CN107384833B - Composite microbial inoculum, preparation method and application thereof, and treatment method of black and odorous river channel - Google Patents
Composite microbial inoculum, preparation method and application thereof, and 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 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 B 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; each strain in the microbial inoculum B combination is bacillus coagulans, silicate bacteria, bacillus firmus and lactobacillus respectively; the strains in the microbial inoculum gamma combination are lactobacillus, bacillus subtilis, saccharomycetes, bacillus licheniformis and bacillus coagulans respectively. The composite microbial composite bacterial agent can be used for treating black and odorous river channels, has a short treatment period, and can effectively solve the problem that nitrogen and phosphorus in black and odorous water exceed standards and eliminate odorous gas hydrogen sulfide in the water.
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 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 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 B combination and a bacterial agent gamma combination, and can be used for treating black and odorous watercourses, effectively reduce pollutant components such as nitrogen, phosphorus, organic matters and the like in the black and odorous water body, and obviously eliminate 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 B 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 B composition comprises bacillus coagulans, silicate bacteria, bacillus firmus and lactic acid bacteria;
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 B combination is bacillus coagulans: silicate bacteria: bacillus firmus: lactic acid bacteria (4.5-5.5): (94.5-110.5): (94.5-110.5): (1.8-2.2).
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).
In the composite microbial combined bacterial agent, the weight ratio of the bacterial agent A combination, the bacterial agent B combination and the bacterial agent gamma combination is 1: 1: 1.
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) after the mass production culture of the step (2)Mixing the strains 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, and respectively obtaining a microbial inoculum A combination, a microbial inoculum B 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 Mn2+、Mg2 +Or Fe2+All used are sulfates thereof;
(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 Mn2+、Mg2+Or Fe2+All used are sulfates thereof;
(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): (2.7-3.3) mixing, carrying out cross antagonistic culture for 48h, and culturingIn the process, sterile air is introduced for 2 minutes every 8 hours, the stirring 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: 15-20, and a bacterium agent A combination is obtained, wherein the viable count of the bacterium agent A combination is 4-6 × 1010cfu/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 Mn2+、Mg2+Or Fe2+All used are their sulfates.
The preparation method of the microbial inoculum B combination comprises the following steps:
(1) respectively inoculating bacillus coagulans, silicate bacteria, bacillus firmus and lactic acid bacteria into a liquid culture medium, and culturing for 36h 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, 3.6-4.4 g of yeast extract, 1.53-1.87 g of calcium carbonate, 18-22 g of glucose, 800.9-1.1 ml of tween, 0.9-1.1 g of monopotassium phosphate, 27-33 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, 9-11 g of peptone, 3.6-4.4 g of yeast extract, 1.53-1.87 g of calcium carbonate, 18-22 g of glucose, 800.9-1.1 ml of tween, 0.9-1.1 g of monopotassium phosphate, 27-33 g 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 bacillus coagulans: silicate bacteria: bacillus firmus: mixing lactobacillus (4.5-5.5): (94.5-110.5): 1.8-2.2), cross-antagonistic culturing for 36h, and the weight ratio of the mixed strain to the cross-antagonistic culture medium1: 15-20, obtaining a microbial inoculum B combination, wherein the viable count is 7-9 × 1010cfu/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, 3.6-4.4 g of yeast extract, 1.53-1.87 g of calcium carbonate, 18-22 g of glucose, 800.9-1.1 ml of tween, 0.9-1.1 g of monopotassium phosphate, 27-33 g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2.
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, 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;
(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-antagonistic 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 and 1.1-78 m of tween 800.91.35-1.65 g of magnesium sulfate heptahydrate, 18-22 g of glucose, 45-55 g of cane sugar 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.
A treatment method of 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 B combination, and putting the mixture into a black and odorous river;
C. after 5-6 months, putting the microbial inoculum gamma combination into a river channel, stocking 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.
The adding amount of the microbial inoculum A combination, the microbial inoculum B 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;
silicate bacteria: the insoluble potassium and phosphorus are dissolved out to be utilized by crops and thalli, and substances such as hormone, amino acid, polysaccharide and the like are generated to promote the growth of plants;
bacillus firmus: amylase and protease are secreted, and the growth of harmful bacteria is effectively inhibited;
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 B combination is obtained by cross antagonistic culture of bacillus coagulans, silicate bacteria, bacillus firmus and lactic acid bacteria according to the weight ratio of (4.5-5.5): (94.5-110.5): 1.8-2.2, and the strains act together, so that the microbial inoculum B combination can rapidly convert and degrade ammonia nitrogen substances in water, can remove eutrophication substances in water, and can improve the water quality of a river and a microbial system.
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 B 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 B 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 B combination and a bactericide gamma combination, the treatment period is short, the problems of excessive nitrogen and phosphorus and over-high COD (chemical oxygen demand) 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 microbial inoculum A combination and the microbial inoculum B combination are mixed and restored to restore the water quality of the black and odorous riverway, a microbial inoculum gamma combination is put into the water 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 B 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 Mn2+、Mg2+Or Fe2+All used are sulfates thereof;
(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 Mn2+、Mg2+Or Fe2+All used are sulfates thereof;
(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. The cross-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 steamed extractDistilled water with the pH value of 6.8-7.2; the trace element is Mn2+、Mg2 +Or Fe2+All used are their sulfates.
The preparation method of the microbial inoculum B combination comprises the following steps:
(1) respectively inoculating bacillus coagulans, silicate bacteria, bacillus firmus and lactic acid bacteria into a liquid culture medium, and culturing for 36h at 28-32 ℃; the liquid culture medium comprises the following components in each liter of culture medium: 8g of beef extract, 10g of peptone, 4g of yeast extract, 1.7g of calcium carbonate, 20g of glucose, 801.0ml of tween, 1.0g of monopotassium phosphate, 30g 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, 10g of peptone, 4g of yeast extract, 1.7g of calcium carbonate, 20g of glucose, 801.0ml of tween, 1.0g of monopotassium phosphate, 30g 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 manner according to the weight ratio of bacillus coagulans to silicate bacteria to bacillus firmus to lactobacillus which is 5: 105: 105: 2, 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 bacterium agent B combination with the viable count of 8 × 1010cfu/ml. The cross-antagonistic culture medium comprises the following components in each liter of culture medium: 8g of beef extract, 10g of peptone, 4g of yeast extract, 1.7g of calcium carbonate, 20g of glucose, 801.0ml of tween, 1.0g of monopotassium phosphate, 30g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2.
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, 801.0ml 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, 801.0ml 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, 801.0ml 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.
The composite microbial composite bacterial agent is used for treating black and odorous riverways, the riverway in a certain development area is about 3000 meters long, 15-35 meters wide, 1-3.5 meters deep, and 0.5-0.7 meter thick bottom mud. River water has been seriously polluted for a long time due to natural discharge of a large amount of domestic sewage and industrial sewage. The river water is seriously bloom, eutrophicated, black and smelly, low in transparency, black in silt and more in white garbage and oil stains on the river surface.
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 B 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 B 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 B, 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 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 91.7 percent and NH is reduced by the treatment of the composite microbial composite inoculant for 6 months3The N content is reduced by 88.9 percent, the total phosphorus content is reduced by 92.2 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 B 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 Mn2+、Mg2+Or Fe2+All used are sulfates thereof;
(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: 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 Mn2+、Mg2+Or Fe2+All used are sulfates thereof;
(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: 1.8: 0.55: 1.65: 3.3 mixing, cross-antagonistic culturing for 48h, and stirring and introducing sterile air for 2 min every 8h in the culturing processStirring at 70rpm/min, flow rate of the sterile air being 90L/min, weight ratio of mixed strain to cross antagonistic culture medium being 1: 20 to obtain bacterial agent A composition with viable count of 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 Mn2+、Mg2+Or Fe2+All used are their sulfates.
The preparation method of the microbial inoculum B combination comprises the following steps:
(1) respectively inoculating bacillus coagulans, silicate bacteria, bacillus firmus and lactic acid bacteria into a liquid culture medium, and culturing for 36h 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, 4.4g of yeast extract, 1.87g of calcium carbonate, 22g of glucose, 801.1ml of tween, 1.1g of monopotassium phosphate, 33g 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, 11g of peptone, 4.4g of yeast extract, 1.87g of calcium carbonate, 22g of glucose, 801.1ml of tween, 1.1g of monopotassium phosphate, 33g 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 bacillus coagulans, silicate bacteria, bacillus firmus and lactobacillus which are 5.5: 110.5: 110.5: 2.2, carrying out cross antagonistic culture for 36h, wherein the weight ratio of the mixed strains to the cross antagonistic culture medium is 1: 20, and obtaining a microbial inoculum B combination with the viable count of 9 × 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, 4.4g of yeast extract, 1.87g of calcium carbonate and grape22g of sugar, 801.1ml of tween, 1.1g of monopotassium phosphate, 33g of sucrose and the balance of distilled water, wherein the pH value is 6.8-7.2.
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) mixing the strains cultured in the step (2) according to the weight ratio of lactobacillus to bacillus subtilis to saccharomycetes to bacillus licheniformis to bacillus coagulans of 3.85: 2.2: 0.55: 1.65: 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: 20, obtaining a microbial inoculum gamma combination with 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, 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.
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 microorganism is adopted to combine with a microbial inoculum pairTreating the black and odorous river channels, wherein the treatment range of the river channels in a certain village is about 1500 meters, the width of the river channels is about 2 to 6 meters, the water depth is 1 to 3 meters, the thickness of bottom mud is 0.5 to 0.8 meter, and the total capacity is about 20000m3. The river mainly takes domestic sewage as main material, and a few industrial sewages are converged to show black color, serious water bloom, eutrophication, extremely low transparency, and strong putrefactive odor emitted by blackening sludge.
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 B 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 B 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 70 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 B, 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 81.1 percent and NH is reduced by the treatment of the composite microbial inoculum for 6 months3The N content is reduced by 94.3 percent, the total phosphorus content is reduced by 92.2 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 B is replaced by the input of a single combination of the microbial inoculum B 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 57.9 percent and NH is reduced by combining the microbial inoculum B for 6 months3The N content is reduced by 92.7 percent, the total phosphorus content is reduced by 72.8 percent, and the treatment effect of the riverway water quality is far lower than that of the mixture of the microbial inoculum A combination and the microbial inoculum B combination although the riverway water quality is close to the national surface water V-type water quality standard.
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 B 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 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 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 39.1% and NH is reduced by the treatment of 6 months by combining the microbial inoculum A3The N content is reduced by 76.2 percent, the total phosphorus content is reduced by 78.1 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 B combination.
The above detailed description of a composite microbial composite inoculum, its preparation method and application, and a treatment method of black and odorous river channels 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 (7)
1. The compound microorganism combined microbial inoculum is characterized by comprising a microbial inoculum A combination, a microbial inoculum B 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 B composition comprises bacillus coagulans, silicate bacteria, bacillus firmus and lactic acid bacteria;
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 B combination are mixed and put into the black and odorous river channel, and after 5-6 months, the microbial inoculum gamma combination is put into the river channel;
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 B combination is bacillus coagulans: silicate bacteria: bacillus firmus: lactic acid bacteria = (4.5-5.5): (94.5-110.5): (94.5-110.5): 1.8-2.2);
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 composite inoculum of claim 1, wherein the weight ratio of the inoculum a combination, the inoculum B combination and the inoculum γ combination in the composite microbial composite inoculum is 1: 1: 1.
3. 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 4-9 × 1010cfu/ml, and respectively obtaining a microbial inoculum A combination, a microbial inoculum B combination and a microbial inoculum gamma combination.
4. The composite microbial composite inoculum of claim 3, wherein 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.
5. The application of the composite microbial composite inoculum according to claim 1 in treatment of black and odorous river channels.
6. The treatment method of the 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 B in the composite microbial combined microbial inoculum according to claim 1, and putting the mixture into a black and odorous river;
C. after 5-6 months, putting the microbial inoculum gamma combination in the composite microbial inoculum of claim 1 into a river channel, breeding aquatic animals and plants, and recovering the self-purification capacity of the water ecosystem;
D. and then the riverway is maintained for a long time.
7. The treatment method according to claim 6, wherein the adding amount of the combination of the microbial inoculum A, the combination of the microbial inoculum B and the combination of the microbial inoculum gamma is 20-50 ppm.
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