CN109626555A - A kind of method of fast degradation water body withered fallen leaf - Google Patents
A kind of method of fast degradation water body withered fallen leaf Download PDFInfo
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- CN109626555A CN109626555A CN201910054238.5A CN201910054238A CN109626555A CN 109626555 A CN109626555 A CN 109626555A CN 201910054238 A CN201910054238 A CN 201910054238A CN 109626555 A CN109626555 A CN 109626555A
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/02—Aerobic processes
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/342—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the enzymes used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/347—Use of yeasts or fungi
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02F2101/30—Organic compounds
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The invention belongs to technical field of water environment treatment, provide a kind of method of fast degradation water body withered fallen leaf.This method, comprising: (1) raw material prepare: prepare the fungi strain for capableing of decomposition of cellulose；Prepare Mandel nutrient solution；Prepare carboxymethyl cellulose plating medium；Prepare potato dextrose agar；Prepare potato dextrose broth；Preparation can decompose the bacteria culture of lignin；Prepare LB solid medium；Prepare LB liquid medium；(2) the fixed fungi and bacterium for producing lignin and cellulase of PP biologic packing material ball: cellulase-producing fungi culture；Produce lignoenzyme Bacteria Culture；PP biologic packing material ball fixes bacterium；(3) water sample is handled.This method has the advantages that fast decoupled bottom withered fallen leaf, solves the disadvantage that withered fallen leaf is not easy to be decomposed by heterotroph in practical water body；Water body withered fallen leaf is quickly reduced after processing, and water body recovery clarification, Water quality can be made to be improved as early as possible.
Description
Technical field
The invention belongs to technical field of water environment treatment, specifically, being related to a kind of side of fast degradation water body withered fallen leaf
Method.
Background technique
Urban landscape water, river, lake because around normal kind have a large amount of plants, have a large amount of fallen leaves during seasonal law
Water body can be entered, by accumulation year in year out, the bottom can deposit a large amount of fallen leaves.Fallen leaves are deposited on water body and are difficult by microorganism point
Solution, water bottom dissolved oxygen deficiency, which is easy anaerobic fermentation, causes water body nigrescence smelly.The general water quality of urban landscape water is good, various fingers
Mark is all very low, but a large amount of green plants are had around urban landscape water, is easier to accumulate withered fallen leaf compared to lake, river, in addition trees
Concealment be difficult to see sunlight, the submerged plant in water body be easy death, exacerbate the deficiency of water body dissolved oxygen, lead to water ecology
The destruction of balance.Water body accumulates a large amount of litters, and i.e. water body clarity can be seriously affected by passing through anaerobic fermentation, become water body sense organ
Difference influences its ornamental value.
Therefore, water body ornamental value can not only be improved by inventing a kind of method for capableing of fast degradation water body withered fallen leaf, may be used also
With alleviate water body dissolved oxygen it is insufficient caused by aquatic animals and plants it is dead, caused by aquatic ecological balance destroy and water body self-regeneration
The problems such as ability reduces.
Summary of the invention
For deficiency above-mentioned in the prior art, the object of the present invention is to provide a kind of fast degradation water body withered fallen leafs
Method；This method has the advantages that fast decoupled bottom withered fallen leaf, solves withered fallen leaf and is not easy in practical water body by heterotrophism
The shortcomings that bacterium is decomposed；Water body withered fallen leaf is quickly reduced after processing, and water body recovery clarification, Water quality can be made to be changed as early as possible
It is kind.
In order to achieve the above object, the solution that the present invention uses is:
A kind of method of fast degradation water body withered fallen leaf, comprising:
(1) raw material prepare: preparing the fungi strain for capableing of decomposition of cellulose；Prepare Mandel nutrient solution；Prepare carboxylic first
Base cellulose (CMC) plating medium；Prepare potato dextrose agar (PDA) culture medium；Prepare potato glucose liquid
(PDB) culture medium；Preparation can decompose the bacteria culture of lignin；Prepare LB solid medium；Prepare LB liquid medium；
(2) the fixed fungi and bacterium for producing lignin and cellulase of PP biologic packing material ball:
Cellulase-producing fungi culture: fungi strain is inoculated on CMC plating medium, and after cultivating 3-5d, picking is long
On the good mycelia of gesture to new CMC plating medium, gained strain is inoculated into PDA base culture medium after repeating this operation 2-3 times
On, 28 DEG C of constant incubator culture 3-5d, then gained strain is inoculated into PDB culture medium, 28 DEG C of constant incubator culture 3-5d,
Obtain fungi bacterium solution.
It produces lignoenzyme Bacteria Culture: bacteria culture being inoculated on LB solid medium, is cultivated at 37 DEG C and selects length afterwards for 24 hours
The good bacterium colony of gesture is inoculated on new LB solid medium, after repeating this operation 2-3 times, by colony inoculation to LB Liquid Culture
On base, 24-48h is cultivated, obtains bacterial solution；
PP biologic packing material ball fixes bacterium: PP biologic packing material ball being put into fungi bacterium solution, 28 DEG C are further cultured for 4-6d, are adhered to
The biologic packing material ball of fungi strain；PP biologic packing material filling ball is put into bacterial solution, 3-5d is further cultured at 37 DEG C, is adhered to
There is the biologic packing material ball of bacteria culture；
(3) water sample is handled: fungi and bacterium can generate the enzyme of a variety of can degrade leaf and organic matters, first complete at the bottom
The PP biologic packing material ball for being attached with fungi strain is added in aerator；Aerator is opened after it sinks naturally, aeration is strong
Degree adjusts minimum value；After 5d, the PP biologic packing material ball for being attached with bacteria culture is added；Aerator interval 1-2d opens 22-26h, holds
Continuous 2-3 months.For fungi, bacterium easily breeds and growth cycle is short, easily survives in water body.
A kind of beneficial effect of the method for fast degradation water body withered fallen leaf provided by the invention is:
The method of this kind of fast degradation water body withered fallen leaf provided by the invention has the advantages that fast decoupled bottom withered fallen leaf,
Solve the disadvantage that withered fallen leaf is not easy to be decomposed by heterotroph in practical water body.Utilize efficient cellulase-producing and lignoenzyme
Microorganism carries out PP biologic packing material ball biofilm, then the withered fallen leaf of water body is handled with the biologic packing material ball handled well, substantially reduces
Leaf-litter decomposing course, and handle Yi Huishou and reusable after water sample.Water body withered fallen leaf is quickly reduced after processing, Ke Yijin
Fastly improved water body recovery clarification, Water quality, provides a kind of new thinking to water environment treatment.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
A kind of method of fast degradation water body withered fallen leaf provided in an embodiment of the present invention is specifically described below.
A kind of method of fast degradation water body withered fallen leaf, it is characterised in that: include:
(1) raw material prepare:
Prepare the fungi strain for capableing of decomposition of cellulose；In the present embodiment, further, fungi strain includes that spine spore is bent
At least one of rotten mould and trichoderma of mould, sharp spore reaping hook enzyme, type.
Wherein, (Aspergillus aculeatus is bought in German microbial preservation microorganism Aspergillus aculeatus DSM-63261
The heart)；Sharp spore reaping hook enzyme DSM-63310 (Fusarium sp. is bought in German Organism Depositary)；Between the rotten mould DSM- of type
62950 (Pythium intermedium is bought in German Culture Collection Center)；Trichoderma (Trichoderma sp., commercially available).
Prepare Mandel nutrient solution: according to parts by weight, including NaNO32 parts, K2HPO41.5 parts, CaCl21.5 parts,
MgSO40.3 part, FeSO4·7H20.005 part of O, MnSO4·H20.0016 part of O, ZnSO4·H20.0014 part of O, CoCl2
0.0005 part, H2O100 parts；pH5.5.
Prepare CMC plating medium: including Mandel nutrient solution, 2% carboxymethyl cellulose, 0.08% deoxycholic acid
Sodium, 1.5% agar, 121 DEG C of sterilizing 20min；It is cooled to inverted plate after non-scald on hand, is inverted after plate solidification spare.
Prepare PDA culture medium: according to parts by weight, including 150-200 parts of potato, 15-20 parts of glucose, agar 15-
20 parts, H21000 parts of O；115 DEG C of high pressure sterilization 15min.
Prepare PDB culture medium: according to parts by weight, including 150-200 parts of potato, 15-20 parts of glucose, H2O 1000
Part；115 DEG C of high pressure sterilization 15min.
Preparation can decompose the bacteria culture of lignin: in the present embodiment, further, bacteria culture includes withered grass bud
At least one of spore bacillus Pseudomonas, bacillus cereus category and Pseudomonas chlororaphis.
Wherein, (Bacillus subtilis is bought in China General Microbiological culture presevation bacillus subtilis Pseudomonas
Center)；Bacillus cereus category (Bacillus；FGcereus is bought in China General Microbiological Culture Collection Center)；Green needle
Pseudomonad (Pseudomonas chlororaphis is bought in China General Microbiological Culture Collection Center).
Prepare LB solid medium: according to parts by weight, including 10 parts of tryptone, 5 parts of yeast powder, 10 parts of sodium chloride,
10-15 parts of agar powder, 1000 parts of distilled water；Tune pH is 7.0,121 DEG C of sterilizing 20min, is cooled to inverted plate after non-scald on hand, is solidified
It is inverted afterwards stand-by.
Prepare LB liquid medium: according to parts by weight, including 10 parts of tryptone, 5 parts of yeast powder, 10 parts of sodium chloride,
10-15 parts of agar powder, 1000 parts of distilled water, 7.0,121 DEG C of sterilizing 20min of pH, 4 DEG C of placements are spare.
The pretreatment of PP biologic packing material ball: PP biologic packing material ball first impregnates 6h with alcohol, and taking-up is dried spare.
(2) the fixed fungi and bacterium for producing lignin and cellulase of PP biologic packing material ball:
Cellulase-producing fungi culture: fungi strain is inoculated on CMC plating medium, and after cultivating 3-5d, picking is long
On the good mycelia of gesture to new CMC plating medium, gained strain is inoculated into PDA culture medium training after repeating this operation 2-3 times
It supports on base, 28 DEG C of constant incubator culture 3-5d, then gained strain is inoculated into PDB culture medium, 28 DEG C of constant incubator cultures
3-5d obtains fungi bacterium solution.
It produces lignoenzyme Bacteria Culture: bacteria culture being inoculated on LB solid medium, is cultivated at 37 DEG C and selects length afterwards for 24 hours
The good bacterium colony of gesture is inoculated on new LB solid medium, after repeating this operation 2-3 times, by colony inoculation to LB Liquid Culture
On base, 24-48h is cultivated, obtains bacterial solution.
PP biologic packing material ball fixes bacterium: PP biologic packing material ball being put into fungi bacterium solution, 28 DEG C are further cultured for 4-6d, are adhered to
The biologic packing material ball of fungi strain；PP biologic packing material filling ball is put into bacterial solution, 3-5d is further cultured at 37 DEG C, is adhered to
There is the biologic packing material ball of bacteria culture；
(3) water sample is handled: first completing aerator at the bottom, the PP biologic packing material ball for being attached with fungi strain is added；To
It opens aerator after sinking naturally, by aeration intensity tune minimum value；After 5d, the PP biology that addition is attached with bacteria culture is filled out
Pellet；Aerator interval 1-2d opens 22-26h, continues 2-3 months.
Litter mainly nature difficult to degrade lacks the microorganism that can generate cellulase and lignoenzyme, lacks at the bottom
Few oxygen, anaerobic fermentation lead to water quality deterioration, and bed mud blackening is smelly.
The leaf natural degradation period is long, and to improve litter degradation speed, the present embodiment, which is taken, fixes aerobic microbiological
On PP biologic packing material ball, the aerobic microbiological being attached on PP biologic packing material ball is set to sink to water body naturally, due to degradation process
Need to expend a large amount of oxygen, the present embodiment is aided with aerator, can achieve the purpose of fast degradation water body withered fallen leaf.
Feature and performance of the invention are described in further detail with reference to embodiments.
Embodiment 1
A kind of method for present embodiments providing fast degradation water body withered fallen leaf, comprising:
(1) raw material prepare:
Prepare the fungi strain for capableing of decomposition of cellulose: microorganism Aspergillus aculeatus, sharp spore reaping hook enzyme, the rotten mould and trichoderma of type.
Prepare Mandel nutrient solution: according to parts by weight, including NaNO32g, K2HPO41.5g, CaCl21.5g, MgSO4
0.3g, FeSO4·7H2O 0.005g, MnSO4·H2O 0.0016g, ZnSO4·H2O 0.0014g, CoCl20.0005g,
H2O100g；pH5.5.
Prepare CMC plating medium: including Mandel nutrient solution, 2% carboxymethyl cellulose, 0.08% deoxycholic acid
Sodium, 1.5% agar, 121 DEG C of sterilizing 20min；It is cooled to inverted plate after non-scald on hand, is inverted after plate solidification spare.
Prepare PDA culture medium: including potato 200g, glucose 20g, agar 20g, H2O 1000ml；115 DEG C of high pressures are gone out
Bacterium 15min, cooling are spare.
Prepare PDB culture medium: according to parts by weight, including potato 200g, glucose 20g, H2O 1000ml；115℃
High pressure sterilization 15min, cooling are spare.
Preparation can decompose the bacteria culture of lignin: bacillus subtilis Pseudomonas, bacillus cereus category and green needle are false
Monad.
Prepare LB solid medium: including tryptone 10g, yeast powder 5g, sodium chloride 10g, agar powder 15g, distilled water
1000mL adjusts pH7.0,121 DEG C of high-temperature sterilization 20min to be cooled to inverted plate after non-scald on hand, is inverted after solidification stand-by.
Prepare LB liquid medium: including tryptone 10g, yeast powder 5g, sodium chloride 10g, agar powder 15g, distilled water
1000mL adjusts pH7.0, and 121 DEG C of high-temperature sterilization 20min, 4 DEG C of refrigerators are placed spare.
(2) the fixed fungi and bacterium for producing lignin and cellulase of PP biologic packing material ball:
Cellulase-producing fungi culture: by aspergillus echinulatus, sharp spore reaping hook is mould, white rot is mould and trichoderma is inoculated into the training of CMC plate
It supports on base, after cultivating 4d, on the good mycelia of picking growing way to new CMC plating medium, by gained bacterium after repeating this operation 2 times
Kind is inoculated into PDA culture medium, 28 DEG C of constant incubator culture 4d, obtains aspergillus echinulatus strain, sharp spore sickle mycete kind, white rot
Mould species and trichoderma strain, then the above strain is inoculated on PDB culture medium, 28 DEG C of constant incubator culture 4d, obtain thorn spore
Aspergillus bacterium solution, sharp spore sickle mycete liquid, white rot mould liquid and Trichoderma liquor.
It produces lignoenzyme Bacteria Culture: bacillus subtilis, bacillus cereus, Pseudomonas chlororaphis is inoculated with LB solid
It on culture medium, is cultivated at 37 DEG C and selects the good bacterium colony of growing way afterwards for 24 hours, be inoculated on new LB solid medium, repeat this operation 2
After secondary, by colony inoculation to LB liquid medium, culture for 24 hours, obtains bacillus subtilis bacterium solution, bacillus cereus bacterium solution
With Pseudomonas chlororaphis bacterium solution.
PP biologic packing material ball fixes bacterium: PP biologic packing material ball is put into aspergillus echinulatus bacterium solution, sharp spore sickle mycete liquid, white rot
Mould liquid and Trichoderma liquor, 28 DEG C are further cultured for 5d, obtain be attached with that aspergillus echinulatus, sharp spore reaping hook is mould, white rot is mould and trichoderma respectively
Biologic packing material ball；It is false that PP biologic packing material filling ball is put into bacillus subtilis bacterium solution, bacillus cereus bacterium solution and green needle
Monad bacterium solution, 3d is further cultured at 37 DEG C, and acquisition is attached with bacillus subtilis, bacillus cereus and Pseudomonas chlororaphis
Biologic packing material ball.
(3) water sample is handled: the first step, first completes aerator at the bottom, preparation 6 is withered to pocket, and each withered pocket fills 10g
Dried leaf is put into the bottom；Second step is first added and adheres to that aspergillus echinulatus, sharp spore reaping hook is mould, white rot is mould and the PP biology of trichoderma is filled out
Pellet, opens aerator after it sinks naturally, aeration intensity tune minimum value, be added after 5d be attached with bacillus subtilis,
The PP biologic packing material ball of bacillus cereus and Pseudomonas chlororaphis, aerator interval 1d are opened for 24 hours, continue 2 months；The
Three steps, data test, every 10d take one it is withered pocket, survey its dry weight after drying.
Embodiment 2
A kind of method for present embodiments providing fast degradation water body withered fallen leaf, comprising:
(1) raw material prepare:
Prepare the fungi strain for capableing of decomposition of cellulose: microorganism Aspergillus aculeatus, sharp spore reaping hook enzyme, the rotten mould and trichoderma of type.
Prepare Mandel nutrient solution: according to parts by weight, including NaNO32g, K2HPO41.5g, CaCl21.5g, MgSO4
0.3g, FeSO4·7H2O 0.005g, MnSO4·H2O 0.0016g, ZnSO4·H2O 0.0014g, CoCl20.0005g,
H2O100g；pH5.5.
Prepare CMC plating medium: including Mandel nutrient solution, 2% carboxymethyl cellulose, 0.08% deoxycholic acid
Sodium, 1.5% agar, 121 DEG C of sterilizing 20min；It is cooled to inverted plate after non-scald on hand, is inverted after plate solidification spare.
Prepare PDA culture medium: including potato 150g, glucose 15g, agar 15g, H2O 1000ml；115 DEG C of high pressures are gone out
Bacterium 15min, cooling are spare.
Prepare PDB culture medium: according to parts by weight, including potato 150g, glucose 15g, H2O 1000ml；115℃
High pressure sterilization 15min, cooling are spare.
Preparation can decompose the bacteria culture of lignin: bacillus subtilis Pseudomonas, bacillus cereus category and green needle are false
Monad.
Prepare LB solid medium: including tryptone 10g, yeast powder 5g, sodium chloride 10g, agar powder 10g, distilled water
1000mL adjusts pH7.0,121 DEG C of high-temperature sterilization 20min to be cooled to inverted plate after non-scald on hand, is inverted after solidification stand-by.
Prepare LB liquid medium: including tryptone 10g, yeast powder 5g, sodium chloride 10g, agar powder 10g, distilled water
1000mL adjusts pH7.0, and 121 DEG C of high-temperature sterilization 20min, 4 DEG C of refrigerators are placed spare.
(2) the fixed fungi and bacterium for producing lignin and cellulase of PP biologic packing material ball:
Cellulase-producing fungi culture: by aspergillus echinulatus, sharp spore reaping hook is mould, white rot is mould and trichoderma is inoculated into the training of CMC plate
It supports on base, after cultivating 3d, on the good mycelia of picking growing way to new CMC plating medium, by gained bacterium after repeating this operation 3 times
Kind is inoculated on PDA culture medium culture medium, 28 DEG C of constant incubator culture 3d, obtains aspergillus echinulatus strain, sharp spore sickle mycete
Kind, white rot mould species and trichoderma strain, then the above strain is inoculated on PDB culture medium, 28 DEG C of constant incubator culture 4d are obtained
Obtain aspergillus echinulatus bacterium solution, sharp spore sickle mycete liquid, white rot mould liquid and Trichoderma liquor.
It produces lignoenzyme Bacteria Culture: bacillus subtilis, bacillus cereus, Pseudomonas chlororaphis is inoculated with LB solid
It on culture medium, is cultivated at 37 DEG C and selects the good bacterium colony of growing way afterwards for 24 hours, be inoculated on new LB solid medium, repeat this operation 3
After secondary, by colony inoculation to LB liquid medium, 48h is cultivated, obtains bacillus subtilis bacterium solution, bacillus cereus bacterium solution
With Pseudomonas chlororaphis bacterium solution.
PP biologic packing material ball fixes bacterium: PP biologic packing material ball is put into aspergillus echinulatus bacterium solution, sharp spore sickle mycete liquid, white rot
Mould liquid and Trichoderma liquor, 28 DEG C are further cultured for 4d, obtain be attached with that aspergillus echinulatus, sharp spore reaping hook is mould, white rot is mould and trichoderma respectively
Biologic packing material ball；It is false that PP biologic packing material filling ball is put into bacillus subtilis bacterium solution, bacillus cereus bacterium solution and green needle
Monad bacterium solution, 5d is further cultured at 37 DEG C, and acquisition is attached with bacillus subtilis, bacillus cereus and Pseudomonas chlororaphis
Biologic packing material ball.
(3) water sample is handled: the first step, first completes aerator at the bottom, preparation 6 is withered to pocket, and each withered pocket fills 10g
Dried leaf is put into the bottom；Second step is first added and adheres to that aspergillus echinulatus, sharp spore reaping hook is mould, white rot is mould and the PP biology of trichoderma is filled out
Pellet, opens aerator after it sinks naturally, aeration intensity tune minimum value, be added after 5d be attached with bacillus subtilis,
The PP biologic packing material ball of bacillus cereus and Pseudomonas chlororaphis, aerator interval 2d open 26h, continue 2 months；The
Three steps, data test, every 10d take one it is withered pocket, survey its dry weight after drying.
Embodiment 3
A kind of method for present embodiments providing fast degradation water body withered fallen leaf, comprising:
(1) raw material prepare:
Prepare the fungi strain for capableing of decomposition of cellulose: microorganism Aspergillus aculeatus, sharp spore reaping hook enzyme, the rotten mould and trichoderma of type.
Prepare Mandel nutrient solution: according to parts by weight, including NaNO32g, K2HPO41.5g, CaCl21.5g, MgSO4
0.3g, FeSO4·7H2O 0.005g, MnSO4·H2O 0.0016g, ZnSO4·H2O 0.0014g, CoCl20.0005g,
H2O100g；pH5.5.
Prepare CMC plating medium: including Mandel nutrient solution, 2% carboxymethyl cellulose, 0.08% deoxycholic acid
Sodium, 1.5% agar, 121 DEG C of sterilizing 20min；It is cooled to inverted plate after non-scald on hand, is inverted after plate solidification spare.
Prepare PDA culture medium: including potato 180g, glucose 18g, agar 18g, H2O 1000ml；115 DEG C of high pressures are gone out
Bacterium 15min, cooling are spare.
Prepare PDB culture medium: according to parts by weight, including potato 150g, glucose 15g, H2O 1000ml；115℃
High pressure sterilization 15min, cooling are spare.
Preparation can decompose the bacteria culture of lignin: bacillus subtilis Pseudomonas, bacillus cereus category and green needle are false
Monad.
Prepare LB solid medium: including tryptone 10g, yeast powder 5g, sodium chloride 10g, agar powder 12g, distilled water
1000mL adjusts pH7.0,121 DEG C of high-temperature sterilization 20min to be cooled to inverted plate after non-scald on hand, is inverted after solidification stand-by.
Prepare LB liquid medium: including tryptone 10g, yeast powder 5g, sodium chloride 10g, agar powder 12g, distilled water
1000mL adjusts pH7.0, and 121 DEG C of high-temperature sterilization 20min, 4 DEG C of refrigerators are placed spare.
(2) the fixed fungi and bacterium for producing lignin and cellulase of PP biologic packing material ball:
Cellulase-producing fungi culture: by aspergillus echinulatus, sharp spore reaping hook is mould, white rot is mould and trichoderma is inoculated into the training of CMC plate
It supports on base, after cultivating 5d, on the good mycelia of picking growing way to new CMC plating medium, by gained bacterium after repeating this operation 3 times
Kind is inoculated on PDA culture medium culture medium, 28 DEG C of constant incubator culture 5d, obtains aspergillus echinulatus strain, sharp spore sickle mycete
Kind, white rot mould species and trichoderma strain, then the above strain is inoculated on PDB culture medium, 28 DEG C of constant incubator culture 4d are obtained
Obtain aspergillus echinulatus bacterium solution, sharp spore sickle mycete liquid, white rot mould liquid and Trichoderma liquor.
It produces lignoenzyme Bacteria Culture: bacillus subtilis, bacillus cereus, Pseudomonas chlororaphis is inoculated with LB solid
It on culture medium, is cultivated at 37 DEG C and selects the good bacterium colony of growing way afterwards for 24 hours, be inoculated on new LB solid medium, repeat this operation 3
After secondary, by colony inoculation to LB liquid medium, 35h is cultivated, obtains bacillus subtilis bacterium solution, bacillus cereus bacterium solution
With Pseudomonas chlororaphis bacterium solution.
PP biologic packing material ball fixes bacterium: PP biologic packing material ball is put into aspergillus echinulatus bacterium solution, sharp spore sickle mycete liquid, white rot
Mould liquid and Trichoderma liquor, 28 DEG C are further cultured for 6d, obtain be attached with that aspergillus echinulatus, sharp spore reaping hook is mould, white rot is mould and trichoderma respectively
Biologic packing material ball；It is false that PP biologic packing material filling ball is put into bacillus subtilis bacterium solution, bacillus cereus bacterium solution and green needle
Monad bacterium solution, 4d is further cultured at 37 DEG C, and acquisition is attached with bacillus subtilis, bacillus cereus and Pseudomonas chlororaphis
Biologic packing material ball.
(3) water sample is handled: the first step, first completes aerator at the bottom, preparation 6 is withered to pocket, and each withered pocket fills 10g
Dried leaf is put into the bottom；Second step is first added and adheres to that aspergillus echinulatus, sharp spore reaping hook is mould, white rot is mould and the PP biology of trichoderma is filled out
Pellet, opens aerator after it sinks naturally, aeration intensity tune minimum value, be added after 5d be attached with bacillus subtilis,
The PP biologic packing material ball of bacillus cereus and Pseudomonas chlororaphis, aerator interval 1.5d open 25h, continue 2 months；
Third step, data test, every 10d take one it is withered pocket, survey its dry weight after drying.
Experimental example 1
Experimental method: the dried leaf of embodiment 1-3 is weighed respectively in weight in different time periods, the results are shown in Table 1:
Table 1
| Time | 0d | 10d | 20d | 30d | 40d | 50d | 60d |
| 1 leaf dry weight (g) of embodiment | 10 | 9.12 | 8.03 | 6.56 | 4.54 | 3.23 | 1.52 |
| 2 leaf dry weight (g) of embodiment | 10 | 9.08 | 8.01 | 6.50 | 4.49 | 3.19 | 1.48 |
| 3 leaf dry weight (g) of embodiment | 10 | 9.14 | 7.98 | 6.49 | 4.51 | 3.18 | 1.45 |
From the data in table 1, it can be seen that can be at 2 months by the method for fast degradation water body withered fallen leaf provided in an embodiment of the present invention
Dead leaf degradation 85% or so can be greatly shortened natural degradation spent time by left and right, solved leaf and rotted to lead to water at the bottom
The problem of body anoxic.
In conclusion using the method for fast degradation water body withered fallen leaf provided by the invention；This method has fast decoupled
The advantages of water-bed withered fallen leaf, solves the disadvantage that withered fallen leaf is not easy to be decomposed by heterotroph in practical water body；Water body is withered after processing
Fallen leaves are quickly reduced, and water body recovery clarification, Water quality can be made to be improved as early as possible.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, made any to repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of method of fast degradation water body withered fallen leaf, it is characterised in that: include:
(1) raw material prepare: preparing the fungi strain for capableing of decomposition of cellulose；Prepare Mandel nutrient solution；It is fine to prepare carboxymethyl
Tie up plain plating medium；Prepare potato dextrose agar；Prepare potato dextrose broth；Preparation can
Decompose the bacteria culture of lignin；Prepare LB solid medium；Prepare LB liquid medium；
(2) the fixed fungi and bacterium for producing lignin and cellulase of PP biologic packing material ball:
Cellulase-producing fungi culture: the fungi strain is inoculated on the carboxymethyl cellulose plating medium, culture
It, will after repeating this operation 2-3 times on the good mycelia of picking growing way to the new carboxymethyl cellulose plating medium after 3-5d
Gained strain is inoculated on the potato dextrose agar culture medium, 28 DEG C of constant incubator culture 3-5d, then will
Gained strain is inoculated into potato dextrose broth, and 28 DEG C of constant incubator culture 3-5d obtain fungi bacterium solution；
It produces lignoenzyme Bacteria Culture: the bacteria culture being inoculated on the LB solid medium, cultivates at 37 DEG C and chooses afterwards for 24 hours
The bacterium colony for selecting growing way good is inoculated on the new LB solid medium, after repeating this operation 2-3 times, by the colony inoculation
Onto the LB liquid medium, 24-48h is cultivated, obtains bacterial solution；
PP biologic packing material ball fixes bacterium: PP biologic packing material ball being put into the fungi bacterium solution, 28 DEG C are further cultured for 4-6d, are adhered to
The biologic packing material ball of the fungi strain；PP biologic packing material filling ball is put into the bacterial solution, is further cultured for 3-5d at 37 DEG C,
Obtain the biologic packing material ball for being attached with the bacteria culture；
(3) water sample is handled: first completing aerator at the bottom, the PP biologic packing material ball for being attached with the fungi strain is added；To
It opens the aerator after sinking naturally, by aeration intensity tune minimum value；After 5d, addition is attached with the bacteria culture
PP biologic packing material ball；Aerator interval 1-2d opens 22-26h, continues 2-3 months.
2. the method for fast degradation water body withered fallen leaf according to claim 1, it is characterised in that: the fungi strain includes
At least one of microorganism Aspergillus aculeatus, sharp spore reaping hook enzyme, the rotten mould and trichoderma of type.
3. the method for fast degradation water body withered fallen leaf according to claim 1, it is characterised in that: the bacteria culture includes
At least one of bacillus subtilis Pseudomonas, bacillus cereus category and Pseudomonas chlororaphis.
4. the method for fast degradation water body withered fallen leaf according to claim 1, it is characterised in that: further include the PP biology
The pretreatment of filling ball: the PP biologic packing material ball first impregnates 6h with alcohol, and taking-up is dried spare.
5. the method for fast degradation water body withered fallen leaf according to claim 1, it is characterised in that: the Mandel nutrition
Liquid: according to parts by weight, including NaNO32 parts, K2HPO41.5 parts, CaCl21.5 parts, MgSO40.3 part, FeSO4·7H2O
0.005 part, MnSO4·H20.0016 part of O, ZnSO4·H20.0014 part of O, CoCl20.0005 part, H2100 parts of O；pH5.5.
6. the method for fast degradation water body withered fallen leaf according to claim 5, it is characterised in that: the carboxymethyl cellulose
Plating medium: including the Mandel nutrient solution, 2% carboxymethyl cellulose, 0.08% NaTDC, 1.5%
Agar, 121 DEG C of sterilizing 20min；It is cooled to inverted plate after non-scald on hand, is inverted after plate solidification spare.
7. the method for fast degradation water body withered fallen leaf according to claim 1, it is characterised in that: prepare the LB solid training
Support base: according to parts by weight, including 10 parts of tryptone, 5 parts of yeast powder, 10 parts of sodium chloride, 10-15 parts of agar powder, distilled water
1000 parts；Tune pH is 7.0,121 DEG C of sterilizing 20min, is cooled to inverted plate after non-scald on hand, is inverted after solidification stand-by.
8. the method for fast degradation water body withered fallen leaf according to claim 1, it is characterised in that: prepare the LB liquid training
Support base: according to parts by weight, including 10 parts of tryptone, 5 parts of yeast powder, 10 parts of sodium chloride, 10-15 parts of agar powder, distilled water
1000 parts, 7.0,121 DEG C of sterilizing 20min of pH, 4 DEG C of placements are spare.
9. the method for fast degradation water body withered fallen leaf according to claim 1, it is characterised in that: prepare the potato Portugal
Grape sugar agar medium: according to parts by weight, including 150-200 parts of potato, 15-20 parts of glucose, 15-20 parts of agar, H2O
1000 parts；115 DEG C of high pressure sterilization 15min.
10. the method for fast degradation water body withered fallen leaf according to claim 1, it is characterised in that: prepare the potato
Dextrose broth: according to parts by weight, including 150-200 parts of potato, 15-20 parts of glucose, H21000 parts of O；
115 DEG C of high pressure sterilization 15min.
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| CN114368833A (en) * | 2021-12-27 | 2022-04-19 | 江鹰 | Nitrogen and phosphorus removal composite filler and purification device containing same |
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