CN107400648B - Composite microbial inoculum, preparation method and application thereof, and comprehensive treatment method of black and odorous river channel - Google Patents
Composite microbial inoculum, preparation method and application thereof, and comprehensive treatment method of black and odorous river channel Download PDFInfo
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Abstract
The invention discloses a composite microbial inoculum, a preparation method and application thereof, and a comprehensive treatment method of a black and odorous river channel. The composite microbial combined bacterial agent comprises three bacterial agent combinations of a bacterial agent A combination, a bacterial agent C combination and a bacterial agent gamma combination, wherein each strain in the bacterial agent A combination is bacillus coagulans, lactic acid bacteria, saccharomycetes, aspergillus sojae and thiobacillus denitrificans; the strains in the fungicide C combination are respectively bacillus coagulans, rhizopus, saccharomycetes, aspergillus sojae and penicillium chrysogenum; the strains in the microbial inoculum gamma combination are respectively lactobacillus, bacillus subtilis, saccharomycetes, bacillus licheniformis and bacillus coagulans. The composite microbial composite bacterial agent can be used for comprehensive treatment of black and odorous watercourses, has short treatment period, and can effectively solve the problem that nitrogen and phosphorus of black and odorous water bodies exceed standards and eliminate odorous gas hydrogen sulfide in the water bodies.
Description
Technical Field
The invention belongs to the field of sewage treatment, and particularly relates to a composite microbial inoculum, a preparation method and application thereof, and a comprehensive treatment method of a black and odorous river.
Background
Urban water bodies such as rivers and the like are important components of urban ecological systems, have the characteristics of small water environment capacity, poor water body self-purification capacity, easiness in pollution and the like, are often used as important sewage discharge and sewage receiving channels in cities, and are easy to deteriorate water quality. The inflow of a large amount of pollutants causes serious eutrophication of urban water bodies, causes rapid consumption of dissolved oxygen of the water bodies, and causes the water bodies to lack oxygen for a long time to present a black and odorous state. The water bodies in cities such as rivers and the like are black and odorous, so that the water bodies become a common phenomenon in cities in China, and the living and urban image of coastal residents are seriously influenced.
The black and odorous water in rivers and other cities is mainly caused by ecological imbalance of water due to excessive sewage containing, conversion of pollutants under anoxic and even anaerobic conditions of water and generation of odorous substances such as ammonia nitrogen, hydrogen sulfide, volatile organic acid and the like and black substances such as iron and manganese sulfides and the like.
At present, the treatment method of urban black and odorous water bodies comprises a physical method, a chemical method and a biological method. Wherein, the biological method is often applied to the treatment of urban black and odorous water body by using microbial preparations, and a certain amount of microbial preparations are added into the water body, so that the degradation of pollutants in the water body is accelerated, and the self-cleaning function of the water body is enhanced.
The application process of the single microbial preparation in treating the black and odorous water body often has the problems of weak degradation capability on pollutants such as nitrogen, phosphorus, organic matters and the like, low treatment efficiency, unobvious elimination effect on the odorous gas hydrogen sulfide in the black and odorous water body and the like.
Disclosure of Invention
In order to solve the technical problems, the invention provides a composite microbial inoculum, a preparation method and application thereof, and a comprehensive treatment method of a black and odorous river channel. The composite microbial combined bacterial agent comprises three bacterial agent combinations of a bacterial agent A combination, a bacterial agent C combination and a bacterial agent gamma combination, and the bacterial agent combination can be used for treating black and odorous watercourses, can effectively reduce pollutant components such as nitrogen, phosphorus, organic matters and the like in the black and odorous water body, and has an obvious eliminating effect on odorous gas hydrogen sulfide in the black and odorous water body, so that the aims of quickly improving the water quality of the black and odorous watercourses and recovering the ecological balance of the water body are fulfilled.
The technical scheme adopted by the invention is as follows:
a composite microbial combined bacterial agent comprises a bacterial agent A combination, a bacterial agent C combination and a bacterial agent gamma combination;
the microbial inoculum A combination comprises bacillus coagulans, lactic acid bacteria, saccharomycetes, aspergillus sojae and thiobacillus denitrificans;
the microbial inoculum C combination comprises rhizopus, saccharomycetes, aspergillus sojae and penicillium chrysogenum;
the microbial inoculum gamma combination comprises lactobacillus, bacillus subtilis, saccharomycetes, bacillus licheniformis and bacillus coagulans.
The weight ratio of each strain in the microbial inoculum A combination is bacillus coagulans: lactic acid bacteria: yeast: soy sauce aspergillus: thiobacillus denitrificans ═ (2.7-3.3): (1.8-2.2): (0.45-0.55): (1.35-1.65): (2.7-3.3).
The weight ratio of each strain in the microbial inoculum C combination is bacillus coagulans: rhizopus: yeast: soy sauce aspergillus: penicillium chrysogenum (1.8-2.2), 3.6-4.4, (1.35-1.65) and (2.25-2.75).
The weight ratio of each strain in the microbial inoculum gamma combination is lactobacillus: b, bacillus subtilis: yeast: b, bacillus licheniformis: bacillus coagulans ═ (3.15-3.85): (1.8-2.2): (0.45-0.55): (1.35-1.65): (2.25-2.75).
The preparation method of each microbial inoculum combination comprises the following steps:
(1) respectively inoculating each strain to a liquid culture medium for culturing for 36-48 h;
(2) respectively placing the strains cultured in the step (1) into a fermentation tank, and carrying out mass production culture for 36-48 h, wherein the weight ratio of the strains to the fermentation medium is 1: 9-15;
(3) mixing the strains cultured in the step (2) according to the weight ratio of the strains in each microbial inoculum combination, and performing cross antagonistic culture for 36-48 h until the viable count is 4-9 × 1010cfu/mL, respectively obtaining a microbial inoculum A combination, a microbial inoculum C combination and a microbial inoculum gamma combination. The weight ratio of the mixed strain to the cross antagonistic culture medium is 1: 15 to 20.
In the step (2), sterile air is introduced for 2 minutes every 8 hours during the fermentation process, and the stirring speed is 70 rpm/min. The flow rate of the sterile air is 90L/min.
The preparation method of the microbial inoculum A combination comprises the following steps:
(1) respectively inoculating bacillus coagulans, lactobacillus, saccharomycetes, aspergillus sojae and thiobacillus denitrificans in a liquid culture medium, and culturing for 48h at 28-32 ℃; the liquid culture medium comprises the following components in each liter of culture medium: 7.2-8.8 g of beef extract, 9-11 g of peptone, 2.7-3.3 g of yeast extract, 4.5-5.5 g of sodium chloride, 0.9-1.1 g of monopotassium phosphate, 0.09-0.11 g of trace elements, 27-33 g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2; the trace element is Mg2+Or Fe2+The corresponding sulfates are used;
(2) respectively placing the strains cultured in the step (1) in a fermentation tank, carrying out mass production culture at 28-32 ℃ for 48h, and introducing sterile air for 2 minutes every 8 hours in the fermentation process, wherein the flow rate of the sterile air is 90L/min, the rotating speed is 70rpm/min, and the weight ratio of the strains to the fermentation medium is 1: 9-15; the fermentation medium comprises the following components in each liter of medium: 7.2-8.8 g of beef extract, 9-11 g of peptone, 2.7-3.3 g of yeast extract, 4.5-5.5 g of sodium chloride, 0.9-1.1 g of monopotassium phosphate, 0.09-0.11 g of trace elements, 27-33 g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2; the trace element is Mg2+Or Fe2+The corresponding sulfates are used;
(3) and (3) performing mass production culture on the strains obtained in the step (2) according to the weight ratio of bacillus coagulans: lactic acid bacteria: yeast: soy sauce aspergillus: thiobacillus denitrificans ═ (2.7-3.3): (1.8-2.2): (0.45-0.55): (1.35-1.65): (2.7E &3.3) mixing, and carrying out cross antagonistic culture for 48h, wherein sterile air is introduced for 2 minutes every 8 hours in the culture process, the stirring speed is 70rpm/min, the flow rate of the sterile air is 90L/min, the weight ratio of the mixed strain to the cross antagonistic culture medium is 1: 15-20, and a bacterium agent A combination with the viable count of 4-6 × 10 is obtained10cfu/mL. The cross-antagonistic culture medium comprises the following components in each liter of culture medium: 7.2-8.8 g of beef extract, 9-11 g of peptone, 2.7-3.3 g of yeast extract, 4.5-5.5 g of sodium chloride, 0.9-1.1 g of monopotassium phosphate, 0.09-0.11 g of trace elements, 27-33 g of sucrose, and the balance of distilled water, wherein the pH value is 6.8-7.2. The trace element is Mg2+Or Fe2+The corresponding sulfates were used.
The preparation method of the microbial inoculum C combination comprises the following steps:
(1) inoculating rhizopus, saccharomycetes, aspergillus sojae and penicillium chrysogenum into a liquid culture medium, and culturing for 48h at 28-32 ℃; the liquid culture medium comprises the following components in each liter of culture medium: 4.5-5.5 g of beef extract, 9-11 g/L of peptone, 180-220 g of potato extract, 4.5-5.5 g of sodium chloride, 18-22 g of glucose, 0.45-0.55 g of monopotassium phosphate, 45-55 g of cane sugar and the balance of distilled water, wherein the pH value is 6.8-7.2;
(2) respectively placing the strains cultured in the step (1) in a fermentation tank, carrying out mass production culture at 28-32 ℃ for 48h, and introducing sterile air for 2 minutes every 8 hours in the fermentation process, wherein the flow rate of the sterile air is 90L/min, the rotating speed is 70rpm/min, and the weight ratio of the strains to the fermentation medium is 1: 9-15; the fermentation medium comprises the following components in each liter of medium: 4.5-5.5 g of beef extract, 9-11 g of peptone, 180-220 g of potato extract, 4.5-5.5 g of sodium chloride, 18-22 g of glucose, 0.45-0.55 g of monopotassium phosphate, 45-55 g of cane sugar and the balance of distilled water, wherein the pH value is 6.8-7.2;
(3) and (3) performing mass production culture on the strains obtained in the step (2) according to the weight ratio of rhizopus: yeast: soy sauce aspergillus: mixing (1.8-2.2) Penicillium chrysogenum, (3.6-4.4) and (1.35-1.65) and (2.25-2.75), performing cross antagonistic culture at 28-32 ℃ for 48h, wherein the weight ratio of the mixed strain to the cross antagonistic culture medium is 1: 15-20 to obtain a microbial inoculum CThe combination has a viable count of 1-3 × 1010cfu/mL. The cross-antagonistic culture medium comprises the following components in each liter of culture medium: 4.5-5.5 g of beef extract, 9-11 g of peptone, 180-220 g of potato extract, 4.5-5.5 g of sodium chloride, 18-22 g of glucose, 0.45-0.55 g of monopotassium phosphate, 45-55 g of cane sugar and the balance of distilled water, wherein the pH value is 6.8-7.2.
The preparation method of the potato extracting solution comprises the following steps: peeling and cutting rhizoma Solani Tuber osi, squeezing to obtain rhizoma Solani Tuber osi juice, filtering the lower layer precipitate to obtain supernatant, and sterilizing at 121 deg.C.
The preparation method of the microbial inoculum gamma combination comprises the following steps:
(1) respectively inoculating lactobacillus, bacillus subtilis, saccharomycetes, bacillus licheniformis and bacillus coagulans in a liquid culture medium, and culturing at 28-32 ℃ for 36 h; the liquid culture medium comprises the following components in each liter of culture medium: 7.2-8.8 g of beef extract, 2.7-3.3 g of peptone, 3.15-3.85 g of yeast extract, 0.9-1.1 mL of Tween, 1.35-1.65 g of magnesium sulfate heptahydrate, 18-22 g of glucose, 45-55 g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2;
(2) respectively placing the strains cultured in the step (1) in a fermentation tank, carrying out mass production culture at 28-32 ℃ for 36h, and introducing sterile air for 2 minutes every 8 hours in the fermentation process, wherein the flow rate of the sterile air is 90L/min, the rotating speed is 70rpm/min, and the weight ratio of the strains to the fermentation medium is 1: 9-15; the fermentation medium comprises the following components in each liter of medium: 7.2-8.8 g of beef extract, 2.7-3.3 g of peptone, 3.15-3.85 g of yeast extract, 800.9-1.1 mL of Tween, 1.35-1.65 g of magnesium sulfate heptahydrate, 18-22 g of glucose, 45-55 g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2;
(3) mixing the strains cultured in the step (2) according to the weight ratio of lactic acid bacteria to bacillus subtilis to saccharomycetes to bacillus licheniformis to bacillus coagulans (3.15-3.85) to (1.8-2.2) to (0.45-0.55) to (1.35-1.65) to (2.25-2.75), carrying out cross antagonistic culture for 36h, wherein the weight ratio of the mixed strains to the cross antagonistic culture medium is 1: 15-20, and obtaining a microbial inoculum gamma combination with the viable count of 4-7 × 1010cfu/mL. The cross armThe anti-culture medium comprises the following components in each liter of culture medium: 7.2-8.8 g of beef extract, 2.7-3.3 g of peptone, 3.15-3.85 g of yeast extract, 800.9-1.1 mL of Tween, 1.35-1.65 g of magnesium sulfate heptahydrate, 18-22 g of glucose, 45-55 g of sucrose, and the balance of distilled water, wherein the pH value is 6.8-7.2.
The invention also provides application of the composite microbial inoculum in treatment of black and odorous watercourses. The combination of the microbial inoculum A and the combination of the microbial inoculum C are mixed and then put into a black and odorous river channel, and the combination of the microbial inoculum gamma is put into the river channel after the water quality reaches the standard.
A comprehensive treatment method for a black and odorous river channel comprises the following steps:
A. controlling the exogenous pollution of the black and odorous river channel, and well performing the work of a sewage interception and storage pipe;
B. mixing the microbial inoculum A combination and the microbial inoculum C combination, and putting the mixture into a black and odorous river;
C. after the water quality reaches the standard, putting the microbial inoculum gamma combination into a river channel, breeding aquatic animals and plants, and recovering the self-purification capacity of the water ecological system;
D. and then the riverway is maintained for a long time.
In the treatment process, the water quality needs to reach the national IV-class water quality standard of surface water.
The adding amount of the microbial inoculum A combination, the microbial inoculum C combination and the microbial inoculum gamma combination is 20-50 ppm. Within the range of the dosage, the strain can grow rapidly, the growth of harmful bacteria is inhibited by the advantages of flora, pollutants can be rapidly decomposed, and the effect is rapid and obvious. The strains with too little dosage are fed, the strains grow and reproduce slowly without the advantages of flora, the treatment effect is not ideal, and the effect of too much feeding is the same as that of the dosage, so that the waste is caused by too much feeding.
The functions of each strain in the composite microbial combined bacterial agent disclosed by the invention are as follows:
bacillus coagulans: amylase and protease are secreted, and the growth of harmful bacteria is effectively inhibited;
lactic acid bacteria: decomposing protein in the water body and providing nutrition for the aquatic east plants;
yeast: decomposing polysaccharides such as starch in water;
soy sauce aspergillus: absorbing heavy metals, dissolving phosphorus and providing nutrition for aquatic plants;
thiobacillus denitrificans: converting sulfide into sulfate or elemental sulfur, and converting nitrate and nitrite into nitrogen;
rhizopus: absorbing heavy metals, dissolving phosphorus and providing nutrition for aquatic plants;
producing penicillium chrysogenum: absorbing heavy metals, dissolving phosphorus and providing nutrition for aquatic plants;
b, bacillus subtilis: secreting amylase, lipase, protease, cellulase;
b, bacillus licheniformis: decomposing organic matters and secreting amylase.
The microbial inoculum A combination is obtained by cross antagonistic culture of bacillus coagulans, lactic acid bacteria, saccharomycetes, aspergillus sojae and thiobacillus denitrificans according to the weight ratio of (2.7-3.3), (1.8-2.2), (0.45-0.55), (1.35-1.65) and (2.7-3.3), and the strains act together to enable the microbial inoculum A combination to rapidly decompose black and odorous substances such as ammonia gas, methyl mercaptan, hydrogen sulfide and the like in water.
The microbial inoculum C combination is obtained by cross antagonistic culture of rhizopus, saccharomycetes, aspergillus sojae and penicillium chrysogenum according to the weight ratio of (1.8-2.2) to (3.6-4.4) to (1.35-1.65) to (2.25-2.75), and the strains act together, so that the microbial inoculum C combination can rapidly remove water eutrophication substances and convert and degrade phosphorus compounds.
The microbial agent gamma combination is obtained by cross antagonistic culture of lactic acid bacteria, bacillus subtilis, saccharomycetes, bacillus licheniformis and bacillus coagulans according to the weight ratio of (3.15-3.85), (1.8-2.2), (0.45-0.55) and (1.35-1.65) and (2.25-2.75), the strains play a role together, so that the microbial agent gamma combination has the effects of adjusting the pH value of a water body, adjusting the microbial environment of the water body and improving the transparency of the water body, the ecology of the water body can be recovered, and when the microbial agent gamma combination is put into a black and odorous water body repaired by the microbial agent A combination and the microbial agent C combination, aquatic animals are released to regulate, monitor and repair the components of the system, so that river channel indexes can be optimized, the water quality is balanced, the harmony and stability of various biological floras are maintained, the biological diversity of the system is increased, the aquatic ecosystem is recovered, and the self-cleaning capability of the water body is recovered.
The dosage of each strain in the microbial inoculum combination needs to be strictly controlled within the range disclosed by the invention, and the strains can be mutually antagonistic and can grow together within the range, so that the death of a single strain can not occur, and the use effect is influenced. During the fixed culture process, the bacteria are promoted to secrete more and more effective enzymes more quickly due to the antagonism, so that the effect of the enzyme is far greater than that of a single strain.
In the treatment process of the black and odorous river channel, the microbial inoculum A combination and the microbial inoculum C combination are firstly used for restoring the water body environment and reducing the sludge thickness of the river bottom, and after all indexes of the water body meet the requirements, the microbial inoculum gamma combination is added to restore the ecological balance in the water body.
Compared with the prior art, the invention has the following advantages:
1. the strains in each microbial inoculum combination are cultured by adopting a uniform culture medium, the preparation steps of the microbial inoculum combination are simplified on the premise of ensuring the optimal propagation conditions of the strains, and the cost is saved;
2. each bactericide composition consists of a plurality of strains, and each strain plays a synergistic effect in sewage treatment to quickly form dominant flora, quickly decompose various rich nutrients in black and odorous water, eliminate peculiar smell and degrade harmful substances;
3. the pollution problem of the black and odorous river channel is treated by combining three bactericides, namely a bactericide A combination, a bactericide C combination and a bactericide gamma combination, the treatment period is short, the problems of excessive nitrogen and phosphorus and over-high COD value of the black and odorous water body can be effectively solved, odorous gas hydrogen sulfide in the water body can be effectively eliminated, and the thickness of river bottom sludge is reduced;
4. after the water quality of the black and odorous riverway is restored by mixing the microbial inoculum A combination and the microbial inoculum C combination, a microbial inoculum gamma combination is put into the mixture to balance the flora in the water, improve the water quality environment and restore the ecological balance in the water body;
5. according to the invention, the water body is treated by microorganisms, and then the water body is treated by the self-cleaning capacity of the water body, so that the purpose of treating both symptoms and root causes is achieved, and secondary pollution is avoided;
6. has the characteristics of safety, high efficiency, low cost, investment saving, simple operation and the like.
Detailed Description
All strains disclosed in the invention are purchased.
Example 1
A composite microbial combined bacterial agent comprises a bacterial agent A combination, a bacterial agent C combination and a bacterial agent gamma combination;
the preparation method of the microbial inoculum A combination comprises the following steps:
(1) respectively inoculating bacillus coagulans, lactobacillus, saccharomycetes, aspergillus sojae and thiobacillus denitrificans in a liquid culture medium, and culturing for 48h at 28-32 ℃; the liquid culture medium comprises the following components in each liter of culture medium: 8g of beef extract, 10g of peptone, 3g of yeast extract, 5g of sodium chloride, 1g of monopotassium phosphate, 0.1g of trace elements, 30g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2; the trace element is Mg2+Or Fe2+The corresponding sulfates are used;
(2) respectively placing the strains cultured in the step (1) in a fermentation tank, carrying out mass production culture at 28-32 ℃ for 48h, and introducing sterile air for 2 minutes every 8 hours in the fermentation process, wherein the flow rate of the sterile air is 90L/min, the rotating speed is 70rpm/min, and the weight ratio of the strains to the fermentation medium is 1: 10; the fermentation medium comprises the following components in each liter of medium: 8g of beef extract, 10g of protein, 3g of yeast extract, 5g of sodium chloride, 1g of monopotassium phosphate, 0.1g of trace elements, 30g of cane sugar and the balance of distilled water, wherein the pH value is 6.8-7.2; the trace element is Mg2+Or Fe2+The corresponding sulfates were used.
(3) Mixing the strains cultured in the step (2) in a mass production manner according to the weight ratio of bacillus coagulans, lactobacillus, yeast, aspergillus sojae, thiobacillus denitrificans, 3: 2: 0.5: 1.5: 3, and carrying out cross antagonistic culture for 48h, wherein sterile air is introduced for 2 minutes every 8 hours in the culture process, the stirring rotating speed is 70rpm/min, the flow rate of the sterile air is 90L/min, the weight ratio of the mixed strains to the cross antagonistic culture medium is 1: 15, so as to obtain a microbial inoculum A combination, and the viable count of the microbial inoculum A is 5 × 1010cfu/mL. Said crossingThe antagonistic culture medium comprises the following components in each liter of culture medium: 8g of beef extract, 10g of peptone, 3g of yeast extract, 5g of sodium chloride, 1g of monopotassium phosphate, 0.1g of trace elements, 30g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2; the trace element is Mg2+Or Fe2+The corresponding sulfates were used.
The preparation method of the microbial inoculum C combination comprises the following steps:
(1) inoculating rhizopus, saccharomycetes, aspergillus sojae and penicillium chrysogenum into a liquid culture medium, and culturing for 48h at 28-32 ℃; the liquid culture medium comprises the following components in each liter of culture medium: 5g of beef extract, 10g/L of peptone, 200g of potato extract, 5g of sodium chloride, 20g of glucose, 0.5g of monopotassium phosphate, 50g of sucrose and the balance of distilled water;
(2) respectively placing the cultured strains in a fermentation tank, carrying out mass production culture at 28-32 ℃ for 48h, stirring and introducing sterile air for 2 minutes every 8 hours in the fermentation process, wherein the flow of the sterile air is 90L/min, the rotating speed is 70rpm/min, and the weight ratio of the strains to the fermentation medium is 1: 10; the fermentation medium comprises the following components in each liter of medium: 5g of beef extract, 10g of peptone, 200g of potato extract, 5g of sodium chloride, 20g of glucose, 0.5g of monopotassium phosphate, 50g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2;
(3) mixing the strains cultured in the step (2) in a mass production way according to the weight ratio of rhizopus, saccharomycete, aspergillus sojae and penicillium chrysogenum of 2: 4: 1.5: 2.5, carrying out cross antagonistic culture at 28-32 ℃ for 48h, wherein the weight ratio of the mixed strains to the cross antagonistic culture medium is 1: 15, and obtaining a microbial inoculum C combination with the viable count of 2 × 1010cfu/mL. The cross-antagonistic culture medium comprises the following components in each liter of culture medium: 5g of beef extract, 10g of peptone, 200g of potato extract, 5g of sodium chloride, 20g of glucose, 0.5g of monopotassium phosphate, 50g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2.
The preparation method of the potato extracting solution comprises the following steps: peeling and cutting rhizoma Solani Tuber osi, squeezing to obtain rhizoma Solani Tuber osi juice, filtering the lower layer precipitate to obtain supernatant, and sterilizing at 121 deg.C.
The preparation method of the microbial inoculum gamma combination comprises the following steps:
(1) respectively inoculating lactobacillus, bacillus subtilis, saccharomycetes, bacillus licheniformis and bacillus coagulans in a liquid culture medium, and culturing at 28-32 ℃ for 36 h; the liquid culture medium comprises the following components in each liter of culture medium: 8g of beef extract, 3g of peptone, 3.5g of yeast extract, 801.0 mL of tween, 1.5g of magnesium sulfate heptahydrate, 20g of glucose, 50g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2;
(2) respectively placing the strains cultured in the step (1) in a fermentation tank, carrying out mass production culture at 28-32 ℃ for 36h, and introducing sterile air for 2 minutes every 8 hours in the fermentation process, wherein the flow rate of the sterile air is 90L/min, the rotating speed is 70rpm/min, and the weight ratio of the strains to the fermentation medium is 1: 10; the fermentation medium comprises the following components in each liter of medium: 8g of beef extract, 3g of peptone, 3.5g of yeast extract, 801.0 mL of tween, 1.5g of magnesium sulfate heptahydrate, 20g of glucose, 50g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2;
(3) mixing the strains cultured in the step (2) in a mass production way according to the weight ratio of lactobacillus to bacillus subtilis to saccharomycetes to bacillus licheniformis to bacillus coagulans of 3.5: 2: 0.5: 1.5: 2.5, carrying out cross antagonistic culture for 36h, wherein the weight ratio of the mixed strains to the cross antagonistic culture medium is 1: 15, and obtaining a microbial inoculum gamma combination with the viable count of 6 × 1010cfu/mL. The cross-antagonistic culture medium comprises the following components in each liter of culture medium: 8g of beef extract, 3g of peptone, 3.5g of yeast extract, 801.0 mL of tween, 1.5g of magnesium sulfate heptahydrate, 20g of glucose, 50g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2.
The composite microbial composite bacterial agent can be used for treating black and odorous watercourses, reducing the content of ammonia nitrogen in water bodies, decomposing various eutrophic substances in the black and odorous water bodies, eliminating peculiar smell, degrading harmful substances and specifically solving different pollution problems of the water bodies.
Treating the black and odorous river channel by adopting the composite microbial combined bacterial agent, wherein the treatment range of the river channel in a certain development area is 1740 m; the water surface width is about 12-15 m, and the average width is about 14 m; the water depth is 2-3.7 m, and the average water depth is about 3.0 m; the thickness of the bottom mud is 0.7-1.0 m. For a long time, because a large amount of domestic sewage and industrial sewage are directly discharged, the pollution load is high, and river water is seriously polluted. The black and odorous phenomenon of the water body is obvious, the aquatic animal and plant ecosystem is seriously damaged, and more white garbage and oil stains are arranged on the river surface, so that the surrounding living and production environment is seriously influenced.
The treatment method comprises the following steps:
A. performing touch investigation on exogenous pollution points of the black and odorous riverway, and performing work of a sewage interception receiving pipe;
C. mixing the microbial inoculum A combination and the microbial inoculum C combination, and putting the mixture into a black and odorous river channel, wherein the putting dose of the two is 20-50 ppm;
C. after 6 months, the microbial inoculum gamma combination is put into a river channel according to the dosage of 20-50 ppm, aquatic animals and plants are simultaneously bred, and all the components of the system are regulated, monitored and maintained, so that the river channel index is optimized, the water quality is balanced, the harmony and stability of all biological flora are kept, the biological diversity of the system is increased, the water ecological system is recovered, and the self-cleaning capacity of the water body is recovered.
D. And then the riverway is maintained for a long time.
After the bactericide A combination and the bactericide C combination are mixed and put into the river channel, the monitoring process in the treatment process is as follows:
the first stage is as follows: after 48 hours of putting, the odor on the water surface and the environment is reduced by about 50 percent.
And a second stage: after 72 hours of putting, the river bottom sludge floats after being fermented under the mixing action of the combination of the microbial inoculum A and the combination of the microbial inoculum C, and the river sludge layer floats up to the water surface and enters a water body and a bottom sludge odor elimination and sludge removal period. The stage has different lengths according to the thickness of the river silt and the water flow state, and the period is generally 14 to 20 days.
And a third stage: and (3) putting for 21-62 days, so that the effect is achieved at first, the black and odorous substances are gradually eliminated, the water level of the riverbed is deepened, the transparency of the water body is improved, more microorganisms appear, and the survival conditions of the aquatic animals are improved.
A fourth stage: and (3) putting the mixture for 63-90 days, so that the water quality is obviously improved, various aquatic plants start to breed, various water quality indexes change by about 60%, and the sediment is reduced by about 10 cm.
The fifth stage: and putting for 6 months, reducing 15-30 cm of sludge and greatly improving water quality.
In different stages of river regulation, the results of each index in the water body are shown in table 1:
TABLE 1 results of various indexes in water body at different stages of river regulation
As can be seen from Table 1, the COD value in the water body is reduced by 94.5 percent and NH is reduced by the treatment of the composite microbial composite inoculant for 6 months3the-N content is reduced by 99.1 percent, the total phosphorus content is reduced by 94.7 percent, and the water quality of the riverway reaches the national IV-class water quality standard of surface water.
Example 2
A composite microbial combined bacterial agent comprises a bacterial agent A combination, a bacterial agent C combination and a bacterial agent gamma combination;
the preparation method of the microbial inoculum A combination comprises the following steps:
(1) respectively inoculating bacillus coagulans, lactobacillus, saccharomycetes, aspergillus sojae and thiobacillus denitrificans in a liquid culture medium, and culturing for 48h at 28-32 ℃; the liquid culture medium comprises the following components in each liter of culture medium: 8.8g of beef extract, 11g of peptone, 3.3g of yeast extract, 5.5g of sodium chloride, 1.1g of monopotassium phosphate, 0.11g of trace elements, 33g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2; the trace element is Mg2+Or Fe2+The corresponding sulfates are used;
(2) respectively placing the strains cultured in the step (1) in a fermentation tank, carrying out mass production culture at 28-32 ℃ for 48h, and introducing sterile air for 2 minutes every 8 hours in the fermentation process, wherein the flow rate of the sterile air is 90L/min, the rotating speed is 70rpm/min, and the weight ratio of the strains to the fermentation medium is 1: 15; the fermentation medium comprises the following components in each liter of medium: beef extract 8.8g, peptone 11g, 3.3g of yeast extract, 5.5g of sodium chloride, 1.1g of monopotassium phosphate, 0.11g of trace elements, 33g of cane sugar and the balance of distilled water, wherein the pH value is 6.8-7.2; the trace element is Mg2+Or Fe2+The corresponding sulfates are used;
(3) mixing the strains cultured in the step (2) in a mass production manner according to the weight ratio of bacillus coagulans, lactobacillus, yeast, aspergillus sojae, thiobacillus denitrificans, 2.7: 1.8: 0.55: 1.65: 3.3, carrying out cross antagonistic culture for 48h, stirring and introducing sterile air for 2 minutes every 8 hours in the culture process, wherein the stirring rotating speed is 70rpm/min, the flow rate of the sterile air is 90L/min, and the weight ratio of the mixed strains to the cross antagonistic culture medium is 1: 20 to obtain a microbial inoculum A combination, wherein the viable count of the microbial inoculum A combination is 6 × 1010cfu/mL. The cross-antagonistic culture medium comprises the following components in each liter of culture medium: 8.8g of beef extract, 11g of peptone, 3.3g of yeast extract, 5.5g of sodium chloride, 1.1g of monopotassium phosphate, 0.11g of trace elements, 33g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2; the trace element is Mg2+Or Fe2+The corresponding sulfates were used.
The preparation method of the microbial inoculum C combination comprises the following steps:
(1) inoculating rhizopus, saccharomycetes, aspergillus sojae and penicillium chrysogenum into a liquid culture medium, and culturing for 48h at 28-32 ℃; the liquid culture medium comprises the following components in each liter of culture medium: 5.5g of beef extract, 11g/L of peptone, 220g of potato extract, 5.5g of sodium chloride, 22g of glucose, 0.55g of monopotassium phosphate, 55g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2;
(2) respectively placing the strains cultured in the step (1) in a fermentation tank, carrying out mass production culture at 28-32 ℃ for 48h, and introducing sterile air for 2 minutes every 8 hours in the fermentation process, wherein the flow rate of the sterile air is 90L/min, the rotating speed is 70rpm/min, and the weight ratio of the strains to the fermentation medium is 1: 15; the fermentation medium comprises the following components in each liter of medium: 5.5g of beef extract, 11g of peptone, 220g of potato extract, 5.5g of sodium chloride, 22g of glucose, 0.55g of monopotassium phosphate, 55g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2;
(3) mixing the strains cultured in the step (2) according to the weight ratio of rhizopus to saccharomycete to aspergillus sojae to penicillium chrysogenum to 2.2: 3.6: 1.65: 2.75, performing cross antagonistic culture at 28-32 ℃ for 48h, wherein the weight ratio of the mixed strains to the cross antagonistic culture medium is 1: 20, and obtaining a microbial inoculum C combination with the viable count of 3 × 1010cfu/mL. The cross-antagonistic culture medium comprises the following components in each liter of culture medium: 5.5g of beef extract, 11g of peptone, 220g of potato extract, 5.5g of sodium chloride, 22g of glucose, 0.55g of monopotassium phosphate, 55g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2.
The preparation method of the potato extracting solution comprises the following steps: peeling and cutting rhizoma Solani Tuber osi, squeezing to obtain rhizoma Solani Tuber osi juice, filtering the lower layer precipitate to obtain supernatant, and sterilizing at 121 deg.C.
The preparation method of the microbial inoculum gamma combination comprises the following steps:
(1) respectively inoculating lactobacillus, bacillus subtilis, saccharomycetes, bacillus licheniformis and bacillus coagulans in a liquid culture medium, and culturing at 28-32 ℃ for 36 h; the liquid culture medium comprises the following components in each liter of culture medium: 8.8g of beef extract, 3.3g of peptone, 3.85g of yeast extract, 801.1mL of tween, 1.65g of magnesium sulfate heptahydrate, 22g of glucose, 55g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2;
(2) respectively placing the strains cultured in the step (1) in a fermentation tank, carrying out mass production culture at 28-32 ℃ for 36h, and introducing sterile air for 2 minutes every 8 hours in the fermentation process, wherein the flow rate of the sterile air is 90L/min, the rotating speed is 70rpm/min, and the weight ratio of the strains to the fermentation medium is 1: 15; the fermentation medium comprises the following components in each liter of medium: 8.8g of beef extract, 3.3g of peptone, 3.85g of yeast extract, 801.1mL of tween, 1.65g of magnesium sulfate heptahydrate, 22g of glucose, 55g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2;
(3) and (3) performing mass production culture on the strains obtained in the step (2) according to the weight ratio of lactic acid bacteria: b, bacillus subtilis: yeast: b, bacillus licheniformis: bacillus coagulans 3.85: 2.2: 0.55: 1.65: 2.75 mixing, cross-antagonistic culturing for 36h, wherein the weight ratio of the mixed strain to the cross-antagonistic culture medium is 1: 20,the obtained microbial inoculum gamma combination has the viable count of 7 × 1010cfu/mL. The cross-antagonistic culture medium comprises the following components in each liter of culture medium: 8.8g of beef extract, 3.3g of peptone, 3.85g of yeast extract, 1.1mL of tween, 1.65g of magnesium sulfate heptahydrate, 22g of glucose, 55g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2.
The composite microbial composite bacterial agent can be used for treating black and odorous watercourses, reducing the content of ammonia nitrogen in water bodies, decomposing various eutrophic substances in the black and odorous water bodies, eliminating peculiar smell, degrading harmful substances and specifically solving different pollution problems of the water bodies.
The composite microbial inoculum is adopted to treat the black and odorous river channel, the side ditch of a certain village is about 1000 meters long and 4-6 meters wide, the water depth is 1-2 meters, and the thickness of bottom mud is 0.5-1.0 meter. The river mainly takes domestic sewage as main material, a few industrial sewage is converged, sludge in the river is seriously accumulated, malignant water and grass in partial ditch sections grow excessively, biological phases of periodic excessive reproduction, putrefaction and excessive reproduction are presented, mosquitoes are bred, and odor is emitted.
The treatment method comprises the following steps:
A. controlling the exogenous pollution of the black and odorous river channel, and well performing the work of a sewage interception and storage pipe;
B. mixing the microbial inoculum A combination and the microbial inoculum C combination, and putting the mixture into a black and odorous river channel, wherein the putting dose of the two is 15-40 ppm;
C. after 6 months, the microbial inoculum gamma combination is put into a river channel according to the dosage of 15-40 ppm, aquatic animals and plants are simultaneously bred, and all the components of the system are regulated, monitored and maintained, so that the river channel index is optimized, the water quality is balanced, the harmony and stability of all biological flora are kept, the biological diversity of the system is increased, the water ecological system is recovered, and the self-cleaning capacity of the water body is recovered.
D. And then the riverway is maintained for a long time.
After the bactericide A combination and the bactericide C combination are mixed and put into the river channel, the monitoring process in the treatment process is as follows:
the first stage is as follows: after 48 hours of putting, the odor on the water surface and the environment is reduced by about 50 percent.
And a second stage: after 72 hours of putting, the river bottom sludge floats after being fermented under the mixing action of the combination of the microbial inoculum A and the combination of the microbial inoculum C, and the river sludge layer floats up to the water surface and enters a water body and a bottom sludge odor elimination and sludge removal period. The stage has different lengths according to the thickness of the river silt and the water flow state, and the period is generally 14 to 20 days.
And a third stage: and (3) putting for 21-62 days, so that the effect is achieved at first, the black and odorous substances are gradually eliminated, the water level of the riverbed is deepened, the transparency of the water body is improved, more microorganisms appear, and the survival conditions of the aquatic animals are improved.
A fourth stage: and (3) putting the mixture for 63-90 days, so that the water quality is obviously improved, various aquatic plants start to breed, various water quality indexes change by about 60%, and the sediment is reduced by about 20 cm.
The fifth stage: and putting for 6 months, reducing 30-40 cm of sludge and greatly improving water quality.
In different stages of river regulation, the results of each index in the water body are shown in table 2:
table 2 results of each index in water body at different stages of river regulation
As can be seen from Table 2, the COD value in the water body is reduced by 70.4 percent and NH is reduced by the treatment of the composite microbial composite inoculant for 6 months3The N content is reduced by 77.6 percent, the total phosphorus content is reduced by 93.5 percent, and the water quality of the riverway reaches the national IV-class water quality standard of surface water.
Comparative example 1
In other same examples 1, the mixed input of the combination of the microbial inoculum A and the combination of the microbial inoculum C is replaced by the input of a single combination of the microbial inoculum C in the previous stage of river channel treatment, the input dosage is 40-100 ppm, other conditions are kept unchanged, and in different stages of river channel treatment, the results of all indexes in the water body are shown in table 3:
TABLE 3 results of various indexes in water body at different stages of river regulation
As can be seen from Table 3, the COD value in the water body is reduced by 56.3 percent and NH is reduced by combining the microbial inoculum C for 6 months3The N content is reduced by 78.9 percent, the total phosphorus content is reduced by 86.9 percent, the water quality of the riverway is close to the V-type water quality standard of national surface water, but the treatment effect is far lower than that of the mixture of the microbial inoculum A combination and the microbial inoculum C combination.
Comparative example 2
The other steps are the same as example 2, except that the mixed input of the combination of the microbial inoculum A and the combination of the microbial inoculum C is replaced by the input of a single combination of the microbial inoculum A in the previous stage of river channel treatment, the input dosage is 30-80 ppm, other conditions are kept unchanged, and in different stages of river channel treatment, the results of all indexes in the water body are shown in table 4:
table 4 results of each index in water body at different stages of river regulation
As can be seen from Table 4, the COD value in the water body is reduced by 63.5 percent and NH is reduced by the treatment of 6 months by combining the microbial inoculum A3The N content is reduced by 50.1 percent, the total phosphorus content is reduced by 16.9 percent, the water quality of a riverway is close to the V-type water quality standard of national surface water, but the treatment effect is far lower than that of a mixture of the microbial inoculum A combination and the microbial inoculum C combination.
The above detailed description of a composite microbial composite inoculum, its preparation method and application, and a comprehensive treatment method for black and odorous river channels, which are described with reference to the embodiments, is illustrative and not restrictive, and several embodiments can be cited according to the limited scope, so that changes and modifications without departing from the general concept of the present invention shall fall within the protection scope of the present invention.
Claims (6)
1. The compound microorganism combined microbial inoculum is characterized by comprising a microbial inoculum A combination, a microbial inoculum C combination and a microbial inoculum gamma combination;
the microbial inoculum A combination comprises bacillus coagulans, lactic acid bacteria, saccharomycetes, aspergillus sojae and thiobacillus denitrificans;
the microbial inoculum C combination comprises rhizopus, saccharomycetes, aspergillus sojae and penicillium chrysogenum;
the microbial inoculum gamma combination comprises lactobacillus, bacillus subtilis, saccharomycetes, bacillus licheniformis and bacillus coagulans;
when the black and odorous river channel is treated, the microbial inoculum A combination and the microbial inoculum C combination are mixed and then put into the black and odorous river channel, and the microbial inoculum gamma combination is put into the river channel after the water quality reaches the standard;
the weight ratio of each strain in the microbial inoculum A combination is bacillus coagulans: lactic acid bacteria: yeast: soy sauce aspergillus: thiobacillus denitrificans (2.7-3.3) = (1.8-2.2): 0.45-0.55): 1.35-1.65): 2.7-3.3;
the weight ratio of each strain in the fungicide C combination is rhizopus: yeast: soy sauce aspergillus: penicillium chrysogenum = (1.8-2.2): (3.6-4.4): 1.35-1.65): 2.25-2.75);
the weight ratio of each strain in the microbial inoculum gamma combination is lactobacillus: b, bacillus subtilis: yeast: b, bacillus licheniformis: bacillus coagulans = (3.15-3.85): (1.8-2.2): (0.45-0.55): (1.35-1.65): 2.25-2.75).
2. The composite microbial inoculum of claim 1, wherein the preparation method of each microbial inoculum combination comprises the following steps:
(1) respectively inoculating each strain to a liquid culture medium for culturing for 36-48 h;
(2) respectively placing the strains cultured in the step (1) into a fermentation tank, and carrying out mass production culture for 36-48 h, wherein the weight ratio of the strains to the fermentation medium is 1: 9-15;
(3) mixing the strains cultured in the step (2) according to the weight ratio of the strains in each microbial inoculum combination, and performing cross antagonistic culture for 36-48 h until the viable count is 1-9 × 1010cfu/ml, separately obtaining bacteriaAn agent A combination, a fungicide C combination and a fungicide gamma combination.
3. The composite microbial inoculant according to claim 2, wherein in the step (2), sterile air is introduced for 2 minutes every 8 hours during fermentation, and the stirring speed is 70 rpm/min.
4. The application of the composite microbial composite inoculum according to claim 1 in treatment of black and odorous river channels.
5. A comprehensive treatment method for a black and odorous river channel is characterized by comprising the following steps:
A. controlling the exogenous pollution of the black and odorous river channel, and well performing the work of a sewage interception and storage pipe;
B. mixing the combination of the microbial inoculum A and the combination of the microbial inoculum C in the composite microbial combined microbial inoculum according to claim 1, and putting the mixture into a black and odorous river;
C. after the water quality reaches the standard, putting the microbial inoculum gamma combination in the composite microbial combined microbial inoculum of claim 1 into a river channel, breeding aquatic animals and plants, and recovering the self-purification capacity of the water ecological system;
D. and then the riverway is maintained for a long time.
6. The treatment method according to claim 5, wherein the adding amount of the combination of the microbial inoculum A, the combination of the microbial inoculum C and the combination of the microbial inoculum gamma is 20-50 ppm.
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