CN106148246A - Purify compounding microbial inoculum of black and odorous water and preparation method thereof - Google Patents
Purify compounding microbial inoculum of black and odorous water and preparation method thereof Download PDFInfo
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Abstract
The present invention provides a kind of compounding microbial inoculum purifying black and odorous water and preparation method thereof, compounding microbial inoculum to include following component: actinomycetes, bacillus subtilis, nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa.The weight proportion of each component is: 2:2:2:1:1:1.Advantage is: COD content, ammonia-nitrogen content and the phosphate content of black and odorous water of can effectively degrading, and can eliminate the stink of black and odorous water；It is simultaneously introduced removal ammonia nitrogen strain, water transparency can be stepped up.
Description
Technical field
The invention belongs to environment-protective water treatment and purification technical field, be specifically related to a kind of compounding microbial inoculum purifying black and odorous water and
Its preparation method.
Background technology
In recent years, the water body smelly phenomenon of blackout grows in intensity, and black and odorous water is a kind of extreme table that water body organism pollutes
Existing, major part black and odorous water mechanism is essentially the same, and due to water hypoxia, Organic substance is corrupt and causes.Concrete, when in a large number
Organic pollution enter water body, under the biochemical action of oxygen consumption microorganism, consume substantial amounts of oxygen in water body, make water body turn
Chemical conversion anaerobic condition, causes anaerobic bacteria amount reproduction, and Organic substance is corrupt, decompose, ferment, so that water body blackening fouling.
For black and odorous water, its situation is complicated, depends merely on single a kind of recovery technique and does not often reach ideal effect.Cause
This, extensive microbe microbial inoculum carries out improvement to black and odorous water and becomes focus at present.
But, microbial bacterial agent currently on the market, generally have expensive, black and odorous water regulation effect is undesirable
Problem.Therefore, research and develop a kind of new high efficiency composition microbial inoculum, be thing the most in the urgent need to address.
Summary of the invention
The defect existed for prior art, the present invention provides a kind of compounding microbial inoculum purifying black and odorous water and preparation side thereof
Method, can effectively solve the problems referred to above.
The technical solution used in the present invention is as follows:
An object of the present disclosure provides a kind of compounding microbial inoculum purifying black and odorous water, including following component: actinomycetes, hay
Bacillus cereus, nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa.
Preferably, the weight proportion of each component is: 2:2:2:1:1:1.
The preparation method providing a kind of compounding microbial inoculum purifying black and odorous water of the present invention the second mesh, comprises the following steps:
Step 1, according to the characteristic of aimed strain, enrichment medium from the activated sludge of sewage treatment plant, respectively obtain
Aimed strain orienting enriching culture medium corresponding to every kind of aimed strain；Wherein, described aimed strain includes actinomycetes, hay bud
Spore bacillus, nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa；
Aimed strain is inoculated in the aimed strain orienting enriching culture medium of correspondence, is oriented enrichment culture；Treat it
OD600When=2, stop enrichment culture, obtain the aimed strain after enrichment culture；
Step 2, prepares the aimed strain screening culture medium corresponding to every kind of aimed strain；
Aimed strain after step 1 enrichment culture is carried out gradient dilution, is diluted to 10 respectively3Again, 105Again with 107Times；
Then, use aimed strain screening culture medium that the aimed strain after corresponding dilution is carried out screening and culturing；
Step 3, carries out secondary separation purification by the target list bacterium colony after step 2 screening and culturing, obtains after purification
Target list bacterium colony；
Step 4, carries out qualitative detection experiment to step 3 target list bacterium colony after purification, and the aimed strain that choosing is positive is made
For alternative target bacterial strain；
Step 5, uses and tames culture medium accordingly, alternative target bacterial strain carries out domestication and cultivates；
Step 6, the aimed strain after domestication being cultivated carries out liquid fermentation amplification culture；
Step 7, by the aimed strain after step 6 amplification culture, compounds according to formula proportion, prepares Compound bacterium
Agent.
Preferably, in step 1, the orienting enriching Medium Proportion corresponding to each aimed strain is:
Bacillus subtilis, lactic acid bacteria are identical with the orienting enriching culture medium corresponding to Pseudomonas aeruginosa, all by with
Lower method obtains: river sewage is stood 2～3 days, takes supernatant, adds peptone 8～10g, Carnis Bovis seu Bubali cream 1～3g and sodium chloride
20～30g, with water constant volume to 1L；
Orienting enriching culture medium corresponding to actinomycetes is: add soluble starch 15～20g, dipotassium hydrogen phosphate 0.5～
1g, sodium chloride 0.5～1g, magnesium sulfate 0.2～0.5g, potassium nitrate 1～2g, ferrous sulfate 0.01～0.03g, arrive with water constant volume
1L, then to regulate pH be 7.2～7.4；
Producing orienting enriching culture medium corresponding to Ruan's candidiasis is: add glucose 80～100g, peptone 5～
10g, yeast extract 10g, potassium dihydrogen phosphate 1～2g, magnesium sulfate 0.2～0.5g, with water constant volume to 1L, then regulate pH be 4.0～
6.5；
Orienting enriching culture medium corresponding to nitrifier is: add peptone 10～15g, sodium chloride 5～7g, Carnis Bovis seu Bubali cream 3
～5g, potassium nitrate 1～2g, with water constant volume to 1L, then to regulate pH be 7.0～7.5.
Preferably, in step 2, the aimed strain screening culture medium proportioning corresponding to various aimed strains is:
The screening culture medium of bacillus subtilis, lactic acid bacteria and Pseudomonas aeruginosa is identical, is: add glucose 50～
60g, ammonium chloride 0.5～1g and yeast extract 10g, with water constant volume to 1L, then to regulate pH be 7.0～7.5；
Actinomycetes screening culture medium is: add soluble starch 20～25g, dipotassium hydrogen phosphate 1～1.5g, sodium chloride 0.5
～1g, magnesium sulfate 0.2～0.3g, potassium nitrate 0.5～1g, ferrous sulfate 0.01～0.03g, agar 15～20g, arrive with water constant volume
1L, then to regulate pH be 7.2～7.4；Additionally, every 1 actinomycetes screening culture medium adds the potassium dichromate that mass fraction is 3%
1.5mL, suppression antibacterial and the interference of mycete；
Producing Ruan's candidiasis screening culture medium is: claims Rhizoma Solani tuber osi 180～200g, adds 1L boiling tap water 30min, mistake
After filtering residue, constant volume to 1L, add glucose 20～25g, lactic acid 5～6mL, agar 15～20g, regulation pH be 4.0～
6.5, thus suppress the growth of other miscellaneous bacteria；
Nitrifier screening culture medium: add sodium succinate 8～10g, altheine acid 1～1.5g, sodium nitrate 0.5～
1g, potassium dihydrogen phosphate 1～2g, ferrous chloride 0.02～0.05g, calcium chloride 0.2～0.3g, magnesium sulfate 1～2g, BTB solution 1mL
(1% ethanol) and agar 15～20g, with water constant volume to 1L, then to regulate pH be 7.0～7.3；Wherein, BTB solution refers to: BTB according to
The volume fraction of 1% is dissolved in ethanol, takes weighing 1ml.
Preferably, step 4 particularly as follows:
Step 4.1, prepares following three class Qualitative Identification culture medium:
A () protease production measures culture medium: take the skim milk that mass fraction is 10～15%, in 110～113 DEG C
Sterilizing 30min, with the beef extract-peptone ratio mixing with mass ratio as 1:8, the mass fraction by 15～20g/L adds fine jade
Fat, adjusting pH is 7.0～7.2, makes protease production and measures culture medium；
(b) produce amylase activity measure culture medium: take soluble starch 1～2g, peptone 3～6g, potassium chloride 0.5～
1g, Magnesium sulfate heptahydrate 0.5～1g, agar 15～20g, with water constant volume to 1L, then to regulate pH be 7.0～7.2, makes product amylase
Determination of activity culture medium；
C () denitrogenates qualitative determination culture medium: take potassium nitrate 1～2g, sodium citrate 5～7g, ammonium sulfate 0.3～0.5g, phosphorus
Acid hydrogen dipotassium 1～3g, potassium dihydrogen phosphate 2～3g, Magnesium sulfate heptahydrate 0.1～0.2g, agar 15～20g, with water constant volume to 1L, then
Regulation pH is 7.0～7.2, makes and denitrogenates qualitative determination culture medium；
Step 4.2, for step 3 every kind of target list bacterium colony after purification, is respectively adopted protease production and measures cultivation
Base, produce amylase activity and measure culture medium and denitrogenate qualitative determination culture medium it is carried out qualitative detection, select to produce protease activity
Property and/or produce amylase activity and/or denitrogenate the aimed strain alternately aimed strain that activity is positive.
Preferably, step 5 particularly as follows:
Step 5.1, prepare following three classes domestication culture medium:
A () denitrogenates domestication culture medium: take potassium nitrate 1～2g, sodium citrate 5～7g, ammonium sulfate 0.3～0.5g, phosphoric acid hydrogen
Dipotassium 1～3g, potassium dihydrogen phosphate 2～3g, Magnesium sulfate heptahydrate 0.1～0.2g, stirring mixing, with water constant volume to 1L；
(b) protease production domestication culture medium: configuration quality mark is the skim milk of 10～15%, adds dilution
In the beef extract-peptone Carnis Bovis seu Bubali cream culture medium of 10 times of volumes, milk and Carnis Bovis seu Bubali cream culture medium mass ratio are 1:1, reconcile pH and are
7.2～7.4；
(c) produce amylase activity domestication culture medium: take soluble starch 1～5g, peptone 10～15g, potassium chloride 0.1～
1g and magnesium sulfate 0.2～0.5g, with water constant volume to 1L, reconciling pH is 7.2～7.4；
Step 5.2, the aimed strain that protease production is positive, use protease production domestication culture medium to mesh
Mark bacterial strain carries out domestication and cultivates；
To producing the aimed strain that amylase activity is positive, use product amylase activity domestication culture medium that aimed strain is entered
Row domestication is cultivated；
To denitrogenating the aimed strain that activity is positive, employing is denitrogenated domestication culture medium and aimed strain is carried out domestication cultivation.
Preferably, step 6 particularly as follows:
Aimed strain after domestication being cultivated activates, it may be assumed that is respectively adopted the domestication culture medium identical with step 5 and carries out
Activation, treats its OD600When reaching 1.5, stop activation, obtain the aimed strain after amplification culture.
Preferably, in step 7, also include:
By the aimed strain after step 6 amplification culture, following methods is used to preserve:
For bacillus subtilis, Pseudomonas aeruginosa, nitrifier and lactic acid bacteria, the method for diatomite adsorption is used to protect
Deposit, it may be assumed that first the kieselguhr containing mass fraction 10～15% Semen Maydis powder and mass fraction 10～20% glucose is mixed all
Even, it is placed in 121 DEG C of high temperature sterilize 30min, is layered in solid fermentation dish；Solid fermentation dish is placed in air dry oven in
30～35 DEG C of constant temperature dryings, obtain the adsorption substrates being dried；Then, aimed strain is adsorbed onto in described adsorption substrates；
For producing Ruan's candidiasis and actinomycetes, the method using Testa Tritici absorption, it may be assumed that first will be containing mass fraction 10
～15% Semen Maydis powder and mass fraction 10～the Testa Tritici mix homogeneously of 20% glucose, it is placed in 121 DEG C of high temperature sterilize 30min, will
It is layered in solid fermentation dish；Solid fermentation dish is placed in air dry oven in 30～35 DEG C of constant temperature dryings, obtains being dried
Adsorption substrates；Then, aimed strain is adsorbed onto in described adsorption substrates.
Compounding microbial inoculum purifying black and odorous water that the present invention provides and preparation method thereof has the advantage that
The compounding microbial inoculum purifying black and odorous water that the present invention provides, effectively purifies mainly for black-odor riverway sewage,
In this compounding microbial inoculum rich in the bacterium producing multi enzyme preparation such as bacillus cereus, the organic material decomposition in water body can be become little by these bacterium producing multi enzyme preparations
Molecular substance/carbon dioxide and water；Small-molecule substance further decomposes.Meanwhile, aimed strain is riotous growth in water body, has
The growth of effect suppression miscellaneous bacteria, forms dominant strain；Additionally compound in microbial inoculum and screen the actinomyces strain obtained, can effective degradation water
Organic substance in body, thus the COD in water body that degrades；The nitrification bacterial strain that obtains of screening can be degraded the ammonia nitrogen in water body；Yeast
Bacterium can be that other bacterial strain provides nutrient substance.Therefore, can effectively degrade COD content, ammonia-nitrogen content and the phosphoric acid of black and odorous water
Salt content, and the stink of black and odorous water can be eliminated；It is simultaneously introduced removal ammonia nitrogen strain, water transparency can be stepped up.
Accompanying drawing explanation
The schematic flow sheet of the preparation method of the compounding microbial inoculum purifying black and odorous water that Fig. 1 provides for the present invention.
Detailed description of the invention
Below in conjunction with accompanying drawing, the present invention is described in detail:
The present invention provides a kind of compounding microbial inoculum purifying black and odorous water, this product microorganism belonging to genus microbial inoculum, is added to black smelly river
In water, malodorous black water is processed, COD content, ammonia-nitrogen content and the phosphate content of black and odorous water of can effectively degrading, and
The stink of black and odorous water can be eliminated；It is simultaneously introduced removal ammonia nitrogen strain, water transparency can be stepped up.
The compounding microbial inoculum purifying black and odorous water that the present invention provides, mainly uses by efficient degrading bacterial strain in sewage
Screening, purification and enrichment, through combine experiment, obtain the composite bacteria agent capable high to black-odor riverway treatment effeciency.The compounding bar of this microbial inoculum
Part is flexible, can meet the process of different river water body environment, and it has with low cost, widely used advantage.
It addition, proposing of also innovating of the present invention aimed strain orienting enriching culture medium the most corresponding with every kind of single bacterium,
Aimed strain screening culture medium, target list bacterium colony qualitative detection culture medium, aimed strain domestication culture medium, uses the present invention to provide
Various culture medium, after respectively single bacterium being oriented enrichment, screens and tame, prepare single bacterium of excellent performance；Then
After compounding for 6 kinds of single bacterium, obtain compounding microbial inoculum；Each single bacterium of compounding microbial inoculum cooperates and plays a role, thus can effectively degrade
COD content, ammonia-nitrogen content and the phosphate content of black and odorous water, and the stink of black and odorous water can be eliminated, effect is the most aobvious
Write.
Concrete, the present invention provides a kind of compounding microbial inoculum purifying black and odorous water, including following component: actinomycetes, hay
Bacillus cereus, nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa.The weight proportion of each component is: 2:2:2:
1:1:1。
The present invention also provides for the preparation method of a kind of compounding microbial inoculum purifying black and odorous water, comprises the following steps:
Step 1, according to the characteristic of aimed strain, enrichment medium from the activated sludge of sewage treatment plant, respectively obtain
Aimed strain orienting enriching culture medium corresponding to every kind of aimed strain；Wherein, described aimed strain includes actinomycetes, hay bud
Spore bacillus, nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa；
Concrete, the orienting enriching Medium Proportion corresponding to each aimed strain is:
Bacillus subtilis, lactic acid bacteria are identical with the orienting enriching culture medium corresponding to Pseudomonas aeruginosa, all by with
Lower method obtains: river sewage is stood 2～3 days, takes supernatant, adds peptone 8～10g, Carnis Bovis seu Bubali cream 1～3g and sodium chloride
20～30g, with water constant volume to 1L；
Orienting enriching culture medium corresponding to actinomycetes is: add soluble starch 15～20g, dipotassium hydrogen phosphate 0.5～
1g, sodium chloride 0.5～1g, magnesium sulfate 0.2～0.5g, potassium nitrate 1～2g, ferrous sulfate 0.01～0.03g, arrive with water constant volume
1L, then to regulate pH be 7.2～7.4；
Producing orienting enriching culture medium corresponding to Ruan's candidiasis is: add glucose 80～100g, peptone 5～
10g, yeast extract 10g, potassium dihydrogen phosphate 1～2g, magnesium sulfate 0.2～0.5g, with water constant volume to 1L, then regulate pH be 4.0～
6.5；
Orienting enriching culture medium corresponding to nitrifier is: add peptone 10～15g, sodium chloride 5～7g, Carnis Bovis seu Bubali cream 3
～5g, potassium nitrate 1～2g, with water constant volume to 1L, then to regulate pH be 7.0～7.5.
Then, aimed strain is inoculated in the aimed strain orienting enriching culture medium of correspondence, is oriented enrichment culture；
Treat its OD600When=2, stop enrichment culture, obtain the aimed strain after enrichment culture；
Step 2, prepares the aimed strain screening culture medium corresponding to every kind of aimed strain；
Aimed strain after step 1 enrichment culture is carried out gradient dilution, is diluted to 10 respectively3Again, 105Again with 107Times；
Then, use aimed strain screening culture medium that the aimed strain after corresponding dilution is carried out screening and culturing；
Wherein, the aimed strain screening culture medium proportioning corresponding to various aimed strains is:
The screening culture medium of bacillus subtilis, lactic acid bacteria and Pseudomonas aeruginosa is identical, is: add glucose 50～
60g, ammonium chloride 0.5～1g and yeast extract 10g, with water constant volume to 1L, then to regulate pH be 7.0～7.5；
Actinomycetes screening culture medium is: add soluble starch 20～25g, dipotassium hydrogen phosphate 1～1.5g, sodium chloride 0.5
～1g, magnesium sulfate 0.2～0.3g, potassium nitrate 0.5～1g, ferrous sulfate 0.01～0.03g, agar 15～20g, arrive with water constant volume
1L, then to regulate pH be 7.2～7.4；Additionally, every 1 actinomycetes screening culture medium adds the potassium dichromate that mass fraction is 3%
1.5mL, suppression antibacterial and the interference of mycete；
Producing Ruan's candidiasis screening culture medium is: claims Rhizoma Solani tuber osi 180～200g, adds 1L boiling tap water 30min, mistake
After filtering residue, constant volume to 1L, add glucose 20～25g, lactic acid 5～6mL, agar 15～20g, regulation pH be 4.0～
6.5, thus suppress the growth of other miscellaneous bacteria；
Nitrifier screening culture medium: add sodium succinate 8～10g, altheine acid 1～1.5g, sodium nitrate 0.5～
1g, potassium dihydrogen phosphate 1～2g, ferrous chloride 0.02～0.05g, calcium chloride 0.2～0.3g, magnesium sulfate 1～2g, BTB solution 1mL
(1% ethanol) and agar 15～20g, with water constant volume to 1L, then to regulate pH be 7.0～7.3；Wherein, BTB solution refers to: BTB according to
The volume fraction of 1% is dissolved in ethanol, takes weighing 1ml.
Step 3, carries out secondary separation purification by the target list bacterium colony after step 2 screening and culturing, obtains after purification
Target list bacterium colony；
Step 4, carries out qualitative detection experiment to step 3 target list bacterium colony after purification, and the aimed strain that choosing is positive is made
For alternative target bacterial strain；
Step 4 particularly as follows:
Step 4.1, prepares following three class Qualitative Identification culture medium:
A () protease production measures culture medium: take the skim milk that mass fraction is 10～15%, in 110～113 DEG C
Sterilizing 30min, with the beef extract-peptone ratio mixing with mass ratio as 1:8, the mass fraction by 15～20g/L adds fine jade
Fat, adjusting pH is 7.0～7.2, makes protease production and measures culture medium；
(b) produce amylase activity measure culture medium: take soluble starch 1～2g, peptone 3～6g, potassium chloride 0.5～
1g, Magnesium sulfate heptahydrate 0.5～1g, agar 15～20g, with water constant volume to 1L, then to regulate pH be 7.0～7.2, makes product amylase
Determination of activity culture medium；
C () denitrogenates qualitative determination culture medium: take potassium nitrate 1～2g, sodium citrate 5～7g, ammonium sulfate 0.3～0.5g, phosphorus
Acid hydrogen dipotassium 1～3g, potassium dihydrogen phosphate 2～3g, Magnesium sulfate heptahydrate 0.1～0.2g, agar 15～20g, with water constant volume to 1L, then
Regulation pH is 7.0～7.2, makes and denitrogenates qualitative determination culture medium；
Step 4.2, for step 3 every kind of target list bacterium colony after purification, is respectively adopted protease production and measures cultivation
Base, produce amylase activity and measure culture medium and denitrogenate qualitative determination culture medium it is carried out qualitative detection, select to produce protease activity
Property and/or produce amylase activity and/or denitrogenate the aimed strain alternately aimed strain that activity is positive.
Step 5, uses and tames culture medium accordingly, alternative target bacterial strain carries out domestication and cultivates；
Step 5 particularly as follows:
Step 5.1, prepare following three classes domestication culture medium:
A () denitrogenates domestication culture medium: take potassium nitrate 1～2g, sodium citrate 5～7g, ammonium sulfate 0.3～0.5g, phosphoric acid hydrogen
Dipotassium 1～3g, potassium dihydrogen phosphate 2～3g, Magnesium sulfate heptahydrate 0.1～0.2g, stirring mixing, with water constant volume to 1L；
(b) protease production domestication culture medium: configuration quality mark is the skim milk of 10～15%, adds dilution
In the beef extract-peptone Carnis Bovis seu Bubali cream culture medium of 10 times of volumes, milk and Carnis Bovis seu Bubali cream culture medium mass ratio are 1:1, reconcile pH and are
7.2～7.4；
(c) produce amylase activity domestication culture medium: take soluble starch 1～5g, peptone 10～15g, potassium chloride 0.1～
1g and magnesium sulfate 0.2～0.5g, with water constant volume to 1L, reconciling pH is 7.2～7.4；
Step 5.2, the aimed strain that protease production is positive, use protease production domestication culture medium to mesh
Mark bacterial strain carries out domestication and cultivates；
To producing the aimed strain that amylase activity is positive, use product amylase activity domestication culture medium that aimed strain is entered
Row domestication is cultivated；
To denitrogenating the aimed strain that activity is positive, employing is denitrogenated domestication culture medium and aimed strain is carried out domestication cultivation.
Step 6, the aimed strain after domestication being cultivated carries out liquid fermentation amplification culture；
Step 6 is particularly as follows: the aimed strain after domestication being cultivated activates, it may be assumed that be respectively adopted identical the taming and dociling with step 5
Change culture medium to activate, treat its OD600When reaching 1.5, stop activation, obtain the aimed strain after amplification culture.
Step 7, by the aimed strain after step 6 amplification culture, compounds according to formula proportion, prepares Compound bacterium
Agent.
In step 7, also include:
By the aimed strain after step 6 amplification culture, following methods is used to preserve:
For bacillus subtilis, Pseudomonas aeruginosa, nitrifier and lactic acid bacteria, the method for diatomite adsorption is used to protect
Deposit, it may be assumed that first the kieselguhr containing mass fraction 10～15% Semen Maydis powder and mass fraction 10～20% glucose is mixed all
Even, it is placed in 121 DEG C of high temperature sterilize 30min, is layered in solid fermentation dish；Solid fermentation dish is placed in air dry oven in
30～35 DEG C of constant temperature dryings, obtain the adsorption substrates being dried；Then, aimed strain is adsorbed onto in described adsorption substrates；
For producing Ruan's candidiasis and actinomycetes, the method using Testa Tritici absorption, it may be assumed that first will be containing mass fraction 10
～15% Semen Maydis powder and mass fraction 10～the Testa Tritici mix homogeneously of 20% glucose, it is placed in 121 DEG C of high temperature sterilize 30min, will
It is layered in solid fermentation dish；Solid fermentation dish is placed in air dry oven in 30～35 DEG C of constant temperature dryings, obtains being dried
Adsorption substrates；Then, aimed strain is adsorbed onto in described adsorption substrates.
Embodiment one:
Step 1, according to the characteristic of aimed strain, enrichment medium from the activated sludge of sewage treatment plant, respectively obtain
Aimed strain orienting enriching culture medium corresponding to every kind of aimed strain；Wherein, described aimed strain includes actinomycetes, hay bud
Spore bacillus, nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa；
Concrete, the orienting enriching Medium Proportion corresponding to each aimed strain is:
Bacillus subtilis, lactic acid bacteria are identical with the orienting enriching culture medium corresponding to Pseudomonas aeruginosa, all by with
Lower method obtains: river sewage is stood 2～3 days, takes supernatant, adds peptone 8g, Carnis Bovis seu Bubali cream 3g and sodium chloride 20g, uses
Water constant volume is to 1L；
Orienting enriching culture medium corresponding to actinomycetes is: add soluble starch 15g, dipotassium hydrogen phosphate 0.5g, chlorination
Sodium 1g, magnesium sulfate 0.2g, potassium nitrate 1g, ferrous sulfate 0.01g, with water constant volume to 1L, then to regulate pH be 7.2；
Producing the orienting enriching culture medium corresponding to Ruan's candidiasis is: add glucose 100g, peptone 10g, yeast
Cream 10g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.2g, with water constant volume to 1L, then to regulate pH be 4.0；
Orienting enriching culture medium corresponding to nitrifier is: add peptone 10g, sodium chloride 7g, Carnis Bovis seu Bubali cream 3g, potassium nitrate
1g, with water constant volume to 1L, then to regulate pH be 7.0.
Then, aimed strain is inoculated in the aimed strain orienting enriching culture medium of correspondence, is oriented enrichment culture；
Treat its OD600When=2, stop enrichment culture, obtain the aimed strain after enrichment culture；
Step 2, prepares the aimed strain screening culture medium corresponding to every kind of aimed strain；
Aimed strain after step 1 enrichment culture is carried out gradient dilution, is diluted to 10 respectively3Again, 105Again with 107Times；
Then, use aimed strain screening culture medium that the aimed strain after corresponding dilution is carried out screening and culturing；
Wherein, the aimed strain screening culture medium proportioning corresponding to various aimed strains is:
The screening culture medium of bacillus subtilis, lactic acid bacteria and Pseudomonas aeruginosa is identical, is: addition glucose 50g,
Ammonium chloride 0.5g and yeast extract 10g, with water constant volume to 1L, then to regulate pH be 7.1；
Actinomycetes screening culture medium is: add soluble starch 20g, dipotassium hydrogen phosphate 1g, sodium chloride 0.5g, magnesium sulfate
0.2g, potassium nitrate 0.8g, ferrous sulfate 0.01g, agar 20g, with water constant volume to 1L, then to regulate pH be 7.2；Additionally, every 1 unwrapping wire
Bacterium screening culture medium adds the potassium dichromate 1.5mL that mass fraction is 3%, suppression antibacterial and the interference of mycete；
Producing Ruan's candidiasis screening culture medium is: claims Rhizoma Solani tuber osi 180g, adds 1L boiling tap water 30min, crosses and filter
After removing residue, constant volume to 1L, add glucose 20g, lactic acid 5mL, agar 20g, regulation pH is 4.0, thus suppresses other miscellaneous bacteria
Growth；
Nitrifier screening culture medium: add sodium succinate 8g, altheine acid 1g, sodium nitrate 0.5g, potassium dihydrogen phosphate
1g, ferrous chloride 0.05g, calcium chloride 0.2g, magnesium sulfate 2g, BTB solution 1mL (1% ethanol) and agar 15g, arrive with water constant volume
1L, then to regulate pH be 7.0；Wherein, BTB solution refers to: BTB dissolves in ethanol according to the volume fraction of 1%, takes weighing 1ml.
Step 3, carries out secondary separation purification by the target list bacterium colony after step 2 screening and culturing, obtains after purification
Target list bacterium colony；
Step 4, carries out qualitative detection experiment to step 3 target list bacterium colony after purification, and the aimed strain that choosing is positive is made
For alternative target bacterial strain；
Step 4 particularly as follows:
Step 4.1, prepares following three class Qualitative Identification culture medium:
A () protease production measures culture medium: take the skim milk that mass fraction is 15%, in 110 DEG C of sterilizings
30min, with the beef extract-peptone ratio mixing with mass ratio as 1:8, adds agar by the mass fraction of 15g/L, and tune pH is
7.0, make protease production and measure culture medium；
B () is produced amylase activity and is measured culture medium: take soluble starch 2g, peptone 3g, potassium chloride 0.9g, seven water sulphuric acid
Magnesium 1g, agar 15g, with water constant volume to 1L, then to regulate pH be 7.0, makes product amylase activity and measures culture medium；
C () denitrogenates qualitative determination culture medium: take potassium nitrate 1g, sodium citrate 5g, ammonium sulfate 0.3g, dipotassium hydrogen phosphate 1g,
Potassium dihydrogen phosphate 3g, Magnesium sulfate heptahydrate 0.1g, agar 15g, with water constant volume to 1L, then to regulate pH be 7.0, makes and denitrogenates qualitative survey
Determine culture medium；
Step 4.2, for step 3 every kind of target list bacterium colony after purification, is respectively adopted protease production and measures cultivation
Base, produce amylase activity and measure culture medium and denitrogenate qualitative determination culture medium it is carried out qualitative detection, select to produce protease activity
Property and/or produce amylase activity and/or denitrogenate the aimed strain alternately aimed strain that activity is positive.
Step 5, uses and tames culture medium accordingly, alternative target bacterial strain carries out domestication and cultivates；
Step 5 particularly as follows:
Step 5.1, prepare following three classes domestication culture medium:
A () denitrogenates domestication culture medium: take potassium nitrate 1g, sodium citrate 5g, ammonium sulfate 0.5g, dipotassium hydrogen phosphate 1g, phosphoric acid
Potassium dihydrogen 3g, Magnesium sulfate heptahydrate 0.1g, stirring mixing, with water constant volume to 1L；
(b) protease production domestication culture medium: configuration quality mark is the skim milk of 15%, adds 10 times of dilution
In the beef extract-peptone Carnis Bovis seu Bubali cream culture medium of volume, milk and Carnis Bovis seu Bubali cream culture medium mass ratio are 1:1, and reconciling pH is 7.2；
C () produces amylase activity domestication culture medium: take soluble starch 5g, peptone 10g, potassium chloride 0.1g and magnesium sulfate
0.2g, with water constant volume to 1L, reconciling pH is 7.2；
Step 5.2, the aimed strain that protease production is positive, use protease production domestication culture medium to mesh
Mark bacterial strain carries out domestication and cultivates；
To producing the aimed strain that amylase activity is positive, use product amylase activity domestication culture medium that aimed strain is entered
Row domestication is cultivated；
To denitrogenating the aimed strain that activity is positive, employing is denitrogenated domestication culture medium and aimed strain is carried out domestication cultivation.
Step 6, the aimed strain after domestication being cultivated carries out liquid fermentation amplification culture；
Step 6 is particularly as follows: the aimed strain after domestication being cultivated activates, it may be assumed that be respectively adopted identical the taming and dociling with step 5
Change culture medium to activate, treat its OD600When reaching 1.5, stop activation, obtain the aimed strain after amplification culture.
Step 7, by the aimed strain after step 6 amplification culture, compounds according to formula proportion, prepares Compound bacterium
Agent.Formula proportion is: actinomycetes, bacillus subtilis, nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa.
The weight proportion of each component is: 2:2:2:1:1:1.
In step 7, also include:
By the aimed strain after step 6 amplification culture, following methods is used to preserve:
For bacillus subtilis, Pseudomonas aeruginosa, nitrifier and lactic acid bacteria, the method for diatomite adsorption is used to protect
Deposit, it may be assumed that first by containing mass fraction 10% Semen Maydis powder and the kieselguhr mix homogeneously of mass fraction 10% glucose, be placed in
121 DEG C of high temperature sterilize 30min, are layered in solid fermentation dish；Solid fermentation dish is placed in air dry oven in 30 DEG C of perseverances
Temperature is dried, and obtains the adsorption substrates being dried；Then, aimed strain is adsorbed onto in described adsorption substrates；
For producing Ruan's candidiasis and actinomycetes, the method using Testa Tritici absorption, it may be assumed that first will be containing mass fraction
10% Semen Maydis powder and the Testa Tritici mix homogeneously of mass fraction 20% glucose, be placed in 121 DEG C of high temperature sterilize 30min, be layered on
In solid fermentation dish；Solid fermentation dish is placed in air dry oven in 35 DEG C of constant temperature dryings, obtains the adsorption substrates being dried；So
After, aimed strain is adsorbed onto in described adsorption substrates.
Embodiment two:
Step 1, according to the characteristic of aimed strain, enrichment medium from the activated sludge of sewage treatment plant, respectively obtain
Aimed strain orienting enriching culture medium corresponding to every kind of aimed strain；Wherein, described aimed strain includes actinomycetes, hay bud
Spore bacillus, nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa；
Concrete, the orienting enriching Medium Proportion corresponding to each aimed strain is:
Bacillus subtilis, lactic acid bacteria are identical with the orienting enriching culture medium corresponding to Pseudomonas aeruginosa, all by with
Lower method obtains: river sewage is stood 2～3 days, takes supernatant, adds peptone 10g, Carnis Bovis seu Bubali cream 1g and sodium chloride 30g, uses
Water constant volume is to 1L；
Orienting enriching culture medium corresponding to actinomycetes is: add soluble starch 20g, dipotassium hydrogen phosphate 1g, sodium chloride
0.5g, magnesium sulfate 0.4g, potassium nitrate 2g, ferrous sulfate 0.02g, with water constant volume to 1L, then to regulate pH be 7.4；
Producing the orienting enriching culture medium corresponding to Ruan's candidiasis is: add glucose 80g, peptone 6g, yeast extract
10g, potassium dihydrogen phosphate 2g, magnesium sulfate 0.4g, with water constant volume to 1L, then to regulate pH be 6.5；
Orienting enriching culture medium corresponding to nitrifier is: add peptone 15g, sodium chloride 5g, Carnis Bovis seu Bubali cream 5g, potassium nitrate
2g, with water constant volume to 1L, then to regulate pH be 7.5.
Then, aimed strain is inoculated in the aimed strain orienting enriching culture medium of correspondence, is oriented enrichment culture；
Treat its OD600When=2, stop enrichment culture, obtain the aimed strain after enrichment culture；
Step 2, prepares the aimed strain screening culture medium corresponding to every kind of aimed strain；
Aimed strain after step 1 enrichment culture is carried out gradient dilution, is diluted to 10 respectively3Again, 105Again with 107Times；
Then, use aimed strain screening culture medium that the aimed strain after corresponding dilution is carried out screening and culturing；
Wherein, the aimed strain screening culture medium proportioning corresponding to various aimed strains is:
The screening culture medium of bacillus subtilis, lactic acid bacteria and Pseudomonas aeruginosa is identical, is: addition glucose 60g,
Ammonium chloride 0.9g and yeast extract 10g, with water constant volume to 1L, then to regulate pH be 7.4；
Actinomycetes screening culture medium is: add soluble starch 25g, dipotassium hydrogen phosphate 1.5g, sodium chloride 0.9g, magnesium sulfate
0.3g, potassium nitrate 0.5g, ferrous sulfate 0.03g, agar 15g, with water constant volume to 1L, then to regulate pH be 7.4；Additionally, every 1 unwrapping wire
Bacterium screening culture medium adds the potassium dichromate 1.5mL that mass fraction is 3%, suppression antibacterial and the interference of mycete；
Producing Ruan's candidiasis screening culture medium is: claims Rhizoma Solani tuber osi 200g, adds 1L boiling tap water 30min, crosses and filter
After removing residue, constant volume to 1L, add glucose 24g, lactic acid 6mL, agar 15g, regulation pH is 5.5, thus suppresses other miscellaneous bacteria
Growth；
Nitrifier screening culture medium: add sodium succinate 10g, altheine acid 1.5g, sodium nitrate 1g, potassium dihydrogen phosphate
2g, ferrous chloride 0.02g, calcium chloride 0.3g, magnesium sulfate 1g, BTB solution 1mL (1% ethanol) and agar 20g, arrive with water constant volume
1L, then to regulate pH be 7.2；Wherein, BTB solution refers to: BTB dissolves in ethanol according to the volume fraction of 1%, takes weighing 1ml.
Step 3, carries out secondary separation purification by the target list bacterium colony after step 2 screening and culturing, obtains after purification
Target list bacterium colony；
Step 4, carries out qualitative detection experiment to step 3 target list bacterium colony after purification, and the aimed strain that choosing is positive is made
For alternative target bacterial strain；
Step 4 particularly as follows:
Step 4.1, prepares following three class Qualitative Identification culture medium:
A () protease production measures culture medium: take the skim milk that mass fraction is 10%, in 113 DEG C of sterilizings
30min, with the beef extract-peptone ratio mixing with mass ratio as 1:8, adds agar by the mass fraction of 20g/L, and tune pH is
7.2, make protease production and measure culture medium；
B () is produced amylase activity and is measured culture medium: take soluble starch 1g, peptone 5g, potassium chloride 0.5g, seven water sulphuric acid
Magnesium 0.5g, agar 20g, with water constant volume to 1L, then to regulate pH be 7.2, makes product amylase activity and measures culture medium；
C () denitrogenates qualitative determination culture medium: take potassium nitrate 2g, sodium citrate 6g, ammonium sulfate 0.4g, dipotassium hydrogen phosphate 3g,
Potassium dihydrogen phosphate 2g, Magnesium sulfate heptahydrate 0.2g, agar 20g, with water constant volume to 1L, then to regulate pH be 7.2, makes and denitrogenates qualitative survey
Determine culture medium；
Step 4.2, for step 3 every kind of target list bacterium colony after purification, is respectively adopted protease production and measures cultivation
Base, produce amylase activity and measure culture medium and denitrogenate qualitative determination culture medium it is carried out qualitative detection, select to produce protease activity
Property and/or produce amylase activity and/or denitrogenate the aimed strain alternately aimed strain that activity is positive.
Step 5, uses and tames culture medium accordingly, alternative target bacterial strain carries out domestication and cultivates；
Step 5 particularly as follows:
Step 5.1, prepare following three classes domestication culture medium:
A () denitrogenates domestication culture medium: take potassium nitrate 2g, sodium citrate 7g, ammonium sulfate 0.3g, dipotassium hydrogen phosphate 3g, phosphoric acid
Potassium dihydrogen 2g, Magnesium sulfate heptahydrate 0.2g, stirring mixing, with water constant volume to 1L；
(b) protease production domestication culture medium: configuration quality mark is the skim milk of 10%, adds 10 times of dilution
In the beef extract-peptone Carnis Bovis seu Bubali cream culture medium of volume, milk and Carnis Bovis seu Bubali cream culture medium mass ratio are 1:1, and reconciling pH is 7.4；
C () produces amylase activity domestication culture medium: take soluble starch 2g, peptone 15g, potassium chloride 0.8g and magnesium sulfate
0.5g, with water constant volume to 1L, reconciling pH is 7.4；
Step 5.2, the aimed strain that protease production is positive, use protease production domestication culture medium to mesh
Mark bacterial strain carries out domestication and cultivates；
To producing the aimed strain that amylase activity is positive, use product amylase activity domestication culture medium that aimed strain is entered
Row domestication is cultivated；
To denitrogenating the aimed strain that activity is positive, employing is denitrogenated domestication culture medium and aimed strain is carried out domestication cultivation.
Step 6, the aimed strain after domestication being cultivated carries out liquid fermentation amplification culture；
Step 6 is particularly as follows: the aimed strain after domestication being cultivated activates, it may be assumed that be respectively adopted identical the taming and dociling with step 5
Change culture medium to activate, treat its OD600When reaching 1.5, stop activation, obtain the aimed strain after amplification culture.
Step 7, by the aimed strain after step 6 amplification culture, compounds according to formula proportion, prepares Compound bacterium
Agent.Formula proportion is: actinomycetes, bacillus subtilis, nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa.
The weight proportion of each component is: 2:2:2:1:1:1.
In step 7, also include:
By the aimed strain after step 6 amplification culture, following methods is used to preserve:
For bacillus subtilis, Pseudomonas aeruginosa, nitrifier and lactic acid bacteria, the method for diatomite adsorption is used to protect
Deposit, it may be assumed that first by containing mass fraction 15% Semen Maydis powder and the kieselguhr mix homogeneously of mass fraction 20% glucose, be placed in
121 DEG C of high temperature sterilize 30min, are layered in solid fermentation dish；Solid fermentation dish is placed in air dry oven in 35 DEG C of perseverances
Temperature is dried, and obtains the adsorption substrates being dried；Then, aimed strain is adsorbed onto in described adsorption substrates；
For producing Ruan's candidiasis and actinomycetes, the method using Testa Tritici absorption, it may be assumed that first will be containing mass fraction
15% Semen Maydis powder and the Testa Tritici mix homogeneously of mass fraction 10% glucose, be placed in 121 DEG C of high temperature sterilize 30min, be layered on
In solid fermentation dish；Solid fermentation dish is placed in air dry oven in 30 DEG C of constant temperature dryings, obtains the adsorption substrates being dried；So
After, aimed strain is adsorbed onto in described adsorption substrates.
Inspection example
Compounding microbial inoculum embodiment one prepared, for Dezhou City Shandong Province, long village ditch malodorous black water is administered.This is black
Smelly quality of river water is: COD content is 230mg/L, and ammonia-nitrogen content is 95mg/L, and phosphate content is 6mg/L.In malodorous black water
Add the compounding microbial inoculum that embodiment one prepares, add according to the ratio that adds of 1:200；Carry out without any simultaneously
The control experiment group (CK) of material, after adding 5 days, testing result such as table 1:
Table 1: testing result table
| Component | Add compounding microbial inoculum group | It is not added with microbial inoculum matched group (CK) |
| COD content | 75mg/L | 200mg/L |
| Ammonia-nitrogen content | 25mg/L | 85mg/L |
| Phosphate content | 3mg/L | 6mg/L |
| Water body color | Clarification | Black |
| Water body taste | Tasteless | Stink |
The compounding microbial inoculum preparing embodiment two, has carried out same test.Result of the test is identical.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For Yuan, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improvements and modifications also should
Depending on protection scope of the present invention.
Claims (9)
1. the compounding microbial inoculum purifying black and odorous water, it is characterised in that include following component: actinomycetes, bacillus subtilis,
Nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa.
The compounding microbial inoculum of purification black and odorous water the most according to claim 1, it is characterised in that the weight proportion of each component
For: 2:2:2:1:1:1.
3. the preparation method of the compounding microbial inoculum purifying black and odorous water, it is characterised in that comprise the following steps:
Step 1, according to the characteristic of aimed strain, enrichment medium from the activated sludge of sewage treatment plant, respectively obtain every kind
Aimed strain orienting enriching culture medium corresponding to aimed strain；Wherein, described aimed strain includes actinomycetes, bacillus subtilis
Bacterium, nitrifier, lactic acid bacteria, product Ruan's candidiasis and Pseudomonas aeruginosa；
Aimed strain is inoculated in the aimed strain orienting enriching culture medium of correspondence, is oriented enrichment culture；Treat its OD600
When=2, stop enrichment culture, obtain the aimed strain after enrichment culture；
Step 2, prepares the aimed strain screening culture medium corresponding to every kind of aimed strain；
Aimed strain after step 1 enrichment culture is carried out gradient dilution, is diluted to 10 respectively3Again, 105Again with 107Times；Then,
Use aimed strain screening culture medium that the aimed strain after corresponding dilution is carried out screening and culturing；
Step 3, carries out secondary separation purification by the target list bacterium colony after step 2 screening and culturing, obtains target after purification
Single bacterium colony；
Step 4, carries out qualitative detection experiment to step 3 target list bacterium colony after purification, and the aimed strain that choosing is positive is as standby
Select aimed strain；
Step 5, uses and tames culture medium accordingly, alternative target bacterial strain carries out domestication and cultivates；
Step 6, the aimed strain after domestication being cultivated carries out liquid fermentation amplification culture；
Step 7, by the aimed strain after step 6 amplification culture, compounds according to formula proportion, prepares compounding microbial inoculum.
The preparation method of the compounding microbial inoculum of purification black and odorous water the most according to claim 3, it is characterised in that in step 1,
Orienting enriching Medium Proportion corresponding to each aimed strain is:
Bacillus subtilis, lactic acid bacteria are identical with the orienting enriching culture medium corresponding to Pseudomonas aeruginosa, all by with lower section
Method obtains: river sewage is stood 2～3 days, takes supernatant, add peptone 8～10g, Carnis Bovis seu Bubali cream 1～3g and sodium chloride 20～
30g, with water constant volume to 1L；
Orienting enriching culture medium corresponding to actinomycetes is: add soluble starch 15～20g, dipotassium hydrogen phosphate 0.5～1g, chlorine
Change sodium 0.5～1g, magnesium sulfate 0.2～0.5g, potassium nitrate 1～2g, ferrous sulfate 0.01～0.03g, with water constant volume to 1L, then adjust
Joint pH is 7.2～7.4；
Producing the orienting enriching culture medium corresponding to Ruan's candidiasis is: add glucose 80～100g, peptone 5～10g, ferment
Female cream 10g, potassium dihydrogen phosphate 1～2g, magnesium sulfate 0.2～0.5g, with water constant volume to 1L, then to regulate pH be 4.0～6.5；
Orienting enriching culture medium corresponding to nitrifier is: add peptone 10～15g, sodium chloride 5～7g, Carnis Bovis seu Bubali cream 3～5g,
Potassium nitrate 1～2g, with water constant volume to 1L, then to regulate pH be 7.0～7.5.
The preparation method of the compounding microbial inoculum of purification black and odorous water the most according to claim 3, it is characterised in that in step 2,
Aimed strain screening culture medium proportioning corresponding to various aimed strains is:
The screening culture medium of bacillus subtilis, lactic acid bacteria and Pseudomonas aeruginosa is identical, is: addition glucose 50～60g,
Ammonium chloride 0.5～1g and yeast extract 10g, with water constant volume to 1L, then to regulate pH be 7.0～7.5；
Actinomycetes screening culture medium is: add soluble starch 20～25g, dipotassium hydrogen phosphate 1～1.5g, sodium chloride 0.5～1g,
Magnesium sulfate 0.2～0.3g, potassium nitrate 0.5～1g, ferrous sulfate 0.01～0.03g, agar 15～20g, with water constant volume to 1L, then
Regulation pH is 7.2～7.4；Additionally, every 1 actinomycetes screening culture medium adds the potassium dichromate that mass fraction is 3%
1.5mL, suppression antibacterial and the interference of mycete；
Producing Ruan's candidiasis screening culture medium is: claims Rhizoma Solani tuber osi 180～200g, adds 1L boiling tap water 30min, crosses and filter
After removing residue, constant volume to 1L, add glucose 20～25g, lactic acid 5～6mL, agar 15～20g, regulation pH is 4.0～6.5,
Thus suppress the growth of other miscellaneous bacteria；
Nitrifier screening culture medium: add sodium succinate 8～10g, altheine acid 1～1.5g, sodium nitrate 0.5～1g, phosphorus
Acid dihydride potassium 1～2g, ferrous chloride 0.02～0.05g, calcium chloride 0.2～0.3g, magnesium sulfate 1～2g, BTB solution 1mL (1%
Ethanol) and agar 15～20g, with water constant volume to 1L, then to regulate pH be 7.0～7.3；Wherein, BTB solution refers to: BTB is according to 1%
Volume fraction dissolve in ethanol, take weighing 1ml.
The preparation method of the compounding microbial inoculum of purification black and odorous water the most according to claim 3, it is characterised in that step 4 has
Body is:
Step 4.1, prepares following three class Qualitative Identification culture medium:
A () protease production measures culture medium: take the skim milk that mass fraction is 10～15%, in 110～113 DEG C of sterilizings
30min, with the beef extract-peptone ratio mixing with mass ratio as 1:8, the mass fraction by 15～20g/L adds agar, adjusts
PH is 7.0～7.2, makes protease production and measures culture medium；
B () is produced amylase activity and is measured culture medium: take soluble starch 1～2g, peptone 3～6g, potassium chloride 0.5～1g, seven
Water magnesium sulfate 0.5～1g, agar 15～20g, with water constant volume to 1L, then to regulate pH be 7.0～7.2, makes product amylase activity
Measure culture medium；
C () denitrogenates qualitative determination culture medium: take potassium nitrate 1～2g, sodium citrate 5～7g, ammonium sulfate 0.3～0.5g, phosphoric acid hydrogen
Dipotassium 1～3g, potassium dihydrogen phosphate 2～3g, Magnesium sulfate heptahydrate 0.1～0.2g, agar 15～20g, with water constant volume to 1L, then regulate
PH is 7.0～7.2, makes and denitrogenates qualitative determination culture medium；
Step 4.2, for step 3 every kind of target list bacterium colony after purification, is respectively adopted protease production and measures culture medium, product
Amylase activity measures culture medium and denitrogenates qualitative determination culture medium it is carried out qualitative detection, select protease production and/
Or produce amylase activity and/or denitrogenate the aimed strain alternately aimed strain that activity is positive.
The preparation method of the compounding microbial inoculum of purification black and odorous water the most according to claim 6, it is characterised in that step 5 has
Body is:
Step 5.1, prepare following three classes domestication culture medium:
A () denitrogenates domestication culture medium: take potassium nitrate 1～2g, sodium citrate 5～7g, ammonium sulfate 0.3～0.5g, dipotassium hydrogen phosphate 1
～3g, potassium dihydrogen phosphate 2～3g, Magnesium sulfate heptahydrate 0.1～0.2g, stirring mixing, with water constant volume to 1L；
(b) protease production domestication culture medium: configuration quality mark is the skim milk of 10～15%, adds 10 times of dilution
In the beef extract-peptone Carnis Bovis seu Bubali cream culture medium of volume, milk and Carnis Bovis seu Bubali cream culture medium mass ratio are 1:1, and reconciling pH is 7.2
～7.4；
(c) produce amylase activity domestication culture medium: take soluble starch 1～5g, peptone 10～15g, potassium chloride 0.1～1g and
Magnesium sulfate 0.2～0.5g, with water constant volume to 1L, reconciling pH is 7.2～7.4；
Step 5.2, the aimed strain that protease production is positive, use protease production domestication culture medium to object bacteria
Strain carries out domestication and cultivates；
To producing the aimed strain that amylase activity is positive, use product amylase activity domestication culture medium that aimed strain is tamed and dociled
Change and cultivate；
To denitrogenating the aimed strain that activity is positive, employing is denitrogenated domestication culture medium and aimed strain is carried out domestication cultivation.
The preparation method of the compounding microbial inoculum of purification black and odorous water the most according to claim 7, it is characterised in that step 6 has
Body is:
Aimed strain after domestication being cultivated activates, it may be assumed that is respectively adopted the domestication culture medium identical with step 5 and lives
Change, treat its OD600When reaching 1.5, stop activation, obtain the aimed strain after amplification culture.
The preparation method of the compounding microbial inoculum of purification black and odorous water the most according to claim 3, it is characterised in that in step 7,
Also include:
By the aimed strain after step 6 amplification culture, following methods is used to preserve:
For bacillus subtilis, Pseudomonas aeruginosa, nitrifier and lactic acid bacteria, the method for diatomite adsorption is used to preserve, it may be assumed that
First by containing mass fraction 10～15% Semen Maydis powder and mass fraction 10～the kieselguhr mix homogeneously of 20% glucose, it is placed in
121 DEG C of high temperature sterilize 30min, are layered in solid fermentation dish；Solid fermentation dish is placed in air dry oven in 30～35
DEG C constant temperature drying, obtains the adsorption substrates being dried；Then, aimed strain is adsorbed onto in described adsorption substrates；
For producing Ruan's candidiasis and actinomycetes, the method using Testa Tritici absorption, it may be assumed that first will containing mass fraction 10～
15% Semen Maydis powder and mass fraction 10～the Testa Tritici mix homogeneously of 20% glucose, be placed in 121 DEG C of high temperature sterilize 30min, by it
It is layered in solid fermentation dish；Solid fermentation dish is placed in air dry oven in 30～35 DEG C of constant temperature dryings, obtains the suction being dried
Attached base material；Then, aimed strain is adsorbed onto in described adsorption substrates.
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| CN110104762A (en) * | 2019-05-22 | 2019-08-09 | 江苏中百洲环境科技有限公司 | A kind of floatation type sewage water treatment method |
| CN110117063A (en) * | 2019-05-22 | 2019-08-13 | 江苏中百洲环境科技有限公司 | A kind of floatation type sewage-treatment plant |
| CN116656528A (en) * | 2023-03-15 | 2023-08-29 | 天津科技大学 | Pseudomonas and method for preparing deodorizing nitrogen-fixing bacteria agent by utilizing same |
| CN116656528B (en) * | 2023-03-15 | 2024-06-04 | 天津科技大学 | Pseudomonas and method for preparing deodorizing nitrogen-fixing bacteria agent by utilizing same |
| CN117187088A (en) * | 2023-11-08 | 2023-12-08 | 吉林省中科兆翔生态科技集团有限公司 | Microbial preparation for surface water pollutant treatment, preparation method and application |
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