CN102703420A - Microbial degradation preparation for municipal garbage and sewage and preparation method thereof - Google Patents
Microbial degradation preparation for municipal garbage and sewage and preparation method thereof Download PDFInfo
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- CN102703420A CN102703420A CN2010105625881A CN201010562588A CN102703420A CN 102703420 A CN102703420 A CN 102703420A CN 2010105625881 A CN2010105625881 A CN 2010105625881A CN 201010562588 A CN201010562588 A CN 201010562588A CN 102703420 A CN102703420 A CN 102703420A
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Abstract
The present invention relates to a microbial degradation preparation and a preparation method thereof, and the purpose is to provide the microbial preparation for degrading the municipal garbage and sewage and the preparation method thereof. After being applied into the garbage and the sewage, the preparation can degrade garbage pollutants such as carbohydrates, proteins, lipids, celluloses and other organic substances to reduce the impacts of the bad smell generated by the garbage and the sewage on human life. The microbial degradation preparation is prepared by mixing a fermentation broth of bacillus subtilis strain BS76, turfy soil, diatomaceous earth and the like together. The preparation method comprises the following steps: (1) activating bacteria; (2) preparing a seed liquid and fermenting in a fermentation tank; (3) preparing a culture medium; (4) inoculating and culturing the bacteria; (5) fermenting in another fermentation tank; (6) disinfecting the turfy soil and the diatomaceous earth; (7) mixing the bacteria liquid with the adsorbents to obtain a living bacteria dry powder preparation of the microbial degradation preparation. The product can be used in the municipal garbage and the sewage disposal, has positive industrial applicability, and is simple, low in cost and easy to implement.
Description
Technical field
The present invention relates to belong to biotechnology, production in light industry technical field, relate in particular to a kind of municipal wastes and sewage microbiological deterioration preparation and preparation method thereof.
Background technology
Along with the raising of the level of urbanization, the rubbish and the waste water that produce in the city increase day by day, cause the pollution of environment, produce the orthobiosis that bad smell has influenced people.Municipal wastes and sewage microbiological deterioration preparation are to be made rubbish and the sewage products similar that obtains degrading by the microbial life activity; Be that effective mushroom and sorbing material are mixed the composite biological agent of processing, the use of municipal wastes and sewage microbiological deterioration preparation is the central glucide of degradation of contaminant effectively, protein; Amino acid; Lipid, organic substances such as Mierocrystalline cellulose are purified environment.
Summary of the invention
Goal of the invention of the present invention will provide a kind of municipal wastes and sewage microbiological deterioration preparation and preparation method thereof exactly; After said preparation applies in rubbish and the sewage, the glucide in the middle of can degradation of contaminant, protein; Amino acid; Lipid, organic substances such as Mierocrystalline cellulose are purified environment.
The present invention obtains subtilis BS76 bacterial strain by separating in the soil of area, Qingdao.This bright employing nutrient agar medium solid medium flat board :/1000 milliliters of Carnis Bovis seu Bubali cream 3 grams ,/1000 milliliters of peptone 10 grams ,/1000 milliliters of sodium-chlor 5 grams ,/1000 milliliters of agar 15-20 grams, pH7.4-7.6 sterilized 20-30 minute for 121 ℃; Rich soil is done 10 times of serial dilutions with saline water; Each extent of dilution is got the 0.1-0.2 milliliter and is coated on this nutrient agar medium solid medium flat board; Cultivated 36-48 hour down at 37 ℃ again; Picking list bacterium colony, the purifying of on nutrient agar plate, ruling again is up to obtaining purebred subtilis BS76 bacterial strain.
Task of the present invention is accomplished by following technical scheme:
A kind of municipal wastes and sewage microbiological deterioration preparation, said preparation are by subtilis BS76 bacterial strain, add what sorbent material was prepared from, and the viable count of this bacterium in municipal wastes and sewage microbiological deterioration preparation is >=2 hundred million viable bacteria/grams;
The making method of a kind of municipal wastes and sewage microbiological deterioration preparation may further comprise the steps successively:
1) activation of bacterial classification calibrating:
The freeze-drying lactobacillus of aseptic unlatching subtilis BS76 bacterial strain; Be inoculated in the nutrient broth liquid tube substratum; Cultivated 18-48 hour at 20 ℃-45 ℃, streak inoculation is dull and stereotyped in the nutrient agar medium solid medium then, cultivates 18-48 hour at 20 ℃-45 ℃; Choose colonies typical, carry out the calibrating of morphology and characteristic;
2) preparation of seed liquor:
Pure growth with the subtilis BS76 bacterial strain bacterial classification of assay approval is inoculated in the nutrient broth liquid nutrient medium, 20 ℃-45 ℃ shake-flask culture 18-48 hour, carry out pure bacterium and bacterium property flag check;
3) preparing culture medium:
Nutrient broth medium: Carnis Bovis seu Bubali cream, peptone, sodium-chlor are added in the zero(ppm) water; Above-mentioned each component is prepared by the component of the contained weight (g) of 1000 milliliters solution in zero(ppm) water :/1000 milliliters of Carnis Bovis seu Bubali cream 2-8 grams ,/1000 milliliters of peptone 8-12 grams ,/1000 milliliters of sodium-chlor 3-7 grams; After the said mixture heating for dissolving; Adjust pH 7.2-7.6 sterilized 20-30 minute down for 121 ℃, promptly processed the cultivation subtilis and used liquid nutrient medium.
4) fermentation culture:
The bacteria suspension of the subtilis BS76 of above-mentioned assay approval is inoculated the nutrient solution that nutrient broth medium is housed by the 5-15% inoculum size in fermentor tank, cultivate, culture temperature is 25-45 ℃, and incubation time is 48-72 hour;
5) the nutrient solution cell concentration detects:
Aseptic technique is got and is cultivated the nutrient broth medium that subtilis B81 is arranged, and in microscopically counting thalline quantity, requires thalline quantity to reach 2,000,000,000/milliliter with blood cell counting plate; The ratio that the inspection gemma generates, and the spore rate of will seeking survival reaches more than 90%;
6) collect thalline:
With the overdue culture of above-mentioned cultivation, after pure bacterium experimental verification did not have living contaminants, the centrifugal supernatant that goes obtained spissated wet thallus;
7) carrier sterilization:
Turfy soil and zeyssatite were sterilized 100 ℃-160 ℃ heating in 1-4 hour.
8) preparation mikrobe municipal wastes and sewage microbiological deterioration preparation bacterium powder:
With the turfy soil and the zeyssatite of sterilization, ratio is 10%-90%: 90%-10%, mixes stirring with above-mentioned spissated wet thallus, and is dry under 40 ℃-70 ℃, processes dry powder, is municipal wastes and sewage microbiological deterioration preparation;
Municipal wastes according to the invention and sewage microbiological deterioration preparation and preparation method thereof; Said sorbent material is turfy soil and zeyssatite; The sorbent material proportion is 95%-99% (weight fraction), and all the other are thalline and substratum, and the viable count of said preparation is >=2 hundred million viable bacteria/grams.
The beneficial effect of municipal wastes of the present invention and sewage microbiological deterioration preparation and preparation method thereof is: this product can be applicable to treatment of urban garbage, is applied to municipal sewage treatment, as using product in the middle of the sewage work.This product has positive industrial applicibility, and simple, cost is low, be prone to implement.
Embodiment
Be described further in the face of municipal wastes of the present invention and sewage microbiological deterioration preparation and preparation method thereof down.
Municipal wastes of the present invention and sewage microbiological deterioration preparation; This municipal wastes and sewage microbiological deterioration preparation are by subtilis BS76 bacterial strain; Add what sorbent material was prepared from, the viable count of bacterium in municipal wastes and sewage microbiological deterioration preparation is >=2 hundred million viable bacteria/grams;
Sorbent material is turfy soil and zeyssatite, and the sorbent material proportion is 98% (weight fraction), and all the other are thalline and substratum, and the viable count of said preparation is >=2 hundred million viable bacteria/grams.
Described preparation method is following:
1) activation of bacterial classification calibrating:
The freeze-drying lactobacillus of aseptic unlatching subtilis BS76 bacterial strain is inoculated in the nutrient broth liquid tube substratum, cultivates 18-48 hour at 30 ℃; Streak inoculation is dull and stereotyped in the nutrient agar medium solid medium then; Cultivated 30 hours at 30 ℃, choose colonies typical, carry out the calibrating of morphology and characteristic;
2) preparation of seed liquor:
Pure growth with the subtilis BS76 bacterial strain bacterial classification of assay approval is inoculated in the nutrient broth liquid nutrient medium, 30 ℃ of shake-flask culture 30 hours, carries out pure bacterium and bacterium property flag check;
3) preparing culture medium:
Nutrient broth medium: Carnis Bovis seu Bubali cream, peptone, sodium-chlor are added in the zero(ppm) water; Above-mentioned each component is prepared by the component of the contained weight (g) of 1000 milliliters solution in zero(ppm) water :/1000 milliliters of Carnis Bovis seu Bubali cream 4 grams ,/1000 milliliters of peptone 10 grams ,/1000 milliliters of sodium-chlor 5 grams; After the said mixture heating for dissolving; Adjust pH 7.2-7.6 sterilized 30 minutes down for 121 ℃, promptly processed the cultivation subtilis and used liquid nutrient medium.
4) fermentation culture:
The bacteria suspension of the subtilis BS76 of above-mentioned assay approval is inoculated the nutrient solution that nutrient broth medium is housed by the 5-15% inoculum size in fermentor tank, cultivate, culture temperature is 25 ℃, and incubation time is 72 hours;
5) the nutrient solution cell concentration detects:
Aseptic technique is got and is cultivated the nutrient broth medium that subtilis BS76 is arranged, and in microscopically counting thalline quantity, requires thalline quantity to reach 2,000,000,000/milliliter with blood cell counting plate; The ratio that the inspection gemma generates, and the spore rate of will seeking survival reaches more than 90%;
6) collect thalline:
With the overdue culture of above-mentioned cultivation, after pure bacterium experimental verification did not have living contaminants, the centrifugal supernatant that goes obtained spissated wet thallus;
7) carrier sterilization:
Turfy soil and zeyssatite were sterilized 130 ℃ of heating in 4 hours.
8) preparation municipal wastes and sewage microbiological deterioration preparation bacterium powder:
(ratio is 10%-90%: 90%-10%) mix stirring with above-mentioned spissated wet thallus, 40 ℃-70 ℃ times dryings, process dry powder, be municipal wastes and sewage microbiological deterioration preparation with turfy soil and the zeyssatite of sterilization;
The genus bacillus strain separating:
This bright employing nutrient agar medium solid medium flat board :/1000 milliliters of Carnis Bovis seu Bubali cream 3 grams ,/1000 milliliters of peptone 10 grams ,/1000 milliliters of sodium-chlor 5 grams ,/1000 milliliters of agar 15-20 grams, pH7.4-7.6 sterilized 20-30 minute for 121 ℃; Rich soil is done 10 times of serial dilutions with saline water; Each extent of dilution is got the 0.1-0.2 milliliter and is coated on this nutrient agar medium solid medium flat board; Cultivated 36-48 hour down at 37 ℃ again; Picking list bacterium colony, the purifying of on nutrient agar plate, ruling again is up to obtaining purebred subtilis BS76 bacterial strain.
Bacterial strain screening:
1) screening of decomposing protein bacterium:
Dull and stereotyped primary dcreening operation: casein is dull and stereotyped :/1000 milliliters of casein 5 grams ,/1000 milliliters of sodium-chlor 5 grams ,/1000 milliliters of Carnis Bovis seu Bubali cream 0.1 grams ,/1000 milliliters of glucose 0.05 grams ,/1000 milliliters of agar 15-20 grams, pH7.0-7.2; On casein screening flat board, place 37 ℃ to cultivate 2-7 days bacterial classification inoculation (dibbling), on flat board, add 5% trichoroacetic acid(TCA) solution, observe the situation that transparent circle produces, choose and produce the big bacterial strain of transparent circle.
Proteinase activity is measured: get the zymocyte liquid of cultivating 2-4 days; Centrifuging and taking supernatant in whizzer is got supernatant and is measured proteinase activity, and enzyme activity adopts Filon-phenol method to measure; Casein with 2% is a substrate; Act on 1 hour with supernatant at 37 ℃, it is big or small with how many definite enzymes of generation tyrosine to live, and chooses enzyme big bacterial strain alive.
2) screening of decompose lipide bacterium:
Plate screening: separate fat screening culture medium flat board :/1000 milliliters of peptone 10 grams ,/1000 milliliters of Carnis Bovis seu Bubali cream 5 grams ,/1000 milliliters of sodium-chlor 5 grams, 10 milliliters/1000 milliliters of tween 80s ,/1000 milliliters of agar 15-20 grams, pH7.0-7.2; In separating on the fat screening culture medium flat board, place 37 ℃ to cultivate 2-7 days microbionation (dibbling), observe periphery of bacterial colonies haloing production, choose and produce the big bacterial strain of haloing.
3) screening of decomposition of cellulose bacterium:
Cellulase activity is measured: get the zymocyte liquid of cultivating 2-4 days; Centrifuging and taking supernatant in whizzer is got supernatant and is measured cellulase activity, uses 2% Xylo-Mucine to be substrate; Act on 1 hour with last clear enzyme solution at 37 ℃; With the glucose of DNS reagent mensuration generation, it is big or small with glucogenic how many definite enzymes to live, and chooses enzyme big bacterial strain alive.
4) screening of starch-splitting bacterium:
Plate screening: starch is dull and stereotyped :/1000 milliliters of peptone 10 grams ,/1000 milliliters of Carnis Bovis seu Bubali cream 5 grams ,/1000 milliliters of sodium-chlor 5 grams ,/1000 milliliters of Zulkovsky starch 2 grams ,/1000 milliliters of agar 15-20 grams, pH7.0-7.2; On the starch flat board, place 37 ℃ to cultivate 2-7 days bacterial classification inoculation (dibbling), on flat board, add Lu Ge Shi iodine liquid, observe the situation that transparent circle produces, choose and produce the big bacterial strain of transparent circle.
Fungus characteristic:
The characteristic of subtilis:
Affiliated subtilis single bacterium colony on the nutrient agar flat board is irregular, opaque, white, the edge is irregular; Examine under a microscope, shaft-like, single or one-tenth catenation that thalline is, Gram-positive, size is 0.4-0.7 * 1.2-2.3 micron; Peritrichous, the central spore ovalize, the starch hydrolysis is positive; Protease-producing is positive, and lipase is positive, through being accredited as subtilis.
The subtilis that the present invention uses has the ability of stronger decomposing protein, lipid, starch.
The composition of the nutrient broth medium that described fermentor tank is equipped with is following: Carnis Bovis seu Bubali cream 2 grams, 4 grams, 5 grams, 8 grams; Peptone 8 grams, 10 grams, 12 grams; Sodium-chlor 3 grams, 5 grams, 7 grams; All the other add to 1000 milliliters for water.
Concrete fermentation condition of nutrient broth medium of the present invention and result see table one.
Table one, the concrete fermentation condition of nutrient broth medium and result:
Best zymotechnique instance of the present invention: the top condition of applied nutrition broth culture fermentative prodn subtilis is: the initial pH7.4 of substratum; 35 ℃ of culture temperature; Fermentor tank rotating speed 140rpm; Air flow 5l/min, under this top condition, can ferment produces the viable bacteria greater than 2,000,000,000/milliliter
The municipal wastes of preparation of the present invention and sewage microbiological deterioration preparation application example:
(1) is applied to the processing of changing food waste: the microbiological deterioration preparation that in 100 kilograms of changing food wastes, adds 5 kilograms; Preparation and an amount of water mixing; Be sprayed in the changing food waste; Spray the concentration that detects stink substance after 3-5 days, wherein the clearance of ammonia can reach 82.3%, and the clearance of hydrogen sulfide can reach 75.6%.
(2) be applied to the processing of Wastewater from Pig Farm: get Wastewater from Pig Farm; Ratio with 5 kilograms of every cube of water body addings adds microbiological deterioration preparation of the present invention; Room temperature treatment under aeration condition; Carry out the detection of COD (COD) and ammonia-nitrogen content after 3-10 days, detected result shows that the clearance of the waste water COD after the processing reaches 81.4%, and ammonia-N removal rate can reach 76.5%.
(3) be applied to the processing of sewage: get sewage; Ratio with 2 kilograms of every cubic metre of water body addings adds microbiological deterioration preparation of the present invention; Room temperature treatment under aeration condition is carried out the detection of COD (COD) and ammonia-nitrogen content after 2-9 days, detected result shows that the clearance of the waste water COD after the processing reaches 79.5%; Ammonia-N removal rate can reach 73.5%, and the clearance of hydrogen sulfide can reach 81.7%.
Above embodiment describes preferred implementation of the present invention; Be not that scope of the present invention is limited; Design under the prerequisite of spirit not breaking away from the present invention; Various distortion and improvement that the common engineering technical personnel in this area make technical scheme of the present invention all should fall in the definite protection domain of claims of the present invention.
Claims (3)
1. municipal wastes and sewage microbiological deterioration preparation, it is characterized in that: this municipal wastes and sewage microbiological deterioration preparation are by bacillus subtilis strain BS76 fermented liquid, mix with sorbent material;
The viable count of said subtilis in municipal wastes and sewage microbiological deterioration preparation is >=2 hundred million viable bacteria/grams.
2. the making method of municipal wastes and sewage microbiological deterioration preparation is characterized in that, may further comprise the steps successively:
1) activation of bacterial classification calibrating:
The freeze-drying lactobacillus of aseptic unlatching subtilis BS76 bacterial strain; Be inoculated in the nutrient broth liquid tube substratum; Cultivated 18-48 hour at 20 ℃-45 ℃, streak inoculation is dull and stereotyped in the nutrient agar medium solid medium then, cultivates 18-48 hour at 20 ℃-45 ℃; Choose colonies typical, carry out the calibrating of morphology and characteristic;
2) preparation of seed liquor:
Pure growth with the subtilis BS76 bacterial strain bacterial classification of assay approval is inoculated in the nutrient broth liquid nutrient medium, 20 ℃-45 ℃ shake-flask culture 18-48 hour, carry out pure bacterium and bacterium property flag check;
3) preparing culture medium:
Nutrient broth medium: Carnis Bovis seu Bubali cream, peptone, sodium-chlor are added in the zero(ppm) water; Above-mentioned each component is prepared by the component of the contained weight (g) of 1000 milliliters solution in zero(ppm) water :/1000 milliliters of Carnis Bovis seu Bubali cream 2-8 grams ,/1000 milliliters of peptone 8-12 grams ,/1000 milliliters of sodium-chlor 3-7 grams; After the said mixture heating for dissolving; Adjust pH 7.2-7.6 sterilized 20-30 minute down for 121 ℃, promptly processed the cultivation subtilis and used liquid nutrient medium.
4) fermentation culture:
The bacteria suspension of the subtilis BS76 of above-mentioned assay approval is inoculated the nutrient solution that nutrient broth medium is housed by the 5-15% inoculum size in fermentor tank, cultivate, culture temperature is 25-45 ℃, and incubation time is 48-72 hour;
5) the nutrient solution cell concentration detects:
Aseptic technique is got and is cultivated the nutrient broth medium that subtilis BS76 is arranged, and in microscopically counting thalline quantity, requires thalline quantity to reach 2,000,000,000/milliliter with blood cell counting plate; The ratio that the inspection gemma generates, and the spore rate of will seeking survival reaches more than 90%;
6) collect thalline:
With the overdue culture of above-mentioned cultivation, after pure bacterium experimental verification did not have living contaminants, the centrifugal supernatant that goes obtained spissated wet thallus;
7) carrier sterilization:
Turfy soil and zeyssatite were sterilized 100 ℃-160 ℃ heating in 1-4 hour.
8) preparation municipal wastes and sewage microbiological deterioration preparation bacterium powder:
With the turfy soil and the zeyssatite of sterilization, ratio is 10%-90%: 90%-10%, mixes stirring with above-mentioned spissated wet thallus, and is dry under 40 ℃-70 ℃, processes dry powder, is municipal wastes and sewage microbiological deterioration preparation;
3. according to said municipal wastes of claim 1 and sewage microbiological deterioration preparation; It is characterized in that: said sorbent material is turfy soil and zeyssatite; The sorbent material proportion is 95%-99% (weight fraction), and all the other are thalline and substratum, and the viable count of said preparation is >=2 hundred million viable bacteria/grams.
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| CN2010105625881A CN102703420A (en) | 2010-11-29 | 2010-11-29 | Microbial degradation preparation for municipal garbage and sewage and preparation method thereof |
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| CN2010105625881A CN102703420A (en) | 2010-11-29 | 2010-11-29 | Microbial degradation preparation for municipal garbage and sewage and preparation method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103241842A (en) * | 2013-05-14 | 2013-08-14 | 四川清和科技有限公司 | Method for controlling water pollution by using EPSB (Especial Photosynthetic Bacteria) |
| CN104961244A (en) * | 2015-07-02 | 2015-10-07 | 山东阜丰发酵有限公司 | Treatment method for L-arginine fermentation waste liquid |
| CN105441352A (en) * | 2015-09-16 | 2016-03-30 | 广州金水动物保健品有限公司 | Compound microorganism water stabilizing preparation |
| CN109266590A (en) * | 2018-11-27 | 2019-01-25 | 合肥市东方美捷分子材料技术有限公司 | A kind of denitrification compound bacteria agent and its preparation method and application |
| CN113755482A (en) * | 2021-08-16 | 2021-12-07 | 广西荣烁环境工程有限公司 | Efficient denitrification microbial preparation for sewage treatment and preparation method thereof |
| CN114540240A (en) * | 2022-03-10 | 2022-05-27 | 皖西学院 | Microecological preparation for sewage treatment of farm, preparation method and application |
-
2010
- 2010-11-29 CN CN2010105625881A patent/CN102703420A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103241842A (en) * | 2013-05-14 | 2013-08-14 | 四川清和科技有限公司 | Method for controlling water pollution by using EPSB (Especial Photosynthetic Bacteria) |
| CN103241842B (en) * | 2013-05-14 | 2014-11-26 | 四川清和科技有限公司 | Method for controlling water pollution by using EPSB (Especial Photosynthetic Bacteria) |
| CN104961244A (en) * | 2015-07-02 | 2015-10-07 | 山东阜丰发酵有限公司 | Treatment method for L-arginine fermentation waste liquid |
| CN105441352A (en) * | 2015-09-16 | 2016-03-30 | 广州金水动物保健品有限公司 | Compound microorganism water stabilizing preparation |
| CN109266590A (en) * | 2018-11-27 | 2019-01-25 | 合肥市东方美捷分子材料技术有限公司 | A kind of denitrification compound bacteria agent and its preparation method and application |
| CN113755482A (en) * | 2021-08-16 | 2021-12-07 | 广西荣烁环境工程有限公司 | Efficient denitrification microbial preparation for sewage treatment and preparation method thereof |
| CN114540240A (en) * | 2022-03-10 | 2022-05-27 | 皖西学院 | Microecological preparation for sewage treatment of farm, preparation method and application |
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Application publication date: 20121003 |