CN108624538A - A kind of composite bacteria agent and preparation method thereof for efficient process offshore oil sewage - Google Patents
A kind of composite bacteria agent and preparation method thereof for efficient process offshore oil sewage Download PDFInfo
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02F3/00—Biological treatment of water, waste water, or sewage
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Abstract
The invention belongs to field of environment microorganism, and in particular to a kind of offshore oil sewage disposal microbial bacterial agent and preparation method thereof.The composite bacteria agent composition includes 4 kinds of bacterial strains, respectively Ya De alkane eating bacteria (Alcanivorax jadensis), graceful alkane eating bacteria (Alcanivorax venustensis), Venice acinetobacter calcoaceticus (Acinetobacter venetianus), Maxwell Alteromonad (Alteromonas macleodii).Different strains play different function in handling petroleum waste water in microbial inoculum of the present invention, synergistic effect, it mutually promotes, the oil content being substantially reduced in petroleum waste water, including being a large amount of existing various straight-chain hydrocarbons, branched-chain hydrocarbons and aromatic hydrocarbon in oil, the industrialization and commercialization of the microbial bacterial agent of offshore oil wastewater efficient processing are helped to realize.
Description
Technical field
The invention belongs to field of environment microorganism, and in particular to a kind of offshore oil sewage disposal microbial bacterial agent and its system
Preparation Method.
Background technology
Oil pollution mostly occurs in marine environment, extremely serious to the harm of ocean and its periphery ecological environment,
Not only cause large area marine site anoxic, cause fish, shrimp, shellfish and seabird dead, oil slick pollutes seabeach, can also cause seabeach waste
Grassland destroys mariculture and Solar pond production, damages coastal recreational area, reduces the service function and value of marine ecosystems；This
Outside, many petroleum hydrocarbons especially polycyclic aromatic hydrocarbon can also be entered fish, shrimp by food chain, in body of shellfish, influence its economic valence
Value, and enter human body eventually by food enrichment, endanger human health.Because of its strong lasting harmfulness, oil pollution becomes
The biggest threat of the marine eco-environment.
Oil pollution is mainly due to the discharge of marine oil overflow oil accident and shop sump oil water.Ship oil wastewater mainly by
Ship ballast water, oil-containing tank washings and engine room water are constituted, and all kinds of harbours in current China are produced ship oil wastewater per year and are up to according to statistics
20000000 tons, IMO statistics, since petroleum transportation enters in 1,470,000 tons of oil of ocean every year, 700,000 tons are with ship
Oceangoing ship water ballast, tank washings and engine room water, which enter sea, to be caused, wherein 300,000 tons are with caused by engine room water discharge into sea.Shop sump oil water
Whereabouts be can not be ignored the problem of, development shop sump oil water technology is to reduce arranging at all for petroleum pollution in ocean
It applies.
Not only pollutant component is complicated for offshore oil sewage, also contains a large amount of salinity, and concentration variation is frequent, this is dirt
The processing of water adds many uncertain factors.Currently, the repair mode of petroleum-contaminated water is divided into two kinds, one is places in situ
It manages (in situ), i.e., the water body of pollution is not carried or conveyed, in the method that former place is repaired.For by petroleum hydrocarbon
The marine site of pollution, in-situ treatment method have the sides such as the oil fence, oil skimmer, oil absorption material, combustion (of oil) insitu, dispensing chemical oxidizing agent
Formula.There are oily waste treatments to be not thorough for such methods, be easy to cause secondary pollution problems.Another kind is dystopy processing (ex
Situ), i.e., petroleum waste water is transferred to sewage disposal place to handle, is discharged after up to standard.Dystopy processing method mainly passes through
Physiochemical mutagens, chemical emulsification and biodegradable coordinated are completed.Though the method for physics and chemistry is in oil pollution in-situ treatment
Aspect be easy to cause secondary pollution using more but limited to the Scavenging activity of petroleum hydrocarbon, and microbial method removes oil
Hydrocarbon is then that the specified microorganisms microbial inoculum that can carry out growth metabolism using petroleum hydrocarbon as carbon source is used to absorb and by Self substances generation
It thanks to decomposing petroleum hydrocarbon, is translated into thalline constituent and free of contamination CO2, to achieve the purpose that administer oil pollution.
During microbiological treatment oil pollution, petroleum hydrocarbon is oxidized to alcohol first, and phase is oxidized under the action of alcohol dehydrogenase
The aldehyde answered, aldehyde are then further oxidized to aliphatic acid by the effect of aldehyde dehydrogenase, and then participate in TCA cycles, are gradually degraded.Ring
Since the biodegradation of alkane then being oxidized to monohydric alcohol；The katabolism approach of compound fragrant hydrocarbon is in oxygen molecule
And the common metabolic intermediate of catechol derivatives is formed under the action of enzyme, then further pass through oxygen molecule and ring-opening enzyme
Effect, form it into straight chain molecule, finally decompose again into TCA recycle, realize the degradation of long-chain.For marine ship greasy dirt
The dystopy of water is handled, and petroleum waste water is handled using microbial method, is the generally acknowledged ideal for capturing oil pollution problem in the world today
Approach, advantage are embodied in：(1) petroleum pollution degradation is thorough, and will not cause secondary pollution, and (2) use peace without combustibility
Complete convenient, easy to operate, (3) are of low cost, non-corrosive, and (4) environmental suitability is strong, is applicable to different temperature conditions,
And tolerable hypersaline environment, (5) are applied widely.
Currently, finding that the microorganism of energy decomposing petroleum hydrocarbon there are more than 100 more than 200 kinds of categories or more, it is capable of the thin of decomposing petroleum hydrocarbon
Bacterium mainly has：Pseudomonas (Pseudomonas), Arthrobacter (Arthrobacter), acinetobacter
(Acinetobacter), alcaligenes (Alcaligennes), Micrococcus (Microcoecus), corynebacterium
(Corynebacterium), Flavobacterium (Flavobacterium), Rhod (Rhodococcus), achromobacter
(Acheromobacter), bacillus (Bacillus), Xanthomonas (Xanthomonas) etc..These petroleum hydrocarbons
Bacterium for degrading both is from fresh water or soil, it is difficult to the hypersaline environment for adapting to ocean or harbour petroleum waste water, for handling ship
Oceangoing ship oily water has comparable limitation.On the other hand, the country is for petroleum hydrocarbon under offshore oil sewage disposal and hypersaline environment
Certain work has also been made in the screening of degrading microorganism.Zheng Zhou etc. filters out 1 plant of false alternately list from 385 plants of South Pole marine bacterias
Born of the same parents bacterium and 1 plant of Colwell Salmonella, the two respectively reach 43.95% and 62.47% at 15 DEG C to the degradation rate of diesel oil.Forestry
Political affairs 26 plants of oil degradation bacterias of separation screening from the marine sediment that second of arctic scientific investigation of China acquires, the results showed that wherein
Three plants can respectively reach 30%-50% to the degradation rate of crude oil, and the petroleum hydrocarbon degradation bacterium being separated to is with Pseudoalteromonas
Dominant microflora accounts for 42%.
Petroleum hydrocarbon ingredient is extremely complex, without all components in any type microorganism energy degraded oil.It is international
Studies have shown that compared with single bacterium is degraded, composite flora can degrade the petroleum hydrocarbon of wider range.Deppe etc. has studied Chukchi Sea
Degradation potentiality of the homobium of 9 plants of bacteriums of middle separation to crude oil, the results showed that bacterial symbiont can degrade different alkane,
Including linear paraffin, branched paraffin and aromatic hydrocarbon.This research illustrates that composite flora is the ideal work of petroleum hydrocarbon contaminant processing
Tool.Compared with single culture system, artificial synthesized composite flora has better collaborative, suitable for being completed at the same time multinomial complexity
Work, and interaction is in dynamic equilibrium between flora, has stronger environmental suitability and functional stabilization；With the natural of complexity
Flora system is compared, then has better handling and safety.
Therefore, be based on systems biology and ecological theory, the functional characteristic for having grasped different oil degradation bacterias with
And on the basis of the action rule between flora, engineer is carried out to the composite flora suitable for oil polluted environment reparation, is opened
Definite functions, reliable and stable, easily controllable composite bacteria agent product are sent out, is ocean and harbour petroleum waste water biochemical processing hair
The new trend of exhibition.
Invention content
In consideration of it, it is necessary to provide a kind of efficiently reply ocean and harbour petroleum waste water microbial bacterial agent regarding to the issue above
Preparation method.This microbial bacterial agent preparation process is simple, and device requirement is brief, and operation is simple, and the environmental condition needed is mild.
It can effective degraded oil sewage largely existing straight-chain hydrocarbons, branched-chain hydrocarbons and aromatic hydrocarbon simultaneously.
The present invention is achieved by the following technical solutions：
A kind of composite bacteria agent for efficient process offshore oil sewage, composition includes 4 kinds of bacterial strains, and respectively Ya De is eaten
Alkane bacterium (Alcanivorax jadensis, strain number 216L-S13-9), graceful alkane eating bacteria (Alcanivorax
Venustensis, strain number 216L-S19-12), Venice acinetobacter calcoaceticus (Acinetobacter venetianus, bacterium
Strain number be WP02421), Maxwell Alteromonad (Alteromonas macleodii, strain number MA-S13-6)；Institute
It is respectively 1A08173,1A08174,1A00294,1A08617 to state 4 kinds of bacterial strain deposit numbers.
Further, the Ya De alkane eating bacteria bacterial strain in microbial inoculum a concentration of 1 × 107-109CFU/mL；The graceful food
Alkane bacteria strain in microbial inoculum a concentration of 1 × 107-109CFU/mL；Concentration of Venice acinetobacter calcoaceticus bacterial strain in microbial inoculum
It is 1 × 107-109CFU/mL；The Maxwell Alteromonas bacteria strain in microbial inoculum a concentration of 1 × 107-109CFU/mL.Four kinds
The concentration values of bacterium can appoint in affiliated range to be taken, the concentration values of 4 kinds of bacterium can it is identical, part is identical or complete
Difference, and it is suitable with when taking same concentrations value to the degradation effect of sewage through practical proof.
Further, the storage conditions of the bacterial strain are respectively：Ya De alkane eating bacteria (216L-S13-9) and graceful food alkane
Bacterium (216L-S19-12) used medium is No. 1001 culture mediums, and Venice acinetobacter calcoaceticus (WP02421) culture medium is No. 0033
Culture medium, Maxwell Alteromonad (MA-S13-6) used medium are No. 0471 culture medium.
Further, the method for preserving of the bacterial strain is bacterium solution and 40% glycerine (v/v) 1:1-1:1.5 mixing, set
In -80 DEG C of Storage in refrigerator.
Further, the production method of the complex microbial inoculum for efficient process offshore oil sewage of the invention,
Including：4 kinds of strains, are then inoculated with by the pure seed liquor that the progress seed liquor pure culture of each bacterial strain is first obtained to each plant of bacterium simultaneously again
It cultivates and is formed in microbial inoculum culture medium (S culture mediums).
Further, the preparation method of the microbial bacterial agent, sodium citrate is sole carbon wherein in seed liquid culture medium
Source.
Further, the preparation method of the microbial bacterial agent, wherein the carbon source of the microbial inoculum culture medium is sodium citrate
With 0# diesel oil.
Further, the preparation method of the microbial bacterial agent, wherein bacterial strain seed culture medium and microbial inoculum Medium's PH Value
Starting is 5.8-7.8.
Advantageous effect of the present invention：
Different strains play different function in handling petroleum waste water in microbial inoculum of the present invention, and synergistic effect is mutually promoted, from
And the treatment effect reached.The oil content that can be substantially reduced in petroleum waste water, including be a large amount of existing various in oil
Straight-chain hydrocarbons, branched-chain hydrocarbons and aromatic hydrocarbon help to realize industrialization and the quotient of the microbial bacterial agent of offshore oil wastewater efficient processing
Product.
Description of the drawings
Fig. 1：3 days variation tendencies of Fuel Oil Remaining in the oil-containing culture medium of 216L-S13-9 processing；Wherein 1-13 is straight chain
Hydrocarbon (C11-C24 is free of C23)；14-15 is branched-chain hydrocarbons (C15 and C19)；16-27 is aromatic hydrocarbon (C9-C12).
Fig. 2：3 days variation tendencies of Fuel Oil Remaining in the oil-containing culture medium of 216L-S19-12 processing；Wherein 1-13 is straight chain
Hydrocarbon (C11-C24 is free of C23)；14-15 is branched-chain hydrocarbons (C15 and C19)；16-27 is aromatic hydrocarbon (C9-C12).
Fig. 3：3 days variation tendencies of Fuel Oil Remaining in the oil-containing culture medium of WP02421 processing；Wherein 1-13 is straight-chain hydrocarbons
(C11-C24 is free of C23)；14-15 is branched-chain hydrocarbons (C15 and C19)；16-27 is aromatic hydrocarbon (C9-C12).
Fig. 4：3 days variation tendencies of Fuel Oil Remaining in the oil-containing culture medium of MA-S13-6 processing；1-13 is straight-chain hydrocarbons (C11-
C24 is free of C23)；14-15 is branched-chain hydrocarbons (C15 and C19)；16-27 is aromatic hydrocarbon (C9-C12).
Fig. 5：3 days variation tendencies of Fuel Oil Remaining in the oil-containing culture medium of composite bacteria agent processing of the present invention；1-13 is straight chain
Hydrocarbon (C11-C24 is free of C23)；14-15 is branched-chain hydrocarbons (C15 and C19)；16-27 is aromatic hydrocarbon (C9-C12).
Fig. 6：Fuel Oil Remaining 3 days in the oil-containing culture medium of 216L-S13-9,216L-S19-12 and WP02421 bacterium processing
Variation tendency；Wherein 1-13 is straight-chain hydrocarbons (C11-C24 is free of C23)；14-15 is branched-chain hydrocarbons (C15 and C19)；16-27 is virtue
Fragrant hydrocarbon (C9-C12).
Specific implementation mode
In order to better illustrate technical solution of the present invention problem to be solved, the technical solution of use and reach beneficial
Effect is expanded on further in conjunction with specific implementation mode.It is worth noting that technical solution of the present invention is including but not limited to following
Embodiment.
Particular technique or condition are not specified in the embodiment of the present invention, according to technology described in document in the art or
Condition is carried out according to product description.Reagents or instruments used without specified manufacturer, being can be by purchased in market etc.
The conventional products that approach obtains.
Culture medium composition according to the present invention is as follows：
Activated strains fluid nutrient medium (referred to as liquid HLB)：Tryptone (8-12g/L), yeast extract (4-6g/
L), NaCl (25-35g/L), water are deionized water.
Activated strains solid medium (referred to as solid HLB)：Tryptone (8-12g/L), yeast extract (4-6g/
L), NaCl (25-35g/L), agar powder (1.2-2.0g/L), water is deionized water.
Seed liquid culture medium (referred to as S)：Trisodium citrate dihydrate (15-25g/L), ammonium chloride (0.8-1.5g/L), three
Hypophosphite monohydrate hydrogen dipotassium (0.2-0.8g/L), potassium dihydrogen phosphate (0.1-0.4g/L), water is artificial seawater.
Microbial inoculum culture medium/microbial inoculum processing assay medium (referred to as D)：Ammonium chloride (0.8-1.5g/L), two citrate hydrates
Sour sodium (2-5g/L), 0# diesel oil (0.2%-1.5% (v/v)), dipotassium hydrogen phosphate trihydrate (0.2-0.8g/L), potassium dihydrogen phosphate
(0.1-0.4g/L) water is artificial synthetic seawater.
No. 1001 culture mediums：Sodium acetate (0.5-1.5g/L), peptone (8-12g/L), yeast powder (1-3g/L), common meat
Juice (0.1-0.8g/L), ammonium nitrate (0.1-0.3g/L), trisodium citrate (0.1-0.8g/L), potassium dihydrogen phosphate (0.2-0.8g/
L), pH 7.0-7.8, water are artificial synthetic seawater.
No. 0033 culture medium：Yeast powder (3-7g/L), peptone (8-12g/L), sodium chloride (5-15g/L), pH 6.5-
7.5 water is deionized water.
No. 0471 culture medium：Yeast powder (0.5-3g/L), peptone (2-8g/L), beef extract (0.1-3g/L), orthophosphoric acid
Iron (0.005-0.3g/L), pH 6.5-7.5, water are to remove artificial synthesized seawater.
Artificial synthesized seawater：Sodium chloride (20-28g/L), six water magnesium sulfates (8-14g/L), sodium sulphate (3-5g/L), six water
Calcium chloride (1-3g/L), potassium chloride (0.5-1.0g/L), potassium bromide (0.05-0.2g/L), boric acid (0.01-0.05g/L), nine water
Sodium metasilicate (4-8mg/L), six water strontium chlorides (0.02-0.08g/L), sodium fluoride (1-5mg/L), ammonium nitrate (1-6mg/L), phosphoric acid
Iron (0.5-1.5mg/L), pH 6.8-7.8, water are deionized water.
Embodiment one prepares petroleum waste water processing microbial bacterial agent of the present invention
Bacterial strain activation culture：Each bacterial strain glycerol tube strain that -80 DEG C preserve is inoculated in equipped with 2-5mL HLB Liquid Cultures
In the 15mL culture tubes of base, it is placed in 20-30 DEG C, cultivates 48-120h in the oscillation shaking table of 180-220r/min, then by activation
Bacterium solution lines respectively is placed in rejuvenation culture 24-96h in 20-30 DEG C of constant incubator in HLB solid mediums, will be on tablet
Bacterium colony is respectively inoculated in the 15mL culture tubes equipped with 2-5mLHLB fluid nutrient mediums in 20-30 DEG C, the oscillation of 180-220r/min
In shaking table overnight (12-16h) culture, be inoculated in S culture mediums (seed liquid culture medium) with 1% inoculum concentration, be placed in 20-30 DEG C,
12-18h is cultivated in the oscillation shaking table of 180-220r/min, then (20- is cultivated in S culture mediums by the switching of 0.5-2.5% inoculum concentrations
30 DEG C, 180-220r/min) 10-15h obtains each bacterial strain seed liquor.
It is prepared by petroleum waste water processing microbial bacterial agent of the present invention：Each bacterial strain seed liquor obtained above is inoculated with and is trained into microbial inoculum
It supports in base (D culture mediums), 20-30 DEG C, culture 8-16h obtains microbial inoculum in the oscillation shaking table of 180-220r/min, wherein four plants of bacterium
Concentration is all 1 × 10 respectively in microbial inoculum7-109CFU/mL。
Two petroleum waste water of the present invention of embodiment handles microbial bacterial agent functional evaluation
Single strain functional evaluation：4 plants of bacterium described in invention are pressed to 1 method of embodiment respectively and carry out bacterial strain activation culture,
It is centrifuged under 5000-8000 revs/min 3-6 minutes and collects thalline, obtained thalline will collected, base weight is cultivated with isometric S respectively
It is outstanding, D culture mediums are inoculated in 0.5-2.5% ratios respectively, the system is placed in 20-30 DEG C, the constant temperature of 180-220r/min
It is cultivated in shaking table.Since 0h, per sampling for 24 hours, it is remaining to carry out GC-MS detection oil for continuous sampling 3 times after being extracted with organic solvent
Amount.
Petroleum waste water of the present invention handles microbial bacterial agent function evaluation methods：By 4 plants of bacterium described in invention
Above-mentioned bacterial strains activation culture is pressed respectively, is centrifuged under 5000-8000 revs/min 3-6 minutes and is collected thalline, the thalline that collection is obtained
It is resuspended into S culture mediums, it is all 1 × 10 respectively to make four plants of bacterium concentration in S culture mediums8CFU/mL, then turned with 0.5%-2.5%
The amount of connecing accesses D culture mediums, as being handled 3 days in 20-30 DEG C, the oscillation shaking table of 180-220r/min.It is every to take for 24 hours since 0h
Sample, continuous sampling 3 times after being extracted with organic solvent, by GC-MS methods to the substance qualitative analysis that is degraded, and measure opposite
Fuel Oil Remaining obtains degradation rate.
Microbial bacterial agent function evaluation methods are handled according to petroleum waste water of the present invention, handle following 2 groups of control groups：
Composite bacteria agent control group 1：Preparation method is identical as composite bacteria agent of the present invention, difference lies in：Four kinds of bacterium in control group 1
The concentration of strain is 1 × 106CFU/mL.And similarly sample, with GC-MS methods to the substance qualitative analysis that is degraded, and measure
Relative surplus oil mass, obtains degradation rate.
Compound bacteria control group 2：Preparation method is identical as composite bacteria agent of the present invention, difference lies in：Only 3 kinds of bacterium in control group 2
Strain, respectively Ya De alkane eating bacteria (216L-S13-9), graceful alkane eating bacteria (216L-S19-12) and Venice acinetobacter calcoaceticus
(WP02421), the concentration of three kinds of bacterial strains is 1 × 108CFU/mL.And similarly sample, with GC-MS methods to the substance that is degraded
Qualitative analysis, and relative surplus oil mass is measured, obtain degradation rate.
Wherein GC-MS conditions are：Gas-chromatography (Agilent 7890A)；Mass detector (Agilent HP5975C)；
Chromatographic column is HP-5MS (30m × 0.25mm × 0.25 μm) capillary column；Injector temperature and detection temperature are 300,230 respectively
DEG C, sample size 1uL takes Splitless injecting samples, and auxiliary heating zone, level four bars temperature are respectively 280,150 DEG C, and carrier gas is nitrogen,
Flow velocity is 1mL/min.Temperature program：60 DEG C of holding 1min are warming up to 300 DEG C of holding 20min with 5 DEG C/min speed.MS is used
Full scan, scanning range (m/z) are 40-600.
It can be obtained from GC-MS analyses：
See Fig. 1,216L-S13-9 bacterium are best to the treatment effect of alkane and aromatic hydrocarbon under the same conditions, connect bacterium 72 hours
92% or more is reached to the removal rate of alkane afterwards, 90% or more is reached to the removal rate of aromatic hydrocarbon.
Seeing Fig. 2,216L-S19-12 bacterium are close to the treatment effect of straight-chain hydrocarbons, branched-chain hydrocarbons, aromatic hydrocarbon under the same conditions,
48 hours latter three removal rates of bacterium are connect 94% or more.
See Fig. 3, WP02421 bacterium are best to the treatment effect of straight-chain hydrocarbons under the same conditions, connect bacterium after 48 hours to straight chain
The removal rate of hydrocarbon reaches 95% or more.
See Fig. 4, MA-S13-6 bacterium are best to the treatment effect of aromatic hydrocarbon under the same conditions, connect bacterium after 72 hours to straight chain
The removal rate of hydrocarbon reaches 92% or more.
3 days post-processing hydro carbons average efficiencys of single strain are all 85% or more, wherein WP02421 and 216L-S13-9 processing
Effect is best, so hydro carbons removal average efficiency may be up to 94% or more.
See Fig. 5, degradation efficiency of the microbial bacterial agent of the present invention to straight-chain hydrocarbons, branched-chain hydrocarbons and aromatic hydrocarbon in petroleum waste water
All it is higher than any independent bacterial strain, it is interior for 24 hours that 95% or more is up to the removal of chain hydrocarbon and aromatic hydrocarbon.
The composite bacteria agent of control group 1 finds that the residual rate of each target compound is both greater than 91% when third day detects,
I.e. removal rate is less than 10%.
See Fig. 6, control group 2,216L-S13-9,216L-S19-12 and WP02421 bacterium (each bacteria concentration is all 1 ×
108CFU/mL) the effect of mixing post-processing petroleum waste water, for 24 hours after, fatty average removal efficiency is about 82%, aromatic hydrocarbon
Average removal efficiency be about 87%；After 3 days, the degradation efficiency of straight-chain hydrocarbons, branched-chain hydrocarbons and aromatic hydrocarbon in petroleum waste water is higher than
94%.
Several embodiments of the invention above described embodiment only expresses, the description thereof is more specific and detailed, but simultaneously
Cannot the limitation to the scope of the claims of the present invention therefore be interpreted as.It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the guarantor of the present invention
Protect range.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (8)
1. a kind of composite bacteria agent for efficient process offshore oil sewage, which is characterized in that the microbial inoculum composition includes 4 kinds of bacterium
Strain, respectively Ya De alkane eating bacteria, graceful alkane eating bacteria, Venice acinetobacter calcoaceticus, Maxwell Alteromonad.
2. composite bacteria agent according to claim 1, which is characterized in that concentration of the Ya De alkane eating bacteria bacterial strain in microbial inoculum
It is 1 × 107-109CFU/mL；It is described gracefulness alkane eating bacteria bacterial strain in microbial inoculum a concentration of 1 × 107-109CFU/mL；The Vinnie
This acinetobacter calcoaceticus bacterial strain in microbial inoculum a concentration of 1 × 107-109CFU/mL；The Maxwell Alteromonas bacteria strain is in microbial inoculum
A concentration of 1 × 107-109CFU/mL。
3. composite bacteria agent according to claim 1, which is characterized in that the storage conditions of the bacterial strain are respectively：Ya De
Alkane eating bacteria and graceful alkane eating bacteria used medium are No. 1001 culture mediums, and Venice acinetobacter calcoaceticus culture medium is No. 0033 culture
Base, Maxwell Alteromonad used medium are No. 0471 culture medium.
4. composite bacteria agent according to claim 1, which is characterized in that the method for preserving of the bacterial strain be bacterium solution and
40% glycerine presses 1:1-1:1.5 (v/v) are mixed, and are placed in -80 DEG C of Storage in refrigerator.
5. the production of the complex microbial inoculum for efficient process offshore oil sewage described in claim 1-4 any one
Method, which is characterized in that including：The progress seed liquor pure culture of each bacterial strain is first obtained to the pure seed liquor of each plant of bacterium, then again
4 kinds of strains are inoculated in microbial inoculum culture medium simultaneously and cultivates and is formed.
6. the preparation method of composite bacteria agent according to claim 5, which is characterized in that wherein, used in seed liquor culture
Sodium citrate is sole carbon source in culture medium.
7. the preparation method of composite bacteria agent according to claim 5, which is characterized in that wherein, the microbial inoculum culture medium
Carbon source is sodium citrate and 0# diesel oil.
8. the preparation method of composite bacteria agent according to claim 5, which is characterized in that the training used in bacterial strain seed liquor culture
The pH value starting for supporting base and microbial inoculum culture medium is 5.8-7.8.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110218689A (en) * | 2019-07-09 | 2019-09-10 | 中国科学院烟台海岸带研究所 | A kind of efficient offshore oil degradation composite bacteria agent and the preparation method and application thereof |
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| CN110699294A (en) * | 2019-11-07 | 2020-01-17 | 自然资源部第三海洋研究所 | Polycyclic aromatic hydrocarbon degrading bacteria and screening method and application thereof |
| CN110699294B (en) * | 2019-11-07 | 2021-06-18 | 自然资源部第三海洋研究所 | Polycyclic aromatic hydrocarbon degrading bacteria and screening method and application thereof |
| CN112574915A (en) * | 2020-12-15 | 2021-03-30 | 哈尔滨工业大学（威海） | Deep sea bacterium eurotium idescens and application thereof in inhibiting aflatoxin |
| CN114032204A (en) * | 2021-12-27 | 2022-02-11 | 郑州云益生物科技有限公司 | Composite microbial inoculum for treating marine petroleum sewage |
| CN114032204B (en) * | 2021-12-27 | 2024-02-20 | 郑州云益生物科技有限公司 | Composite microbial agent for treating offshore oil sewage |
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