CN101230322A - Novel medical stone micro-ecological repairing additive and preparation method thereof - Google Patents
Novel medical stone micro-ecological repairing additive and preparation method thereof Download PDFInfo
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- CN101230322A CN101230322A CNA2007100595568A CN200710059556A CN101230322A CN 101230322 A CN101230322 A CN 101230322A CN A2007100595568 A CNA2007100595568 A CN A2007100595568A CN 200710059556 A CN200710059556 A CN 200710059556A CN 101230322 A CN101230322 A CN 101230322A
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Abstract
The invention relates to a preparation and the preparation method for restoring the microecological bacteria on the interface layer of water-sediment. The microecological bacteria preparation uses 0.2-0.4mm of medical stone as carrier, absorbs and fixes the composite microbe optimally combined by bacillus subtilis, bacillus licheniformis, bacillus megatherium, and thin yellow streptomyces. The amount ratio of bacillus subtilis to bacillus licheniformis to bacillus megatherium approximates to one to one to one; the volume ratio of the mixture liquid of the three kinds of bacillus to the thin yellow streptomyces liquid is two to one. The invention has the advantages that spreading the preparation of the invention into the interface layer of functional water body water-sediment in the spring March to April each year can prevent blue algae (aerugo micro-capsule algae is the most) from fulminantly growing in summer June to August. The preparation can improve the quality of water, can prevent eutrophication development, and can promote the carbon, nitrogen, and oxygen circulation on the interface layer of water-sediment. The microecological bacteria preparation has high effective colony amount and strong adaptability; and is effective to restore the microecological environment at the interface layer of water-sediment.
Description
Technical field
Biological technical field the present invention relates to a kind of micro-ecological bacterial preparation and preparation method thereof.
Background technology
At present surface water such as the freshwater lake of China, river have been in eutrophication or height eutrophication state, and it is exactly clear proof that Taihu Lake, Chaohu and Dian Chi take place in succession that blue-green algae breaks out.Administer or the breaking out of control blue-green algae, except the pollution in the point of strictness control lake, river periphery, face source, to add an amount of micro-ecological bacterial preparation be practical effective measure to machine in due course.About the more existing both at home and abroad researchs of the micro-ecological bacterial preparation of purifying water body, reparation water ecological environment, still, yet there are no with the medical stone is carrier, absorption high-density, highly active bacterium liquid, and the report of the micro-ecological bacterial preparation of energy big area use.
Summary of the invention
Technical problem to be solved by this invention is: a kind of micro-ecological bacterial preparation that is adsorbed in medical stone and preparation method thereof is provided.
The present invention solves this technical problem the technical scheme that is adopted:
Based on subtilis (Bacillus subtilis), Bacillus licheniformis (Bacillus licheniformis) and bacillus megaterium (Bacillus megaterium), be aided with an amount of streptomyces microflavus (Streptomyces microflavus), and adsorption of immobilization is prepared into bacteria preparation on medical stone.Four strain bacterial classifications are all purchased the DSMZ in Institute of Microorganism, Academia Sinica.
The micro-ecological bacterial preparation that is adsorbed on the medical stone is the complex microorganism of above-mentioned three kinds of genus bacillus and streptomyces microflavus optimum combination, and wherein the ratio of three kinds of genus bacillus was near 1: 1: 1; The volume ratio of three kinds of genus bacillus and streptomyces microflavus is 2: 1.
The preparation method of medical stone-micro-ecological bacterial preparation comprises the steps:
1. each slant strains activation, the shake-flask culture respectively that will preserve.Subtilis, Bacillus licheniformis and bacillus megaterium are used the starch nutrient solution that contains 10% humic acid, and difference one-level enlarged culturing when carrying out secondary, three grades of enlarged culturing then, adopts three kinds of bacterium to mix enlarged culturing earlier.The ratio of three kinds of bacterium is near 1: 1: 1, culture condition: 35～37 ℃, cultivate 20～24h.The living bacteria count of every milliliter of bacterium liquid reaches 100 more than hundred million, promptly greater than 10
10Cfu/mL.The streptomyces microflavus starch nutrient solution that contains 10% humic acid, nutrient solution adds 2% yeast powder again, through one, two, three enlarged culturing, culture condition: cultivate 36～42h for 28～32 ℃.The living bacteria count of every milliliter of bacterium liquid reaches 100 more than hundred million, promptly greater than 10
10Cfu/mL.With the bacterium liquid of above-mentioned two kinds of enlarged culturing by 2: 1 (v/v) thorough mixing (three kinds of genus bacillus are 2, and streptomyces microflavus is 1).
2. medical stone is pulverized, particle diameter is controlled at 0.2～0.4mm, absorption or be immobilized onto on the medical stone bacterium liquid adsorptive capacity 10%～20% under abundant agitation condition;
3. with in 40 ℃～50 ℃ hot gas dryings or the moist woven bag that directly is sealed in polyethylene liner (thick).Room temperature preservation.
4. using method is opened the water-settling interlayer that is spread on various surface water bodies after the sealed bag equably.Usage quantity 0.5～1.0kg/m
2
5. be sprinkled into water-settling interlayer in the spring (3～April) in every year, can prevent and treat the fulminant growth in summer (6～August) of blue-green algae (mainly being microcystic aeruginosa).Improve water quality, stop the eutrophication development, promote the carbon nitrogen oxygen cycle of water-settling interlayer, repair water-settling interlayer micro-ecological environment.
Beneficial effect of the present invention
1) composite bacteria is immobilized onto on the medical stone, make composite bacteria preparation be added in water-sedimental interlayer effectively. medical stone can not only comprise that various bacterium provide trace element for all biologies of interlayer, also because its netted, vesicular structure, has bigger specific surface area, so can in unit volume, adhere to the more thalline of absorption.So, in a single day the active bacterium of absorption enters water-sedimental interlayer just can very fast formation dominant microflora, bring into play its degradation of organic substances, effects such as redox, especially promote the balance of nitrated-denitrification microecosystem, make ammonia nitrogen, nitrite nitrogen and nitric nitrogen be in quick running balance, it is low-level that nitrite nitrogen is in, and can not accumulate.Experiment showed, that 0.02～1mg/L nitrite nitrogen has the effect of the growth that promotes microcystic aeruginosa one class algae, and nitrite nitrogen suppresses the growth of other algae in 0.02～1mg/L scope.If the algae of microcystic aeruginosa one class can not form dominant growth, just blue-green algae can not be taken place breaks out incident.
2) subtilis and bacillus megaterium are aerobic bacterias, can secrete multiple protein enzyme, oxide compound enzyme.Be degradation of organic substances, transform mineral substance, the main force bacterium of effect such as redox; Bacillus licheniformis is a facultative anaerobe, can with subtilis, bacillus megaterium and streptomyces microflavus symbiosis, complement each other, repair micro-ecological environment jointly.
Streptomyces microflavus can produce the disease-resistant expelling parasite of microbiotic, and secretion yield stimulant promotes most of algal growns, transforms mineral substance.
Four kinds of bacterium are to form dominant microflora after carrier enters water-settling interlayer with the medical stone, with indigenous bacterium etc. biological at medical stone the surface or innerly form abundant biogenic accumulation island, promote, repair water-sedimental interlayer microecosystem balance with active microbial film.
Specific embodiments
Embodiment 1
Follow these steps to carry out:
1) each slant strains with 4 ℃ of preservations activates, respectively shake-flask culture.Subtilis, Bacillus licheniformis and bacillus megaterium all use the starch culture-medium that contains 10% humic acid to cultivate, and cultivate 20～24h for 37 ℃; Streptomyces microflavus is cultivated with the starch culture-medium that contains 10% humic acid, cultivates 36～42h for 32 ℃.
2) level liquid enlarged culturing (three kinds of genus bacillus and streptomyces microflavus be independent enlarged culturing respectively, and substratum and culture condition are the same).Inoculum size 5%.
3) (three kinds of genus bacillus are by 1: 1: 1 volume ratio, mixed culture for the secondary fluid enlargement culture.The independent enlarged culturing of streptomyces microflavus.Substratum and culture condition are the same).Inoculum size 5%.
4) (three kinds of genus bacillus mix enlarged culturing to three grades of fluid enlargement culture.The independent enlarged culturing of streptomyces microflavus.Substratum and culture condition are the same).Inoculum size 5%.
5) three grades of fluid enlargement culture liquid of three kinds of genus bacillus and three grades of fluid enlargement culture liquid of streptomyces microflavus are pressed 2: 1 (v/v) thorough mixing.
6) be carrier immobilized above-mentioned mixed bacteria liquid with 0.2～0.4mm medical stone.Immobilized bacterium liquid 10%～20%.
7) warm air-dry dry with 35～45 ℃, the control water ratio is sealed in after the drying in the packing bag of lined with polyethylene below 20%.The medical stone that perhaps directly will adsorb bacterium liquid is sealed in the packing bag of lined with polyethylene.
8) using method is opened the water-settling interlayer that is spread on various surface water bodies after the sealed bag equably.Usage quantity 0.5～1.0kg/m
2
9) be sprinkled into water-settling interlayer in the spring (3～April) in every year, can prevent and treat the fulminant growth in summer (6～August) of blue-green algae (mainly being microcystic aeruginosa).Improve water quality, stop the eutrophication development, promote the carbon nitrogen oxygen cycle of water-settling interlayer, repair water-settling interlayer micro-ecological environment.
Embodiment 2
Follow these steps to carry out:
1) each slant strains with 4 ℃ of preservations activates, respectively shake-flask culture.Subtilis, Bacillus licheniformis and bacillus megaterium all use the starch culture-medium that contains 10% humic acid to cultivate, and cultivate 24h for 37 ℃; Streptomyces microflavus is cultivated with the starch culture-medium that contains 10% humic acid, cultivates 42h for 32 ℃.
2) level liquid enlarged culturing (three kinds of genus bacillus and streptomyces microflavus be independent enlarged culturing respectively, and substratum and culture condition are the same).Inoculum size 10%.
3) (three kinds of genus bacillus are by 1: 1: 1 volume ratio, mixed culture for the secondary fluid enlargement culture.The independent enlarged culturing of streptomyces microflavus.Substratum and culture condition are the same).Inoculum size 10%.
4) (three kinds of genus bacillus mix enlarged culturing to three grades of fluid enlargement culture.The independent enlarged culturing of streptomyces microflavus.Substratum and culture condition are the same).Inoculum size 10%.
5) three grades of fluid enlargement culture liquid of three kinds of genus bacillus and three grades of fluid enlargement culture liquid of streptomyces microflavus are pressed 2: 1 (v/v) thorough mixing.
6) be that carrier adsorbs above-mentioned mixed bacteria liquid with 0.2～0.4mm medical stone.Absorption bacterium liquid 10%～20%.
7) warm air-dry dry with 45 ℃, the control water ratio is sealed in after the drying in the packing bag of lined with polyethylene below 20%.The medical stone that perhaps directly will adsorb bacterium liquid is sealed in the packing bag of lined with polyethylene.
8) using method is opened the water-settling interlayer that is spread on various surface water bodies after the sealed bag equably.Usage quantity 0.5～1.0kg/m
2
9) be sprinkled into water-settling interlayer in the spring (3～April) in every year, can prevent and treat the fulminant growth in summer (6～August) of blue-green algae (mainly being microcystic aeruginosa).Improve water quality, stop the eutrophication development, promote the carbon nitrogen oxygen cycle of water-settling interlayer, repair water-settling interlayer micro-ecological environment.
Embodiment 3
Follow these steps to carry out:
1) each slant strains with 4 ℃ of preservations activates, respectively shake-flask culture.Subtilis, Bacillus licheniformis and bacillus megaterium all use the starch culture-medium that contains 10% humic acid to cultivate, and cultivate 22h for 37 ℃; Streptomyces microflavus is cultivated with the starch culture-medium that contains 10% humic acid, cultivates 40h for 32 ℃.
2) level liquid enlarged culturing (three kinds of genus bacillus and streptomyces microflavus be independent enlarged culturing respectively, and substratum and culture condition are the same).Inoculum size 10%.
3) (three kinds of genus bacillus are by 1: 1: 1 volume ratio, mixed culture for the secondary fluid enlargement culture.The independent enlarged culturing of streptomyces microflavus.Substratum and culture condition are the same).Inoculum size 5%.
4) (three kinds of genus bacillus mix enlarged culturing to three grades of fluid enlargement culture.The independent enlarged culturing of streptomyces microflavus.Substratum and culture condition are the same).Inoculum size 5%.
5) three grades of fluid enlargement culture liquid of three kinds of genus bacillus and three grades of fluid enlargement culture liquid of streptomyces microflavus are pressed 2: 1 (v/v) thorough mixing.
6) be carrier immobilized above-mentioned mixed bacteria liquid with 0.2～0.4mm medical stone.Fixedly bacterium liquid 10%～20%.
7) warm air-dry dry with 40 ℃, the control water ratio is sealed in after the drying in the packing bag of lined with polyethylene below 20%.The medical stone that perhaps directly will adsorb bacterium liquid is sealed in the packing bag of lined with polyethylene.
8) using method is opened the water-settling interlayer that is spread on various surface water bodies after the sealed bag equably.Usage quantity 0.5～1.0kg/m
2
9) be sprinkled into water-settling interlayer in the spring (3～April) in every year, can prevent and treat the fulminant growth in summer (6～August) of blue-green algae (mainly being microcystic aeruginosa).Improve water quality, stop the eutrophication development, promote the carbon nitrogen oxygen cycle of water-settling interlayer, repair water-settling interlayer micro-ecological environment.
Claims (2)
1. novel medical stone micro-ecological repairing additive, it is characterized in that: this micro-ecological repairing additive is made up of the medical stone of 0.2～0.4mm and the active bacterium that is adsorbed on the medical stone, based on subtilis (Bacillus subtilis), Bacillus licheniformis (Bacillus licheniformis) and bacillus megaterium (Bacillus megaterium), be aided with an amount of streptomyces microflavus (Streptomyces microflavus), and adsorption of immobilization is prepared into bacteria preparation on medical stone; The micro-ecological bacterial preparation that is adsorbed on the medical stone is the complex microorganism of above-mentioned three kinds of genus bacillus and streptomyces microflavus combination, and wherein the ratio of three kinds of genus bacillus was near 1: 1: 1; The volume ratio of three kinds of genus bacillus and streptomyces microflavus is 2: 1.
2. the preparation method of micro-ecological repairing additive as claimed in claim 1 is characterized in that it comprises the steps:
1) medical stone is pulverized, particle diameter is controlled at 0.2～0.4mm;
2) form active bacterium liquid by subtilis, Bacillus licheniformis, bacillus megaterium and four kinds of bacterium optimum combinations of streptomyces microflavus; Subtilis, Bacillus licheniformis and bacillus megaterium are used the starch nutrient solution that contains 10% humic acid, and the one-level enlarged culturing when secondary, three grades of enlarged culturing, adopts three kinds of bacterium to mix enlarged culturing respectively; The ratio of three kinds of bacterium is near 1: 1: 1, culture condition: 35～37 ℃, cultivate 20～24h; The living bacteria count of every milliliter of bacterium liquid reaches 100 more than hundred million, and promptly 10
10Cfu/mL; The streptomyces microflavus starch nutrient solution that contains 10% humic acid, nutrient solution adds 2% yeast powder again, carries out one, two, three enlarged culturing, culture condition: cultivate 36～42h for 28～30 ℃; The living bacteria count of every milliliter of bacterium liquid reaches 100 more than hundred million, and promptly 10
10Cfu/mL; The bacterium liquid of the two kinds of enlarged culturing in above-mentioned front and back is pressed 2: 1 volume ratio thorough mixing;
3) fully be adsorbed on 0.2～0.4mm medical stone bacterium liquid adsorptive capacity 10%～20% under the agitation condition; In 40 ℃～50 ℃ hot gas dryings or the moist woven bag that directly is sealed in the thick liner of polyethylene, room temperature preservation;
Be spread on the water-settling interlayer of various surface water bodies when 4) using equably; Usage quantity 0.5～1.0kg/m
2
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| CNA2007100595568A CN101230322A (en) | 2007-09-11 | 2007-09-11 | Novel medical stone micro-ecological repairing additive and preparation method thereof |
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