CN106422149A - Biological preparation of degrading organophosphorus pesticide residues - Google Patents
Biological preparation of degrading organophosphorus pesticide residues Download PDFInfo
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
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- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/04—Pesticides, e.g. insecticides, herbicides, fungicides or nematocides
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/26—Organic substances containing nitrogen or phosphorus
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Abstract
The invention relates to a biological preparation of degrading organophosphorus pesticide residues. The biological preparation is prepared from the following components: a biosorbent, a composite microbial agent, lignosulfonate, polyvinyl alcohol, fatty alcohol polyoxyethylen ether, glycerol and Tween. According to the biological preparation, by utilizing compounding of strains of Bacillus amyloliquefaciens, Paracoccus sp. strain, Trichoderma harzianum, Burkholderia and Aeruginosa, the degrading capability of degrading organophosphorus pesticides and strains with different degradation characteristics can be fully exerted, and the biological preparation can be suitable for the demands on governing environments with different pollution degrees and has a good application prospect.
Description
Technical field
The present invention relates to the pesticide engineering bacterium of degradable organophosphors is and in particular to the biological of degrading organic phosphor pesticides residual is made
Agent.
Background technology
Pesticide residues are not to be decomposed in the period after Pesticide use and residue in organism, cutting, soil, water
Trace pesticide substance in body, air, the general name of toxic metabolite, degradation product and impurity.It is applied to the pesticide on crop, wherein
A part is attached on crop, and a part is scattered in the environment such as soil, air and water, the part in the pesticide of environment remaining
Can be absorbed by plants again.Residual pesticide directly passes through fruit or water, air reach in people, animal body, or passes through environment, food
Chain is ultimately transferred to people, animal.
At present, its function difference pressed by the main chemical pesticide using, and is broadly divided into insecticide (Insecticide), weeding
Agent (Herbicide) and antibacterial (Fungicide) three major types.Different according to chemical constitution, organochlorine class, organophosphors can be divided into
The polytypes such as class, pyrethroid lipid, carbamic acid lipid and inorganic pesticide.The common feature of these chemical pesticides is:①
Toxic, and targeting is poor；2. residual, is generally divided into high residue, middle residual and low-residual etc.；3. animal migration and cumulative bad,
Can be migrated by air, soil or subsoil water, or even accumulate in vivo.Organophosphorus insecticides becomes current use
One of maximum pesticide of amount, belongs to highly effective pesticide more.At present, organophosphorus pesticide include thimet, disulfoton, demeton, to sulfur
Phosphorus, Bayer 71628, orthene, fenifrothion, Mortopl, metrifonate, Rogor, Malathion, parathion-methyl, dimethyl sulfide are inhaled
Phosphorus, parathion-methyl, demeton-methyl, omethoate, Azodrin etc..Organochlorine pesticide has clofenotane, Gamma Hexaochlorocyclohexane, gamma hch, first
Oxygen, high residual hazard DDT, 5a,6,9,9a-hexahydro-6,9-methano-2,4.Conventional pyrethroid lipid pesticide has decamethrin, cypermethrin, permethrin, tetramethrin, first
Ether chrysanthemum fat etc..Carbamatess have sevin, isoprocarb, Aldicarb, Furadan and isolan etc..
Organophosphorus pesticide is so that it is wide in variety, drug effect is high, prevention and treatment range wide spectrum the features such as, be still so far in pesticide consumption most
Classification, play the important and pivotal role in agricultural production, it is total that the production of China's organophosphorus pesticide and usage amount account for pesticide
More than half of production and usage amount.But, organophosphorus pesticide has stronger toxicity, and after long-term, a large amount of uses, it is in farming
Residual on thing can cause food-safety problem, also have impact on agricultural products in China outlet, and the pesticide being scattered also can make water and
Soil is contaminated.
With the biological renovation method of Microbial Degradation of Organophosphates residual, there is inexpensive, nontoxic, non-secondary pollution etc.
Advantage, is one of method of reduction pollution by pesticides of most application prospect.The side that organic phosphorus pesticide degradation microbe to screen is commonly used
Method is collection sample from the soil suffering from pollution by pesticides for a long time or water body, and enriched culture, plate streaking etc. operate, and separate
To single bacterium colony, then through domestication culture, or the method such as ultraviolet chemomorphosises obtains efficient degrading bacterial strain, or by cell engineering,
The technological means such as genetic engineering build engineered strain.
But there are a lot of problems in the biological preparation of the pesticide residues such as existing degrading organic phosphor, thus limit modern its
Development.The principal element of developmental limitation has：Biological high specificity except residual, it is to the environment bar such as temperature, humidity and soil
Part is bordering on the demand of harshness, not the matching of Industrialized processing technique and equipment, the backwardness of formulating technology etc..
Currently in order to improving the efficiency of degraded residual, people are generally compounded using multiple bacterial strains, and from suitable
Bacterial strain is compounded, and reaches mutual supplement with each other's advantages, stablizes the problem that the purpose degraded is that needs solve.
Content of the invention
It is an object of the invention to provide a kind of biological preparation of degrading organic phosphor pesticides residual, described biological preparation is by biology
Adsorbent, composite bacteria agent capable, lignosulfonates, polyvinyl alcohol, fatty alcohol-polyoxyethylene ether, glycerol, tween composition.
Preferably, described biological preparation its by following weight portion raw material preparation：
Biological adsorption agent 10-12 part, composite bacteria agent capable 20-30 part, lignosulfonates 2-3 part, polyvinyl alcohol 2-4 part, fatty alcohol
Polyoxyethylene ether 3-5 part, glycerol 0.5-0.7 part, tween 0.5-0.7 part.
Described biological adsorption agent is shitosan and kieselguhr compares 1 according to weight:2 are mixed with；
Described composite bacteria agent capable is made up of bacillus amyloliquefaciens, secondary coccus, trichoderma harzianum, Burkholderia, pseudomonass；
Described bacillus amyloliquefaciens are（Bacillus amyloliquefaciens）ATCC 23843；
Described pair coccus（Paracoccus sp.）For CGMCC NO.3639 (CN101899414)；
Described trichoderma harzianum（Trichoderma harzianum）For CGMCC NO.4963 (CN102505010);
Described Burkholderia（Burkholderia jiangsuensis）CGMCC9028（CN103952354）；
Described pseudomonass（Pseudomonas aeruginosa）For ATCC 15442.
The preparation method of described composite bacteria agent capable is：By bacillus amyloliquefaciens, secondary coccus, trichoderma harzianum, Burkholderia,
It is 1 × 10 that pseudomonass are cultivated respectively to concentration7The bacterium solution of individual/ml, then according to 5:4:3:3:2 volume ratio mixing, obtains final product；
The preparation method of described biological preparation comprises the steps：
（1）Prepare biological adsorption agent：Shitosan and kieselguhr are compared 1 according to weight:Ethanol at a temperature of 150 DEG C after 2 mixing
Solution（20% ethanol solution）Middle heating was dried after 4 hours, and pulverized, and obtained biological adsorption agent；
（2）By biological adsorption agent, lignosulfonates, polyvinyl alcohol, fatty alcohol-polyoxyethylene ether, glycerol, tween mix homogeneously,
1000 revs/min of rotating speed stirs 20 minutes, obtains suspension；
（3）In step（2）The suspension of preparation is stirred, is slowly added composite bacteria agent capable, all with 800 revs/min of rotating speeds
Continue stirring after having added to obtain final product for 10 minutes.
Strain of the present invention all can be from China Committee for Culture Collection of Microorganisms's common micro-organisms center
And American Type Culture collection warehousing (CGMCC)（ATCC）It is commercially available.
Strain of the present invention all can obtain the bacterium solution of desired concn by conventional cultural method, this and non-invention
Innovative point, as space is limited, do not repeat one by one.
The having the beneficial effect that of hinge structure of the present invention：
The host speciality of the pesticide residues such as microbial degradation organophosphors is its advantage, is also that it mainly lacks simultaneously
Fall into.The specialization characteristic of single microorganism preparation so as to be difficult to all farmlands are effectively prevented and kill off, thus with regard to restriction micro-organisms
Preparation popularization and application.And there is following subject matter in the biological restoration field of organophosphorus pesticide.1., in varying environment
The concentration of organophosphorus pesticide has differences, and the degradation capability of the bacterial strain that the screening of different experiments room obtains there is also difference, has
Bacterial strain the organophosphorus residue of low concentration is degraded very fast, some bacterial strains are degraded very fast to the organophosphorus residue of higher concentration, make
With the preparation of this kind of single bacterial strain, it is difficult quickly to remove the organophosphorus pesticide in environment.2., the general strains for degrading adopting
Speed is slower, needs the abundant degradation effect of process time competence exertion of 7 days～more than 30 days.Some researchs show, using many
It is the effective way solving excessively single defect that individual biocontrol bacterial strain mixed developing goes out complex microorganism preparations, the study show that, profit
With compounding and can fully send out between strains B. amyloliquefaciens, secondary coccus, trichoderma harzianum, Burkholderia, pseudomonass
Wave the degradation capability of the degrading organic phosphor pesticides degradation bacteria strains with different degradation characteristics, be suitable for different pollution level environment and control
The needs of reason, the biological preparation of preparation can in 3-7 days the different degrees of organophosphorus pesticide of fast degradation, degradation rate reaches
More than 95%, effectively remove organophosphorus pesticide pollution.
It is pure biological preparation that invention formulation does not contain chemical productss, it is to avoid the accumulation of chemical residue amount protects soil
And environment, combine, with specific surfactant, synergist, dispersant, wetting agent etc., the biological preparation made and there is adaptability
By force, good stability, non-aging the features such as, and rationally utilize microorganism between synergism, nontoxic, ecological, environmental protective, show
Work reduces dosage, spraying times and drug cost, and biological preparation of the present invention is prepared conveniently, and the holding time is long, has
Very strong practical value.
Specific embodiment
Form is described in further detail to the above of the present invention again by the following examples, but should not manage this
Solve the scope for the above-mentioned theme of the present invention and be only limitted to below example, all technology realized based on the above of the present invention are equal
Belong to the scope of the present invention.
Embodiment 1
A kind of biological preparation of degrading organic phosphor pesticides residual, described biological preparation its by following weight portion raw material preparation：
10 parts of biological adsorption agent, 20 parts of composite bacteria agent capable, 2 parts of lignosulfonates, 2 parts of polyvinyl alcohol, fatty alcohol-polyoxyethylene ether 3
Part, 0.5 part of glycerol, 0.5 part of tween.
Described biological adsorption agent is shitosan and kieselguhr compares 1 according to weight:2 are mixed with；
Described composite bacteria agent capable is made up of bacillus amyloliquefaciens, secondary coccus, trichoderma harzianum, Burkholderia, pseudomonass；
Described bacillus amyloliquefaciens are（Bacillus amyloliquefaciens）ATCC 23843；
Described pair coccus（Paracoccus sp.）For CGMCC NO.3639 (CN101899414)；
Described trichoderma harzianum（Trichoderma harzianum）For CGMCC NO.4963 (CN102505010);
Described Burkholderia（Burkholderia jiangsuensis）CGMCC9028（CN103952354）；
Described pseudomonass（Pseudomonas aeruginosa）For ATCC 15442.
The preparation method of described composite bacteria agent capable is：By bacillus amyloliquefaciens, secondary coccus, trichoderma harzianum, Burkholderia,
It is 1 × 10 that pseudomonass are cultivated respectively to concentration7The bacterium solution of individual/ml, then according to 5:4:3:3:2 volume ratio mixing, obtains final product；
The preparation method of described biological preparation comprises the steps：
（1）Prepare biological adsorption agent：Shitosan and kieselguhr are compared 1 according to weight:Ethanol at a temperature of 150 DEG C after 2 mixing
Solution（20% ethanol solution）Middle heating was dried after 4 hours, and pulverized, and obtained biological adsorption agent；
（2）By biological adsorption agent, lignosulfonates, polyvinyl alcohol, fatty alcohol-polyoxyethylene ether, glycerol, tween mix homogeneously,
1000 revs/min of rotating speed stirs 20 minutes, obtains suspension；
（3）In step（2）The suspension of preparation is stirred, is slowly added composite bacteria agent capable, all with 800 revs/min of rotating speeds
Continue stirring after having added to obtain final product for 10 minutes.
Using method：Amount of application 5-10ml/m2.
Embodiment 2
A kind of biological preparation of degrading organic phosphor pesticides residual, it is by the raw material preparation of following weight portion：
12 parts of biological adsorption agent, 30 parts of composite bacteria agent capable, 3 parts of lignosulfonates, 4 parts of polyvinyl alcohol, fatty alcohol-polyoxyethylene ether 5
Part, 0.7 part of glycerol, 0.7 part of tween.
Described biological adsorption agent is shitosan and kieselguhr compares 1 according to weight:2 are mixed with；
Described composite bacteria agent capable is made up of bacillus amyloliquefaciens, secondary coccus, trichoderma harzianum, Burkholderia, pseudomonass；Described
Bacillus amyloliquefaciens are（Bacillus amyloliquefaciens）ATCC 23843；
Described pair coccus（Paracoccus sp.）For CGMCC NO.3639 (CN101899414)；
Described trichoderma harzianum（Trichoderma harzianum）For CGMCC NO.4963 (CN102505010);
Described Burkholderia（Burkholderia jiangsuensis）CGMCC9028（CN103952354）；
Described pseudomonass（Pseudomonas aeruginosa）For ATCC 15442.
The preparation method of described composite bacteria agent capable is：By bacillus amyloliquefaciens, secondary coccus, trichoderma harzianum, Burkholderia,
It is 1 × 10 that pseudomonass are cultivated respectively to concentration7The bacterium solution of individual/ml, then according to 5:4:3:3:2 volume ratio mixing, obtains final product；
The preparation method of described biological preparation comprises the steps：
（1）Prepare biological adsorption agent：Shitosan and kieselguhr are compared 1 according to weight:Ethanol at a temperature of 150 DEG C after 2 mixing
Solution（20% ethanol solution）Middle heating was dried after 4 hours, and pulverized, and obtained biological adsorption agent；
（2）By biological adsorption agent, lignosulfonates, polyvinyl alcohol, fatty alcohol-polyoxyethylene ether, glycerol, tween mix homogeneously,
1000 revs/min of rotating speed stirs 20 minutes, obtains suspension；
（3）In step（2）The suspension of preparation is stirred, is slowly added composite bacteria agent capable, all with 800 revs/min of rotating speeds
Continue stirring after having added to obtain final product for 10 minutes.
Using method：Amount of application 5-10ml/m2.
Embodiment 3
The Degradation to parathion-methyl, Malathion and orthene for the biological preparation of the present invention
All add 20mL minimal medium (g.L in the triangular flask of three 100mL sterilizings-1, NH4NO3 0.5,
Na2HPO41.19, KH2PO40.45, MgSO40.5, deionized water 1000ml, pH6.8), then add parathion-methyl, horse respectively
Draw sulfur phosphorus and orthene concentration to 300mg/L, add the biological preparation of embodiment 1 preparation.
Timing sampling, measures parathion-methyl, Malathion and orthene concentration respectively by high performance liquid chromatography.
Efficient liquid phase measures Malathion chromatographic condition：Chromatographic column Hypersil C18 post；Mobile phase:70% methanol (contains
2mM ammonium formate+30% water；Flow velocity：0.3mL/min；Detection wavelength 220nm, column temperature:Room temperature；Sample size:15μ1.
Efficient liquid phase measures orthene chromatographic condition：Chromatographic column Hypersil C18 post；Mobile phase:V (methanol)：V
(water)：=15：85；Flow velocity：1.5mL/min；Detection wavelength 215nm, column temperature:30℃；Sample size:15μ1.
Degradation rate (%)=(1- processes sample carryover amount/control sample residual quantity) × 100%.
Result is as shown in table 1
Table 1
| Degradation rate | 5h | 10h | 15h |
| Parathion-methyl | 73.45% | 93.27% | 99.33% |
| Malathion | 80.03% | 90.54% | 99.81% |
| Orthene | 68.77% | 89.73% | 99.75% |
Embodiment 4
The degradation effect to Fructus Cucumidis sativi surface dichlorvos pesticide residues for the biological preparation of the present invention
1) two identical Fructus Cucumidis sativi plot, each 50m are chosen2, uniformly spray 80% DDT EC agriculture of 1000 times of dilutions
Medicine；
2), after 8h, a plot uniformly sprays the biological preparation of the degrading organic phosphor pesticides residual of embodiment 1 preparation as treatment group,
Amount of application 5ml/m2, another plot compares group with spray clear water；
3) after 24h, five point sampling detection Fructus Cucumidis sativi surface dichlorvos residual, result shows, treatment group is residual compared with matched group dichlorvos
Allowance reduces 90%.
Although, above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the scope of protection of present invention.
Claims (7)
1. a kind of biological preparation of degrading organic phosphor pesticides residual is it is characterised in that described biological preparation is by biological adsorption agent, multiple
Close microbial inoculum, lignosulfonates, polyvinyl alcohol, fatty alcohol-polyoxyethylene ether, glycerol, tween composition.
2. biological preparation according to claim 1 is it is characterised in that described biological adsorption agent is shitosan and kieselguhr is pressed
Compare 1 according to weight:2 are mixed with and obtain.
3. the biological preparation according to claim 1-2 it is characterised in that described biological preparation by following weight portion raw material
Preparation：
10 parts of biological adsorption agent, 20 parts of composite bacteria agent capable, 2 parts of lignosulfonates, 2 parts of polyvinyl alcohol, fatty alcohol-polyoxyethylene ether 3
Part, 0.5 part of glycerol, 0.5 part of tween.
4. the preparation according to claim 1-3 is it is characterised in that described composite bacteria agent capable is by bacillus amyloliquefaciens, secondary ball
Bacterium, trichoderma harzianum, Burkholderia, pseudomonass are prepared.
5. the preparation according to claim 1-4 it is characterised in that
Described bacillus amyloliquefaciens（Bacillus amyloliquefaciens）For ATCC 23843；
Described pair coccus（Paracoccus sp.）For CGMCC NO.3639；
Described trichoderma harzianum（Trichoderma harzianum）For CGMCC NO.4963；
Described Burkholderia（Burkholderia jiangsuensis）For CGMCC NO.9028；
Described pseudomonass（Pseudomonas aeruginosa）For ATCC 15442；
The preparation method of described composite bacteria agent capable is：By bacillus amyloliquefaciens, secondary coccus, trichoderma harzianum, Burkholderia, false list
It is 1 × 10 that born of the same parents bacterium is cultivated respectively to concentration7The bacterium solution of individual/ml, then according to 5:4:3:3:2 volume ratio mixing, obtains final product.
6. preparation described in claim 1-5 is used for the purposes of degrading pesticide residues.
7. preparation described in claim 1-6 is used for the purposes of degrading organic phosphor residual.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106244502A (en) * | 2016-09-27 | 2016-12-21 | 湖北大学 | One strain efficient dephosphorization and the pseudomonas of degraded lecithin |
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| CN101637779A (en) * | 2008-07-30 | 2010-02-03 | 中国科学院沈阳应用生态研究所 | Organic phosphorus degrading preparation and preparation method thereof |
| CN102296043A (en) * | 2010-09-17 | 2011-12-28 | 安徽农业大学 | Multi-strain composition for high-efficiency degradation of organic phosphorus pesticides in environment |
| CN104525557A (en) * | 2014-12-15 | 2015-04-22 | 上海创博生态工程有限公司 | Ecological restoration agent for fallow ginseng soil and preparation method of ecological restoration agent |
| CN105483066A (en) * | 2016-02-25 | 2016-04-13 | 北京联合大学 | Organophosphorus pesticide degrading bacterium agent and preparation method thereof |
| CN105950172A (en) * | 2016-06-17 | 2016-09-21 | 战锡林 | Organophosphorus pesticide contaminated soil remediation agent |
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2016
- 2016-09-23 CN CN201610841011.1A patent/CN106422149A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101637779A (en) * | 2008-07-30 | 2010-02-03 | 中国科学院沈阳应用生态研究所 | Organic phosphorus degrading preparation and preparation method thereof |
| CN102296043A (en) * | 2010-09-17 | 2011-12-28 | 安徽农业大学 | Multi-strain composition for high-efficiency degradation of organic phosphorus pesticides in environment |
| CN104525557A (en) * | 2014-12-15 | 2015-04-22 | 上海创博生态工程有限公司 | Ecological restoration agent for fallow ginseng soil and preparation method of ecological restoration agent |
| CN105483066A (en) * | 2016-02-25 | 2016-04-13 | 北京联合大学 | Organophosphorus pesticide degrading bacterium agent and preparation method thereof |
| CN105950172A (en) * | 2016-06-17 | 2016-09-21 | 战锡林 | Organophosphorus pesticide contaminated soil remediation agent |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106244502A (en) * | 2016-09-27 | 2016-12-21 | 湖北大学 | One strain efficient dephosphorization and the pseudomonas of degraded lecithin |
| CN106244502B (en) * | 2016-09-27 | 2019-06-18 | 湖北大学 | The pseudomonad of one plant of efficient dephosphorization and degradation lecithin |
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Application publication date: 20170222 |