CN106424125B - A kind of microbial inoculum and the preparation method and application thereof handling heavy metal pollution of soil - Google Patents
A kind of microbial inoculum and the preparation method and application thereof handling heavy metal pollution of soil Download PDFInfo
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- CN106424125B CN106424125B CN201610570018.4A CN201610570018A CN106424125B CN 106424125 B CN106424125 B CN 106424125B CN 201610570018 A CN201610570018 A CN 201610570018A CN 106424125 B CN106424125 B CN 106424125B
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Abstract
The present invention relates to a kind of microbial bacterial agents and the preparation method and application thereof for handling heavy metal pollution of soil.The microbial bacterial agent for handling heavy metal pollution of soil, is made of the following raw material: rice husk, pig manure, chicken manure, soy meal, single cell protein, Mortierella alpina fermentation liquid, and trichoderma asperellum fermentation liquid rolls up branch Mucor fermentation liquid.The microbial bacterial agent for the processing heavy metal pollution of soil that the present invention uses Mortierella alpina, trichoderma asperellum and volume branch Mucor to be mixed to prepare in proportion for the first time, the microbial inoculum can be applied among environment complicated and changeable, in face of poor soil environment, it can still play a role, functional microorganism is high to the tolerance of heavy metal, and activity is still able to maintain in the soil of heavy metal serious pollution and carries out efficient repairing and is answered.
Description
Technical field
The present invention relates to a kind of microbial inoculums and the preparation method and application thereof for handling heavy metal pollution of soil, belong to soil remediation
Technical field.
Background technique
Ecological disruption problem has become the critical issue for restricting human development.Mining activities are formed by refuse dump, tailing
Often pollutant component is complicated for library and ambient contamination place, contains heavy metal and Some Organic Pollutants, the dirt of these heavy metals
Concentration height, pollution level weight are contaminated, causes animals and plants to be difficult to grow wherein, seriously affects ecological environment.Heavy metal in soil,
The soil, waters and atmosphere of surrounding can be polluted by surface water body runoff, atmosphere floating dust, influencing face will be considerably beyond useless
The region and space of gurry storeyard, pollution effect will spend a large amount of human and material resources, financial resources by being lot more time to restore,
And it is difficult to be restored to original level.
The soil restoring technology that modern technologies cope with heavy metal pollution is relatively deficient, is concentrated mainly on hyperaccumulative plant to spy
Determine the enrichment of heavy metal, this method can only cope with more single heavy metal pollution, and requirement of the plant to soil environment compared with
Height limits its scope of application.
Chinese patent literature CN104070062A (application number 201310108243.2) discloses a kind of heavy metal polluted soil
The microorganism in situ restorative procedure of earth, the steps include: (1) bacteria group culture: (2) flora activates；(3) microorganism growth stimulator is prepared；(4) seek
Nutrient solution is prepared；(5) it just throws；(6) repair.By weight in the microorganism in situ restorative procedure for the heavy-metal contaminated soil that the invention provides
Metal microbial degradation flora, microorganism Acceleration of growth agent supplement certain microbial nutrition element as main component,
While putting into preferred heavy metal microorganism conversion flora into soil, increases the agent of microorganism Acceleration of growth and promote its high speed numerous
It grows, to greatly improve conversion and the degradation efficiency of heavy metals in soil, solves heavy-metal contaminated soil microorganism
The problem of time in in-situ remediation method is long, slow effect, make microorganism in situ recovery technique become it is reliable, efficiently, can operate
Heavy-metal contaminated soil governing measure.
The toxicity that can quickly and effectively reduce the heavy metal in polluted soil how is found, current research heat is become
Point.
Summary of the invention
In view of the deficiencies of the prior art, the present invention provides a kind of microbial bacterial agents and its system for handling heavy metal pollution of soil
Preparation Method and application.
Technical solution of the present invention is as follows:
A kind of microbial bacterial agent handling heavy metal pollution of soil, is composed of the following raw materials in parts by weight:
50~80 parts of rice husk, 20~50 parts of pig manure, 20~50 parts of chicken manure, 30~60 parts of soy meal, single cell protein 4~6
Part, 2.5-3.5 parts of fermentation liquid of Mortierella alpina (Mortierella alpina), trichoderma asperellum (Trichoderma
Asperellum) 1.5~2.5 parts of fermentation liquid, 0.5~1.5 part of fermentation liquid of volume branch Mucor (Mucor circinelloides)；
Cell concentration in Mortierella alpina (Mortierella alpina) fermentation liquid is 0.4 × 108cfu/ml；
Cell concentration in trichoderma asperellum (Trichoderma asperellum) fermentation liquid is 1.0 × 108cfu/ml；Roll up branch Mucor
Cell concentration in (Mucor circinelloides) fermentation liquid is 3.6 × 108cfu/ml；Thallus always contains in microbial bacterial agent
Amount is 6.8 × 108cfu/g。
According to the present invention preferably, the Mortierella alpina (Mortierella alpina), trichoderma asperellum
(Trichoderma asperellum) and volume branch Mucor (Mucor circinelloides) viable bacteria body ratio of number are 1:
(1.0~2.5): (1.8~3.6).
According to the present invention it is further preferred that the Mortierella alpina (Mortierella alpina), trichoderma asperellum
(Trichoderma asperellum) and volume branch Mucor (Mucor circinelloides) viable bacteria body ratio of number are 1:
1.5:3.
Preferred according to the present invention, the thallus total content is 6.8 × 108cfu/g。
Preferred according to the present invention, the Mortierella alpina (Mortierella alpina) is Mortierella alpina
(Mortierella alpina) ME-AA01 is derived from China typical culture collection center (CCTCC), bacterium numbering CCTCC
NO:M207067。
Preferred according to the present invention, the trichoderma asperellum (Trichoderma asperellum) is trichoderma asperellum
(Trichoderma asperellum) PD-19 is derived from China typical culture collection center (CCTCC), bacterium numbering
CCTCC NO:M2013113。
Preferred according to the present invention, the volume branch Mucor (Mucor circinelloides) derives from Dutch microbial bacteria
Kind collection (CBS), bacterium numbering CBS131818.
The preparation method of the microbial bacterial agent of above-mentioned processing heavy metal pollution of soil, steps are as follows:
(1) by Mortierella alpina (Mortierella alpina) through first order seed culture, secondary seed culture and expansion
Culture is made 1.2 × 107~1.6 × 107Mortierella alpina (Mortierella alpina) fermentation liquid of cfu/ml；
(2) by trichoderma asperellum (Trichoderma asperellum) through first order seed culture, secondary seed culture and expansion
Big culture, is made 0.6~0.8 × 108Trichoderma asperellum (Trichoderma asperellum) fermentation liquid of cfu/ml；
(3) branch Mucor (Mucor circinelloides) will be rolled up through first order seed culture, secondary seed culture and expansion
Culture is made 1.4 × 108~1.8 × 108Volume branch Mucor (Mucor circinelloides) fermentation liquid of cfu/ml；
(4) Mortierella alpina made from step (1) (Mortierella alpina) fermentation liquid, step (2) is obtained
Trichoderma asperellum (Trichoderma asperellum) fermentation liquid, step (3) are obtained to roll up branch Mucor (Mucor
Circinelloides) fermentation liquid and rice husk, pig manure, chicken manure, soy meal and single cell protein are uniformly mixed, room temperature condition decentralization
It sets 4~6 weeks, stirs weekly 2~4 times, it is dry to be lower than 20% to moisture content, microbial bacterial agent is made.
It is preferred according to the present invention, in the step (1), (2) and step (3), first order seed condition of culture are as follows: illumination
1000~3000LX, 25~30 DEG C of cultivation temperature, Anaerobic culturel 3~4 days；
Every liter of component of the primary-seed medium is as follows:
15~20g of glucose, 6~10g of peptone, 1~3g of yeast extract, 0.2~0.8g of dipotassium hydrogen phosphate, magnesium sulfate
0.01~0.03g, 0.1~0.29g of calcium chloride.
It is preferred according to the present invention, in the step (1), (2) and step (3), secondary seed condition of culture are as follows: illumination
1000~3000LX, 25~30 DEG C of cultivation temperature, Anaerobic culturel 3~4 days；
Every liter of component of the secondary seed medium is as follows:
15~20g of glucose, 6~10g of peptone, 1~3g of yeast extract, 0.2~0.8g of dipotassium hydrogen phosphate, magnesium sulfate
0.01~0.03g, 0.1~0.29g of calcium chloride.
It is preferred according to the present invention, in the step (1), (2) and step (3), expand condition of culture are as follows: cultivation temperature 28
~30 DEG C, dark aerobic culture 6~8 days, ratio of ventilating per hour are liquid and gas volume ratio 1:0.5, speed of agitator 200~
300r/min；
Described every liter of component of culture medium for expanding culture is as follows:
15~20g of glucose, 6~10g of peptone, 1~3g of yeast extract, 0.2~0.8g of dipotassium hydrogen phosphate, magnesium sulfate
0.01~0.03g, 0.1~0.29g of calcium chloride.
The application of the microbial bacterial agent of above-mentioned processing heavy metal pollution of soil, steps are as follows:
The microbial bacterial agent for handling heavy metal pollution of soil is directly applied in the soil of heavy metal pollution as base manure,
Mixing is turned over, amount of application is 8~12g/ square metres, is handled 33~38 days under conditions of 29~31 DEG C of temperature, pH7.5~8.5.
Above-mentioned rice husk, pig manure, chicken manure, soy meal and single cell protein etc. are this field ordinary commercial products.
Beneficial effect
1, the present invention uses Mortierella alpina (Mortierella alpina), trichoderma asperellum (Trichoderma for the first time
Asperellum) the processing heavy metal pollution of soil being mixed to prepare in proportion with volume branch Mucor (Mucor circinelloides)
Microbial bacterial agent, which can apply among environment complicated and changeable, in face of poor soil environment, can still send out
The effect of waving, functional microorganism are high to the tolerance of heavy metal, and activity is still able to maintain in the soil of heavy metal serious pollution simultaneously
Efficiently repaired；
2, the microbial bacterial agent preparation process of processing heavy metal pollution of soil of the present invention is simple, applied widely, at
This low, strong operability.Functional microorganism can not only accumulate the repair mechanisms multiplicity of heavy metal to heavy metal adsorption, moreover it is possible to
It is enough that heavy metal is converted into stable state, it is administered to greatest extent on the basis of not causing secondary destruction to environment heavy metal-polluted
It contaminates, the functional microorganism in composite bacteria agent is to the adaptable of environment, to considerably increase the application range of composite bacteria agent.
Specific embodiment
Below with reference to embodiment, technical scheme is described further, but institute's protection scope of the present invention is not limited to
This.
Biological material source:
Mortierella alpina (Mortierella alpina) is Mortierella alpina (Mortierella alpina) ME-
AA01 is derived from China typical culture collection center (CCTCC), bacterium numbering CCTCC NO:M207067；
Trichoderma asperellum (Trichoderma asperellum) is trichoderma asperellum (Trichoderma asperellum) PD-
19, it derives from China typical culture collection center (CCTCC), bacterium numbering CCTCC NO:M2013113；
It rolls up branch Mucor (Mucor circinelloides) and derives from Dutch Culture Collection (CBS), strain
Number CBS131818.
Rice husk described in embodiment, pig manure, chicken manure, soy meal and single cell protein are this field ordinary commercial products.
Embodiment 1
A kind of microbial bacterial agent handling heavy metal pollution of soil, is composed of the following raw materials in parts by weight:
80 parts of rice husk, 20 parts of pig manure, 50 parts of chicken manure, 30 parts of soy meal, 6 parts of single cell protein, Mortierella alpina
2.5 parts of fermentation liquid of (Mortierella alpina), 2.5 parts of fermentation liquid of trichoderma asperellum (Trichoderma asperellum),
0.5 part of fermentation liquid of volume branch Mucor (Mucor circinelloides)；
Thallus total content is 5.3 × 10 in microbial bacterial agent8cfu/g；Mortierella alpina (the Mortierella
Alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor circinelloides) viable bacteria
Body ratio of number is 1:1.8:2.5.
The preparation method of the microbial bacterial agent of above-mentioned processing heavy metal pollution of soil, steps are as follows:
(1) by Mortierella alpina (Mortierella alpina) through first order seed culture, secondary seed culture and expansion
Culture is made 0.4 × 108Mortierella alpina (Mortierella alpina) fermentation liquid of cfu/ml；
(2) by trichoderma asperellum (Trichoderma asperellum) through first order seed culture, secondary seed culture and expansion
Big culture, is made 1.0 × 108Trichoderma asperellum (Trichoderma asperellum) fermentation liquid of cfu/ml；
(3) branch Mucor (Mucor circinelloides) will be rolled up through first order seed culture, secondary seed culture and expansion
Culture is made 3.6 × 108Volume branch Mucor (Mucor circinelloides) fermentation liquid of cfu/ml；
(4) Mortierella alpina made from step (1) (Mortierella alpina) fermentation liquid, step (2) is obtained
Trichoderma asperellum (Trichoderma asperellum) fermentation liquid, step (3) are obtained to roll up branch Mucor (Mucor
Circinelloides) fermentation liquid and rice husk, pig manure, chicken manure, soy meal and single cell protein are uniformly mixed, room temperature condition decentralization
It sets 4 weeks, stirs weekly 4 times, it is dry to be lower than 20% to moisture content, microbial bacterial agent is made.
In the step (1), (2) and step (3), first order seed condition of culture are as follows: illumination 2000LX, cultivation temperature 27
DEG C, Anaerobic culturel 3 days；Every liter of component of the primary-seed medium is as follows:
Glucose 15g, peptone 10g, yeast extract 1g, dipotassium hydrogen phosphate 0.8g, magnesium sulfate 0.01g, calcium chloride
0.29g。
In the step (1), (2) and step (3), secondary seed condition of culture are as follows: illumination 2000LX, cultivation temperature 27
DEG C, Anaerobic culturel 3 days；Every liter of component of the secondary seed medium is as follows:
Glucose 15g, peptone 10g, yeast extract 1g, dipotassium hydrogen phosphate 0.8g, magnesium sulfate 0.01g, calcium chloride
0.29g。
In the step (1), (2) and step (3), expand condition of culture are as follows: 28 DEG C of cultivation temperature, dark aerobic culture 8
It, ratio of ventilating per hour is liquid and gas volume ratio 1:0.5, speed of agitator 300r/min；
Described every liter of component of culture medium for expanding culture is as follows:
Glucose 15g, peptone 10g, yeast extract 1g, dipotassium hydrogen phosphate 0.8g, magnesium sulfate 0.01g, calcium chloride
0.29g。
Embodiment 2
A kind of microbial bacterial agent handling heavy metal pollution of soil, is composed of the following raw materials in parts by weight:
50 parts of rice husk, 50 parts of pig manure, 20 parts of chicken manure, 60 parts of soy meal, 4 parts of single cell protein, Mortierella alpina
3.5 parts of fermentation liquid of (Mortierella alpina), 1.5 parts of fermentation liquid of trichoderma asperellum (Trichoderma asperellum),
1.5 parts of fermentation liquid of volume branch Mucor (Mucor circinelloides)；
Thallus total content is 8.3 × 10 in microbial bacterial agent8cfu/g；Mortierella alpina (the Mortierella
Alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor circinelloides) viable bacteria
Body ratio of number is 1:1:3.6.
The preparation method is the same as that of Example 1.
Embodiment 3
A kind of microbial bacterial agent handling heavy metal pollution of soil, is composed of the following raw materials in parts by weight:
65 parts of rice husk, 35 parts of pig manure, 35 parts of chicken manure, 45 parts of soy meal, 5 parts of single cell protein, Mortierella alpina
3 parts of fermentation liquid of (Mortierella alpina), 2 parts of fermentation liquid of trichoderma asperellum (Trichoderma asperellum) roll up branch
1 part of fermentation liquid of Mucor (Mucor circinelloides)；
Thallus total content is 6.8 × 10 in microbial bacterial agent8cfu/g；Mortierella alpina (the Mortierella
Alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor circinelloides) viable bacteria
Body ratio of number is 1:1.5:3.
The preparation method is the same as that of Example 1.
Comparative example 1
The system of microbial bacterial agent as described in Chinese patent literature CN104070062A (application number 201310108243.2)
Preparation Method, steps are as follows:
(i) bacteria group culture: selection can convert the microbial degradation flora body of heavy metal characteristic, using the side of solid culture
Formula carries out culture domestication by carrier of wheat bran, the powdered substance for taking in-between bacterial product to mix and formed after desiccation；
(ii) flora activates: into the water by the powdered substance in step (i), impregnating 0.5~2h, removal wheat bran to obtain the final product
Flora liquid after activation；
(iii) microorganism growth stimulator is prepared: compound amino acid, composite trace element, activating enzymes and humic acid are put into water
In, it stirs evenly, obtains microorganism growth stimulator；
(iv) compounding: microbial nutrition element is added in microorganism growth stimulator, water is added to be diluted to get microorganism
Growth stimulator nutrient solution；
Comparative example 2
The microbial bacterial agent of processing heavy metal pollution of soil as described in Example 3, the difference is that,
Trichoderma asperellum is replaced with trichoderma reesei (Trichoderma reesei), the bacterium source is in Chinese common micro- life
Object culture presevation administrative center, bacterium numbering CGMCC No:9644.
Comparative example 3
The microbial bacterial agent of processing heavy metal pollution of soil as described in Example 3, the difference is that,
Volume branch Mucor is replaced with into Mucor racemosus (Mucor racemosus), the bacterium source is in Chinese Typical Representative culture
Collection, bacterium numbering CCTCC NO:M201026.
Experimental example 1
After 2/3 soil moved in improve the original and 1/3 Nutrition Soil are mixed, add respectively heavy metal contaminants background value 10,100,500,
After 1000 times, analog composite heavy-metal contaminated soil is balanced 2-3 days.It is applied described in embodiment 1-3 and comparative example 1-3 respectively
Composite bacteria agent, amount of application are 6kg/ mus, are spaced application in 7 days once after turning over mixing, continuous administration 5 times.Every seven days observing earths
The physicochemical property of the variation of content of beary metal and soil in earth.After applying composite bacteria agent, heavy metal in soil content is significantly reduced.
As a result as shown in the table:
Table 1
After soil improvement, black nightshade, summer cypress, the fluffy three kinds of plants of alkali, every physical signs of periodic monitoring plant, plant are planted
It grows fine.The composite bacteria agent can not only effectively administer heavy metal pollution, and can promote the growth of plant.
Experimental example 2
Yellow triangle possesses about 8,100,000 mu of Unutilized Energy, and wherein salt-soda soil just accounts for 2,700,000 mu, northern in Dongying Lijin,
Kenli northeast and extensively there are also the soil on the ground such as Hekou District all by salinization of soil around east.Not only soil environment is disliked in Dongying salt-soda soil
It is bad, and there are more serious heavy metal pollutions.By surface water body runoff, atmosphere floating dust, soil, the waters of surrounding are polluted
And atmosphere, ambient enviroment is caused to seriously affect.Composite bacteria agent described in embodiment 1-3 and comparative example 1-3, application are applied respectively
Amount is 7kg/ mus, is spaced application in 7 days once after turning over mixing, continuous administration 5 times.After applying composite bacteria agent, the gas permeability of soil
It dramatically increases, the soil weight is from 1.49g/m3Drop to 1.24g/m3, reached normal Soil standard (1.1-1.4g/m3).Soil
Water-retaining property it is aobvious improve, the water content of soil is increased to 21% from 15%.In the comparative example without significant changes.A huge sum of money in soil
Belonging to content significantly reduces, and the 30th day when detects heavy metal removing rate, as a result as described in Table 2:
Table 2
The result shows that microorganism can adapt to alkaline environment after applying the composite bacteria agent, good survival state is kept.
After applying composite bacteria agent, content of beary metal is significantly reduced in tailing soil, and the physical and chemical parameter of soil obtains certain raising.
Experimental example 3
What the waste (tailing, removing rock etc.) during mining needed large area banks up place, in Anshan city
City periphery forms 7,910,000 m of land occupation2Discarded refuse dump and 2,520,000 m2Discarded tailing library.Cause to occupy the excess in soil
With the destruction to the original ecosystem of storeyard；It influences face will be considerably beyond the region and space of waste storeyard.Tailing
Area's soil is in alkalinity, and soil depletion, poor water retention property, heavy metal pollution is more serious, it is difficult to repair.Reality is applied respectively in tailing area
Composite bacteria agent described in a 1-3 and comparative example 1-3 is applied, amount of application is 8kg/ mus, is spaced application in 7 days once after turning over mixing, even
Continuous application 5 times.Heavy metal removing rate is detected at the 30th day.After applying composite bacteria agent, the gas permeability of soil is dramatically increased, and soil holds
Again from 1.57g/m3Drop to 1.33g/m3, reached normal Soil standard (1.1-1.4g/m3).The water-retaining property of soil is aobvious to be improved,
The water content of soil is increased to 18% from 9%.The index is in the comparative example without significant changes.Heavy metal in soil content is significant
It reduces, the 30th day when detects heavy metal removing rate, as a result as described in Table 3:
Table 3
The result shows that microbe survival in order, can adapt to rugged environment after applying the composite bacteria agent.Application
After composite bacteria agent, content of beary metal is significantly reduced in tailing soil.The composite bacteria agent, which has the soil of heavy metal pollution, preferably to be repaired
Multiple effect.
Interpretation of result
Find that the composite bacteria agent has preferable removal effect to five heavy metal species by the comparison of embodiment.High mountain quilt
Spore mould (Mortierella alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor
Circinelloides) optimal proportion is 1:1.5:3.
By comparative example it can be found that Mortierella alpina (Mortierella alpina), trichoderma asperellum in the patent
There is the work mutually cooperateed between (Trichoderma asperellum) and volume branch Mucor (Mucor circinelloides)
With the removal rate of heavy metal can be significantly improved.
Described is only presently preferred embodiments of the present invention, and implementation condition and practical range of the invention cannot be limited with this.According to
Modification and variation made by scope of the present invention patent and invention description content, still belong to the range that the invention patent covers.
Claims (11)
1. a kind of microbial bacterial agent for handling heavy metal pollution of soil, which is characterized in that be composed of the following raw materials in parts by weight:
50~80 parts of rice husk, 20~50 parts of pig manure, 20~50 parts of chicken manure, 30~60 parts of soy meal, 4~6 parts of single cell protein are high
3~5 parts of fermentation liquid of mountain Mortierella (Mortierella alpina), trichoderma asperellum (Trichoderma asperellum) fermentation
1~3 part of liquid, 0.5~1.5 part of fermentation liquid of volume branch Mucor (Mucor circinelloides)；
Cell concentration in Mortierella alpina (Mortierella alpina) fermentation liquid is 0.4 × 108cfu/ml；Spine spore
Cell concentration in trichoderma (Trichoderma asperellum) fermentation liquid is 1.0 × 108cfu/ml；Roll up branch Mucor (Mucor
Circinelloides) cell concentration in fermentation liquid is 3.6 × 108cfu/ml；Thallus total content is 6.8 in microbial bacterial agent
×108cfu/g。
2. the microbial bacterial agent of processing heavy metal pollution of soil as described in claim 1, which is characterized in that the high mountain quilt
Spore mould (Mortierella alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor
Circinelloides) viable bacteria body ratio of number is 1:(1.0~2.5): (1.8~3.6).
3. the microbial bacterial agent of processing heavy metal pollution of soil as claimed in claim 2, which is characterized in that the high mountain quilt
Spore mould (Mortierella alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor
Circinelloides) viable bacteria body ratio of number is 1:1.5:3.
4. the microbial bacterial agent of processing heavy metal pollution of soil as described in claim 1, which is characterized in that the Mortierella alpine
Mould (Mortierella alpina) is Mortierella alpina (Mortierella alpina) ME-AA01, derives from Chinese Typical Representative
Culture collection (CCTCC), bacterium numbering CCTCC NO:M207067.
5. the microbial bacterial agent of processing heavy metal pollution of soil as described in claim 1, which is characterized in that the trichoderma asperellum
(Trichoderma asperellum) is trichoderma asperellum (Trichoderma asperellum) PD-19, from Chinese allusion quotation
Type culture collection (CCTCC), bacterium numbering CCTCC NO:M2013113.
6. the microbial bacterial agent of processing heavy metal pollution of soil as described in claim 1, which is characterized in that the volume branch Mucor
(Mucor circinelloides) derives from Dutch Culture Collection (CBS), bacterium numbering CBS131818.
7. handling the preparation method of the microbial bacterial agent of heavy metal pollution of soil described in claim 1, which is characterized in that step is such as
Under:
(1) Mortierella alpina (Mortierella alpina) through first order seed culture, secondary seed culture and is expanded into culture,
It is made 0.4 × 108Mortierella alpina (Mortierella alpina) fermentation liquid of cfu/ml；
(2) trichoderma asperellum (Trichoderma asperellum) through first order seed culture, secondary seed culture and is expanded into training
It supports, is made 1.0 × 108Trichoderma asperellum (Trichoderma asperellum) fermentation liquid of cfu/ml；
(3) volume branch Mucor (Mucor circinelloides) through first order seed culture, secondary seed culture and is expanded into culture,
It is made 3.6 × 108Volume branch Mucor (Mucor circinelloides) fermentation liquid of cfu/ml；
(4) by spine spore made from Mortierella alpina made from step (1) (Mortierella alpina) fermentation liquid, step (2)
Trichoderma (Trichoderma asperellum) fermentation liquid, step (3) are obtained to roll up branch Mucor (Mucor
Circinelloides) fermentation liquid and rice husk, pig manure, chicken manure, soy meal and single cell protein are uniformly mixed, room temperature condition decentralization
It sets 4~6 weeks, stirs weekly 2~4 times, it is dry to be lower than 20% to moisture content, microbial bacterial agent is made.
8. preparation method as claimed in claim 7, which is characterized in that in the step (1), (2) and step (3), level-one kind
Sub- condition of culture are as follows: 1000~3000LX of illumination, 25~30 DEG C of cultivation temperature, Anaerobic culturel 3~4 days；
Every liter of component of culture medium of the first order seed culture is as follows:
15~20g of glucose, 6~10g of peptone, 1~3g of yeast extract, 0.2~0.8g of dipotassium hydrogen phosphate, magnesium sulfate 0.01~
0.03g, 0.1~0.29g of calcium chloride.
9. preparation method as claimed in claim 7, which is characterized in that in the step (1), (2) and step (3), second level kind
Sub- condition of culture are as follows: 1000~3000LX of illumination, 25~30 DEG C of cultivation temperature, Anaerobic culturel 3~4 days；
Every liter of component of culture medium of the secondary seed culture is as follows:
15~20g of glucose, 6~10g of peptone, 1~3g of yeast extract, 0.2~0.8g of dipotassium hydrogen phosphate, magnesium sulfate 0.01~
0.03g, 0.1~0.29g of calcium chloride.
10. preparation method as claimed in claim 7, which is characterized in that in the step (1), (2) and step (3), expand training
The condition of supporting are as follows: 28~30 DEG C of cultivation temperature, dark aerobic culture 6~8 days, ratio of ventilating per hour is liquid and gas volume ratio
1:0.5,200~300r/min of speed of agitator；
Described every liter of component of culture medium for expanding culture is as follows:
15~20g of glucose, 6~10g of peptone, 1~3g of yeast extract, 0.2~0.8g of dipotassium hydrogen phosphate, magnesium sulfate 0.01~
0.03g, 0.1~0.29g of calcium chloride.
11. handling the application of the microbial bacterial agent of heavy metal pollution of soil described in claim 1, which is characterized in that steps are as follows:
The microbial bacterial agent for handling heavy metal pollution of soil is directly applied in the soil of heavy metal pollution as base manure, is turned over mixed
Even, amount of application is 8~12g/ square metres, is handled 33~38 days under conditions of 29~31 DEG C of temperature, pH7.5~8.5.
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| CN110814003B (en) * | 2019-11-19 | 2020-07-28 | 江苏省地质调查研究院 | In-situ passivation method for fermented organic fertilizer in heavy metal polluted farmland |
| CN114700362A (en) * | 2022-03-08 | 2022-07-05 | 山东大学 | Method for preventing and treating soil heavy metal pollution by using microbial agent |
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