CN108251330A - Complex micro organism fungicide for administering soil pollution and its preparation method and application - Google Patents
Complex micro organism fungicide for administering soil pollution and its preparation method and application Download PDFInfo
- Publication number
- CN108251330A CN108251330A CN201711454685.7A CN201711454685A CN108251330A CN 108251330 A CN108251330 A CN 108251330A CN 201711454685 A CN201711454685 A CN 201711454685A CN 108251330 A CN108251330 A CN 108251330A
- Authority
- CN
- China
- Prior art keywords
- culture
- atcc
- cultures
- micro organism
- complex micro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Soil Sciences (AREA)
- Materials Engineering (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a kind of complex micro organism fungicide, including the following raw material component：Japanese pseudomonad culture, Comamonas testosteroni culture, microbacterium culture and De Shi acidovorax facilis cultures；The present invention to various bacteria culture mixed fermentation by being made complex micro organism fungicide, to in soil because the organic substance residues such as nitrobenzene compound formed when insecticide or pesticide has effective degradation, while the mixed culture of special strain and multiple bacteria compound fermentation provide a kind of simple and practicable preparation method.
Description
Technical field
It is dirty for administering soil the present invention relates to a kind of microbial bacterial agent and its preparation method and application more particularly to one kind
Complex micro organism fungicide of dye and its preparation method and application.
Background technology
Nitrobenzene compounds are widely used in the production nitro of pesticide, dyestuff, explosive, rubber and other chemical products
Benzene-like compounds are widely used, and environment can be entered by waste water and gas, also can because transport and production process in contingency and
The improper disposition of memory tank, and largely enter environment, the nitrobenzene class pollutant in environment mainly includes nitrobenzene, nitroxyl chloride
Benzene, nitrotoleune, nitrophenol, nitroaniline and nitrobenzoic acid etc., most of nitrobenzene compounds have chemical property
The characteristics of stable, high toxicity and easy biological concentration, is put into the screen priority pollutants in US Gov Env Protection Agency and China
In list.
In the prior art, the restorative procedure of nitrobenzene compounds pollution mainly includes Physical, chemical method and bioanalysis；
Physical mainly adsorbs pollutant using porous resin and activated carbon etc.；Chemical method mainly eliminates dirt using oxidation reaction
Contaminate object；Bioanalysis come degradation of contaminant, finally makes pollutant mineralising mainly using the engineering bacteria of domestication or structure.With change
The advantages of method is compared with Physical, biological treatment is that expense is low, does not easily cause secondary pollution, can farthest reduce dirt
The concentration of object is contaminated, is had a wide range of application.Meanwhile the soil containing nitrobenzene compounds is often that multiple pollutant coexists, and is utilized
Synergistic effect and microorganism between microorganism can remove multiple pollutant, accelerate nitre simultaneously to the Co metabolism effect of substrate
The degradation of based compound.Therefore, with the extensive use and development of biological reinforcing technology and technique for gene engineering, in prevention nitro
In terms of the environmental pollution of benzene-like compounds, biological treatment has broad application prospects.
But there are the treatment effect of p-nitrophenyl class compound is poor, and makes for composite microbial bacteria of the prior art
Standby process is complicated, and thalline is easy to run off, so as to cause the wasting of resources.
Invention content
Goal of the invention：The first object of the present invention is to provide a kind of complex micro organism fungicide.
The second object of the present invention is to provide the preparation method of the complex micro organism fungicide.
The third object of the present invention is to provide the application of the complex micro organism fungicide.
Technical solution：In order to achieve the above-mentioned object of the invention, the present invention provides a kind of complex micro organism fungicide, including as follows
Raw material components：Japanese pseudomonad culture, Comamonas testosteroni culture, microbacterium culture and the training of De Shi acidovorax facilis
Support object.
Wherein, Japanese pseudomonad culture, Comamonas testosteroni culture, microbacterium culture and De Shi food acid
The ratio of bacterium culture is 1:(2~3):(1.5~2.5):(3~4).
Preferably, the Japanese pseudomonad culture is Japanese aeruginosa atcc 33616 or ATCC33660 cultures
Object；The Comamonas testosteroni culture is 39523 culture of ATCC 700441 or ATCC；The microbacterium culture
For 31001 cultures of ATCC；The De Shi acidovorax facilis culture is 49664 culture of DSM 50403 or ATCC.
Further, the raw material components of complex micro organism fungicide further include bacillus subtilis culture, fixed nitrogen actinomyces
Culture, hydrogenlike silicon ion culture and candidiasis culture.
Wherein, bacillus subtilis culture is 21556 cultures of bacillus subtilis ATCC；Fixed nitrogen actinomyces are cultivated
Object is 33255 cultures of fixed nitrogen actinomyces ATCC；Hydrogenlike silicon ion culture is hydrogenlike silicon ion ATCC 49419, ATCC
17023rd, 33575 cultures of ATCC；Candidiasis culture is cultivated for candidiasis CICC 33050, NYNU 14772
Object.
Preferably, Japanese pseudomonad culture, Comamonas testosteroni culture, microbacterium culture, De Shi foods
Sour bacterium culture, bacillus subtilis culture, fixed nitrogen actinomyces culture, hydrogenlike silicon ion culture and candidiasis training
Support object ratio be：1:(2~3):(1.5~2.5):(3~4):(2.5~3.5):(0.5~1):(1~1.5):(2~3).
A kind of preparation method of complex micro organism fungicide, includes the following steps：
(1) using mixed culture medium A to Japanese pseudomonad culture, Comamonas testosteroni culture, microbacterium
Culture and De Shi acidovorax facilis culture are mixed to obtain mixed culture A by weight；
(2) nitrobenzene compound is added in the mixed culture A of step (1), the mixed culture B after being tamed；
(3) bacillus subtilis culture in the mixed culture B and raw material components after the domestication for obtaining step (2),
Fixed nitrogen actinomyces culture, hydrogenlike silicon ion culture and candidiasis culture are inoculated into level-one training according to parts by weight respectively
It supports in case, aeration culture obtains level-one bacterium solution, then level-one bacterium solution is inoculated into two level incubator and obtains two level bacterium solution；
(4) the two level bacterium solution for each strain for obtaining step (3) uniformly mixes, and bacterial suspension is made, and controls effectively living
Bacterium number >=1.5 × 1010/g obtain the complex micro organism fungicide.
Wherein, in step (1), the acquisition pattern of each microbial strain culture is：Each strain in the raw material components is inoculated with
In slant medium, expand culture respectively, obtain the culture of each strain；
Preferably, each slant medium and condition of culture are respectively：Japanese aeruginosa atcc 33616 or
ATCC33660 cultures：Nutrient agar culture medium, pH 6.8 ± 0.2, cultivation temperature：28℃；Comamonas testosteroni
39523 culture of ATCC 700441 or ATCC：Nutrient agar (NA), pH 6.8 ± 0.2, cultivation temperature:30.0℃；Microbacterium
31001 cultures of ATCC：Nutrient agar, pH 6.8 ± 0.2, cultivation temperature：37℃；De Shi acidovorax facilis DSM 50403 or
49664 cultures of ATCC：Nutrient agar (NA), pH 6.8 ± 0.2, cultivation temperature:30.0℃；Bacillus subtilis ATCC
21556 cultures：Nutrient agar (NA) plus 1% potato starch, 6.8 ± 0.2 cultivation temperatures of pH：37℃；Fixed nitrogen actinomyces
33255 cultures of ATCC：Malt extract powder 130g/L, agar 15g/L, chloramphenicol 0.1g/L, pH 6.0 ± 0.2, cultivation temperature:
28.0℃；Hydrogenlike silicon ion ATCC 49419, ATCC 17023,33575 cultures of ATCC：Model Neil yeast agar culture
Base, pH 7.0 ± 0.2, cultivation temperature:24.0℃；Candidiasis CICC 33050,14772 cultures of NYNU：YCM is cultivated
Base, cultivation temperature：25℃.
In step (1), the mixed culture medium A includes the component of following concentration of component：8~12g/L of tryptone, soybean
10~20g/L of peptone, 2~5g/L of beef leaching object, H3BO32~5g/L, NaH2PO4·H2O2~4g/L, 5~8g/L of NaCl, fine jade
Fat 10~20g/L and MnCl20.2~0.3g/L of 4H2O, pH value are 6.5~6.8.
Preferably, it is the culture for the mixed culture A that will be in exponential phase the step of the domestication in step (2)
Nitrobenzene compound is added in base, respectively the secondary culture at 2~3 days, 4~6 days and 7~9 days, make strain density reach 4 ×
106~5 × 106/ml, bacterium survival rate >=85%.
Further, it in step (3), is inoculated into level-one incubator, aeration culture obtains level-one bacterium solution, then by level-one bacterium
Liquid be inoculated into two level incubator the specific steps are：The incubator of 5~8L is inoculated by 2~4% inoculum concentration of culture medium
In, temperature is 20~25 DEG C, and pH value of solution is neutral, aeration culture 40h, then 5~8L level-one bacterium solutions are transferred in two level incubator,
Obtain 20~30L of two level bacterium solution；Wherein, nutrient media components are：Corn flour 10g/L, 5 g/L of glucose, beancake powder 10g/L, fish
Powder 7g/L, CaCO3 7g/L、(NH4)2SO4 10g/L、K2HPO4 0.4g/L、 MgSO4.7H2O 0.3g/L and MnSO4·H2O
0.2g/L。
Preferably, in step (4), the preparation method of bacterial suspension：It is dilute that 3.0ml~5.0ml is drawn with 5.0ml suction pipes
It releases liquid to add in the culture medium of each microbial strain culture, pressure-vaccum, washes lower lawn repeatedly；Then, washing lotion is moved to separately with 5.0ml suction pipes
In one sterile test tube, mixed several seconds with motorized agitator or shaken on palm and struck several times, so that each microbial strain culture suspends
Uniformly obtained multi strain co cultivation object suspension.
The content of present invention further includes a kind of application of complex micro organism fungicide in soil pollution is administered.
Further, application of the complex micro organism fungicide in soil pollution is administered is being administered for the complex micro organism fungicide
Application in by nitrobenzene compound contaminated soil.
Advantageous effect：Compared with prior art, the present invention is compound micro- by the way that various bacteria culture mixed fermentation is made
Bacteria agent, to effectively being dropped in soil because the organic substance residues such as nitrobenzene compound formed when insecticide or pesticide has
Solution acts on, while the mixed culture of special strain and multiple bacteria compound fermentation provide a kind of simple and practicable preparation method.
Specific embodiment
Technical scheme of the present invention is described in detail below, unless otherwise specified, following raw materials according can be obtained from commercially available
.
Embodiment 1
Raw material components：Japanese 33616 culture of aeruginosa atcc, Comamonas testosteroni ATCC 700441 are cultivated
The ratio of 50403 culture of object, 31001 cultures of microbacterium ATCC and De Shi acidovorax facilis DSM is 1:2:1.5:3.
It prepares and applies：(1) Japanese pseudomonad culture, Comamonas testosteroni are trained using mixed culture medium A
Object, microbacterium culture and De Shi acidovorax facilis culture is supported to be mixed to obtain mixed culture A, mixing training by weight
Support the component that base A includes following concentration of component：Tryptone 8g/L, soya peptone 10g/L, beef leaching object 2g/L, H3BO3 2g/
L、NaH2PO4·H2O 2g/L, NaCl 5g/L, agar 10g/L and MnCl2.4H20.2 g/L of O, pH value 6.5；
(2) nitrobenzene compound is added in the mixed culture A of step (1), the mixed culture B after being tamed,
The step of domestication, adds nitrobenzene compound will to be in the culture medium of mixed culture A, respectively at 2 days, 4 days and 7 days
Secondary culture makes strain density reach 4 × 106A/ml, bacterium survival rate 85%.
(3) the mixed culture B after the domestication for obtaining step (2) is inoculated into level-one incubator, and aeration culture obtains
Level-one bacterium solution, then level-one bacterium solution is inoculated into two level incubator.
(4) zymotic fluid that step (3) obtains uniformly is mixed, is made bacterial suspension, control living bacteria count 1.5 ×
1010A/g obtains the complex micro organism fungicide.
Embodiment 2
Raw material components：Japanese 33660 culture of aeruginosa atcc, Comamonas testosteroni ATCC 39523 are cultivated
The ratio of 49664 culture of object, 31001 cultures of microbacterium ATCC and De Shi acidovorax facilis ATCC is 1:2.5:2: 3.5.
It prepares and applies：(1) Japanese pseudomonad culture, Comamonas testosteroni are trained using mixed culture medium A
Object, microbacterium culture and De Shi acidovorax facilis culture is supported to be mixed to obtain mixed culture A, mixing training by weight
Support the component that base A includes following concentration of component：Tryptone 10g/L, soya peptone 15g/L, beef leaching object 3g/L, H3BO3
4g/L、NaH2PO4·H2O 3g/L, NaCl 7g/L, agar 15g/L and MnCl2.4H20.25 g/L of O, pH value 6.7；
(2) nitrobenzene compound is added in the mixed culture A of step (1), the mixed culture B after being tamed,
The step of domestication, adds nitrobenzene compound will to be in the culture medium of mixed culture A, respectively at 3 days, 5 days and 8 days
Secondary culture makes strain density reach 4.5 × 106A/ml, bacterium survival rate 90%；
(3) the mixed culture B after the domestication for obtaining step (2) is inoculated into level-one incubator, and aeration culture obtains
Level-one bacterium solution, then level-one bacterium solution is inoculated into two level incubator；
(4) zymotic fluid that step (3) obtains uniformly is mixed, is made bacterial suspension, control living bacteria count for 2 ×
1010A/g obtains the complex micro organism fungicide.
Embodiment 3
Raw material components：Japanese 33660 culture of aeruginosa atcc, Comamonas testosteroni ATCC 39523 are cultivated
The ratio of 49664 culture of object, 31001 cultures of microbacterium ATCC and De Shi acidovorax facilis ATCC is 1:3:2.5:4.
It prepares and applies：(1) Japanese pseudomonad culture, Comamonas testosteroni are trained using mixed culture medium A
Object, microbacterium culture and De Shi acidovorax facilis culture is supported to be mixed to obtain mixed culture A, mixing training by weight
Support the component that base A includes following concentration of component：Tryptone 12g/L, soya peptone 20g/L, beef leaching object 5g/L, H3BO3 5g/
L、NaH2PO4·H2O 4g/L, NaCl 8g/L, agar 20g/L and MnCl2.4H2O 0.3g/L, pH value 6.8；
(2) nitrobenzene compound is added in the mixed culture A of step (1), the mixed culture B after being tamed,
The step of domestication, adds nitrobenzene compound will to be in the culture medium of mixed culture A, respectively at 3 days, 6 days and 9 days
Secondary culture makes strain density reach 5 × 106A/ml, bacterium survival rate 90%；
(3) the mixed culture B after the domestication for obtaining step (2) is inoculated into level-one incubator, and aeration culture obtains
Level-one bacterium solution, then level-one bacterium solution is inoculated into two level incubator；
(4) zymotic fluid that step (3) obtains uniformly is mixed, is made bacterial suspension, control living bacteria count 2.0 ×
1010A/g obtains the complex micro organism fungicide.
Embodiment 4
Raw material components：Japanese 33660 culture of aeruginosa atcc, Comamonas testosteroni ATCC 39523 are cultivated
Object, 31001 cultures of microbacterium ATCC, 49664 cultures of De Shi acidovorax facilis ATCC, bacillus subtilis ATCC 21556 are trained
Support object, 33255 cultures of fixed nitrogen actinomyces ATCC, 49419 cultures of hydrogenlike silicon ion ATCC and candidiasis CICC
The ratio of 33050 cultures is：1:2:1.5:3:2.5:0.5:1:2.
It prepares and applies：(1) Japanese pseudomonad culture, Comamonas testosteroni are trained using mixed culture medium A
Object, microbacterium culture and De Shi acidovorax facilis culture is supported to be mixed to obtain mixed culture A, mixing training by weight
Support the component that base A includes following concentration of component：Tryptone 8g/L, soya peptone 10g/L, beef leaching object 2g/L, H3BO3 2g/
L、NaH2PO4·H2O2g/L, NaCl 5g/L, agar 10g/L and MnCl2.4H2O 0.2g/L, pH value 6.5；
(2) nitrobenzene compound is added in the mixed culture A of step (1), the mixed culture B after being tamed,
The step of domestication, adds nitrobenzene compound will to be in the culture medium of mixed culture A, respectively at 2 days, 4 days and 7 days
Secondary culture makes strain density reach 4 × 106A/ml, bacterium survival rate are 85%；
(3) bacillus subtilis culture in the mixed culture B and raw material components after the domestication for obtaining step (2),
Fixed nitrogen actinomyces culture, hydrogenlike silicon ion culture and candidiasis culture are inoculated into level-one training according to parts by weight respectively
It supports in case, aeration culture obtains level-one bacterium solution, then level-one bacterium solution is inoculated into two level incubator and obtains two level bacterium solution；
(4) zymotic fluid that step (3) obtains uniformly is mixed, bacterial suspension is made, control living bacteria count is 1.5
×1010A/g obtains the complex micro organism fungicide.
Embodiment 5
Raw material components：Japanese 33616 culture of aeruginosa atcc, Comamonas testosteroni ATCC 700441 are cultivated
Object, 31001 cultures of microbacterium ATCC, 50403 cultures of De Shi acidovorax facilis DSM, bacillus subtilis ATCC 21556 are trained
Support object, 33255 cultures of fixed nitrogen actinomyces ATCC, 17023 cultures of hydrogenlike silicon ion ATCC and candidiasis NYNU
The ratio of 14772 cultures is：1:2.5:2:3.5:3:0.75:1.25:2.5.
It prepares and applies：(1) Japanese pseudomonad culture, Comamonas testosteroni are trained using mixed culture medium A
Object, microbacterium culture and De Shi acidovorax facilis culture is supported to be mixed to obtain mixed culture A, mixing training by weight
Support the component that base A includes following concentration of component：Tryptone 10g/L, soya peptone 15g/L, beef leaching object 4g/L, H3BO3 3g/
L、NaH2PO4·H2O 3g/L, NaCl 7g/L, agar 15g/L and MnCl2.4H2O 0.25g/L, pH value 6.7；
(2) nitrobenzene compound is added in the mixed culture A of step (1), the mixed culture B after being tamed,
The step of domestication, adds nitrobenzene compound will to be in the culture medium of mixed culture A, respectively at 2 days, 5 days and 8 days
Secondary culture makes strain density reach 4.5 × 106A/ml, bacterium survival rate are 90%；
(3) bacillus subtilis culture in the mixed culture B and raw material components after the domestication for obtaining step (2),
Fixed nitrogen actinomyces culture, hydrogenlike silicon ion culture and candidiasis culture are inoculated into level-one training according to parts by weight respectively
It supports in case, aeration culture obtains level-one bacterium solution, then level-one bacterium solution is inoculated into two level incubator and obtains two level bacterium solution；
(4) zymotic fluid that step (3) obtains uniformly is mixed, is made bacterial suspension, control living bacteria count 2.0 ×
1010A/g obtains the complex micro organism fungicide.
Embodiment 6
Raw material components：Japanese 33616 culture of aeruginosa atcc, Comamonas testosteroni ATCC 700441 are cultivated
Object, 31001 cultures of microbacterium ATCC, 50403 cultures of De Shi acidovorax facilis DSM, bacillus subtilis ATCC 21556 are trained
Support object, 33255 cultures of fixed nitrogen actinomyces ATCC, 33575 cultures of hydrogenlike silicon ion ATCC and candidiasis NYNU
The ratio of 14772 cultures is：1:3:2.5:4:3.5:1:1.5:3.
It prepares and applies：(1) Japanese pseudomonad culture, Comamonas testosteroni are trained using mixed culture medium A
Object, microbacterium culture and De Shi acidovorax facilis culture is supported to be mixed to obtain mixed culture A, mixing training by weight
Support the component that base A includes following concentration of component：Tryptone 12g/L, soya peptone 20g/L, beef leaching object 5g/L, H3BO3 5g/
L、NaH2PO4·H2O4g/L, NaCl 8g/L, agar 20g/L and MnCl2.4H2O 0.3g/L, pH value 6.8；
(2) nitrobenzene compound is added in the mixed culture A of step (1), the mixed culture B after being tamed,
The step of domestication, adds nitrobenzene compound will to be in the culture medium of mixed culture A, respectively at 3 days, 6 days and 9 days
Secondary culture makes strain density reach 5 × 106A/ml, bacterium survival rate 93%；
(3) bacillus subtilis culture in the mixed culture B and raw material components after the domestication for obtaining step (2),
Fixed nitrogen actinomyces culture, hydrogenlike silicon ion culture and candidiasis culture are inoculated into level-one training according to parts by weight respectively
It supports in case, aeration culture obtains level-one bacterium solution, then level-one bacterium solution is inoculated into two level incubator and obtains two level bacterium solution；
(4) zymotic fluid that step (3) obtains uniformly is mixed, is made bacterial suspension, control living bacteria count 2.5 ×
1010A/g obtains the complex micro organism fungicide.
Comparative example 1
Raw material components：21556 cultures of bacillus subtilis ATCC, 33255 cultures of fixed nitrogen actinomyces ATCC, class ball
The ratio of red 33575 cultures of bacterium ATCC and 14772 cultures of candidiasis NYNU is 3.5:1:1.5: 3.
It prepares and applies：Other specific steps and implementation condition are same as Example 6.
Comparative example 2
Raw material components：700441 cultures of Comamonas testosteroni ATCC, 31001 cultures of microbacterium ATCC, moral
50403 cultures of family name's acidovorax facilis DSM, 21556 cultures of bacillus subtilis ATCC, fixed nitrogen actinomyces ATCC 33255 are cultivated
The ratio of 14772 culture of object, 33575 cultures of hydrogenlike silicon ion ATCC and candidiasis NYNU is 3:2.5:4:3.5:
1:1.5:3.(lacking Japanese pseudomonad culture)
It prepares and applies：Other specific steps and implementation condition are same as Example 6.
Comparative example 3
Raw material components：Japanese 33616 culture of aeruginosa atcc, 31001 cultures of microbacterium ATCC, De Shi food acid
50403 cultures of bacterium DSM, 21556 cultures of bacillus subtilis ATCC, 33255 cultures of fixed nitrogen actinomyces ATCC, class
The ratio of red 33575 cultures of bacterium ATCC of ball and 14772 cultures of candidiasis NYNU is： 1：2.5:4:3.5:1:
1.5:3.(lacking Comamonas testosteroni culture)
It prepares and applies：Other specific steps and implementation condition are same as Example 6.
Comparative example 4
Raw material components：Japanese 33616 culture of aeruginosa atcc, Comamonas testosteroni ATCC 700441 are cultivated
Object, 50403 cultures of De Shi acidovorax facilis DSM, 21556 cultures of bacillus subtilis ATCC, fixed nitrogen actinomyces ATCC
The ratio of 14772 culture of 33255 cultures, 33575 cultures of hydrogenlike silicon ion ATCC and candidiasis NYNU is：1:
3:4:3.5:1:1.5:3.(lacking microbacterium culture)
It prepares and applies：Other specific steps and implementation condition are same as Example 6.
Comparative example 5
Raw material components：Japanese 33616 culture of aeruginosa atcc, Comamonas testosteroni ATCC 700441 are cultivated
Object, 31001 cultures of microbacterium ATCC, 21556 cultures of bacillus subtilis ATCC, fixed nitrogen actinomyces ATCC 33255 are trained
The ratio for supporting object, 33575 cultures of hydrogenlike silicon ion ATCC and candidiasis NYNU 14772 culture is：1:3:2.5:
3.5:1:1.5:3.(lacking De Shi acidovorax facilis culture)
It prepares and applies：Other specific steps and implementation condition are same as Example 6.
Using the complex micro organism fungicide in above-described embodiment 1~6 and comparative example 1~5 to by nitrobenzene compound dirt
Pollutant in the soil of dye is handled, by being detected to the nitrobenzene compound in soil, as a result such as table 1：
Influence of the complex micro organism fungicide of 1 different material component of table to nitrobenzene compound degradation rate in polluted soil
As shown in Table 1, it is red thin to only have bacillus subtilis culture, fixed nitrogen actinomyces culture, class ball for the explanation of comparative example 1
When bacterium culture and candidiasis culture act on polluted soil nitrobenzene compound, effect is smaller, about
9.54%；Comparative example 2~5 illustrate when the complex micro organism fungicide that act on soil lack respectively Japanese pseudomonad culture,
When Comamonas testosteroni culture, microbacterium culture or De Shi acidovorax facilis cultures, pollutant nitrobenzene compound
Degradation rate is no more than 40.57%；Examples 1 to 3 explanation only has bacillus subtilis culture, fixed nitrogen actinomyces culture, class
When polluted soil, p-nitrophenyl compound can play for the red bacterial cultures of ball and candidiasis culture collective effect
Good degradation, more than 72.44%；Especially, when Japanese pseudomonad culture, C testosteroni in embodiment 4~6
Pseudomonas culture, microbacterium culture, De Shi acidovorax facilis culture, bacillus subtilis culture, the culture of fixed nitrogen actinomyces
Object, hydrogenlike silicon ion culture and candidiasis culture collective effect when polluted soil, p-nitrophenyl compound
Degradation rate is excellent, minimum more than 80%；Wherein, the effect of 6 degrading nitrobenzene compound of embodiment is best, p-nitrophenyl chemical combination
The degradation rate of object can reach 88.51%.It can be seen that the raw material components in complex micro organism fungicide in the present invention lack one
Can not, considerable effect is played to administering one of soil pollutant nitrobenzene compound.
Claims (10)
1. a kind of complex micro organism fungicide, it is characterised in that：Including the following raw material component：Japanese pseudomonad culture, testis
Ketone comamonas culture, microbacterium culture and De Shi acidovorax facilis cultures.
2. complex micro organism fungicide according to claim 1, it is characterised in that：In the raw material components, Japanese pseudomonad
Culture, Comamonas testosteroni culture, microbacterium culture and De Shi acidovorax facilis cultures ratio be 1:(2~3):
(1.5~2.5):(3~4).
3. complex micro organism fungicide according to claim 1, it is characterised in that：Japan's pseudomonad culture is Japan
Aeruginosa atcc 33616 or ATCC33660 cultures；The Comamonas testosteroni culture for ATCC 700441 or
39523 cultures of ATCC；The microbacterium culture is 31001 cultures of ATCC；The De Shi acidovorax facilis culture is DSM
50403 or ATCC, 49664 cultures.
4. complex micro organism fungicide according to claim 1, it is characterised in that：The raw material components of the complex micro organism fungicide
It further includes：Bacillus subtilis culture, fixed nitrogen actinomyces culture, hydrogenlike silicon ion culture and candidiasis culture
Object.
5. complex micro organism fungicide according to claim 4, it is characterised in that：In the raw material components, bacillus subtilis
Culture is 21556 cultures of bacillus subtilis ATCC；Fixed nitrogen actinomyces culture is trained for fixed nitrogen actinomyces ATCC 33255
Support object；Hydrogenlike silicon ion culture is hydrogenlike silicon ion ATCC 49419, ATCC 17023,33575 cultures of ATCC；False silk
Yeast culture is candidiasis CICC 33050,14772 cultures of NYNU；Wherein, Japanese pseudomonad culture,
Comamonas testosteroni culture, microbacterium culture, De Shi acidovorax facilis culture, bacillus subtilis culture, fixed nitrogen
The ratio of actinomyces culture, hydrogenlike silicon ion culture and candidiasis culture is：1:(2~3):(1.5~2.5):
(3~4):(2.5~3.5):(0.5~1):(1~1.5):(2~3).
6. the preparation method of the complex micro organism fungicide of claim 1 or 4, it is characterised in that：Include the following steps：
(1) using mixed culture medium A to Japanese pseudomonad culture, Comamonas testosteroni culture, microbacterium culture
Object and De Shi acidovorax facilis culture are mixed to obtain mixed culture A by weight；
(2) nitrobenzene compound is added in the mixed culture A of step (1), the mixed culture B after being tamed；
(3) bacillus subtilis culture, fixed nitrogen in the mixed culture B and raw material components after the domestication for obtaining step (2)
Actinomyces culture, hydrogenlike silicon ion culture and candidiasis culture are inoculated into level-one incubator according to parts by weight respectively
In, aeration culture obtains level-one bacterium solution, then level-one bacterium solution is inoculated into two level incubator and obtains two level bacterium solution；
(4) the two level bacterium solution for each strain for obtaining step (3) uniformly mixes, and bacterial suspension is made, and controls living bacteria count
≥1.5×1010A/g obtains the complex micro organism fungicide.
7. the preparation method of complex micro organism fungicide according to claim 6, it is characterised in that：In step (1), the mixing
Culture medium A includes the component of following concentration of component：8~12g/L of tryptone, 10~20g/L of soya peptone, beef leaching object 2~
5g/L、H3BO32~5g/L, NaH2PO4·H2O2~4g/L, 5~8g/L of NaCl, agar 10~20g/L and MnCl2.4H2O
0.2~0.3g/L, pH value are 6.5~6.8.
8. the preparation method of complex micro organism fungicide according to claim 6, it is characterised in that：In step (2), the domestication
The step of will to be in the culture medium of the mixed culture A of exponential phase add nitrobenzene compound, respectively 2~3 days,
Secondary culture at 4~6 days and 7~9 days, makes strain density reach 4 × 106~5 × 106A/ml, bacterium survival rate >=85%.
9. application of the complex micro organism fungicide in soil pollution is administered described in claim 1 or 4.
10. complex micro organism fungicide is administering the application in by nitrobenzene compound contaminated soil according to claim 9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711454685.7A CN108251330B (en) | 2017-12-28 | 2017-12-28 | Compound microbial agent for treating soil pollution and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711454685.7A CN108251330B (en) | 2017-12-28 | 2017-12-28 | Compound microbial agent for treating soil pollution and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108251330A true CN108251330A (en) | 2018-07-06 |
| CN108251330B CN108251330B (en) | 2021-03-23 |
Family
ID=62724254
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711454685.7A Active CN108251330B (en) | 2017-12-28 | 2017-12-28 | Compound microbial agent for treating soil pollution and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108251330B (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101665801A (en) * | 2005-04-26 | 2010-03-10 | 清华大学 | Method for enhancing reverse resistance of microorganisms |
| CN101688213A (en) * | 2007-04-27 | 2010-03-31 | 陶氏环球技术公司 | Be used for rapidly screening microbial hosts to identify some method at the bacterial strain of output that has improvement aspect the expressing heterologous protein and/or quality |
| WO2011134998A8 (en) * | 2010-04-27 | 2011-12-15 | Lysando Holding Ag | Method of reducing biofilms |
| CN102515366A (en) * | 2011-12-23 | 2012-06-27 | 重庆文泰节能环保科技有限公司 | Microbiological degradation method for nitrobenzene industrial wastewater |
| CN102757896A (en) * | 2012-07-10 | 2012-10-31 | 徐州奇缘生态农业发展有限公司 | Efficient microbial soil remediation agent |
| AU2011347152A1 (en) * | 2010-12-23 | 2013-07-11 | Katholieke Universiteit Leuven | Antimicrobial agents |
| SE1351168A1 (en) * | 2013-10-03 | 2015-04-04 | Nordic Chemquest Ab | Process for chemical and / or biological transformation |
| WO2015067619A2 (en) * | 2013-11-05 | 2015-05-14 | Carbios | A method for degrading a plastic |
| CN106715702A (en) * | 2014-09-30 | 2017-05-24 | 巴斯夫欧洲公司 | Method for preparing aqueous acrylamide solution having low acrylic acid concentration |
-
2017
- 2017-12-28 CN CN201711454685.7A patent/CN108251330B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101665801A (en) * | 2005-04-26 | 2010-03-10 | 清华大学 | Method for enhancing reverse resistance of microorganisms |
| CN101688213A (en) * | 2007-04-27 | 2010-03-31 | 陶氏环球技术公司 | Be used for rapidly screening microbial hosts to identify some method at the bacterial strain of output that has improvement aspect the expressing heterologous protein and/or quality |
| WO2011134998A8 (en) * | 2010-04-27 | 2011-12-15 | Lysando Holding Ag | Method of reducing biofilms |
| AU2011347152A1 (en) * | 2010-12-23 | 2013-07-11 | Katholieke Universiteit Leuven | Antimicrobial agents |
| CN102515366A (en) * | 2011-12-23 | 2012-06-27 | 重庆文泰节能环保科技有限公司 | Microbiological degradation method for nitrobenzene industrial wastewater |
| CN102757896A (en) * | 2012-07-10 | 2012-10-31 | 徐州奇缘生态农业发展有限公司 | Efficient microbial soil remediation agent |
| SE1351168A1 (en) * | 2013-10-03 | 2015-04-04 | Nordic Chemquest Ab | Process for chemical and / or biological transformation |
| WO2015067619A2 (en) * | 2013-11-05 | 2015-05-14 | Carbios | A method for degrading a plastic |
| CN106715702A (en) * | 2014-09-30 | 2017-05-24 | 巴斯夫欧洲公司 | Method for preparing aqueous acrylamide solution having low acrylic acid concentration |
Non-Patent Citations (6)
| Title |
|---|
| DONGFENG ZHAO等: "Biodegradation of nitrobenzene by aerobic granular sludge in a sequencing batch reactor (SBR)", 《DESALINATION》 * |
| HUI LIN 等: "Isolation and characterization of Rhodococcus sp. NB5 capable of degrading a high concentration of nitrobenzene", 《JOURNAL OF BASIC MICROBIOLOGY》 * |
| PIYUSH CHANDNA等: "Assessment of bacterial diversity in agricultural by-product compost by sequencing of cultivated isolates and amplified compost by sequencing of cultivated isolates and amplified", 《APPL MICROBIOL BIOTECHNOL》 * |
| ZHENG CHUNLI 等: "Isolation and characterization of a nitrobenzene degrading yeast strain from activated sludge", 《JOURNAL OF HAZARDOUS MATERIALS》 * |
| 韦朝海 等: "硝基苯好氧降解的共基质及生物协同作用", 《中国环境科学》 * |
| 马汐平: "硝基苯好氧降解菌的驯化和筛选", 《辽宁大学学报》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108251330B (en) | 2021-03-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ye et al. | Microbial resources and ecology-microbial degradation of pesticides | |
| Rani et al. | Bioremediation of dyes by fungi isolated from contaminated dye effluent sites for bio-usability | |
| Esposito et al. | Biodegradation of the herbicide diuron in soil by indigenous actinomycetes | |
| Nishioka et al. | Development of culture medium for the isolation of Flavobacterium and Chryseobacterium from rhizosphere soil | |
| CN108546665A (en) | A kind of Degradation of Antibiotics mix bacterium agent and its application | |
| CN107287137B (en) | Pesticide residue broad-spectrum degradation strain DS3, and microbial inoculum produced by same and application thereof | |
| Zhu et al. | Characteristics of an atrazine degrading bacterium and the construction of a microbial agent for effective atrazine degradation | |
| CN102250808B (en) | Endophytic azotobacter of wheat producing ACC (1-aminocyclopropane-1-carboxylate) deaminase and application thereof | |
| CN108396005A (en) | Raw growth-promoting rhizobium and application thereof in one plant of rice | |
| CN101735996A (en) | Mixed bactericide for restoring places polluted by polynuclear aromatic hydrocarbons, preparation method and application method | |
| CN108410773B (en) | Complex microbial inoculant and application thereof in remediation of organochlorine pesticide polluted water body | |
| US6383797B1 (en) | Bacterial consortium EBC1000 and a method using the bacterial consortium EBC1000 for remedying biologically recalcitrant toxic chemicals contained in industrial wastewater, waste materials and soils | |
| CN113862199B (en) | Degrading strain of benzonitrile herbicide and microbial inoculum produced by degrading strain | |
| CN104560777B (en) | High-tolerance aniline-degrading bacterium and application thereof | |
| CN117701479B (en) | Bacterial strain capable of degrading benzonitrile herbicide, microbial agent and application thereof | |
| KR20140093988A (en) | Cleaning fluid | |
| CN107541479A (en) | A kind of insecticide d-phenothrin degradation bacteria strains and its microbial inoculum and degradation technique | |
| CN105238729B (en) | A kind of anthropi and its biological agent and preparation method and application | |
| CN108707559B (en) | A kind of tetracycline antibiotics degradation arthrobacterium and its application | |
| CN106434442A (en) | Preparation method of degrading restoring agent for organic phosphorus pesticide | |
| CN108723084A (en) | Microorganism formulation and its preparation method and application for low temperature soil reparation | |
| CN102286400A (en) | Rice endotrophic azotobacter for improving disease resistance and stress resistance of crops and purpose thereof | |
| CN107586747B (en) | Composite biological agent and application thereof in remediation of polychlorinated biphenyl polluted soil | |
| CN108251330A (en) | Complex micro organism fungicide for administering soil pollution and its preparation method and application | |
| CN102648714B (en) | Method for preparing biopesticide through liquid fermentation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |