CN113755482A - Efficient denitrification microbial preparation for sewage treatment and preparation method thereof - Google Patents

Efficient denitrification microbial preparation for sewage treatment and preparation method thereof Download PDF

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CN113755482A
CN113755482A CN202110945679.1A CN202110945679A CN113755482A CN 113755482 A CN113755482 A CN 113755482A CN 202110945679 A CN202110945679 A CN 202110945679A CN 113755482 A CN113755482 A CN 113755482A
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pectin
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李正山
郑茂盛
刘薇薇
黄中满
李正优
苏成标
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Guangxi Rongshuo Environmental Engineering Co ltd
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Abstract

The invention discloses a high-efficiency denitrification microbial preparation for sewage treatment, which is characterized in that an aerobic denitrifying bacterium No. 144 strain is loaded on a carrier material by adopting an adsorption method, the preservation number of the aerobic denitrifying bacterium No. 144 strain is CGMCC No.22897, and the carrier material is activated turfy soil. The invention adopts the adsorption method to directly load the bacterial strain, thereby avoiding complex manufacturing steps and having simple and convenient operation process. The No. 144 strain provided by the invention is aerobic denitrifying bacteria, is obtained by enrichment screening through a biological culture means, has high-efficiency aerobic denitrification performance, and can obviously remove total nitrogen in black and odorous water under aerobic conditions.

Description

Efficient denitrification microbial preparation for sewage treatment and preparation method thereof
Technical Field
The invention relates to the field of sewage environment restoration, in particular to a high-efficiency denitrification microbial preparation for sewage treatment and a preparation method thereof.
Background
In the actual production, the high-concentration ammonia nitrogen wastewater has complex components and poor biodegradability, the effect of the traditional biological denitrification method needs to be further improved, and along with the increasing severity of the water eutrophication problem and the increasing strictness of the nitrogen discharge standard in the wastewater, how to economically and effectively remove ammonia nitrogen in the wastewater becomes one of the problems to be solved urgently in the treatment of the high-concentration ammonia nitrogen wastewater.
The microbial agent technology has no special requirements on environmental conditions in the process of treating water environmental pollution, has strong adaptability, resists water quality and water quantity fluctuation impact, and can remove NH3-N in a water body. Compared with other physical chemistry and other technologies, the technology has the advantage of being beneficial to ecological environment protection, and can gradually improve the composition and the function of an ecological system of a water body, so that the ecological structure is gradually healthy, and the water quality gradually becomes better.
The immobilization technology is a novel method for preparing the microbial inoculum, and compared with the common microbial inoculum, the immobilized microbial inoculum has the characteristics of high tolerance concentration, high degradation rate, strong processing capacity and favorable reutilization. The microbial immobilization is a technology which uses a selected carrier to adsorb and immobilize bacteria with special functions, so that the bacteria grow and reproduce on the carrier and keep activity, and the bacteria can quickly reproduce and treat sewage after a microbial inoculum is added, thereby having very high practical value. The immobilization technology used in the wastewater treatment can obviously improve the bacterial concentration, reduce the influence of adverse environment on thalli, promote the solid-liquid separation after the reaction and shorten the time required by the wastewater treatment. However, the immobilization technology also has the problems that the bacterial cells are easy to inactivate and the sewage treatment efficiency is low at present.
Disclosure of Invention
Aiming at the problems of easy inactivation of thalli and low sewage treatment efficiency in the prior immobilization technology, the invention provides a high-efficiency denitrification microbial preparation for sewage treatment and a preparation method thereof.
The purpose of the invention is realized by adopting the following technical scheme:
in a first aspect, the invention provides a high-efficiency denitrification microbial preparation for sewage treatment, wherein the high-efficiency denitrification microbial preparation is prepared by loading an aerobic denitrifying bacterium No. 144 strain onto a carrier material by an adsorption method, the preservation number of the aerobic denitrifying bacterium No. 144 strain is CGMCC No.22897, and the carrier material is.
In a second aspect, the invention provides a method for preparing a high-efficiency denitrifying microorganism preparation for sewage treatment, which comprises the following steps:
step 1, culturing strains:
inoculating the aerobic denitrifying bacteria No. 144 strain into a denitrifying culture medium from an inclined plane by using an inoculating ring, culturing in a shaking table, and culturing to a logarithmic phase to obtain a bacterial suspension;
step 2, preparation of a denitrifying microbial preparation:
weighing carrier materials in a beaker, sealing with tin foil paper, and sterilizing in an autoclave; and then under the aseptic condition, adding the sterilized carrier material into the bacterial suspension cultured to the logarithmic phase in the step 1, mixing until the carrier material is completely wetted, filling into an aseptic container, and drying at room temperature to obtain the denitrifying microorganism preparation.
Preferably, in step 1, the temperature of the shaker is 30 ℃ and the rotation speed is 150 rpm.
Preferably, in step 1, the volume of the denitrification medium is 150 mL.
Preferably, in step 1, the formulation (DM) of the denitrification medium comprises: distilled water, 6g/L K2HPO41.5g/L KH2PO40.1g/L MgSO4·7H2O, KNO of 0.6g/L35.64g/L succinic acidSodium and 0.1mL/L trace elements.
Preferably, in step 1, the pH of the culture medium is adjusted to 7.5 after the preparation is completed, and the culture medium is sterilized at the temperature of 118-125 ℃ for at least 30 min.
Preferably, in step 1, the trace elements include at least one of calcium agent, iron agent, zinc agent, selenium agent and chromium agent.
Preferably, in step 1, the determination of the logarithmic phase is to use an ultraviolet spectrophotometer to determine an absorbance value of the bacterial liquid at a wavelength of 600nm, and record the absorbance value as OD600, and the OD600 value verifies the content of the bacterial species in the bacterial liquid, so as to control whether the bacterial liquid is in the logarithmic phase.
Preferably, the OD600 of the bacterial suspension is 0.60-0.63.
More preferably, through experimental comparison of concentration of the bacteria liquid or dilution by 2 times or 4 times, the denitrification effect of the immobilized bacteria is optimal when the growth amount (OD600) of the aerobic denitrifying bacteria No. 144 strain is 0.613.
Preferably, in the step 2, the carrier material is activated turfy soil, the activated turfy soil is obtained by mixing modified calcium pectin into turfy soil and homogenizing, and the mixing amount of the modified calcium pectin is 10-30% of the mass of the turfy soil.
Preferably, the preparation method of the modified calcium pectin comprises the following steps:
a. weighing lignans, mixing the lignans with 30-70% ethanol solution by mass percent, and stirring uniformly to obtain lignan solution; wherein the mass ratio of the lignans to the ethanol solution is 1: 10-15;
b. weighing pectin, adding into the lignan solution, stirring and mixing uniformly to obtain a lignan/pectin mixed solution; wherein the mass ratio of pectin to the lignan solution is 1: 12-18;
c. dropwise adding a calcium hydroxide solution into the lignan/pectin mixed solution, continuously stirring at the speed of 500-600 rpm in the dropwise adding process, and continuously stirring for 1-2 hours after dropwise adding to obtain a calcium pectin pretreatment solution; wherein the concentration of the calcium hydroxide solution is 0.02-0.05 mol/L, and the mass ratio of calcium hydroxide in the calcium hydroxide solution to the lignan/pectin mixed solution is 1: 10-15;
d. weighing sodium bicarbonate solution, continuously dropwise adding the sodium bicarbonate solution into the calcium pectin pretreatment solution, stirring for 2-4 h after dropwise adding, decompressing the reaction solution to remove ethanol, standing in a refrigerating chamber at 3-10 ℃ for 5-8 h, filtering while cold, collecting solid, and freeze-drying to obtain modified calcium pectin; wherein the concentration of the sodium bicarbonate solution is 0.01-0.02 mol/L; the mass ratio of the sodium bicarbonate in the sodium bicarbonate solution to the pectin calcium pretreatment solution is 1: 15-20.
Preferably, the lignan is extracted from flax.
Preferably, in step 2, the support material is used in an amount of 50 g.
Preferably, the temperature of the sterilization treatment in the step 2 is 118-125 ℃, and the sterilization time is at least 30 min.
Preferably, based on experimental data, the peatmoss water absorption is 100%, so that the carrier material is mixed with the bacterial suspension at the same mass.
Preferably, the room temperature environment is 25-35 ℃.
In the step 2, the immobilized carrier turfy soil is sterilized, so that other bacteria are not introduced in the later process, the activity of aerobic denitrifying bacteria is influenced, and even the effect of the microbial inoculum on treating nitrogen-containing sewage is influenced.
The invention has the beneficial effects that:
the invention adopts the adsorption method to directly load the bacterial strain, thereby avoiding complex manufacturing steps and having simple and convenient operation process. The No. 144 strain provided by the invention is aerobic denitrifying bacteria, is obtained by enrichment screening through a biological culture means, has high-efficiency aerobic denitrification performance, and can obviously remove total nitrogen in black and odorous water under aerobic conditions. The immobilized carrier is activated turfy soil, the adaptation ratio of the activated turfy soil to the bacterial liquid is 1:1, a water absorption rate test reflects that the water absorption rate of the carrier is 100%, and the mixing ratio of the carrier material to the bacterial liquid is determined according to the water absorption rate test.
The turfy soil is used as an immobilized carrier, is soft in texture, easy to break, low in price and easy to obtain, but has weaker water absorption performance, particularly the water absorption of dry turfy soil is poorer, and the turfy soil can float on the water surface in sewage directly serving as a carrier, so that the sewage treatment capacity of later-stage microorganisms is influenced. According to research, the factors influencing the water absorption of the turfy soil are mainly the content of organic matters, the porosity and the specific surface area of the organic matters, the turfy soil contains more minerals and less organic matters, and has subacid or neutral reaction and low water absorption, so that the turfy soil is activated to improve the water absorption, the activated treatment process is to mix modified calcium pectin with high organic matter content and large specific surface area into the turfy soil, and the water absorption of the activated turfy soil is greatly improved by detecting that the water absorption of the turfy soil is about 100%.
The growth speed of the microorganism in the logarithmic growth phase is the maximum, the number of cells is increased exponentially, and the selection of the bacterial suspension in the logarithmic growth phase can ensure that the prepared immobilized microbial inoculum has high growth speed and high biological activity.
The immobilized microbial agent prepared by the invention effectively solves the problem that the strain adapts to adverse environment, and has obvious effect on removing total nitrogen.
Drawings
The invention is further illustrated by means of the attached drawings, but the embodiments in the drawings do not constitute any limitation to the invention, and for a person skilled in the art, other drawings can be obtained on the basis of the following drawings without inventive effort.
FIG. 1 is a graph showing the results of the degradation of nitrate nitrogen by a denitrifying microbial preparation prepared in comparative example 1 of the present invention;
FIG. 2 is a graph showing the results of the degradation of nitrate nitrogen by the denitrifying microbial preparation prepared in comparative example 2 of the present invention;
FIG. 3 is a graph showing the results of degradation of nitrate nitrogen by the denitrifying microbial preparation prepared in comparative example 3 of the present invention;
FIG. 4 is a graph showing the results of degradation of nitrate nitrogen by the denitrifying microbial preparation prepared in comparative example 4 of the present invention;
FIG. 5 is a graph showing the results of degradation of ammonia nitrogen-simulated wastewater in Experimental example 1 of the present invention;
FIG. 6 is a graph showing the results of degradation of nitrate nitrogen-simulated wastewater in Experimental example 2 of the present invention;
FIG. 7 is a graph showing the results of degradation of ammonia nitrogen-simulated wastewater in Experimental example 3 of the present invention;
FIG. 8 is a graph showing the results of degradation of nitrate nitrogen-simulated wastewater in Experimental example 4 of the present invention.
Detailed Description
For the purpose of more clearly illustrating the present invention and more clearly understanding the technical features, objects and advantages of the present invention, the technical solutions of the present invention will now be described in detail below, but are not to be construed as limiting the implementable scope of the present invention.
The activated turfy soil is obtained by mixing and homogenizing turfy soil and modified calcium pectin, and the modified calcium pectin contains more organic matters and has a larger specific surface area, so that the water absorption of the turfy soil and the dispersibility of the turfy soil in water are improved to a greater extent after the modified calcium pectin is combined with the turfy soil, the turfy soil can exist in water in a suspended manner, and the high dispersibility improves the sewage treatment capability of microorganisms in water. The preparation process of the modified calcium pectin is characterized in that lignans are added in the synthesis process of the calcium pectin, the pectin belongs to polysaccharide, the surface of the pectin contains a large number of hydrophilic groups and can be dissolved after swelling in water, the lignans (Lignan) are also called lignans and are polyphenol compounds naturally existing in some plants, the lignans and the pectin can be combined to form Lignan glycosides, and the existence of the Lignan glycosides not only increases the hydrophilicity of the lignans, but also weakens the solubility of the pectin in water, so that the pectin can keep a gel state in the water.
After the modified calcium pectin is combined with the turfy soil, the modified calcium pectin has the advantages that firstly, the modified calcium pectin has richer hydrophilic groups and is easier to disperse in water; secondly, the turfy soil is light in weight and easy to float in water, can sink underwater due to increased density after being combined with the modified calcium pectin, and can exist in a suspended state in water based on the hydrophilicity of the modified calcium pectin; thirdly, pectin and lignan in the modified calcium pectin can also provide a carbon source for microorganisms, so that the activity of the microorganisms is stronger.
The invention is further described below with reference to the following examples.
Example 1
A high-efficiency denitrifying microbial preparation for sewage treatment comprises aerobic denitrifying bacteria No. 144 bacteria and a carrier material.
The preparation method of the high-efficiency denitrification microbial preparation comprises the following steps:
step 1, culturing strains:
inoculating aerobic denitrifying bacteria No. 144 strain from the inclined plane to 150mL denitrifying culture medium by using an inoculating loop, culturing in a shaking table with the temperature of 30 ℃ and the rotating speed of 150rpm, and culturing to the logarithmic growth phase to obtain bacterial suspension with the OD600 of 0.613; wherein the formula (DM) of the denitrification culture medium comprises: distilled water, 6g/L K2HPO41.5g/L KH2PO40.1g/L MgSO4·7H2O, KNO of 0.6g/L35.64g/L sodium succinate and 0.1mL/L trace elements; the microelements comprise at least one of calcium agent, ferrum agent, zinc agent, selenium agent, and chromium agent; the judgment of the logarithmic growth phase is to adopt an ultraviolet spectrophotometer to measure the absorbance value of the bacterial liquid at the wavelength of 600nm, the absorbance value is recorded as OD600, the OD600 value verifies the content of the strains in the bacterial liquid, and then whether the bacterial liquid is in the logarithmic growth phase is controlled;
step 2, preparation of a denitrifying microbial preparation:
weighing a carrier material in a beaker, sealing the beaker by using tin foil paper, and then sterilizing the beaker in an autoclave at the temperature of 118-125 ℃ for at least 30 min; and then under the aseptic condition, adding 50g of sterilized carrier material into 50g of bacterial suspension cultured to the logarithmic phase in the step 1, mixing until the carrier material is completely wetted, loose, not adhered to the wall and not aggregated, filling into an aseptic container, and drying at the room temperature of 25-35 ℃ to obtain the denitrifying microbial preparation.
In the step 2, the carrier material is activated turfy soil, the activated turfy soil is obtained by mixing modified calcium pectin into turfy soil and homogenizing, and the mixing amount of the modified calcium pectin is 20% of the mass of the turfy soil.
The preparation method of the modified calcium pectin comprises the following steps:
a. weighing lignans extracted from flax, mixing the lignans with an ethanol solution with the mass fraction of 30-70%, and stirring uniformly to obtain a lignan solution; wherein the mass ratio of the lignans to the ethanol solution is 1: 12;
b. weighing pectin, adding into the lignan solution, stirring and mixing uniformly to obtain a lignan/pectin mixed solution; wherein the mass ratio of pectin to the lignan solution is 1: 15;
c. dropwise adding a calcium hydroxide solution into the lignan/pectin mixed solution, continuously stirring at the speed of 500-600 rpm in the dropwise adding process, and continuously stirring for 1-2 hours after dropwise adding to obtain a calcium pectin pretreatment solution; wherein the concentration of the calcium hydroxide solution is 0.03mol/L, and the mass ratio of the calcium hydroxide in the calcium hydroxide solution to the mixed liquid of the lignan and the pectin is 1: 12;
d. weighing sodium bicarbonate solution, continuously dropwise adding the sodium bicarbonate solution into the calcium pectin pretreatment solution, stirring for 2-4 h after dropwise adding, decompressing the reaction solution to remove ethanol, standing in a refrigerating chamber at 3-10 ℃ for 5-8 h, filtering while cold, collecting solid, and freeze-drying to obtain modified calcium pectin; wherein the concentration of the sodium bicarbonate solution is 0.01 mol/L; the mass ratio of the sodium bicarbonate in the sodium bicarbonate solution to the pectin calcium pretreatment solution is 1: 18.
Example 2
A high-efficiency denitrifying microbial preparation for sewage treatment comprises aerobic denitrifying bacteria No. 144 bacteria and a carrier material.
The preparation method of the high-efficiency denitrification microbial preparation comprises the following steps:
step 1, culturing strains:
inoculating aerobic denitrifying bacteria No. 144 strain from the inclined plane to 150mL denitrifying culture medium by using an inoculating loop, culturing in a shaking table with the temperature of 30 ℃ and the rotating speed of 150rpm, and culturing to the logarithmic phase to obtain bacterial suspension with the OD600 of 0.60; wherein the formula (DM) of the denitrification culture medium comprises: distilled water, 6g/L K2HPO41.5g/L KH2PO40.1g/L MgSO4·7H2O, KNO of 0.6g/L35.64g/L sodium succinate and 0.1mL/L trace elements; the microelements comprise at least one of calcium agent, ferrum agent, zinc agent, selenium agent, and chromium agent; the judgment of the logarithmic growth phase is to adopt an ultraviolet spectrophotometer to measure the absorbance value of the bacterial liquid at the wavelength of 600nm, the absorbance value is recorded as OD600, the OD600 value verifies the content of the strains in the bacterial liquid, and then whether the bacterial liquid is in the logarithmic growth phase is controlled;
step 2, preparation of a denitrifying microbial preparation:
weighing a carrier material in a beaker, sealing the beaker by using tin foil paper, and then sterilizing the beaker in an autoclave at the temperature of 118-125 ℃ for at least 30 min; and then under the aseptic condition, adding 50g of sterilized carrier material into 50g of bacterial suspension cultured to the logarithmic phase in the step 1, mixing until the carrier material is completely wetted, loose, not adhered to the wall and not aggregated, filling into an aseptic container, and drying at the room temperature of 25-35 ℃ to obtain the denitrifying microbial preparation.
In the step 2, the carrier material is activated turfy soil, the activated turfy soil is obtained by mixing modified calcium pectin into turfy soil and homogenizing, and the mixing amount of the modified calcium pectin is 10% of the mass of the turfy soil.
The preparation method of the modified calcium pectin comprises the following steps:
a. weighing lignans extracted from flax, mixing the lignans with an ethanol solution with the mass fraction of 30-70%, and stirring uniformly to obtain a lignan solution; wherein the mass ratio of the lignans to the ethanol solution is 1: 10;
b. weighing pectin, adding into the lignan solution, stirring and mixing uniformly to obtain a lignan/pectin mixed solution; wherein the mass ratio of pectin to the lignan solution is 1: 12;
c. dropwise adding a calcium hydroxide solution into the lignan/pectin mixed solution, continuously stirring at the speed of 500-600 rpm in the dropwise adding process, and continuously stirring for 1-2 hours after dropwise adding to obtain a calcium pectin pretreatment solution; wherein the concentration of the calcium hydroxide solution is 0.02mol/L, and the mass ratio of the calcium hydroxide in the calcium hydroxide solution to the mixed liquid of the lignan and the pectin is 1: 10;
d. weighing sodium bicarbonate solution, continuously dropwise adding the sodium bicarbonate solution into the calcium pectin pretreatment solution, stirring for 2-4 h after dropwise adding, decompressing the reaction solution to remove ethanol, standing in a refrigerating chamber at 3-10 ℃ for 5-8 h, filtering while cold, collecting solid, and freeze-drying to obtain modified calcium pectin; wherein the concentration of the sodium bicarbonate solution is 0.01 mol/L; the mass ratio of the sodium bicarbonate in the sodium bicarbonate solution to the pectin calcium pretreatment solution is 1: 15.
Example 3
A high-efficiency denitrifying microbial preparation for sewage treatment comprises aerobic denitrifying bacteria No. 144 bacteria and a carrier material.
The preparation method of the high-efficiency denitrification microbial preparation comprises the following steps:
step 1, culturing strains:
inoculating the aerobic denitrifying bacteria No. 144 strain to 150mL of denitrifying culture medium from the inclined plane by using an inoculating loop, culturing in a shaking table with the temperature of 30 ℃ and the rotating speed of 150rpm, and culturing to the logarithmic phase to obtain a bacterial suspension with the OD600 of 0.63; wherein the formula (DM) of the denitrification culture medium comprises: distilled water, 6g/L K2HPO41.5g/L KH2PO40.1g/L MgSO4·7H2O, KNO of 0.6g/L35.64g/L sodium succinate and 0.1mL/L trace elements; the microelements comprise at least one of calcium agent, ferrum agent, zinc agent, selenium agent, and chromium agent; the judgment of the logarithmic growth phase is to adopt an ultraviolet spectrophotometer to measure the absorbance value of the bacterial liquid at the wavelength of 600nm, the absorbance value is recorded as OD600, the OD600 value verifies the content of the strains in the bacterial liquid, and then whether the bacterial liquid is in the logarithmic growth phase is controlled;
step 2, preparation of a denitrifying microbial preparation:
weighing a carrier material in a beaker, sealing the beaker by using tin foil paper, and then sterilizing the beaker in an autoclave at the temperature of 118-125 ℃ for at least 30 min; and then under the aseptic condition, adding 50g of sterilized carrier material into 50g of bacterial suspension cultured to the logarithmic phase in the step 1, mixing until the carrier material is completely wetted, loose, not adhered to the wall and not aggregated, filling into an aseptic container, and drying at the room temperature of 25-35 ℃ to obtain the denitrifying microbial preparation.
In the step 2, the carrier material is activated turfy soil, the activated turfy soil is obtained by mixing modified calcium pectin into turfy soil and homogenizing, and the mixing amount of the modified calcium pectin is 30% of the mass of the turfy soil.
The preparation method of the modified calcium pectin comprises the following steps:
a. weighing lignans extracted from flax, mixing the lignans with an ethanol solution with the mass fraction of 30-70%, and stirring uniformly to obtain a lignan solution; wherein the mass ratio of the lignans to the ethanol solution is 1: 15;
b. weighing pectin, adding into the lignan solution, stirring and mixing uniformly to obtain a lignan/pectin mixed solution; wherein the mass ratio of pectin to the lignan solution is 1: 18;
c. dropwise adding a calcium hydroxide solution into the lignan/pectin mixed solution, continuously stirring at the speed of 500-600 rpm in the dropwise adding process, and continuously stirring for 1-2 hours after dropwise adding to obtain a calcium pectin pretreatment solution; wherein the concentration of the calcium hydroxide solution is 0.05mol/L, and the mass ratio of the calcium hydroxide in the calcium hydroxide solution to the mixed liquid of the lignan and the pectin is 1: 15;
d. weighing sodium bicarbonate solution, continuously dropwise adding the sodium bicarbonate solution into the calcium pectin pretreatment solution, stirring for 2-4 h after dropwise adding, decompressing the reaction solution to remove ethanol, standing in a refrigerating chamber at 3-10 ℃ for 5-8 h, filtering while cold, collecting solid, and freeze-drying to obtain modified calcium pectin; wherein the concentration of the sodium bicarbonate solution is 0.02 mol/L; the mass ratio of the sodium bicarbonate in the sodium bicarbonate solution to the pectin calcium pretreatment solution is 1: 20.
In order to more clearly illustrate the present invention, the following experiments were also performed.
Four denitrifying microorganism preparations were prepared by compounding activated peatmoss, rice bran, zeolite, activated carbon prepared in example 1 as carrier materials with aerobic denitrifying bacteria No. 144, wherein the addition of aerobic denitrifying bacteria No. 144 was also compared in 5 steps, i.e., the OD600 of the bacterial liquid was 1/4, the OD600 of the bacterial liquid was 1/2, the OD600 of the bacterial liquid was 0.613, the OD600 of the bacterial liquid was 2 times the original OD600, and the OD600 of the bacterial liquid was 4 times the original OD 600.
Comparative example 1
A denitrifying microorganism preparation comprises aerobic denitrifying bacterium No. 144 and a carrier material, wherein the carrier material is activated turfy soil. The denitrifying microorganism preparation was subjected to a degradation test of nitrate nitrogen, and the results are shown in FIG. 1.
The preparation method of the denitrifying microorganism preparation is the same as that of the embodiment 1, and the differences are that:
the preparation of the bacterial liquid is set to 5 concentrations: the OD600 of the bacterial liquid is 1/4 (FIG. 1, charcoal soil 1) of the original OD600, the OD600 of the bacterial liquid is 1/2 (FIG. 1, charcoal soil 2) of the original OD600, the OD600 of the original bacterial liquid is 0.613 (FIG. 1, charcoal soil 3), the OD600 of the bacterial liquid is 2 times (FIG. 1, charcoal soil 4) of the original OD600, and the OD600 of the bacterial liquid is 4 times (FIG. 1, charcoal soil 5) of the original OD 600.
Comparative example 2
A denitrifying microorganism preparation comprises aerobic denitrifying bacterium No. 144 and carrier material. The denitrifying microorganism preparation was subjected to a degradation test of nitrate nitrogen, and the results are shown in FIG. 2.
The preparation method of the denitrifying microorganism preparation is the same as that of example 1, except that:
the carrier material is rice bran, the water absorption of the rice bran is about 150%, and the preparation of the bacterial liquid is set to be 5 concentrations: the OD600 of the bacterial liquid was 1/4 (rice bran 1 in fig. 2) which was the original OD600, 1/2 (rice bran 2 in fig. 2) which was the original OD600, 0.613 (rice bran 3 in fig. 2) which was the original bacterial liquid, 2 times (rice bran 4 in fig. 2) the OD600 of the bacterial liquid which was the original OD600, and 4 times (rice bran 5 in fig. 2) the OD600 of the bacterial liquid which was the original OD 600.
Comparative example 3
A denitrifying microorganism preparation comprises aerobic denitrifying bacterium No. 144 and carrier material. The denitrifying microorganism preparation was subjected to a degradation test of nitrate nitrogen, and the results are shown in FIG. 3.
The preparation method of the denitrifying microorganism preparation is the same as that of example 1, except that:
the carrier material is zeolite, the water absorption of rice bran is about 30%, and the preparation of bacteria liquid is set to 5 concentrations: the OD600 of the bacterial liquid was 1/4 (zeolite 1 in fig. 3) of the original OD600, the OD600 of the bacterial liquid was 1/2 (zeolite 2 in fig. 3) of the original OD600, the OD600 of the original bacterial liquid was 0.613 (zeolite 3 in fig. 3), the OD600 of the bacterial liquid was 2 times the original OD600 (zeolite 4 in fig. 3), and the OD600 of the bacterial liquid was 4 times the original OD600 (zeolite 5 in fig. 3).
Comparative example 4
A denitrifying microorganism preparation comprises aerobic denitrifying bacterium No. 144 and carrier material. The denitrifying microorganism preparation was subjected to a degradation test of nitrate nitrogen, and the results are shown in FIG. 4.
The preparation method of the denitrifying microorganism preparation is the same as that of example 1, except that:
the carrier material is activated carbon, the water absorption of the activated carbon is about 75%, and the preparation of the bacteria liquid is set to 5 concentrations: the OD600 of the bacterial suspension was 1/4 (activated carbon 1 in fig. 4) of the original OD600, 1/2 (activated carbon 2 in fig. 4) of the original OD600, 0.613 (activated carbon 3 in fig. 4) of the original OD600, 2 times (activated carbon 4 in fig. 4) of the OD600 of the bacterial suspension and 4 times (activated carbon 5 in fig. 4) of the OD600 of the bacterial suspension.
As can be seen from FIGS. 1 to 4, the immobilized microbial inoculum provided by the invention has a better denitrification effect by using the activated turfy soil (FIG. 1) as a carrier, wherein the degradation rate of the microbial inoculum prepared from a bacterial solution with the bacterial growth (OD600) of 0.613 which is 4 times of the original OD600 is better, and the immobilized microbial inoculum has a good denitrification effect.
Attached: the detection method of the water absorption rate comprises the following steps:
the microbial inoculum is prepared by adopting an adsorption immobilization technology which takes rice bran, unactivated turfy soil, activated turfy soil, zeolite and activated carbon as carriers. In order to make the bacterial agents made of the four carriers completely adsorb bacterial liquid, the water absorption of the five carrier materials needs to be measured in advance. Weighing 10g of each of five carrier materials of rice bran, unactivated turfy soil, activated turfy soil, zeolite and activated carbon, putting the carrier materials into a beaker, adding bacterial liquid according to a certain gradient volume, thoroughly mixing the bacterial solution and the carrier until the carrier materials are soft, moist, not adhered to the wall of the beaker and not aggregated, and measuring for three times and then taking an average value. The water absorption rate reflects the adsorption capacity of different carriers, and the mixing ratio of the carrier material and the bacterial liquid is determined according to the water absorption rate.
Experiments show that the water absorption of the rice bran is about 150%, the water absorption of the unactivated turfy soil is about 20%, the water absorption of the activated turfy soil is about 100%, the water absorption of the zeolite is about 30%, and the water absorption of the activated carbon is about 75%.
Experimental example 1
0.05g of the denitrifying microorganism preparation prepared in example 1 was added to ammonia nitrogen simulated wastewater, and then samples were taken at regular intervals to determine changes of ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in the simulated wastewater. The ammonia nitrogen simulated wastewater comprises the following formula: sodium succinate 5.64g/L, ammonium chloride 0.35g/L, K2HPO4 6g/L，KH2PO4 1.5g/L，MgSO4·7H20.1g/L of O, 0.1mL/L of trace elements and pH7.5, and sterilizing at 121 ℃ for 30min before adding the denitrifying microorganism preparation. As shown in FIG. 5, it can be seen that the nitrogen concentration in the ammonia nitrogen is reduced to less than 10mg/L at about 30 hours, which indicates that the denitrifying microorganism preparation prepared in example 1 of the present invention has a better ammonia nitrogen degradation effect, and can still maintain a lower nitrogen concentration at 80 hours, which indicates that example 1 has a better effect of maintaining water quality.
Experimental example 2
0.05g of the denitrifying microorganism preparation prepared in example 1 was added to nitrate nitrogen-simulating wastewater, and samples were taken at regular intervals to determine changes in ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in the simulating wastewater. The results are shown in FIG. 6. The formula of the nitrate nitrogen simulated wastewater is as follows: sodium succinate 5.64g/L, KNO3 0.6g/L，K2HPO4 6g/L，KH2PO4 1.5g/L，MgSO4·7H20.1g/L of O, 0.1mL/L of trace elements and pH7.5, and sterilizing at 121 ℃ for 30min before adding the denitrifying microorganism preparation. As a result, as shown in FIG. 6, it can be seen that the nitrogen concentration in the nitrate nitrogen had decreased to less than 10mg/L at about 30 hours, indicating the denitrifying microorganisms prepared in example 1 of the present inventionThe preparation has better nitrate nitrogen degradation effect, and can still keep lower nitrogen concentration at 80 th hour, which shows that the preparation of example 1 has better effect of keeping water quality.
Experimental example 3
The activated peat from example 1 was replaced with non-activated peat as a carrier material, and then a denitrification microbial preparation was prepared according to the method of example 1, and the microbial preparation was placed in ammonia nitrogen simulation wastewater (as shown in fig. 7). The ammonia nitrogen simulated wastewater comprises the following formula: sodium succinate 5.64g/L, ammonium chloride 0.35g/L, K2HPO4 6g/L，KH2PO41.5g/L，MgSO4·7H20.1g/L of O, 0.1mL/L of trace elements and pH7.5, and sterilizing at 121 ℃ for 30min before adding the denitrifying microorganism preparation. As a result, it was found that the nitrogen concentration in the ammonia nitrogen decreased to about 10mg/L at about 50 hours, indicating that the degradation of the ammonia nitrogen by the denitrifying microorganism preparation using the unactivated peatmoss was inferior to that of example 1, and that nitrite and nitrate nitrogen fluctuated during this period, indicating that the denitrifying microorganism preparation had insufficient activity during this process.
Experimental example 4
The activated turfy carbon in example 1 was replaced with non-activated turfy soil as a carrier material, and then a denitrifying microbial preparation was prepared according to the method of example 1, and the microbial preparation was placed in nitrate nitrogen simulated wastewater (as shown in fig. 8), wherein the formulation of the nitrate nitrogen simulated wastewater was as follows: sodium succinate 5.64g/L, KNO3 0.6g/L，K2HPO4 6g/L，KH2PO4 1.5g/L，MgSO4·7H20.1g/L of O, 0.1mL/L of trace elements and pH7.5, and sterilizing at 121 ℃ for 30min before adding the denitrifying microorganism preparation. As a result, it was found that the nitrogen concentration in the nitrate nitrogen was reduced to about 10mg/L at about 50 hours, indicating that the nitrate nitrogen degradation effect of the denitrifying microorganism preparation using the unactivated peatmoss was inferior to that of example 1, and that nitrite nitrogen and ammonia nitrogen fluctuated during this period, indicating that the denitrifying microorganism preparation had insufficient activity during this process.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Figure BDA0003214262170000111
Figure BDA0003214262170000121

Claims (10)

1. The efficient denitrification microbial preparation for sewage treatment is characterized in that an aerobic denitrifying bacterium strain No. 144 is loaded on a carrier material by adopting an adsorption method, the preservation number of the aerobic denitrifying bacterium strain No. 144 is CGMCC No.22897, and the carrier material is activated turfy soil.
2. The efficient denitrifying microbial agent for wastewater treatment according to claim 1, wherein the activated peatmoss is mixed with modified calcium pectin and homogenized, and the mixing amount of the modified calcium pectin is 10-30% of the mass of the peatmoss.
3. The efficient denitrifying microbial preparation for sewage treatment according to claim 2, wherein the preparation method of the modified calcium pectin is as follows:
a. weighing lignans, mixing the lignans with 30-70% ethanol solution by mass percent, and stirring uniformly to obtain lignan solution; wherein the mass ratio of the lignans to the ethanol solution is 1: 10-15;
b. weighing pectin, adding into the lignan solution, stirring and mixing uniformly to obtain a lignan/pectin mixed solution; wherein the mass ratio of pectin to the lignan solution is 1: 12-18;
c. dropwise adding a calcium hydroxide solution into the lignan/pectin mixed solution, continuously stirring at the speed of 500-600 rpm in the dropwise adding process, and continuously stirring for 1-2 hours after dropwise adding to obtain a calcium pectin pretreatment solution; wherein the concentration of the calcium hydroxide solution is 0.02-0.05 mol/L, and the mass ratio of calcium hydroxide in the calcium hydroxide solution to the lignan/pectin mixed solution is 1: 10-15;
d. weighing sodium bicarbonate solution, continuously dropwise adding the sodium bicarbonate solution into the calcium pectin pretreatment solution, stirring for 2-4 h after dropwise adding, decompressing the reaction solution to remove ethanol, standing in a refrigerating chamber at 3-10 ℃ for 5-8 h, filtering while cold, collecting solid, and freeze-drying to obtain modified calcium pectin; wherein the concentration of the sodium bicarbonate solution is 0.01-0.02 mol/L; the mass ratio of the sodium bicarbonate in the sodium bicarbonate solution to the pectin calcium pretreatment solution is 1: 15-20.
4. The method for preparing the high-efficiency denitrifying microorganism preparation for sewage treatment according to claim 1, characterized by comprising the following steps:
step 1, culturing strains:
inoculating the aerobic denitrifying bacteria No. 144 strain into a denitrifying culture medium from an inclined plane by using an inoculating ring, culturing in a shaking table, and culturing to a logarithmic phase to obtain a bacterial suspension;
step 2, preparation of a denitrifying microbial preparation:
weighing carrier materials in a beaker, sealing with tin foil paper, and sterilizing in an autoclave; and then under the aseptic condition, adding the sterilized carrier material into the bacterial suspension cultured to the logarithmic phase in the step 1, mixing until the carrier material is completely wetted, filling into an aseptic container, and drying at room temperature to obtain the denitrifying microorganism preparation.
5. The method for preparing a high efficiency denitrifying microorganism preparation for wastewater treatment according to claim 4, wherein, in step 1, the temperature of the shaking table is 30 ℃ and the rotation speed is 150 rpm.
6. The method for preparing a high efficiency denitrifying microorganism preparation for wastewater treatment according to claim 4, wherein the volume of the denitrifying culture medium in step 1 is 150 mL.
7. The method for preparing a high efficiency denitrifying microorganism preparation for wastewater treatment according to claim 4, wherein in step 1, the formulation of the denitrifying culture medium includes: distilled water, 6g/L K2HPO41.5g/L KH2PO40.1g/L MgSO4·7H2O, KNO of 0.6g/L35.64g/L sodium succinate and 0.1mL/L trace elements.
8. The method for preparing the efficient denitrifying microbial preparation for wastewater treatment according to claim 4, wherein in step 1, the pH of the culture medium is adjusted to 7.5 after the preparation is completed, and the culture medium is sterilized at a temperature of 118-125 ℃ for at least 30 min.
9. The method for preparing the high-efficiency denitrifying microorganism preparation for wastewater treatment according to claim 7, wherein the trace elements include at least one of calcium agent, iron agent, zinc agent, selenium agent and chromium agent.
10. The method for preparing the efficient denitrifying microbial preparation for sewage treatment according to claim 4, wherein the OD600 of the bacterial suspension is 0.60-0.63.
CN202110945679.1A 2021-08-16 2021-08-16 Efficient denitrification microbial preparation for sewage treatment and preparation method thereof Pending CN113755482A (en)

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Application publication date: 20211207