CN111100858A - Immobilized microbial agent, offshore spilled oil treatment device carrying microbial agent and offshore spilled oil treatment method - Google Patents
Immobilized microbial agent, offshore spilled oil treatment device carrying microbial agent and offshore spilled oil treatment method Download PDFInfo
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- CN111100858A CN111100858A CN201911340926.4A CN201911340926A CN111100858A CN 111100858 A CN111100858 A CN 111100858A CN 201911340926 A CN201911340926 A CN 201911340926A CN 111100858 A CN111100858 A CN 111100858A
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- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 20
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- E—FIXED CONSTRUCTIONS
- E02—HYDRAULIC ENGINEERING; FOUNDATIONS; SOIL SHIFTING
- E02B—HYDRAULIC ENGINEERING
- E02B15/00—Cleaning or keeping clear the surface of open water; Apparatus therefor
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- E—FIXED CONSTRUCTIONS
- E02—HYDRAULIC ENGINEERING; FOUNDATIONS; SOIL SHIFTING
- E02B—HYDRAULIC ENGINEERING
- E02B15/00—Cleaning or keeping clear the surface of open water; Apparatus therefor
- E02B15/04—Devices for cleaning or keeping clear the surface of open water from oil or like floating materials by separating or removing these materials
- E02B15/10—Devices for removing the material from the surface
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/32—Hydrocarbons, e.g. oil
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/08—Seawater, e.g. for desalination
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A20/00—Water conservation; Efficient water supply; Efficient water use
- Y02A20/20—Controlling water pollution; Waste water treatment
- Y02A20/204—Keeping clear the surface of open water from oil spills
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The invention discloses an immobilized microbial agent, a marine oil spill treatment device carrying the microbial agent and a marine oil spill treatment method, and belongs to the technical field of marine oil spill treatment. The invention relates to an immobilized microbial agent, which is immobilized microspheres prepared by adding a cross-linking agent solution into a mixed solution of sodium alginate, carbonized dry powder and petroleum degrading bacteria. The offshore spilled oil treatment device carrying the immobilized microbial agent can reduce the resistance of seawater and fully mix oil-containing seawater and the immobilized microbial agent; the treatment tank is internally provided with a plurality of feed bins, and can carry out grading treatment on the oil-containing seawater, thereby improving the treatment efficiency. The invention combines the microorganism restoring technology and the physical restoring technology, can collect, process and discharge the oil-containing seawater at the same time, and has the characteristics of convenience, flexibility, convenient operation, high processing efficiency, no secondary pollution and the like.
Description
Technical Field
The invention belongs to the technical field of offshore oil spill treatment, and particularly relates to an immobilized microbial agent, an offshore oil spill treatment device carrying the microbial agent, and an offshore oil spill treatment method.
Background
Petroleum, as blood in modern industries, plays an increasingly important role in human production and life. With the increase of oil demand, oil spill and oil leakage accidents frequently occur in the processes of exploitation, transportation, processing and use, and great threat is generated to marine environment. The water body pollution is closely related to human activities, and particularly, the pollution of the offshore slow-flow water body brings serious harm to human survival. In the method for remedying the marine oil pollution, compared with physical remediation and chemical remediation, microbial remediation has the characteristics of low cost, few remaining problems, no secondary pollution and the like, and is an ideal remediation method.
Although a plurality of related inventions exist in bioremediation of marine petroleum pollution at home and abroad, the method can not be widely applied in actual pollution treatment events, and the main reasons are as follows: firstly, the marine environment is complex, and microorganisms are difficult to gather due to the action of waves and tides, so that the repairing effect is poor; secondly, most of microbial preparations are added in a delineation area, so that the method is not suitable for the condition of large pollution area; and (III) most of microbial preparations are added at one time, so that the microbial preparations are not convenient to recover, and metabolites can generate unknown influence on the marine environment and change flora communities after microbial remediation. The offshore slow-flow water body has the characteristics of small self-fluidity and weak self-purification capacity, so that petroleum pollution is easy to accumulate, and the problems of difficulty in collecting and treating floating oil and the like in offshore micro-polluted sea area restoration are solved. Therefore, how to solve the technical problems of low microorganism density, small restoration area, inconvenient recovery and the like has important significance for the microorganism restoration of the actual sea pollution.
Disclosure of Invention
In view of the problems in the prior art, the invention aims to provide an immobilized microbial agent, an offshore spilled oil treatment device carrying the microbial agent and an offshore spilled oil treatment method. And placing the prepared immobilized microbial agent in a marine oil spill treatment device, putting the immobilized microbial agent into a petroleum-polluted sea area, adsorbing and fixing floating oil on the sea surface, and then performing biodegradation. Therefore, the combination of microbial remediation and physical remediation is realized, and the problems of low microbial density, small remediation area, inconvenience in recovery and the like in the microbial remediation process of the petroleum-polluted sea area are effectively solved, so that the efficiency of microbial remediation of the petroleum-polluted sea area is greatly improved.
In order to achieve the purpose, the invention adopts the following technical scheme:
an immobilized microbial agent is immobilized microspheres prepared by adding a cross-linking agent solution into a mixed solution of sodium alginate, carbonized dry powder and petroleum degrading bacteria; the petroleum degrading bacteria comprise a petroleum degrading bacterium belonging to the genus Halomonas and a biosurfactant-producing bacterium belonging to the genus Thiamine.
On the basis of the scheme, the petroleumThe ratio of the bacteria of the genus Halomonas to the bacteria of the genus Thiamine in the degrading bacteria is 1:1 (v/v); wherein the effective number of halomonas is 1 × 108-1×109cfu/mL, the effective number of viable bacteria of the thiamine-decomposing Bacillus bacteria is 2 multiplied by 107-2×108cfu/mL。
The bacteria of the genus Halomonas and the genus Bacillus thiamine are all commercially available or disclosed in the prior art.
On the basis of the scheme, the sodium alginate-carbonized dry powder-petroleum degrading bacteria mixed solution is prepared by the following method:
mixing the carbonized dry powder with a sterilized sodium alginate solution with the mass fraction of 3% -5%, wherein the ratio is 1% (m/v), adding a petroleum degradation bacterial suspension, and uniformly stirring to obtain a sodium alginate-carbonized dry powder-petroleum degradation bacterial mixed solution, wherein the ratio of the petroleum degradation bacterial suspension to the sodium alginate solution is 1:3-1:5 (v/v).
On the basis of the scheme, the cross-linking agent solution is CaCl with the mass fraction of 3% -5%2And (3) solution.
On the basis of the scheme, the carbonized dry powder is agricultural plant fibrous waste. And carbonizing to obtain dry powder.
On the basis of the scheme, the preparation method of the carbonized dry powder comprises the following steps:
cleaning the agricultural plant fibrous waste with clear water and ethanol solution, drying at 105 ℃ for 2h, crushing, screening to obtain particles of 2-4mm, introducing nitrogen at 220-280 ℃ and heating for 2h to obtain carbonized particles; and further grinding and crushing the carbonized particles until the particle size is less than 1mm, washing the obtained dry powder material for 2-3 times by using clear water and ethanol, washing off impurities on the surface, and drying for 2 hours at 105 ℃ to obtain the carbonized dry powder.
On the basis of the scheme, the preparation method of the immobilized microbial agent comprises the following steps:
dripping the mixed solution of sodium alginate, carbonized dry powder and petroleum degrading bacteria with cross-linking agent CaCl2Obtaining immobilized microbial agent in the solution, transferring the immobilized microbial agent into a closed bottle, and continuing to crossSoaking in the combined agent for a period of time, cleaning with sterilized normal saline for 2-3 times, soaking in distilled water, refrigerating at 4 deg.C, and taking out directly for use. CaCl2Can rapidly generate ion exchange with sodium alginate solution to generate gel. The principle of sodium alginate gel is that the gel is mixed with Ca2 +The reaction occurs and cannot occur with other substances.
An offshore spill treatment device comprising: the device comprises a treatment tank (1) and bin separation nets (2), wherein a cavity is formed in the treatment tank (1), a water inlet (11) is formed in front of the treatment tank (1), a water outlet (12) is formed in the rear of a ship body, the bin separation nets (2) are detachably connected with the treatment tank (1), and the immobilized microbial agent prepared by the method is arranged between the bin separation nets (2); the bin separation net (2) is used for limiting the immobilized microbial agent to pass through the bin separation net.
On the basis of the scheme, air bags (3) are fixedly installed on two sides of the treatment pool (1) respectively, and a propelling device (4) is installed on each air bag (3).
On the basis of the scheme, the treatment tank (1) comprises a treatment tank bottom plate (13), a side baffle (14), a water inlet baffle (15), an overflow weir baffle (16) and a treatment tank top plate (17), wherein the two sides of the treatment tank bottom plate (13) are fixedly connected with the side baffle (14) respectively, the front of the treatment tank bottom plate (13) is fixedly connected with the water inlet baffle (15), the rear of the treatment tank bottom plate (13) is fixedly connected with the overflow weir baffle (16), and the side baffle (14) is fixedly connected with the treatment tank top plate (17) above.
On the basis of the scheme, a clamping groove is formed in the side baffle (14), and the bin separation net (2) penetrates through the top plate (17) of the treatment tank to be inserted into the clamping groove.
On the basis of the scheme, a feed inlet (171) is formed in the top plate (17) of the treatment tank.
On the basis of the scheme, the feeding hole (171) is detachably connected with the feeding hole plug.
On the basis of the scheme, the water inlet (11) is T-shaped.
On the basis of the scheme, sawteeth are formed above the overflow weir baffle (16).
A method for treating offshore spilled oil is characterized by comprising the following steps:
(1) filling stage
Firstly, placing a stock bin separation net of the offshore oil spilling treatment device into a corresponding clamping groove in a treatment tank; then the immobilized microbial inoculum is filled into a storage bin from a feed inlet, and the filling amount is about two thirds of the volume of the storage bin; finally, plugging the feed inlet and finishing filling;
(2) application phase
Firstly, fixing two sections of elastic ropes on four lifting rings of a top plate of a treatment pool of the offshore oil spill treatment device according to diagonal lines respectively, and fixing a main lifting ring at the intersection of the diagonal lines of the ropes; then, a cantilever crane is used for hooking the main hoisting ring, the device is lifted to keep balance, and the device is slowly placed into an oil leakage point on the sea surface; after the device is stable, the main hoisting ring is separated from the hook, and the water is drained;
(3) adsorption and biodegradation stages
Operating the offshore oil spilling treatment device to enable the offshore oil spilling treatment device to move forward at a constant speed, enabling oil-containing seawater to flow into the treatment pool from an inlet and an outlet, enabling the oil-containing seawater to enter a storage bin after passing through a storage bin separation net, and starting an adsorption stage; the oil-containing seawater can fully contact with the immobilized microbial agent in the storage bin along with the flowing of the oil-containing seawater, and the oil is adsorbed by the immobilized microbial agent and then flows into the subsequent storage bin for further adsorption; the seawater after adsorption treatment flows through the last bin separation net and is discharged from the water outlet of the overflow weir; the adsorbed oil is degraded by the immobilized microbial agent; the hydraulic retention time is preferably ensured to be 0.5-5 hours.
(4) Recovery stage
After running for 1-4 hours, starting a recovery program by observing that the immobilized microbial agent is obviously broken or the degradation efficiency is obviously reduced. Stopping the offshore oil spill treatment device below the cantilever crane, hooking the main hoisting ring, and lifting the device; after the seawater in the treatment tank flows out, lifting to the shore; and taking down the main hoisting ring and the elastic rope, drawing out the bin separation net, and recovering the immobilized microbial agent from the water inlet.
(5) On-land treatment stage
The recovered immobilized microbial agent is subjected to land treatment. Comprises two aspects, if the breakage rate is higher, filtering out the broken immobilized microbial inoculum and discarding; if the degradation efficiency is obviously reduced, filtering a small amount of broken immobilized microbial inoculum and abandoning, putting the intact immobilized microbial inoculum into a terrestrial treatment pool, adding a proper amount of nutrient salt solutions such as nitrate, phosphate and the like (according to the proportion of N to P being within the range of 5: 1-10: 1), controlling the temperature at 30 ℃, stirring at the speed of 160rpm/min, carrying out activation treatment with the activation period being generally 1-2d, measuring the degradation rate and recovering to 70%, filtering out the immobilized microbial inoculum, and cleaning for reuse; and when the immobilized microbial agent cannot recover 70% after being activated, the immobilized microbial agent is discarded.
The technical scheme of the invention has the advantages
(1) The adopted immobilized carrier is a resource utilization of agricultural plant fibrous waste
The immobilized microbial agent is an immobilized carrier which is agricultural plant fibrous waste corn stalks. And N, P element contained in the corn straw can be used for the growth and reproduction of the petroleum degrading bacteria. The corn stalk can still provide nutrition for the growth of the petroleum degrading bacteria after being modified. The corn straw is used as an immobilized carrier of the petroleum degrading bacteria, and the resource utilization of the petroleum degrading bacteria can be realized.
(2) The combination of microbial repair technology and physical repair technology
Common sea surface oil removing devices (such as oil skimming boats) mostly adopt a physical method to recover floating oil and then carry out secondary treatment. The method is mostly suitable for the condition of large amount of floating oil on the sea surface, and is difficult to recover and treat the condition of small amount of floating oil. The device combines the microbial remediation technology and the physical remediation technology, can collect, treat and discharge the oil-containing seawater while aiming at the condition of the petroleum hydrocarbon places which float or emulsify after oil spilling through physical collection, and has the characteristics of convenience, flexibility, convenient operation, high treatment efficiency, no secondary pollution and the like.
(3) The device has advanced structure
From the perspective of improving the treatment efficiency of the oil-containing seawater, the structural design of the treatment device has the following advancement: the baffle at the water inlet of the treatment pool adopts a cambered surface design, and the front ends of the floating bodies on the two sides adopt a conical design, so that the resistance of the treatment device during sea surface advancing is favorably reduced; the water inlet of the treatment tank is designed into a T shape, so that the smooth entering of the oil-containing seawater on the surface layer is ensured, the seawater can uniformly flow up and down in the treatment tank, the transverse water inlet is long, the water inlet is mainly the oil-containing seawater on the surface layer, the vertical water inlet is short, the water on the lower layer can circulate, and the resistance is reduced; the treatment tank is divided into a plurality of bins, and the bins are filled with high-density immobilized microbial agents, so that the oily seawater can be subjected to graded treatment, and the treatment efficiency is improved; two propellers at the rear end of the floating body can be separately controlled, and the advancing and steering of the experimental device can be realized by adjusting the speed of the two propellers, so that the operation is more flexible.
(4) The treatment method has high efficiency
The device and the processing method can remove a certain amount of residual petroleum hydrocarbons such as floating oil and emulsified oil after physical collection, and can remove more than 70% of petroleum hydrocarbons in the petroleum hydrocarbon polluted area. The device and the treatment method are mainly suitable for the oil spill pollution of offshore oil production platforms, and are also suitable for the restoration of the petroleum pollution of coastal and coastal slow-flow water bodies such as seaports, wharfs and the like.
Drawings
FIG. 1 is a diagram showing the treatment effect of the offshore oil spill treatment method of the present invention;
FIG. 2 shows the effect of removing contaminants under different contamination conditions (1# treatment area of 50 m)2The average forward speed is 2m/min, the oil concentration is 5 percent, and the degradation rate is 78 percent; 2# treatment area 50m2The average forward speed is 3m/min, the oil concentration is 5 percent, and the degradation rate is 67 percent; the 3# treatment area was 50m2The average forward speed is 2m/min, the oil concentration is 10 percent, and the degradation rate is 71 percent; )
FIG. 3 is a schematic structural diagram of an offshore oil spill treatment device according to the present invention;
FIG. 4 is a view showing the internal structure of the treatment tank;
FIG. 5 is a bottom view of the device;
fig. 6 a rear view of the device.
The specific implementation mode is as follows:
terms used in the present invention have generally meanings as commonly understood by one of ordinary skill in the art, unless otherwise specified.
The present invention will be described in further detail with reference to the following data in conjunction with specific examples. The following examples are intended to illustrate the invention and are not intended to limit the scope of the invention in any way.
Example 1
Preparation of immobilized microbial agent
A. Carbonization of dry powder
Selecting agricultural plant fibrous waste (such as straw), cleaning with clear water and ethanol solution, drying at 105 deg.C for 2 hr, crushing, sieving to obtain 2-4mm granules, introducing nitrogen at 220-280 deg.C, and heating for 2 hr to obtain carbonized granules.
B. Preparation and storage of microbial inoculum
And further grinding and crushing the carbonized particles until the particle size is less than 1mm, washing the obtained carbonized dry powder material with clear water and ethanol for 2-3 times, then washing with absolute ethanol for 2-3 times, removing impurities on the surface, and drying at 105 ℃ for 2 hours. Mixing the dried carbonized dry powder with sterilized sodium alginate solution with mass fraction of 3% -5%, wherein the ratio of carbonized dry powder to sodium alginate solution is 1% (m/v), adding petroleum degrading bacteria suspension (composed of petroleum degrading halomonas bacteria and biosurfactant producing thiamine decomposing bacillus according to 1:1(v/v), wherein the effective activity number of halomonas bacteria is 1 × 108～1×109cfu/mL, the effective number of viable bacteria of the thiamine-decomposing Bacillus bacteria is 2 multiplied by 107-2×108cfu/mL), the ratio of the petroleum degrading bacteria liquid to the sodium alginate solution is 1:3-1:5(v/v), and the mixture is uniformly stirred to obtain the sodium alginate-carbonized dry powder-petroleum degrading bacteria mixed solution. Dropwise adding the sodium alginate-carbonized dry powder-petroleum degrading bacteria mixed solution into CaCl with the mass fraction of 3% -5%2Obtaining immobilized microorganism bacteria in the solutionAnd (3) preparing. Transferring the immobilized microbial agent into a closed bottle, continuously soaking in a cross-linking agent for a period of time, washing with sterilized normal saline for 2-3 times, finally soaking in distilled water, refrigerating at 4 ℃, and taking out directly for use.
The petroleum degradation experiment was carried out by using free bacteria (free bacteria group), a microbial agent (non-carbonized microsphere group) prepared from non-carbonized agricultural plant fibrous waste, and a microbial agent (carbonized microsphere group) prepared from carbonized agricultural plant fibrous waste, respectively. Wherein, the experiment of the free bacterium group is carried out by using the bacterium of the genus Halomonas and the bacterium of the genus Thiamine with the ratio of 1:1(v/v), diesel oil and seawater are added, and the ratio of the bacterium liquid, the diesel oil and the seawater is 1:5:100 (v/v); the experiment of the non-carbonized immobilized microbial agent group adopts the steps of mixing agricultural plant fibrous waste with a sterilized sodium alginate solution with the mass fraction of 4 percent, wherein the proportion is 1 percent (m/v), then adding petroleum degrading bacteria suspension halomonas and thiamine degrading bacteria, wherein the proportion is 1:1(v/v), the ratio of the petroleum degrading bacteria suspension (the number of the added bacteria solution is about the same as that of free bacteria solution) to the sodium alginate solution is 1:5(v/v), stirring uniformly to obtain a sodium alginate-dry powder-petroleum degrading bacteria mixed solution, and dripping CaCl dropwise2A solution, wherein an unhardened immobilized microbial agent is formed, and diesel oil, seawater and the unhardened immobilized microbial agent are added; the experiment of the carbonized microsphere group adopts the steps of mixing carbonized agricultural plant fibrous waste with a sterilized sodium alginate solution with the mass fraction of 4 percent, the proportion is 1 percent (m/v), then adding petroleum degrading bacteria suspension of halomonas and thiamine degrading bacteria with the proportion of 1:1(v/v), the ratio of the petroleum degrading bacteria suspension (the number of the added bacteria solution is about the same as that of free bacteria solution) to the sodium alginate solution is 1:5(v/v), stirring uniformly to obtain a sodium alginate-dry powder-petroleum degrading bacteria mixed solution, and dripping CaCl dropwise2And (3) forming a carbonized immobilized microbial agent, and adding diesel oil, seawater and the carbonized immobilized microbial agent. The results of the simultaneous measurement of the degradation rates are shown in FIG. 1, in which the degradation rate of free bacteria was 47%, the degradation rate of non-carbonized immobilized microbial agent was 58%, and the degradation rate of carbonized immobilized microbial agent was 58%The immobilized microbial inoculum is 72 percent.
Example 2
An offshore oil spill treatment device is shown in figures 3-6, and is of a catamaran-like structure overall, and is 12m long, 8m wide and 1m high. The main structure consists of a treatment pool and floating bodies on two sides.
(1) Treatment pool
The treatment tank is 12m long, 4m wide and 1m high and mainly comprises a water inlet baffle, a treatment tank bottom plate, an overflow weir baffle, a side baffle, a bin separation net and a treatment tank top plate.
The water inlet baffle is integrally of a cambered surface structure, so that the resistance of the treatment pool in the advancing process on the sea surface can be reduced; the upper end of the water inlet baffle is 0.2m away from the top plate of the treatment tank to form a transverse water inlet; a vertical water inlet is reserved in the center of the water inlet baffle, and the width and the length of the vertical water inlet are 0.4m and 0.7m respectively; the horizontal water inlet is connected with the vertical water inlet to form a T-shaped water inlet, so that not only can the main water inlet be the seawater containing oil on the surface layer, but also the flow of the seawater in the middle and at the bottom of the treatment tank can be ensured.
The bottom plate of the treatment tank is rectangular, the length of the bottom plate is 10m, the width of the bottom plate is 4m, the front end of the bottom plate is connected with the water inlet baffle, the rear end of the bottom plate is connected with the overflow weir baffle, and the two sides of the bottom plate are connected with the side baffles; three buckles are uniformly distributed in the front and at the back of the center of the outer side of the bottom plate of the treatment tank and are used for being fixed with floating bodies on two sides of the treatment tank.
The overflow weir baffle is rectangular, 4m long and 0.8m wide, the overflow weir baffle is serrated on the upper side and 0.2m away from the top plate of the treatment tank, and a water outlet is formed; the zigzag overflow weir can ensure that the water outlet is uniform and the stability is good.
The side baffles are 2 in number, are distributed in bilateral symmetry, have the length of 12m and the width of 1m, have arc-shaped bottom edges at the front ends and are connected with the water inlet baffle, and the rest edges are respectively connected with the bottom plate of the treatment tank, the overflow weir baffle and the top plate of the treatment tank; the inner sides of the baffles on the two sides are provided with 5 groups of clamping grooves which are distributed in pairwise symmetry and used for placing the bin separation net.
The bin separation net consists of a rectangular net frame and a separation net, the length of the net frame is slightly less than 4m, the net frame can be conveniently inserted into the clamping groove of the side baffle, the width of the net frame is slightly more than 1m, the net frame can be conveniently taken out of the clamping groove, and the thickness of the net frame is about 30 mm; the separation net is tightly stretched around the net frame, the diameter of the mesh is less than 5mm, and the immobilized microbial inoculum is prevented from flowing out; the feed bin separates the net totally 5, and the perpendicular to treatment tank bottom plate evenly distributed divides into four feed bins of equidimension with treatment tank inside.
The top plate of the treatment tank is rectangular, 12m in length and 4m in width, and two sides of the top plate are connected with the side baffles; a gap is reserved on the top plate of the treatment tank at the position corresponding to the bin separation net, the width of the gap is about 40mm, and the bin separation net can be conveniently inserted and taken out; a feed hole is reserved in the center of each bin on the top plate of the treatment tank, and the diameter of the feed hole is 50 mm; 3 same buckles are arranged at the outer side of the top plate of the treatment tank at the positions corresponding to the buckles of the bottom plate of the treatment tank and are used for being fixed with floating bodies at two sides of the treatment tank; near four angles of the top plate of the treatment tank, 4 hoisting rings are symmetrically distributed for fixing elastic ropes, so that the device is convenient to hoist.
(2) Floating body
The floating bodies are symmetrically distributed on the two sides of the treatment tank, and the number of the floating bodies is 2, the length of the floating bodies is 12.5m, the diameter of the floating bodies is 1m, and the floating bodies are mainly divided into an air bag and a propeller.
The whole air bag is wedge-shaped, 12m long and 1m in diameter and is divided into a front part and a rear part; the front end is conical and 2m long, and the tip is flush with the water inlet, so that the conical shape can reduce the resistance of the device in advancing on the sea surface; the rear end is cylindrical and 10m long, the top end is provided with 3 same buckles at the position corresponding to the buckles of the treatment tank top plate, and the bottom end is provided with 3 same buckles at the position corresponding to the buckles of the treatment tank bottom plate; the buckles on the two air bags are respectively connected with the buckles of the top plate and the bottom plate of the treatment tank through 6 steel pipes and are fixed.
The whole propeller is 0.5m long and 1m in diameter, and is mainly divided into four parts, namely a propeller, a protective ring, a coupling motor and a storage battery; the propeller provides power for the device to advance, and the rotating speed can be regulated and controlled; the protective ring can prevent impurities on the sea surface from winding the propeller and has a protective effect on the propeller; the coupling motor is fixed at the rear end of the air bag and connected with a propeller as a main power source; the storage battery is arranged inside the air bag for preventing the storage battery from contacting with seawater and supplies power for the coupling motor, and the rear end of the air bag is provided with a charging hole.
Example 3
Method for treating offshore spilled oil
(1) Filling stage
Firstly, placing a stock bin separation net into a corresponding clamping groove in a treatment tank; then the immobilized microbial inoculum is filled into a storage bin from a feed inlet, and the filling amount is about two thirds of the volume of the storage bin; and finally, plugging the feed inlet and finishing filling. The material can be divided into a plurality of sections by a fine mesh screen according to the adding amount. The effect of grading treatment can be achieved in the running process.
(2) Application phase
Firstly, fixing two sections of elastic ropes on four lifting rings of a top plate of a treatment pool according to diagonal lines respectively, and fixing a main lifting ring at the intersection of the diagonal lines of the ropes; then, a cantilever crane is used for hooking the main hoisting ring, the device is lifted to keep balance, and the device is slowly placed into an oil leakage point on the sea surface; after the device is stable, the main hoisting ring is separated from the hook, and the water is drained.
(3) Adsorption and biodegradation stages
Operating the offshore oil spilling treatment device to enable the offshore oil spilling treatment device to move forward at a constant speed, enabling oil-containing seawater to flow into the treatment pool from an inlet and an outlet, enabling the oil-containing seawater to enter a storage bin after passing through a storage bin separation net, and starting an adsorption stage; the oil-containing seawater can fully contact with the immobilized microbial agent in the storage bin along with the flowing of the oil-containing seawater, and the oil is adsorbed by the immobilized microbial agent and then flows into the subsequent storage bin for further adsorption; the seawater after adsorption treatment flows through the last bin separation net and is discharged from the water outlet of the overflow weir; the adsorbed oil is degraded by the immobilized microbial agent; the hydraulic retention time is preferably ensured to be 0.5-5 hours.
(4) Recovery stage
After the operation is carried out for 1-4 hours, a recovery program is started when the immobilized microbial inoculum is obviously crushed or the degradation efficiency is obviously reduced by observation. Stopping the offshore oil spill treatment device below the cantilever crane, hooking the main hoisting ring, and lifting the device; after the seawater in the treatment tank flows out, lifting to the shore; and taking down the main hoisting ring and the elastic rope, drawing out the bin separation net, and recovering the immobilized microbial agent from the water inlet.
(5) On-land treatment stage
The recovered immobilized microbial agent is subjected to land treatment. Comprises two aspects, if the breakage rate is higher, filtering out the broken immobilized microbial inoculum and discarding; if the degradation efficiency is obviously reduced, filtering a small amount of broken immobilized microbial inoculum and abandoning, putting the intact immobilized microbial inoculum into a terrestrial treatment pool, adding a proper amount of nutrient salt solutions such as nitrate, phosphate and the like (according to the proportion of N to P being within the range of 5: 1-10: 1), controlling the temperature at 30 ℃, stirring at the speed of 160rpm/min, carrying out activation treatment with the activation period being generally 1-2d, measuring the degradation rate and recovering to 70%, filtering out the immobilized microbial inoculum, and cleaning for reuse; and when the immobilized microbial agent cannot recover 70% after being activated, the immobilized microbial agent is discarded.
The foregoing is directed to preferred embodiments of the present invention, other and further embodiments of the invention may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention are within the protection scope of the technical solution of the present invention.
Claims (10)
1. An immobilized microbial agent is characterized in that immobilized microspheres are prepared by adding a cross-linking agent solution into a mixed solution of sodium alginate, carbonized dry powder and petroleum degrading bacteria;
the petroleum degrading bacteria comprise a petroleum degrading bacterium belonging to the genus Halomonas and a biosurfactant-producing bacterium belonging to the genus Thiamine.
2. The immobilized microbial agent according to claim 1, wherein the petroleum-degrading bacteria has a ratio of halomonas to thiamine-degrading bacillus of 1:1 (v/v); wherein the effective number of halomonas is 1 × 108-1×109cfu/mL, the effective number of viable bacteria of the thiamine-decomposing Bacillus bacteria is 2 multiplied by 107～2×108cfu/mL。
3. The immobilized microbial agent as claimed in claim 1 or 2, wherein the mixed solution of sodium alginate, carbonized dry powder and petroleum degrading bacteria is prepared by the following method:
mixing carbonized dry powder with a sterilized sodium alginate solution with the mass fraction of 3% -5%, wherein the proportion of the carbonized dry powder to the sodium alginate solution is 1% (m/v); then adding petroleum degrading bacteria suspension, wherein the ratio of the petroleum degrading bacteria suspension to the sodium alginate solution is 1:3-1:5 (v/v); stirring evenly to obtain the sodium alginate-carbonized dry powder-petroleum degrading bacteria mixed solution.
4. The immobilized microbial inoculant according to claim 3, wherein the carbonized dry powder is prepared by the following steps:
cleaning the agricultural plant fibrous waste with clear water and ethanol solution, drying at 105 ℃ for 2h, crushing, screening to obtain particles of 2-4mm, introducing nitrogen at 220-280 ℃ and heating for 2h to obtain carbonized particles; and further grinding and crushing the carbonized particles until the particle size is less than 1mm, washing the obtained dry powder material by using clear water and ethanol, washing to remove impurities on the surface, and drying at 105 ℃ for 2h to obtain the carbonized dry powder.
5. The method for preparing an immobilized microbial agent according to any one of claims 1 to 4, comprising the steps of:
dripping the mixed solution of sodium alginate, carbonized dry powder and petroleum degrading bacteria into a cross-linking agent solution to obtain an immobilized microbial agent; and transferring the prepared immobilized microbial agent into a closed bottle, continuously soaking in a cross-linking agent for a period of time, washing with sterilized normal saline for 2-3 times, finally soaking in distilled water, and refrigerating at 4 ℃ for later use.
6. The method for preparing an immobilized microbial agent according to claim 5, wherein the cross-linking agent solution is CaCl with a mass fraction of 3% -5%2And (3) solution.
7. An offshore oil spill treatment device, comprising: the device comprises a treatment tank (1) and bin separation nets (2), wherein a cavity is formed in the treatment tank (1), a water inlet (11) is formed in front of the treatment tank (1), a water outlet (12) is formed in the rear of a ship body, the bin separation nets (2) are detachably connected with the treatment tank (1), and the immobilized microbial agent prepared by the method of claim 6 is arranged between the bin separation nets (2); the bin separation net (2) is used for limiting the immobilized microbial agent to pass through the bin separation net.
8. Marine oil spill treatment device according to claim 7, characterized in that air bags (3) are fixedly arranged on two sides of the treatment pool (1) respectively, and the air bags (3) are provided with propulsion devices (4).
9. The offshore oil spill treatment device according to claim 7 or 8, wherein the treatment tank (1) comprises a treatment tank bottom plate (13), a side baffle plate (14), a water inlet baffle plate (15), an overflow weir baffle plate (16) and a treatment tank top plate (17), wherein two sides of the treatment tank bottom plate (13) are fixedly connected with the side baffle plate (14) respectively, the front part of the treatment tank bottom plate (13) is fixedly connected with the water inlet baffle plate (15), the rear part of the treatment tank bottom plate (13) is fixedly connected with the overflow weir baffle plate (16), and the upper part of the side baffle plate (14) is fixedly connected with the treatment tank top plate (17); the side baffle (14) is provided with a clamping groove, and the bin separation net (2) penetrates through the top plate (17) of the treatment tank to be inserted into the clamping groove.
10. A method for treating offshore spilled oil is characterized by comprising the following steps:
(1) filling stage
Firstly, installing a bin separation net of the offshore spilled oil treatment device in the treatment pool; then filling the immobilized microbial agent prepared by the method of claim 6 into a storage bin;
(2) application phase
Putting the offshore oil spill treatment device filled with the immobilized microbial agent into a sea surface oil leakage point;
(3) adsorption and biodegradation stages
Operating the offshore oil spill treatment device to enable oil-containing seawater to flow into the treatment pool from the inlet and the outlet, and enter the storage bin after passing through the storage bin separation net, wherein the adsorption stage begins; the oil-containing seawater can fully contact with the immobilized microbial agent in the storage bin along with the flowing of the oil-containing seawater, and the oil is adsorbed by the immobilized microbial agent and then flows into the subsequent storage bin for further adsorption; the seawater after adsorption treatment flows through the last bin separation net and is discharged from the water outlet of the overflow weir; the adsorbed oil is degraded by the immobilized microbial agent;
(4) recovery stage
After the adsorption stage is finished, lifting the offshore oil spill treatment device; after the seawater in the treatment tank flows out, lifting to the shore; recovering the immobilized microbial agent in the storage bin;
(5) on-land treatment stage
Filtering the recovered immobilized microbial agent to remove the broken immobilized microbial agent, activating, and measuring the degradation rate of the immobilized microbial agent to recover to 70% for recycling; if the degradation rate after activation still cannot be recovered to 70%, the material is discarded.
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