation pH is 7.2~7.4.
3. it is according to claim 1 and 2 improve oil-removing bacteria oil removing efficiency method, it is characterized in that：Seed liquid culture medium
It is beef extract 5.0g/L, peptone 10.0g/L and NaCl 5.0g/L to constitute, and regulation pH is 7.2~7.4.
4. it is according to claim 1 and 2 improve oil-removing bacteria oil removing efficiency method, it is characterized in that：The group of degraded culture medium
As NH4NO3 2.0g/L、K2HPO4 1.5g/L、KH2PO4 3.0g/L、MgSO4·7H2O 0.1g/L, anhydrous CaCl2
0.01g/L and Na2EDTA·2H2O 0.01g/L, regulation pH is 7.2~7.4.
5. it is according to claim 3 improve oil-removing bacteria oil removing efficiency method, it is characterized in that：Degrade culture medium composition be
NH4NO3 2.0g/L、K2HPO4 1.5g/L、KH2PO4 3.0g/L、MgSO4·7H2O 0.1g/L, anhydrous CaCl20.01g/L and
Na2EDTA·2H2O 0.01g/L, regulation pH is 7.2~7.4.
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| CN101786772A (en) * | 2010-03-24 | 2010-07-28 | 哈尔滨工业大学 | Efficient biological treatment method for polymer-containing sewage in oil field |
| CN103114084A (en) * | 2013-01-28 | 2013-05-22 | 厦门大学 | Method for fixing oil decomposing bacteria by polyurethane foam |
| CN103803714A (en) * | 2014-02-27 | 2014-05-21 | 常州大学 | Method for synergistically degrading oil producing wastewater by using petroleum degrading bacteria |
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| CN103114084A (en) * | 2013-01-28 | 2013-05-22 | 厦门大学 | Method for fixing oil decomposing bacteria by polyurethane foam |
| CN103803714A (en) * | 2014-02-27 | 2014-05-21 | 常州大学 | Method for synergistically degrading oil producing wastewater by using petroleum degrading bacteria |
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