CN107617639B - Method for treating polychlorinated biphenyl polluted soil by using microorganisms and mineral substances in combined manner - Google Patents
Method for treating polychlorinated biphenyl polluted soil by using microorganisms and mineral substances in combined manner Download PDFInfo
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Abstract
The invention belongs to the technical field of polluted soil treatment, and discloses a method for treating polychlorinated biphenyl polluted soil by using microorganisms and mineral substances in a combined manner, which comprises the following steps: 1) selecting 0-30cm of polychlorinated biphenyl polluted soil as a repairing layer, fully ploughing the soil of the repairing layer, and removing impurities in the soil; 2) taking out the biological preparation, and activating for 6 hours at the temperature of 28-32 ℃; then uniformly sowing the biological agent according to the amount of 10-30kg biological agent per mu, turning over, uniformly mixing, irrigating until saturation, and biologically treating for 2-4 days. The invention adopts the combination of biology and mineral substances to treat polychlorinated biphenyl soil pollution, and has simple and feasible treatment mode, high remediation efficiency and strong environmental friendliness.
Description
Technical Field
The invention belongs to the technical field of polluted soil treatment, and particularly relates to a method for treating polychlorinated biphenyl polluted soil by using microorganisms and mineral substances in a combined manner.
Background
Polychlorinated biphenyls (PCBs) polychlorinated biphenyls are chlorinated aromatic hydrocarbon compounds formed by replacing hydrogen on biphenyl molecules with 1-10 chlorine atoms at different positions through chemical catalytic reaction, the PCBs have excellent physicochemical properties such as thermodynamic stability, chemical inertness, nonflammability, high resistivity and the like, and are widely applied to the field of industrial production. The cleaning and disposal of PCBs has been greatly advanced, but due to the wide pollution range and long pollution time, the future work is still at random, and particularly, an economical and effective method is not available in the disposal technology.
At present, polychlorinated biphenyl polluted soil chemical remediation technologies comprise a chemical remediation technology and a biological remediation technology. The chemical remediation technology is to add chemical oxidants such as hydrogen peroxide, potassium permanganate and Fenton agent into the soil to cause the chemical oxidants to react with pollutants to realize the purpose of oxidizing and decomposing polychlorinated biphenyl in the soil. The chemical repair technology has high repair efficiency, but has higher operation cost, so that the wide application of the chemical repair technology is greatly limited. The bioremediation technology mainly adopts microorganisms for remediation, has little damage to the environment and high economic benefit, but has higher requirements on environmental conditions and long remediation period, and cannot be implemented due to the difficulty in finding efficient PCBs degrading microbial inoculum and the like. Therefore, the biological method which has controllable repairing condition, accords with environment-friendly development and can efficiently repair the soil PCBs pollution is developed, and has important significance and urgency for repairing the polluted PCBs soil.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides a method for treating polychlorinated biphenyl polluted soil by combining microorganisms and minerals.
The invention is realized by adopting the following technical scheme:
a method for treating polychlorinated biphenyl polluted soil by utilizing microorganisms and minerals in a combined mode comprises the following steps:
1) selecting 0-30cm of polychlorinated biphenyl polluted soil as a repairing layer, fully ploughing the soil of the repairing layer, and removing impurities in the soil;
2) taking out the biological preparation, and activating for 6 hours at the temperature of 28-32 ℃; then uniformly sowing the biological agent according to the amount of 10-30kg biological agent per mu, turning over, uniformly mixing, irrigating until saturation, and biologically treating for 2-4 days.
Preferably, the biological agent is uniformly spread in an amount of 20kg biological agent per mu.
Preferably, the biological treatment is for 3-4 days.
Specifically, the biological agent comprises rhodopseudomonas palustris, citrobacter freundii, shewanella, pseudomonas putida and bacillus pumilus.
Further, the air conditioner is provided with a fan,
the biological agent is prepared according to the following steps:
1) adding 8g of corn steep liquor, 6g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.2g of monopotassium phosphate, 0.1g of sodium chloride, 0.1g of calcium chloride, 0.01g of ferrous sulfate and 0.01g of manganese sulfate into water, uniformly stirring, adjusting the pH to 7.0-7.5, and then fixing the volume to 1L to prepare a liquid fermentation culture medium;
2) activating rhodopseudomonas palustris, citrobacter freundii, shewanella, pseudomonas putida and bacillus pumilus, respectively culturing to obtain seed solutions with the concentration of 0.1-0.5 hundred million cfu/ml, uniformly mixing the five seed solutions according to the volume ratio of 4-7:3-5:3-5:2-3:2-3, then inoculating the seed solutions into a fermentation tank containing a liquid fermentation culture medium according to the inoculation amount of 10%, fermenting and culturing for 12 hours, adding 0.1-0.2% of triton-100 in parts by mass, and continuously culturing for 6 hours to obtain mixed fermentation liquor;
3) crushing zeolite, sieving with a 100-mesh sieve, and uniformly mixing with turfy soil and triton-100 according to the mass ratio of 500-800:1000-1500:1-2 to prepare a mineral carrier;
4) adding the mixed fermentation broth into mineral carrier 1.5 times the weight of the mixed fermentation broth, stirring at 20 deg.C and 100rpm for 30min, and standing at 4 deg.C for 24-48 hr.
Preferably, the first and second electrodes are formed of a metal,
the conditions of the fermentation medium are as follows: the temperature is 28-32 deg.C, the pressure is 0.5Mpa, and the ventilation rate is 1: 0.5-1.
Preferably, the first and second electrodes are formed of a metal,
the rhodopseudomonas palustris adopts a strain with a deposit number of ATCC 17001; the Citrobacter freundii strain was the strain deposited under accession number ATCC 8090; the Shewanella is the strain deposited with ATCC 700550; the pseudomonas putida adopts a strain with a deposit number of ATCC 700007; the Bacillus pumilus adopts a strain with a deposit number of ATCC 7061.
The invention effectively shows that each strain has reasonable compatibility, can effectively promote the degradation of polychlorinated biphenyl pollutants, and needs to adjust the input amount and operation details of biological agents according to different polychlorinated biphenyl contents in polluted soil in practical application.
The strains of the present invention belong to known strains and can be purchased from ATCC and other commercial sources. The activation of the various species and seed culture steps of the present invention are conventional in the art and are not innovative in the present invention and will not be described in detail herein. The starting materials or reagents used in the present invention are commercially available unless otherwise specified.
Compared with the prior art, the invention has the advantages that the following aspects are mainly included but not limited:
the invention adopts the combination of biology and mineral substances to treat polychlorinated biphenyl soil pollution, the treatment mode is simple and feasible, the remediation efficiency is high, and the environment friendliness is strong;
the biological preparation has the advantages of reasonable compatibility of all strains, symbiotic coordination, no mutual antagonism, simple and convenient preparation method, easy method, and effective treatment of polychlorinated biphenyl soil pollution by combining with mineral substances;
the invention adopts two minerals of zeolite and turfy soil, which are mixed according to a certain proportion and used as a carrier and an accelerant, the trace elements can be released when the zeolite and the turfy soil are added into soil, and the zeolite and the turfy soil have a certain adsorption effect on pollutants, can also improve the water retention and ventilation performance of biological preparations, and provide good proliferation conditions for microorganisms, thereby improving the removal of polychlorinated biphenyl pollutants by the microorganisms; the addition of the triton-100 in the fermentation process can stimulate the bacterial strains to generate the expression quantity of various enzyme proteins, and based on the fact that polychlorinated biphenyl in the actual polluted soil is difficult to be utilized by microorganisms, the addition of the surfactant triton-100 on the basis of the combined microorganism remediation technology can also effectively improve the desorption rate of pollutants from the soil, and provide conditions for the catalytic degradation of the microorganisms.
The research shows that five strains are adopted for compatibility, the strains are mutually promoted, the degradation capability of the microorganism on the polychlorinated biphenyl is improved, and the effect is superior to that of comparative examples 1-5; the zeolite powder and the triton-100 in the carrier strengthen the removal of polychlorinated biphenyl in soil by microorganisms, and compared with the comparative example 6 (only turfy soil is adopted as the carrier), the effect is greatly improved, and the application prospect is wide.
Drawings
FIG. 1: influence of treatment time on the polychlorinated biphenyl removing effect;
FIG. 2: the influence of the application amount of the biological preparation on the removal effect of the polychlorinated biphenyl;
FIG. 3: different biological agents are used for treating polychlorinated toluene polluted soil.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present application, the technical solutions in the present application will be clearly and completely described below with reference to specific embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
A method for treating polychlorinated biphenyl polluted soil by utilizing microorganisms and minerals in a combined mode comprises the following steps:
1) selecting 0-30cm of polychlorinated biphenyl polluted soil as a repairing layer, fully ploughing the soil of the repairing layer, and removing impurities in the soil;
2) taking out the biological preparation, and activating for 6 hours at the temperature of 28 ℃; then uniformly sowing the biological agent according to the amount of 20kg biological agent per mu, turning over, uniformly mixing, irrigating until saturation, and biologically treating for 3 days.
The biological agent is prepared from five bacteria of rhodopseudomonas palustris, citrobacter freundii, shewanella, pseudomonas putida and bacillus pumilus;
specifically, the biological agent is prepared according to the following steps:
1) adding 8g of corn steep liquor, 6g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.2g of monopotassium phosphate, 0.1g of sodium chloride, 0.1g of calcium chloride, 0.01g of ferrous sulfate and 0.01g of manganese sulfate into water, uniformly stirring, adjusting the pH to 7.0-7.5, and then fixing the volume to 1L to prepare a liquid fermentation culture medium;
2) activating rhodopseudomonas palustris, citrobacter freundii, shewanella, pseudomonas putida and bacillus pumilus, respectively culturing to obtain seed solutions with the concentration of 0.3 hundred million cfu/ml, uniformly mixing the five seed solutions according to the volume ratio of 4:3:3:2:2, inoculating the seed solutions into a fermentation tank containing a liquid fermentation culture medium according to the inoculation amount of 10%, controlling the temperature to be 28-32 ℃, the tank pressure to be 0.5Mpa and the ventilation amount to be 1:0.5-1, culturing for 12 hours, adding 0.1% of triton-100 in parts by mass, and continuously culturing for 6 hours to obtain a mixed fermentation liquid;
3) crushing zeolite, sieving with a 100-mesh sieve, and uniformly mixing with turfy soil and triton-100 according to a mass ratio of 500:1000:1 to prepare a mineral carrier;
4) adding the mixed fermentation broth into mineral carrier 1.5 times the weight of the mixed fermentation broth, stirring at 20 deg.C and 100rpm for 30min, and standing at 4 deg.C for 24 hr.
The rhodopseudomonas palustris adopts a strain with a deposit number of ATCC 17001; the Citrobacter freundii strain was the strain deposited under accession number ATCC 8090; the Shewanella is the strain deposited with ATCC 700550; the pseudomonas putida adopts a strain with a deposit number of ATCC 700007; the Bacillus pumilus adopts a strain with a deposit number of ATCC 7061.
Example 2
A method for treating polychlorinated biphenyl polluted soil by utilizing microorganisms and minerals in a combined mode comprises the following steps:
1) selecting 0-30cm of polychlorinated biphenyl polluted soil as a repairing layer, fully ploughing the soil of the repairing layer, and removing impurities in the soil;
2) taking out the biological preparation, and activating for 6 hours at the temperature of 30 ℃; then uniformly sowing the biological agent according to the amount of 20kg biological agent per mu, turning over, uniformly mixing, irrigating until saturation, and biologically treating for 4 days.
The biological agent is prepared from five bacteria of rhodopseudomonas palustris, citrobacter freundii, shewanella, pseudomonas putida and bacillus pumilus;
specifically, the biological agent is prepared according to the following steps:
1) adding 8g of corn steep liquor, 6g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.2g of monopotassium phosphate, 0.1g of sodium chloride, 0.1g of calcium chloride, 0.01g of ferrous sulfate and 0.01g of manganese sulfate into water, uniformly stirring, adjusting the pH to 7.0-7.5, and then fixing the volume to 1L to prepare a liquid fermentation culture medium;
2) activating rhodopseudomonas palustris, citrobacter freundii, shewanella, pseudomonas putida and bacillus pumilus, respectively culturing to obtain seed solutions with the concentration of 0.5 hundred million cfu/ml, uniformly mixing the five seed solutions according to the volume ratio of 7:5:5:3:3, inoculating the seed solutions into a fermentation tank containing a liquid fermentation culture medium according to the inoculation amount of 10%, controlling the temperature to be 28-32 ℃, the tank pressure to be 0.5Mpa and the ventilation amount to be 1:0.5-1, culturing for 12 hours, adding 0.2% of triton-100 in parts by mass, and continuously culturing for 6 hours to obtain a mixed fermentation liquid;
3) crushing zeolite, sieving with a 100-mesh sieve, and uniformly mixing with turfy soil and triton-100 according to the mass ratio of 800: 1500:1 to prepare a mineral carrier;
4) adding the mixed fermentation broth into mineral carrier 1.5 times the weight of the mixed fermentation broth, stirring at 20 deg.C and 100rpm for 30min, and standing at 4 deg.C for 48 hr.
The rhodopseudomonas palustris adopts a strain with a deposit number of ATCC 17001; the Citrobacter freundii strain was the strain deposited under accession number ATCC 8090; the Shewanella is the strain deposited with ATCC 700550; the pseudomonas putida adopts a strain with a deposit number of ATCC 700007; the Bacillus pumilus adopts a strain with a deposit number of ATCC 7061.
Example 3
Effect of treatment time and applied amount of biological agent on polychlorinated biphenyl removal:
1. the treatment time is respectively set to be 24h, 48h, 72h, 96h and 120h, the contaminated soil with the polychlorinated biphenyl content of 50mg/kg is selected as a sample, and the rest operation procedures are the same as those in the embodiment 2. The results of detecting the removal rate of polychlorinated biphenyl are shown in fig. 1, wherein the removal rate of polychlorinated biphenyl is about 52.1% in 24 hours, 80.3% in 48 hours, 96.5% in 72 hours and more than 99% in 96 hours later.
2. The application rates of the biological agents are respectively set to be 5 kg/mu, 10 kg/mu, 20 kg/mu, 40 kg/mu and 80 kg/mu, the contaminated soil with the polychlorinated biphenyl content of 50mg/kg is selected as a sample, and the rest operation procedures are the same as those in the example 2. The removal rate of polychlorinated biphenyl is detected, and the result is shown in fig. 2, the removal rate reaches 99% when the application amount of the biological agent is 20 kg/mu, and the improvement range of the removal rate by increasing the application amount is not large after the application amount is increased to more than 20 kg/mu.
Example 4
The test for treating polychlorinated biphenyl polluted soil comprises the following steps:
sample selection and operation procedure: selecting polluted soil containing polychlorinated biphenyl of 40mg/kg, wherein each land is 10 mu and divided into 7 lands, and biological agents adopted by each land are different and are respectively embodiment 1 and comparative examples 1-6, and the specific operation flow is the same as that of embodiment 1; wherein, the biological agent is prepared by the following steps of 1, 2, 3, 4 and 1, wherein the biological agent is prepared by the following steps of 1, 2, 3, 4, 1; comparative example 5 (no addition of biological preparation of Bacillus pumilus, other same as example 1); comparative example 6 (peat only was used as mineral carrier, otherwise the same as example 1). The invention also selects the contaminated soil with polychlorinated biphenyl content of 80mg/kg for testing, and the rest is the same as the above.
The detection method comprises the steps of extracting a treated soil sample for 24 hours by using n-hexane and acetone according to the volume ratio of 1:1, concentrating an extracting solution to 1-2m L by using a rotary evaporator, dissolving a concentrated solution in 50m L n-hexane, purifying the solution by using a Florisil column, blowing the purified solution to 1m L by using a nitrogen blowing instrument, and measuring the content of polychlorinated biphenyl in the solution by using gas chromatography.
The results show that: as shown in figure 3, the biological agent can treat high-concentration polychlorinated toluene pollutants, 40mg/kg of polluted soil is treated in 3 days, the concentration of polychlorinated biphenyl is reduced to 1.1mg/kg, the removal rate is more than 97%, 80mg/kg of polluted soil is treated, the concentration of polychlorinated biphenyl is reduced to 2.9mg/kg, the removal rate is more than 96%, and the removal rate is greatly superior to that of comparative examples 1-6. The research shows that five strains are adopted for compatibility, the strains are mutually promoted, the degradation capability of the microorganism on the polychlorinated biphenyl is improved, and the effect is superior to that of comparative examples 1-5; the zeolite powder and the triton-100 in the carrier strengthen the removal of polychlorinated biphenyl in soil by microorganisms, and compared with the comparative example 6 (only turfy soil is adopted as the carrier), the effect is greatly improved, and the method has extremely high popularization and application values and wide application prospects.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (5)
1. A method for treating polychlorinated biphenyl polluted soil by utilizing microorganisms and minerals in a combined mode comprises the following steps:
1) selecting 0-30cm of polychlorinated biphenyl polluted soil as a repairing layer, fully ploughing the soil of the repairing layer, and removing impurities in the soil;
2) taking out the biological preparation, and activating for 6 hours at the temperature of 28-32 ℃; then uniformly sowing the biological agents according to the amount of 10-30kg biological agents per mu, turning over, uniformly mixing, irrigating until saturation, and carrying out biological treatment for 2-4 days;
the biological agent is prepared according to the following steps:
(1) adding 8g of corn steep liquor, 6g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.2g of monopotassium phosphate, 0.1g of sodium chloride, 0.1g of calcium chloride, 0.01g of ferrous sulfate and 0.01g of manganese sulfate into water, uniformly stirring, adjusting the pH to 7.0-7.5, and then fixing the volume to 1L to prepare a liquid fermentation culture medium;
(2) activating rhodopseudomonas palustris, citrobacter freundii, shewanella, pseudomonas putida and bacillus pumilus, respectively culturing to obtain seed solutions with the concentration of 0.1-0.5 hundred million cfu/ml, uniformly mixing the five seed solutions according to the volume ratio of 4-7:3-5:3-5:2-3:2-3, then inoculating the seed solutions into a fermentation tank containing a liquid fermentation culture medium according to the inoculation amount of 10%, fermenting and culturing for 12 hours, adding 0.1-0.2% of triton-100 in parts by mass, and continuously culturing for 6 hours to obtain mixed fermentation liquor;
(3) crushing zeolite, sieving with a 100-mesh sieve, and uniformly mixing with turfy soil and triton-100 according to the mass ratio of 500-800:1000-1500:1-2 to prepare a mineral carrier;
(4) adding the mixed fermentation broth into mineral carrier 1.5 times the weight of the mixed fermentation broth, stirring at 20 deg.C and 100rpm for 30min, and standing at 4 deg.C for 24-48 hr.
2. The method of claim 1, wherein the biological agent is uniformly applied in an amount of 20kg biological agent per acre.
3. The method of claim 1, wherein the biological treatment is for 3 to 4 days.
4. The method of claim 1, wherein the conditions of the fermentation culture are: the temperature is 28-32 deg.C, the pressure is 0.5Mpa, and the ventilation rate is 1: 0.5-1.
5. The method according to claim 1, wherein the Rhodopseudomonas palustris is a strain deposited under accession number ATCC 17001; the Citrobacter freundii adopts a strain with a deposit number of ATCC 8090; the Shewanella is the strain deposited with ATCC 700550; the pseudomonas putida adopts a strain with a deposit number of ATCC 700007; the Bacillus pumilus adopts a strain with a deposit number of ATCC 7061.
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