CN115305216A - Compound microbial agent for denitrification of black and odorous water and preparation and use methods thereof - Google Patents
Compound microbial agent for denitrification of black and odorous water and preparation and use methods thereof Download PDFInfo
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Abstract
The invention discloses a composite microbial agent for denitrification of black and odorous water and a preparation and use method thereof, wherein the composite microbial agent consists of 50-60 parts by weight of indigenous microbial adsorbent, 20-30 parts by weight of microbial strain, 10-20 parts by weight of nutrient and 5-10 parts by weight of enzyme preparation. The method comprises the following steps: firstly, separating and enriching denitrifying indigenous microorganisms from black and odorous water and preparing a microorganism suspension; secondly, placing bentonite and active carbon in the microbial suspension for vibration adsorption to obtain an indigenous microbial adsorbent; and finally, mixing the indigenous microorganism adsorbent with the microorganism strain, the nutrient and the enzyme preparation to obtain the compound microorganism bacterium agent. The composite microbial agent prepared by the invention has good denitrification effect in black and odorous water, is suitable for low dissolved oxygen environment, and can grow, propagate and denitrify the upper layer, the middle layer and the lower layer of the water.
Description
Technical Field
The invention relates to the technical field of water body denitrification, in particular to a composite microbial agent for black and odorous water body denitrification and a preparation method and a use method thereof.
Background
In recent years, with the acceleration of the industrialization process of China and the rapid development of economy, a large amount of untreated or incompletely treated industrial wastewater enters rivers and lakes, meanwhile, chemical fertilizers are generated and used excessively, organic garbage, livestock and poultry manure and the like enter water along with surface runoff, and the nitrogen element in the water exceeds the standard. Excessive nitrogen causes the plants to swell excessively, algae breed massively, aquatic organism residues are accumulated in water continuously, the content of dissolved oxygen in the water is reduced, eutrophication and black and odorous are caused gradually, the survival of animals and plants and the health of people are seriously affected, and the quality of urban ecological environment is reduced.
At present, the control method of nitrogen pollution of black and odorous water mainly comprises physical, chemical and biological ecological methods. The physical method mainly comprises the steps of intercepting a sewage control source, removing bottom mud, conducting water diversion and water exchange, performing artificial aeration and the like, and the chemical method comprises the step of adding a flocculant, an oxidant and other medicaments. Physical methods and chemical methods are complex and complicated to operate, have the risk of secondary pollution and have poor nitrogen control effect. The biological ecological method utilizes the self life activities of organisms, and certain specific organisms absorb and convert pollutants in the water body into nutrient substances required by self growth and metabolism, so that the method is an economic, feasible and efficient treatment method. The microbial agent method is a technology for fixing the cultured and screened microbes on a proper carrier, and the microbial agent is put into a water body, can degrade specific pollutants, accelerate the improvement of water quality and restore the ecological purification capacity of the water body, and has the advantages of low economic cost, high essence efficiency, no toxicity, environmental protection, wide application prospect and the like. The black and odorous water has extremely low dissolved oxygen content, high pollutant concentration, complex water quality condition, poor denitrification effect of the common microbial agent, is not suitable for low dissolved oxygen environment, needs to be combined with aeration, and has poor impact resistance load.
Disclosure of Invention
Based on the defects of the prior art, the technical problem to be solved by the invention is to provide the compound microbial agent which has good denitrification effect, is suitable for low dissolved oxygen environment and can grow, reproduce and denitrify the upper layer, the middle layer and the lower layer of the black and odorous water body, and the preparation and use methods thereof.
In order to solve the technical problems, the invention provides a preparation method of a composite microbial agent for denitrification of black and odorous water, which comprises the following steps:
(1) Taking black and odorous water, separating and enriching denitrifying indigenous microorganisms and preparing a microorganism suspension;
(2) Sterilizing the microbial carrier, placing the microbial carrier in a microbial suspension, vibrating and adsorbing the microbial carrier, and drying the microbial carrier to obtain an indigenous microbial adsorbent;
(3) The indigenous microorganism adsorbent is mixed with microorganism strains, a nutrient and an enzyme preparation to obtain the compound microorganism bacterium agent.
Preferably, the composite microbial agent for denitrification of black and odorous water and the preparation and use methods thereof provided by the invention further comprise part or all of the following technical characteristics:
as an improvement of the technical scheme, the black and odorous water body is taken in the step (1), the denitrification indigenous microorganisms are separated and enriched, and a microorganism suspension is prepared by the following specific method:
1) Adding a bacterial enrichment culture medium into a black and odorous water sample and the bottom mud, and performing shake culture under anaerobic conditions to obtain a bacterial liquid;
2) Taking the bacterial liquid prepared in the step 1), adding a black and odorous water sample and an inorganic salt nutrient solution at intervals of 24 hours for acclimatization and culture, taking 3d as a period, taking the bacterial liquid after the previous week period, repeating the step 2) for 3 periods to obtain bacterial liquid A;
3) Coating the bacterial liquid A in the step 2) on a flat solid culture medium, and culturing until rod-shaped bacterial colonies which are yellowish, regular in edge and transparent and glossy are grown; shake culturing the bacterial colony and sterilized beef high peptone culture solution to obtain logarithmic phase bacterial liquid;
4) Taking a certain amount of logarithmic phase bacteria liquid to centrifuge in a sterile centrifuge tube, removing the upper culture medium, washing, and preparing the thalli into a microorganism suspension by using normal saline.
As an improvement of the technical scheme, in the step 1), the volume of the black and odorous water sample and the volume of the sediment are 5ml, the volume of the bacteria enrichment culture medium is 45ml, and the shaking culture time is 2d in a constant-temperature shaking table at 30 ℃ and 150 r/min.
As an improvement of the technical scheme, the volume of the bacterial liquid obtained in the step 2) is 10ml, the volume of the added black and odorous water sample is 10ml, and the volume of the added inorganic salt nutrient solution is 30ml.
As an improvement of the technical scheme, in the step 3), the volume of the beef high peptone culture liquid is 100ml, and the beef high peptone culture liquid is shake-cultured for 24 hours in a constant temperature shaking table at 30 ℃ and 150 r/min.
As an improvement of the technical scheme, the volume of the logarithmic phase bacterial liquid taken in the step 4) is 50ml, and the logarithmic phase bacterial liquid is centrifuged at 4000r/min for ten minutes to prepare a microbial suspension of 50m.
As an improvement of the technical scheme, the microorganism carrier in the step (2) is bentonite and activated carbon, and the mass ratio of the bentonite to the activated carbon is 7:3 to 7:1, sterilizing for 20min in a high-pressure steam sterilizing pot under the sterilizing condition; the mass volume ratio of the microbial carrier to the microbial suspension is 1g:10 ml-1 g:20ml, shaking and adsorbing for 12-24h in a constant temperature shaking table at 20-30 ℃ and 150r/min, and drying for 1-2 h at 105 ℃.
As an improvement of the technical scheme, the microorganism strains in the step (3) are anammox bacteria, bacillus and pseudomonas; the nutrient agent comprises 5 to 10 portions of glucose, 2 to 4 portions of starch, 0.5 to 1 portion of NH4Cl0.5, 30.5 to 1 portion of NaNO30, 0.5 to 1 portion of methanol, 0.5 to 1 portion of Na2CO30.5, 1 to 5 portions of KH2PO40.5 and 1 portion of trace elements; the enzyme preparation is glucose enzyme and amylase; the compound microbial agent consists of 50-60 parts of indigenous microorganism adsorbent, 20-30 parts of microbial strain, 10-20 parts of nutrient and 5-10 parts of enzyme preparation in parts by mass.
A use method of a compound microbial agent for denitrification of black and odorous water comprises the following steps:
(1) Taking the composite microbial agent and a black and odorous water sample, and mixing the two according to the mass volume ratio of 1kg:10L of the mixture is uniformly mixed, and the mixture is placed for 24 hours at normal temperature under anaerobic condition to obtain bacterial liquid;
(2) Directly splashing the bacterial liquid into the black and odorous water body, wherein the adding amount is 2-2.5L/m 3 。
Preferably, the method for using the composite microbial agent for denitrification of black and odorous water further comprises the following technical characteristics:
as an improvement of the above technical scheme, the complex microbial inoculum is prepared by the method of any one of claims 1 to 8.
The technical principle of the invention is as follows:
the black and odorous water body has extremely low dissolved oxygen content and complex environment, the black and odorous water body and the sediment are taken and the denitrification microorganisms are separated and enriched, the microorganisms adapt to the black and odorous water body environment, and the nitrogen removal rate is high. By adding microbial strains including anaerobic ammonium oxidation bacteria, bacillus and pseudomonas, the bacteria can grow and reproduce in water under the condition of oxygen deficiency or oxygen deficiency. The microorganisms are fixed on bentonite and active carbon, nitrogen pollutants are adsorbed by utilizing the adsorption characteristics of the bentonite and the active carbon, biological membranes grow on the water bottom and the water surface on the bentonite and the active carbon respectively, bacterial liquid is mixed with black and odorous water, nitrogen elements can be removed at the middle layer and any other positions, and the nitrogen elements are converted into nitrogen by utilizing the microorganisms through nitrification, denitrification and anaerobic ammonia oxidation to be removed. When in use, the bacterium solution and the black and odorous water body are mixed and placed in an anoxic manner, so that the microorganisms adapt to the black and odorous water body environment again, and grow and proliferate in a large amount under the nutritional agent, and the enzyme preparation is beneficial to the decomposition of glucose and starch in the nutritional agent.
Compared with the prior art, the technical scheme of the invention has the following beneficial effects:
(1) Strong adaptability and low oxygen content. The denitrifying microorganisms are obtained by separating and enriching from natural black and odorous water and are suitable for the environment of the black and odorous water, and the added microbial strains can grow and reproduce under the condition of oxygen deficiency or oxygen-free environment without aeration and oxygenation and are suitable for the low dissolved oxygen environment of the black and odorous water.
(2) High denitrification efficiency and good effect. The denitrifying microorganisms taken from the black and odorous water body have high treatment efficiency on nitrogen, can enhance the treatment effect of the denitrifying microorganisms with the added microbial strains, remove nitrogen elements through various ways such as nitrification, denitrification, anaerobic ammonia oxidation and the like, and simultaneously have high adsorbability on nitrogen by using carrier bentonite and activated carbon.
(3) Simple operation and impact load resistance. The microbial agent is mixed with black and odorous water and proliferated, and then directly put into the water, so that the operation is simple, the microorganisms grow a biological film on the upper activated carbon layer of the water, grow a biological film on the lower bentonite layer, and resist impact load because the middle layer and the other layers are in a free state.
The foregoing description is only an overview of the technical solutions of the present invention, and in order to make the technical means of the present invention more clearly understood, the present invention may be implemented in accordance with the contents of the description, and in order to make the above and other objects, features, and advantages of the present invention more clearly understood, the following detailed description is given in conjunction with the preferred embodiments.
Detailed Description
Other aspects, features and advantages of the present invention will become apparent from the following detailed description, which, when taken in conjunction with the drawings, illustrate by way of example the principles of the invention.
Example 1
Taking 5ml of black and odorous water sample and sediment, adding 45ml of bacteria enrichment medium into a conical flask, and carrying out anaerobic shake culture in a constant-temperature shaking table at 30 ℃ and 150r/min for 2d. Taking 10ml of the bacterial liquid in a conical flask, adding 10ml of black and odorous water sample and 30ml of inorganic salt nutrient solution at intervals of 24h for acclimatization and culture, taking 3d as a period, taking the bacterial liquid after the previous week period is finished, and repeating the steps for 3 periods. After 3 cycles of culture, the bacterial liquid is smeared in a flat solid culture medium and cultured until rod-shaped bacterial colonies which are yellowish, regular in edges and transparent and glossy are grown. Taking the bacterial colony and sterilized 100ml beef high peptone culture solution, and shake culturing in a constant temperature shaker at 30 ℃ and 150r/min for 24h to obtain logarithmic phase bacterial solution. And (3) centrifuging the logarithmic phase bacterial liquid in a sterile centrifuge tube for ten minutes at 4000r/min, removing the upper culture medium, washing, and preparing the thalli into microbial suspension with the same volume by using normal saline. Taking the mass ratio of 7:3, sterilizing the bentonite and the active carbon in a high-pressure steam sterilizing pot for 20min, wherein the volume ratio of the bentonite to the active carbon is 1g:20ml of the microbial suspension is subjected to shaking adsorption in a constant temperature shaking table at 30 ℃ and 150r/min for 24h, and is dried at 105 ℃ for 2h to obtain the indigenous microbial adsorbent. The nutrient is prepared from 5 parts by mass of glucose, 2 parts by mass of starch, 0.5 part by mass of NH4Cl0.5 part by mass of NaNO30.5 parts by mass of methanol, 0.5 part by mass of Na2CO30.5 parts by mass of KH2PO40.5 parts by mass of trace elements. The composite microbial agent is prepared from 60 parts of indigenous microbial adsorbent, 25 parts of microbial strain, 10 parts of nutrient and 5 parts of enzyme preparation in parts by mass.
Taking 10g of the compound microbial agent in a conical flask, adding 100ml of black and odorous water, sealing, placing at normal temperature for 24h under anaerobic condition, pouring the bacterial liquid into 40L of black and odorous water, and sealing for culture, wherein the adding amount is 2.5L/m < 3 >.
Example 2
Taking 5ml of black and odorous water sample and sediment in a conical flask, adding 45ml of bacteria enrichment medium, and carrying out anaerobic shake culture in a constant temperature shaking table at 30 ℃ and 150r/min for 2d. Taking 10ml of the bacterial liquid in a conical flask, adding 10ml of black and odorous water sample and 30ml of inorganic salt nutrient solution at intervals of 24h for acclimatization and culture, taking 3d as a period, taking the bacterial liquid after the previous week period is finished, and repeating the steps for 3 periods. After 3 cycles of culture, the bacterial liquid is smeared in a flat solid culture medium and cultured until rod-shaped bacterial colonies which are yellowish, regular in edges and transparent and glossy are grown. Taking the bacterial colony and sterilized 100ml beef high peptone culture solution, and shake culturing in a constant temperature shaker at 30 ℃ and 150r/min for 24h to obtain logarithmic phase bacterial solution. Taking the logarithmic phase bacterial liquid, centrifuging for ten minutes at 4000r/min in a sterile centrifugal tube, removing the upper culture medium, washing, and preparing the thalli into microbial suspension with the same volume by using normal saline. Taking the mass ratio of 7:1, sterilizing the bentonite and the active carbon in a high-pressure steam sterilization pot for 20min, wherein the volume ratio of the bentonite to the active carbon is 1g:10ml of the microbial suspension is subjected to shake adsorption for 12h in a constant temperature shaking table at 20 ℃ and 150r/min, and is dried for 1h at 105 ℃ to obtain the indigenous microbial adsorbent. 8 parts of glucose, 3 parts of starch, 1 part of NH4Cl, 301 parts of NaNO, 1 part of methanol, 31 parts of Na2CO, 41 parts of KH2PO and 1 part of trace elements are prepared into the nutrient according to the mass parts. The composite microbial agent is prepared from 60 parts of indigenous microbial adsorbent, 25 parts of microbial strain, 10 parts of nutrient and 5 parts of enzyme preparation in parts by mass.
Taking 10g of the compound microbial inoculum to be put into a conical flask, adding 100ml of black and odorous water, sealing the opening, placing at normal temperature for 24h under anaerobic condition, taking the bacterial liquid, pouring the bacterial liquid into 40L of black and odorous water, and sealing the opening for culturing, wherein the adding amount is 2.5L/m < 3 >.
Comparative example 1
Taking 5ml of black and odorous water sample and sediment, adding 45ml of bacteria enrichment medium into a conical flask, and carrying out anaerobic shake culture in a constant-temperature shaking table at 30 ℃ and 150r/min for 2d. Taking 10ml of the bacterial liquid in a conical flask, adding 10ml of black and odorous water sample and 30ml of inorganic salt nutrient solution at intervals of 24h for acclimatization and culture, taking 3d as a period, taking the bacterial liquid after the previous week period is finished, and repeating the steps for 3 periods. And (3) coating the bacterial liquid after the 3-period culture in a flat solid culture medium, and culturing until rod-shaped colonies which are yellowish, regular in edge and transparent and glossy in colonies grow out. Mixing the above bacterial colony with sterilized 100ml beef high peptone culture solution, shaking in constant temperature shaking table at 30 deg.C and 150r/minCulturing for 24h to obtain logarithmic phase bacterial liquid. Taking the logarithmic phase bacterial liquid, centrifuging for ten minutes at 4000r/min in a sterile centrifugal tube, removing the upper culture medium, washing, and preparing the thalli into microbial suspension with the same volume by using normal saline. Only taking bentonite, sterilizing for 20min in a high-pressure steam sterilization pot, and mixing the bentonite and the high-pressure steam sterilization pot in a volume ratio of 1g:20ml of the microbial suspension is subjected to shaking adsorption in a constant temperature shaking table at 30 ℃ and 150r/min for 24h, and is dried at 105 ℃ for 2h to obtain the indigenous microbial adsorbent. Mixing 5 parts of glucose, 2 parts of starch and NH by mass 4 Cl0.5 parts, naNO 3 0.5 part of methanol, 0.5 part of Na2CO30.5 parts of KH2PO40.5 parts of trace elements and 0.5 part of trace elements. The composite microbial agent is prepared from 60 parts of indigenous microbial adsorbent, 25 parts of microbial strain, 10 parts of nutrient and 5 parts of enzyme preparation in parts by mass.
Taking 10g of the compound microbial inoculum to be put into a conical flask, adding 100ml of black and odorous water, sealing the opening, placing at normal temperature for 24h under anaerobic condition, taking the bacterial liquid, pouring the bacterial liquid into 40L of black and odorous water, and sealing the opening for culturing, wherein the adding amount is 2.5L/m < 3 >.
Comparative example 2
Taking 5ml of black and odorous water sample and sediment in a conical flask, adding 45ml of bacteria enrichment medium, and carrying out anaerobic shake culture in a constant temperature shaking table at 30 ℃ and 150r/min for 2d. Taking 10ml of the bacterial liquid in a conical flask, adding 10ml of black and odorous water sample and 30ml of inorganic salt nutrient solution at intervals of 24h for acclimatization and culture, taking 3d as a period, taking the bacterial liquid after the previous week period is finished, and repeating the steps for 3 periods. After 3 cycles of culture, the bacterial liquid is smeared in a flat solid culture medium and cultured until rod-shaped bacterial colonies which are yellowish, regular in edges and transparent and glossy are grown. Taking the bacterial colony and sterilized 100ml beef high peptone culture solution, and shake culturing in a constant temperature shaker at 30 ℃ and 150r/min for 24h to obtain logarithmic phase bacterial solution. Taking the logarithmic phase bacterial liquid, centrifuging for ten minutes at 4000r/min in a sterile centrifugal tube, removing the upper culture medium, washing, and preparing the thalli into microbial suspension with the same volume by using normal saline. Taking the mass ratio of 7:3, sterilizing the bentonite and the active carbon in a high-pressure steam sterilizing pot for 20min, wherein the volume ratio of the bentonite to the active carbon is 1g:20ml of the microbial suspension is subjected to shaking adsorption in a constant temperature shaking table at 30 ℃ and 150r/min for 24h, and is dried at 105 ℃ for 2h to obtain the indigenous microbial adsorbent. The nutrient is prepared from 5 parts by mass of glucose, 2 parts by mass of starch, 0.5 part by mass of NH4Cl0.5 part by mass of NaNO30.5 parts by mass of methanol, 0.5 part by mass of Na2CO30.5 parts by mass of KH2PO40.5 parts by mass of trace elements. The composite microbial agent is prepared from 50 parts of indigenous microbial adsorbent, 20 parts of microbial strain, 20 parts of nutrient and 10 parts of enzyme preparation in parts by mass.
Taking 10g of the compound microbial inoculum to be put into a conical flask, adding 100ml of black and odorous water, sealing the opening, placing at normal temperature for 24h under anaerobic condition, taking the bacterial liquid, pouring the bacterial liquid into 40L of black and odorous water, and sealing the opening for culturing, wherein the adding amount is 2.5L/m < 3 >.
Comparative example 3
Taking 5ml of black and odorous water sample and sediment, adding 45ml of bacteria enrichment medium into a conical flask, and carrying out anaerobic shake culture in a constant-temperature shaking table at 30 ℃ and 150r/min for 2d. Taking 10ml of the bacterial liquid in a conical flask, adding 10ml of black and odorous water sample and 30ml of inorganic salt nutrient solution at intervals of 24h for acclimatization and culture, taking 3d as a period, taking the bacterial liquid after the previous week period is finished, and repeating the steps for 3 periods. And (3) coating the bacterial liquid after the 3-period culture in a flat solid culture medium, and culturing until rod-shaped colonies which are yellowish, regular in edge and transparent and glossy in colonies grow out. Taking the bacterial colony and sterilized 100ml beef high peptone culture solution, and shake-culturing in a constant temperature shaker at 30 ℃ and 150r/min for 24h to obtain logarithmic phase bacterial solution. And (3) centrifuging the logarithmic phase bacterial liquid in a sterile centrifuge tube for ten minutes at 4000r/min, removing the upper culture medium, washing, and preparing the thalli into microbial suspension with the same volume by using normal saline. Taking the mass ratio of 7:3, sterilizing the bentonite and the active carbon in a high-pressure steam sterilizing pot for 20min, wherein the volume ratio of the bentonite to the active carbon is 1g:20ml of the indigenous microorganism adsorbent is subjected to vibration adsorption in a constant temperature shaking table at 30 ℃ and 150r/min for 24h in a microorganism suspension, and is dried for 2h at 105 ℃ to obtain the indigenous microorganism adsorbent. The nutrient is prepared from 5 parts by mass of glucose, 2 parts by mass of starch, 0.5 part by mass of NH4Cl0.5 part by mass of NaNO30.5 parts by mass of methanol, 0.5 part by mass of Na2CO30.5 parts by mass of KH2PO40.5 parts by mass of trace elements. The composite microbial agent is prepared from 85 parts of indigenous microbial adsorbent, 10 parts of nutrient and 5 parts of enzyme preparation in parts by mass.
Taking 10g of the compound microbial agent in a conical flask, adding 100ml of black and odorous water, sealing, placing at normal temperature for 24h under anaerobic condition, pouring the bacterial liquid into 40L of black and odorous water, and sealing for culture, wherein the adding amount is 2.5L/m < 3 >.
COMPARATIVE EXAMPLE 4 (dosage)
Taking 5ml of black and odorous water sample and sediment in a conical flask, adding 45ml of bacteria enrichment medium, and carrying out anaerobic shake culture in a constant temperature shaking table at 30 ℃ and 150r/min for 2d. Taking 10ml of the bacterial liquid in a conical flask, adding 10ml of black and odorous water sample and 30ml of inorganic salt nutrient solution at intervals of 24h for acclimatization and culture, taking 3d as a period, taking the bacterial liquid after the previous week period is finished, and repeating the steps for 3 periods. And (3) coating the bacterial liquid after the 3-period culture in a flat solid culture medium, and culturing until rod-shaped colonies which are yellowish, regular in edge and transparent and glossy in colonies grow out. Taking the bacterial colony and sterilized 100ml beef high peptone culture solution, and shake-culturing in a constant temperature shaker at 30 ℃ and 150r/min for 24h to obtain logarithmic phase bacterial solution. And (3) centrifuging the logarithmic phase bacterial liquid in a sterile centrifuge tube for ten minutes at 4000r/min, removing the upper culture medium, washing, and preparing the thalli into microbial suspension with the same volume by using normal saline. Taking the mass ratio of 7:3, sterilizing the bentonite and the active carbon in a high-pressure steam sterilization pot for 20min, wherein the volume ratio of the bentonite to the active carbon is 1g:20ml of the indigenous microorganism adsorbent is subjected to vibration adsorption in a constant temperature shaking table at 30 ℃ and 150r/min for 24h in a microorganism suspension, and is dried for 2h at 105 ℃ to obtain the indigenous microorganism adsorbent. The nutrient is prepared by 5 parts of glucose, 2 parts of starch, 0.5 part of NH4Cl0.5 part, 0.5 part of NaNO30.5 part, 0.5 part of methanol, 30.5 parts of Na2CO30, 40.5 parts of KH2PO40 and 0.5 part of trace elements in parts by mass. The composite microbial agent is prepared from 60 parts of indigenous microbial adsorbent, 25 parts of microbial strain, 10 parts of nutrient and 5 parts of enzyme preparation in parts by mass.
Taking 10g of the compound microbial agent in a conical flask, adding 100ml of black and odorous water, sealing, placing at normal temperature for 24h under anaerobic condition, pouring the bacterial liquid into 50L of black and odorous water, and sealing for culture, wherein the adding amount is 2L/m < 3 >.
Experimental data:
different conditions are set according to examples 1-2 and comparative examples 1-4, sampling detection is carried out every 1 day, the total nitrogen concentration (TN) of the overlying black and odorous water body is detected by adopting an alkaline potassium persulfate digestion ultraviolet spectrophotometry (HJ 636-2012), and the average value is repeatedly obtained for three times, and the results are shown in the following table and are in unit of mg/L.
TABLE 1 Denitrification of black and odorous water by different bacterial agents
| Day 0 | 1 day | 2 days | 3 days | 4 days | 5 days | 6 days | 7 days | |
| Example 1 | 17.14 | 13.69 | 9.21 | 5.02 | 2.56 | 1.77 | 1.66 | 1.62 |
| Example 2 | 17.14 | 14.21 | 10.08 | 6.26 | 3.96 | 2.21 | 2.08 | 1.88 |
| Comparative example 1 | 17.14 | 14.88 | 10.42 | 6.55 | 4.16 | 2.56 | 2.18 | 1.91 |
| Comparative example 2 | 17.14 | 15.13 | 10.81 | 6.86 | 4.36 | 2.88 | 2.54 | 2.22 |
| Comparative example 3 | 17.14 | 15.26 | 11.02 | 7.15 | 4.59 | 4.11 | 3.62 | 3.21 |
| Comparative example 4 | 17.14 | 14.23 | 9.44 | 5.34 | 2.82 | 1.87 | 1.74 | 1.69 |
| Blank space | 17.14 | 17.22 | 17.36 | 17.41 | 17.44 | 17.44 | 17.52 | 17.55 |
From the above table it can be seen that: the black and odorous water body is seriously polluted by nitrogen, the blank group TN is inferior to five types, and the TN in the examples 1-2 and the comparative examples 1-4 is greatly reduced by adding the compound microbial agent. The effect of the embodiment 1 is optimal, and TN reaches five types of water quality standards after 5 days of treatment; example 2 is different from example 1 in preparation conditions, and the effect is slightly poor; comparative examples 1 to 4 lack a certain step or change a certain core condition, and although the removal effect is good, the denitrification effect of example 1 is better than that of comparative examples 1 to 4.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.
The raw materials listed in the invention, the upper and lower limits and interval values of the raw materials of the invention, and the upper and lower limits and interval values of the process parameters (such as temperature, time and the like) can all realize the invention, and the examples are not listed.
While the foregoing is directed to the preferred embodiment of the present invention, other and further embodiments of the invention may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow.
Claims (10)
1. A preparation method of a compound microbial agent for denitrification of black and odorous water is characterized by comprising the following steps:
(1) Taking black and odorous water, separating and enriching denitrifying indigenous microorganisms and preparing a microorganism suspension;
(2) Sterilizing the microbial carrier, placing the microbial carrier in a microbial suspension, vibrating, adsorbing and drying to obtain an indigenous microbial adsorbent;
(3) The indigenous microorganism adsorbent is mixed with microorganism strains, a nutrient and an enzyme preparation to obtain the compound microorganism bacterium agent.
2. The method for preparing the composite microbial agent for denitrification of black and odorous water according to claim 1, wherein the method comprises the following steps: the method comprises the following steps of (1) taking the black and odorous water body, separating and enriching the denitrifying indigenous microorganisms and preparing a microorganism suspension, and specifically comprises the following steps:
1) Adding a bacterial enrichment culture medium into a black and odorous water sample and the bottom mud, and performing shake culture under anaerobic conditions to obtain a bacterial liquid;
2) Taking the bacterial liquid prepared in the step 1), adding a black and odorous water sample and an inorganic salt nutrient solution at intervals of 24 hours for acclimatization and culture, taking 3d as a period, taking the bacterial liquid after the previous week period is finished, repeating the step 2), and taking 3 periods to obtain a bacterial liquid A;
3) Coating the bacterial liquid A in the step 2) on a flat solid culture medium, and culturing until rod-shaped bacterial colonies which are yellowish, regular in edge and transparent and glossy are grown; shake culturing the bacterial colony and sterilized beef high peptone culture solution to obtain logarithmic phase bacterial solution;
4) Taking a certain amount of logarithmic phase bacteria liquid to centrifuge in a sterile centrifuge tube, removing the upper culture medium, washing, and preparing the thalli into a microorganism suspension by using normal saline.
3. The method for preparing the composite microbial agent for denitrification of black and odorous water according to claim 2, wherein the method comprises the following steps: in the step 1), the volume of the black and odorous water sample and the sediment is 5ml, the volume of the bacteria enrichment medium is 45ml, and the shaking culture time is 2d in a constant temperature shaking table at 30 ℃ and 150 r/min.
4. The method for preparing the composite microbial agent for denitrification of black and odorous water according to claim 2, wherein the method comprises the following steps: 10ml of the bacterial liquid obtained in the step 2), 10ml of the volume of the black and odorous water sample and 30ml of inorganic salt nutrient solution.
5. The method for preparing the composite microbial agent for denitrification of black and odorous water according to claim 2, wherein the method comprises the following steps: in the step 3), the volume of the beef high peptone culture liquid is 100ml, and the shake culture is carried out in a constant temperature shaking table at 30 ℃ and 150r/min for 24h.
6. The method for preparing the composite microbial agent for denitrification of black and odorous water according to claim 2, wherein the method comprises the following steps: and (3) centrifuging the logarithmic phase bacterial liquid obtained in the step 4) for ten minutes at a speed of 50ml and 4000r/min to prepare a microbial suspension of 50m.
7. The method for preparing the composite microbial agent for denitrification of black and odorous water according to claim 1, wherein the method comprises the following steps: the microbial carrier in the step (2) is bentonite and activated carbon, and the mass ratio is 7:3 to 7:1, sterilizing for 20min in a high-pressure steam sterilizing pot under the sterilizing condition; the mass volume ratio of the microbial carrier to the microbial suspension is 1g:10 ml-1 g:20ml, shaking and adsorbing for 12-24h in a constant temperature shaking table at 20-30 ℃ and 150r/min, and drying for 1-2 h at 105 ℃.
8. The method for preparing the composite microbial agent for denitrification of black and odorous water according to claim 1, wherein the method comprises the following steps: the microbial strains in the step (3) are anammox bacteria, bacillus and pseudomonas; the nutrient comprises 5 to 10 portions of glucose, 2 to 4 portions of starch, 0.5 to 1 portion of NH4Cl0.5, 30.5 to 1 portion of NaNO30.5, 0.5 to 1 portion of methanol, 30.5 to 1 portion of Na2CO30.5, 1 to 5 portions of KH2PO40.5 and 1 portion of trace elements; the enzyme preparation is selected from glucosidase and amylase; the compound microbial agent consists of 50-60 parts of indigenous microorganism adsorbent, 20-30 parts of microbial strain, 10-20 parts of nutrient and 5-10 parts of enzyme preparation in parts by mass.
9. The use method of the compound microbial agent for denitrification of black and odorous water is characterized by comprising the following steps:
(1) Taking the composite microbial agent and a black and odorous water sample, and mixing the two according to the mass volume ratio of 1kg:10L of the mixture is uniformly mixed, and the mixture is placed for 24 hours at normal temperature under anaerobic condition to obtain bacterial liquid;
(2) Directly splashing the bacterial liquid into the black and odorous water body, wherein the adding amount is 2-2.5L/m 3 。
10. The use method of the composite microbial inoculum for denitrification of black and odorous water according to claim 9, characterized in that: the complex microbial inoculant is prepared by the method as described in any one of claims 1 to 8.
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