CN105733599B - A kind of cadmium pollution soil reparation fixative and its methods for making and using same based on microorganism assembling synthesis - Google Patents
A kind of cadmium pollution soil reparation fixative and its methods for making and using same based on microorganism assembling synthesis Download PDFInfo
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Abstract
The invention discloses a kind of cadmium pollution soils based on microorganism assembling synthesis to repair fixative and its methods for making and using same.The nano phosphate class fixatives for cadmium pollution soil reparation many at present cause secondary pollution because it easily disperses, is easy to run off and leads to body of groundwater eutrophication.For these problems, the present invention wraps up nanometer hydroxyapatite using nanometer hydroxyapatite as center substance, using aspergillus niger mycelia, is made for the problem that the cadmium pollution soil of high pollution highly acidity repairs fixative, is excessively dispersed in practical applications with solving it.The product of synthesis is uniformly mixed with cadmium pollution soil by a certain percentage, available Cd content reduces 67-86%.Fixative of the present invention, preparation process is simple, low in cost, high effect nontoxic evil, biodegradable, without secondary pollution, is a kind of environmentally friendly fixative.
Description
Technical field
The present invention relates to micro-nano materials to assemble the technical field in conjunction with cadmium pollution soil reparation, and in particular to one kind is applicable in
In the fixative product and its methods for making and using same of cadmium pollution soil reparation.
Background technique
In recent years, the soil pollutions case such as " cadmium rice ", " heavy metal vegetables ", " blood lead event " is appeared in the newspapers again and again, causes public affairs
Many greatly concerns.Combine " the national soil pollution situation tune of publication in April, 2014 with Ministry of Land and Resources according to Environmental Protection Department
Look into bulletin " it shows, national soil environment situation totally allows of no optimist, and some areas soil pollution is heavier, arable soil environment matter
Amount causes anxiety, industry and mining discarded ground soil environment outstanding problem.Based on inorganic type, organic type takes second place pollution type, inorganic pollution
Exceeded digit of object accounts for the 82.8% of all exceeded points." the environment five poisonous creatures: scorpion, viper, centipede, house lizard, toad " cadmium, lead, arsenic, chromium, mercury inorganic pollution point are exceeded
Rate is respectively 7.0%, 1.5%, 2.7%, 1.1%, 1.6%.Cadmium (Cd) in soil is mainly derived from a large amount of exploitations in mining area
And Wastewater Irrigation in Farmland, agricultural output, quality and entire farmland ecosystem are not only influenced into the heavy metal in soil, and can lead to
Cross the health that food chain more seriously endangers the mankind and other biologies.It can be seen that being closed to China's cadmium (Cd) contaminated soil
The effective reparation of reason is extremely urgent.
Currently, the improvement of cadmium pollution mainly has two thinkings in soil: first is that removalization, that is to say, that by heavy metal from
It is removed in soil environment system；Second is that heavy metal, i.e., be fixed in soil by immobilization using fixative.After all, this two
The purpose of kind mode is provided to the migration reduced heavy metal in the soil and reduces its biological effectiveness.Think around this two
Road, development go out a series of recovery techniques, mainly have engineering technology (including soil replacement method, soil improvement by importing soil from other places etc.), physical method (including
Electro reclamation, electric heating repairing method etc.), chemical method (including chemical leaching method, curing/stabilizing method etc.), bioanalysis (including plant
Object reparation, microorganism remediation etc.) etc..Every kind of recovery technique has its advantage and disadvantage.For example soil replacement method and soil improvement by importing soil from other places are suitable for severe
The small area soil of pollution, project amount is big, costly；Electro reclamation is still in the experimental study stage, and energy consumption is high, cost of investment
It is high；Chemical leaching method destructible soil physico-chemical property；Curing/stabilizing method treatment effect exists certain depending on fixative
Environmental risk；Biological restoration is easy to operate, harmless to soil, also without secondary pollution, but repairing efficiency is long, is not also suitable for
High density pollution soil.Therefore, need to develop that a kind of good fixing effect, lasting effect is strong, it is lesser to influence on soil physico-chemical property
Renovation agent.
It has been reported that nanometer hydroxyapatite (nano-HAP) is to Cd2+There is very high adsorption capacity, and can obviously mention
High soil liquid pH value significantly reduces the biological effectiveness of soil Cd.The nanometer hydroxyapatite of dispersion in the soil can be quick
Fixation is collided and realized with pollutant, however, the shortcomings that this advantage is also exactly superfine powdery material.Nano-grade hydroxy apatite powder
Body in large-scale use one be dispersed, be easy flow away with water, cause water eutrophication.This does not only result in loss of material, increases
Addition sheet, and secondary pollution can be introduced, it is unfavorable for its practical application.Assembling is solve superfine powdery material practical application one
Item possible technique, is combined into macro-size system for nanometer primitive by certain forms, in the hope of showing nanometer ruler in macrocosm
The property of degree.Filamentous fungi is a kind of typical microorganism, is analyzed from appearance structure, and filamentous fungi has very high transverse and longitudinal
Than height is interlaced between mycelia；It is analyzed from composition, extracellular polymeric is contained on filamentous fungi surface, and main component is more
Sugar and protein have certain biological viscosity.Exactly these characteristics can become the load of nano-powder material,
To develop the soil remediation fixative of clean and effective.
Summary of the invention
The purpose of the present invention is to provide it is a kind of based on microorganism assembling synthesis cadmium pollution soil repair fixative and its
Methods for making and using same, the fixed repairing effect of the fixative is good, and cost is relatively low, and methods for making and using same is simple, does not influence soil
Physicochemical property, the soil after reparation can be directly used for planting.
To achieve the above object, the technical solution adopted by the present invention are as follows:
A kind of cadmium pollution soil based on microorganism assembling synthesis repairs the preparation method of fixative, comprising the following steps:
(1) culture filamentous fungi is activated and expanded using culture medium, it is spare；
(2) hydroxyapatite nano particle is added in fluid nutrient medium and is dispersed；
(3) nanoparticle and medium sterilization processing；
(4) in ultra-clean aseptic operating platform, the mycelium for having activated and having cultivated in step (1) is seeded to step (3)
In sterilising medium；
(5) mycelium pellet that vaccinated culture medium shaken cultivation is formed in step (4).
Filamentous fungi described in step (1) be it is a kind of grow thickly in filiform, the fungi with high transverse and longitudinal ratio, including aspergillus and/
Or mould.
Hydroxyapatite nano particle is added in fluid nutrient medium in step (2), solid-to-liquid ratio 0.1-2g:100-
200mL, ultrasonic disperse；The hydroxyapatite nano particle, it is desirable that partial size < 100nm, purity > 97%.
In step (2) use ultrasonic disperse, 50-100 watts of power, ultrasonic oscillation 0.5-3h.
Sterilization treatment described in step (3) is that the culture medium of hydroxyl phosphorite nano particle is put into autoclave,
115 DEG C of setting, sterilize 25min；After autoclave sterilization, the culture medium of hydroxyl phosphorite nano particle is put into ultra-clean sterile behaviour
Make in platform, opens ultraviolet lamp sterilizing 30-90min.
Inoculative proportion is nanometer hydroxyapatite: fluid nutrient medium: bacterium solution=0.1-2g:100-200mL in step (4):
0.5-3mL, mycelium mass concentration is 5.5-6mg/mL in bacterium solution.
Vaccinated culture medium in step (4) is cultivated 2- under the conditions of revolving speed 140-180rpm at 26-30 DEG C by step (5)
4 days.
Step (5) cultivate 2-4 days after, with deionized water rinse formation mycelium pellet 2-5 times after be placed on mass concentration
It is saved at 4 DEG C in the glutaraldehyde of 2.5-5%.
A kind of cadmium pollution soil reparation fixative based on microorganism assembling synthesis, is prepared by the method
's.
The application method of the cadmium pollution soil reparation fixative, the cadmium pollution soil addition fixative, then plus
Enter deionized water, mass ratio is soil: fixative: water=100:(1-5): (100-300) is uniformly mixed, handles the time at least 7
It.
The present invention has the advantage that
1. assembling object of the invention is nanometer hydroxyapatite (n-HAP), because it lives with high-specific surface area, high reaction
Property and the characteristics such as strong absorption, be paid more and more attention in Soil Environmental Pollution improvement.Although the repairing effect of n-HAP is fine,
Because its partial size is too small in large-scale practical application, it is easy to flow away with water, not only its repairing effect is had a greatly reduced quality, but also is caused
Secondary pollution, this seriously constrains its application in soil remediation, needs to find an ideal carrier for it.The present invention
Selecting filamentous fungi mycelia for the first time is load.For example aspergillus niger is a Common Species in aspergillus fungi, it is from a wealth of sources, be
The type specimen of generally acknowledged filamentous fungi research.Mycelia production cost is extremely low, and process cleans are sustainable, and processing is set without expensive
It is standby, the organizer of pore channel prosperity is constituted in addition, aspergillus niger mycelia transverse and longitudinal ratio is very high convenient for large scale processing, is conducive to pass
Matter, cell wall are mainly made of large biological molecule, and property is stablized, and cell surface can make the adhering zone of nanoparticle.This hair
It is bright it is main using the high transverse and longitudinal of aspergillus niger than with the characteristic with biological viscosity etc., using aspergillus niger mycelia as load, package is received
Rice hydroxyapatite, is made soil remediation fixative, solves the problems, such as that it excessively disperses in practical applications.
2. the present invention wraps up nanometer hydroxyapatite using nanometer hydroxyapatite as center substance, using aspergillus niger mycelia,
The cadmium pollution soil being made for high pollution highly acidity repairs fixative, and feature essentially consists in fixative and adds into cadmium pollution
When soil, the part cadmium in soil is adsorbed by hypha,hyphae, and on the other hand, fixative, which will be slow, releases effective component --- n-
HAP, n-HAP gradually dissolve in the soil liquid, Cd2+With the Ca in hydroxyapatite2+Ion exchange occurs, or by hydroxyl phosphorus
Adsorption of metal ions in lime stone lattice surface turns cadmium from the biologically effective state for being easy to be absorbed by organism to low activity form
Become, fixative better effect in high pollution highly acidity contaminated soil, because the increase of acidity can promote to dissolve nano hydroxyapatite
Lime stone.But nano-grade hydroxy apatite powder release simultaneously is slowly, and burst size is little, not will cause soil or water eutrophication
The consequence of change.
3. essential element group becomes calcium, phosphorus, carbon, hydrogen, oxygen in the fixative that the present invention synthesizes, these elements are that plant is necessary
Nutrient, therefore fixative synthesized by the present invention be applied to cadmium pollution soil fixation, nonhazardous is biodegradable,
It is without secondary pollution, it is a kind of environmentally friendly fixative.
4. fixative synthesized by the present invention and many common phosphorous-based materials (phosphoric acid, potassium dihydrogen phosphate, biphosphates
Calcium, rock phosphate in powder etc.) it compares, it is highly efficient in application process, and not will lead to underground water eutrophication, avoid secondary dirt
The risk of dye will not influence the stabilization of soil texture, more conducively later period farming object.
Deposit number of the present invention: CGMCC No.5599 name: the bacterium of Penicillium Chrysogenum F1
Strain, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: the Chaoyang District, Beijing City North Star
The institute 3 of West Road 1；Preservation date: on December 15th, 2011.
Detailed description of the invention
Fig. 1 is the process flow chart of 1 fixative of embodiment of the present invention synthesis；
Fig. 2 is the scanning electron microscope (SEM) photograph (SEM) of pure mycelium pellet before present invention package nanoparticle；
Fig. 3 is the scanning electron microscope (SEM) photograph (SEM) of fixative of the present invention；
Fig. 4 is the XRD diagram of fixative of the present invention；
Fig. 5 is the XRD diagram after the fixed cadmium of fixative of the present invention.
Specific embodiment
It is intended to be described in further details the present invention in the following with reference to the drawings and specific embodiments, be not intended to limit the present invention.
Embodiment 1
Prepare cadmium pollution soil repair fixative of the present invention:
(1) culture aspergillus niger (Aspergillus niger) is activated and expanded using PDA culture medium, is purchased from Chinese Typical Representative
Culture collection, bacterium numbering CCTCC AF 91006；
(2) hydroxyapatite nano particle (partial size < 100nm, purity > 97%) of 1.0g is added to the liquid of 100mL
In PDA culture medium, disperse 0.5h using 100 watts of power of ultrasonic echographies；
(3) PDA culture medium containing nanometer hydroxyapatite is put into high temperature high-pressure sterilizing pot, is arranged 115 DEG C, sterilizing
25min.After autoclave sterilization, in media transfer to ultra-clean aseptic operating platform, ultraviolet lamp sterilizing 30min is opened；
(4) in ultra-clean aseptic operating platform, the sterilizing that the mycelium activated in step (1) is seeded to step (3) is trained
It supports in base, inoculum concentration 1mL；Mycelium mass concentration is 5.89mg/mL in bacterium solution；
(5) vaccinated culture medium is placed in gas bath shaking table and vibrates in step (4), cultivates 2-3 days at 30 DEG C, revolving speed is
160rpm；
(6) after 2-3 days, with mycelium pellet 2-3 times formed in deionized water rinse step (5), after be placed on mass concentration
It is saved at 4 DEG C in 2.5% glutaraldehyde.
Embodiment 2
Prepare cadmium pollution soil repair fixative of the present invention:
(1) using the LB culture medium (every 100mL culture medium: LB culture medium 2.5g, glucose 9g, sodium nitrate 0.3g) of improvement
Activation and expansion culture penicillium chrysogenum (Penicillium Chrysogenum F1), bacterial strain is voluntarily collected in zhuzhou, hunan province
City is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number: CGMCC No.5599,
And patent was applied for；
(2) hydroxyapatite nano particle (partial size < 100nm, purity > 97%) of 1.0g is added to the liquid of 200mL and changes
In good LB culture medium, disperse 1.5h using 100 watts of power of ultrasonic echographies；
(3) the improvement LB culture medium containing nanometer hydroxyapatite is put into high temperature high-pressure sterilizing pot, is arranged 115 DEG C,
Sterilize 25min.After autoclave sterilization, in media transfer to ultra-clean aseptic operating platform, ultraviolet lamp sterilizing 30min is opened；
(4) in ultra-clean aseptic operating platform, the sterilizing that the mycelium activated in step (1) is seeded to step (3) is trained
It supports in base, inoculum concentration 1mL；Mycelium mass concentration is 5.89mg/mL in bacterium solution；
(5) vaccinated culture medium is placed in shaking bath and vibrates in step (4), cultivates 2-3 days at 28 DEG C, revolving speed is
150rpm；
(6) after 2-3 days, with mycelium pellet 2-3 times formed in deionized water rinse step (5), after be placed on mass concentration
It is saved at 4 DEG C in 2.5% glutaraldehyde.
Embodiment 3
Soil is derived from certain Pb-Zn tailings library periphery low pollution rice soil.For natural wind after examination soil rejecting gravel and broken
Dry, quartering mixes, and pulverizes 18 mesh nylon mesh.Soil pH value is 6.58, weakly acidic.Soil available Cd content is
7.01mg/kg。
Respectively by 0.2g embodiment 1 fixative (amount containing n-HAP is equal with weighed pure n-HAP amount in fixative) with
Pure n-HAP addition enters in 5.0g low pollution soil, is uniformly mixed, the water of 10mL is added, after fixing 7 days, with 25mL DTPA
Extract the available state of cadmium in soil.Through detecting, cadmium bio-available Zn concentration is such as before and after the processing through fixative or pure n-HAP for low pollution soil
Shown in the following table 1.
Available state Cd content before and after 1 low pollution soil treatment of table
| Available state Cd (mg/kg) | Fixed rate | |
| Low pollution soil | 7.01 | — |
| After the processing of 1 fixative of embodiment | 2.32 | 66.9% |
| After pure n-HAP processing | 3.85 | 45.1% |
Embodiment 4
Soil is derived from the rice soil of certain Pb-Zn tailings library periphery intermediate pollution.For natural after examination soil rejecting gravel and broken
It air-dries, quartering mixes, and pulverizes 18 mesh nylon mesh.Soil pH value is 6.52, weakly acidic.Soil available Cd content is
36.77mg/kg。
Respectively by fixative (amount containing n-HAP and the weighed pure n-HAP amount phase in fixative in 0.2g Examples 1 and 2
Deng) enter in 5.0g in contaminated soil with pure n-HAP addition, it is uniformly mixed, the water of 10mL is added, after fixing 7 days, uses 25mL
DTPA extracts the available state of cadmium in soil.Through detecting, middle contaminated soil through embodiment 1,2 fixatives or pure n-HAP before and after the processing
Cadmium bio-available Zn concentration is as shown in table 2 below, it can be seen that, the fixative that embodiment 1 produces is than 2 better effect of embodiment.
Contaminated soil available state Cd content before and after the processing in table 2
Embodiment 5
Soil is derived from certain Pb-Zn tailings library periphery high pollution rice soil.For natural wind after examination soil rejecting gravel and broken
Dry, quartering mixes, and pulverizes 18 mesh nylon mesh.Soil pH value is 5.90, in acidity.Soil available Cd content is respectively
127.0mg/kg。
Respectively by 0.2g embodiment 1 fixative (amount containing n-HAP is equal with weighed pure n-HAP amount in fixative) with
Pure n-HAP addition enters in 5.0g high pollution soil, is uniformly mixed, the water of 10mL is added, after fixing 7 days, is mentioned with 25mL DTPA
Take the available state of cadmium in soil.Through detecting, high pollution soil is through 1 fixative of embodiment or pure n-HAP cadmium available state before and after the processing
Content is as shown in table 3 below, it is seen that the better effect of present invention processing high pollution highly acidity contaminated soil.
Available state Cd content before and after 3 high pollution soil treatment of table
| Available state Cd (mg/kg) | Fixed rate Cd | |
| High pollution soil | 127.0 | — |
| After the processing of 1 fixative of embodiment | 17.2 | 86.5% |
| After pure n-HAP processing | 93.6 | 26.3% |
Claims (8)
1. the preparation method that a kind of cadmium pollution soil based on microorganism assembling synthesis repairs fixative, which is characterized in that including
Following steps:
(1) culture filamentous fungi is activated and expanded using culture medium, it is spare；
(2) hydroxyapatite nano particle is added in fluid nutrient medium and is dispersed；
(3) nanoparticle and medium sterilization processing；
(4) in ultra-clean aseptic operating platform, the mycelium for having activated and having cultivated in step (1) is seeded to the sterilizing of step (3)
In culture medium；
(5) mycelium pellet that vaccinated culture medium shaken cultivation is formed in step (4)；
It is in filiform that filamentous fungi described in step (1), which is a kind of, is grown thickly, the fungi with high transverse and longitudinal ratio, including aspergillus and/or blueness
It is mould；
Step (5) cultivate 2-4 days after, with deionized water rinse formation mycelium pellet 2-5 times after be placed on mass concentration 2.5-5%
Glutaraldehyde in saved at 4 DEG C.
2. preparation method according to claim 1, which is characterized in that step adds hydroxyapatite nano particle in (2)
Enter in fluid nutrient medium, solid-to-liquid ratio 0.1-2g:100-200mL, ultrasonic disperse；The hydroxyapatite nano particle, it is desirable that
Partial size < 100nm, purity > 97%.
3. preparation method according to claim 2, which is characterized in that use ultrasonic disperse, power 50-100 in step (2)
Watt, ultrasonic oscillation 0.5-3h.
4. preparation method according to claim 1, which is characterized in that sterilization treatment described in step (3) is by hydroxyl
The culture medium of phosphorite nano particle is put into autoclave, is arranged 115 DEG C, and sterilize 25min；After autoclave sterilization, contain hydroxyl
The culture medium of base phosphorite nano particle is put into ultra-clean aseptic operating platform, opens ultraviolet lamp sterilizing 30-90min.
5. preparation method according to claim 1, which is characterized in that inoculative proportion is nano-hydroxy-apatite in step (4)
Stone: fluid nutrient medium: bacterium solution=0.1-2g:100-200mL:0.5-3mL, mycelium mass concentration is 5.5-6mg/mL in bacterium solution.
6. preparation method according to claim 1, which is characterized in that step (5) is by vaccinated culture medium in step (4)
It is cultivated 2-4 days under the conditions of revolving speed 140-180rpm at 26-30 DEG C.
7. a kind of cadmium pollution soil based on microorganism assembling synthesis repairs fixative, which is characterized in that be by claim 1-6
What described in any item methods were prepared.
8. cadmium pollution soil as claimed in claim 7 repairs the application method of fixative, for repairing for paddy field cadmium pollution soil
Multiple, the fixative is added in soil, adds deionized water, mass ratio is soil: fixative: water=100:(1-5): (100-
300) it, is uniformly mixed, handles the time at least 7 days.
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