CN113337434A - Microbial agent for treating high-salinity wastewater and preparation method thereof - Google Patents
Microbial agent for treating high-salinity wastewater and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a microbial agent for treating high-salinity wastewater and a preparation method thereof, wherein the active ingredients of the microbial agent for treating the high-salinity wastewater comprise: the bacillus licheniformis, the bacillus subtilis, the microzyme, the lactobacillus and the pseudomonas stutzeri have the effective viable count ratio of 8: 2: 3: 3: 1; according to the invention, the bacillus licheniformis, the bacillus subtilis, the saccharomycetes, the lactobacillus and the pseudomonas stutzeri are acclimated to adapt to a high-salt environment, no protective agent is required to be added, and the high-salt environment cannot be inhibited or even poisoned by high salinity, so that the biomass is reduced, and the treatment efficiency of high-salt wastewater is reduced; and then, the bacillus licheniformis, the bacillus subtilis, the saccharomycetes, the lactobacillus and the pseudomonas stutzeri are used for synergistically removing organic matters, SS, ammonia nitrogen and salts in the high wastewater, so that the removal efficiency of TOC is improved.
Description
Technical Field
The invention relates to the field of industrial wastewater biochemical treatment, in particular to a microbial agent for treating high-salinity wastewater and a preparation method thereof.
Background
The high-salinity wastewater refers to total salt (such as Na)+、K+、Cl－And SO4 2－Etc.) waste water with the mass fraction more than or equal to 1 percent. In recent years, direct utilization of seawater (such as seawater used for flushing toilets, roads, fire fighting and industrial cooling water) is actively developed in coastal cities, and in 2020, the direct utilization amount of seawater in China reaches 1 × 109m3·a－1The rapid increase in the amount of direct seawater use is a significant cause of the large discharge of high salinity wastewater. At present, the most main treatment mode of high-salinity wastewater is to enter a municipal sewage treatment system through a municipal pipe network. High salinity wastewater typically contains Na+、K+、Cl－And SO4 2－And the like, and the high concentration of the ions can quickly increase the osmotic pressure of cells to destroy the somatic cells, and simultaneously produce salting-out action to reduce the activity of dehydrogenase to inhibit the growth of bacteria, and the high concentration of chloride ions has certain toxic action on the bacteria. Therefore, the high-salinity wastewater can generate obvious inhibiting effect on the traditional biological sewage treatment process, and particularly in the starting period of the biological sewage treatment process, the inhibition of the activated sludge can cause the quality deterioration of process effluent and the sharp increase of COD and SS concentration; the biggest problem of the biological process for treating the high-salinity wastewater is that the metabolic function of microorganisms is damaged, the number of primary and secondary animals is greatly reduced, and the removal efficiency of TOC is low.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a microbial agent for treating high-salinity wastewater and a preparation method thereof, so that the removal efficiency of TOC is greatly improved.
The purpose of the invention is realized by the following technical scheme: the microbial agent for treating the high-salinity wastewater comprises active ingredients, wherein the active ingredients comprise: the bacillus licheniformis, the bacillus subtilis, the microzyme, the lactobacillus and the pseudomonas stutzeri have the effective viable count ratio of 8: 2: 3: 3: 1.
further, the total effective viable count of the active ingredients is 1 × 1010-1×1012CFU/ml。
Further, the microbial inoculum also comprises a culture medium mixed with the active ingredients, and the volume of the microbial inoculum is 1-3% of the volume of the culture medium.
Furthermore, the formula of the culture medium is 30-40g of glucose, 10-16g of tryptone, 50-63g of sodium chloride, 15-18g of agar and 4-8gKH2PO4And 1L of water.
The preparation method of the microbial agent for treating the high-salinity wastewater comprises the following steps
S1: respectively culturing the domesticated bacillus licheniformis, bacillus subtilis, saccharomycetes, lactobacillus and pseudomonas stutzeri to a plateau stage;
s2: preparing the culture medium, uniformly mixing the raw materials for preparing the culture medium according to a proportion, adjusting the pH value to be 6.0-7.2, sterilizing at the temperature of 110-;
s3: uniformly mixing the bacillus licheniformis, the bacillus subtilis, the yeast, the lactobacillus and the pseudomonas stutzeri which are cultured to a plateau stage according to a proportion, then adding the mixture into the culture medium at 37 ℃, wherein the volume of the microbial inoculum is 1-3% of the volume of the culture medium, and culturing for 68-75h after uniform mixing.
The acclimatization step comprises the steps of respectively putting the bacillus licheniformis, the bacillus subtilis, the microzyme, the lactobacillus and the pseudomonas stutzeri into LB liquid culture medium with the salinity of 6 percent, and carrying out acclimatization at the temperature of 25 ℃ and at the temperature of 150 r.min－1Cultured under the conditions of (1), and OD was measured every 2 hours600When OD of the strain600Respectively inoculating strains 1 to LB liquid culture medium with salinity of 8% at 25 deg.C and 150r min to obtain strains 1 respectively－1Under the conditions ofCulturing, determining OD every 2h600When OD of the strain600When the values respectively reach 1.2-1.6, the domesticated bacillus licheniformis, bacillus subtilis, saccharomycete, lactobacillus and pseudomonas stutzeri are obtained.
According to the invention, the bacillus licheniformis, the bacillus subtilis, the saccharomycetes, the lactobacillus and the pseudomonas stutzeri are acclimated to adapt to a high-salt environment, no protective agent is required to be added, and the high-salt environment cannot be inhibited or even poisoned by high salinity, so that the biomass is reduced, and the treatment efficiency of high-salt wastewater is reduced; and then, the bacillus licheniformis, the bacillus subtilis, the saccharomycetes, the lactobacillus and the pseudomonas stutzeri are used for synergistically removing organic matters, SS, ammonia nitrogen and salts in the high wastewater, so that the removal efficiency of TOC is improved.
Further, the formula of the LB liquid culture medium is tryptone 10g, NaCl50g, agar 15g, glucose 20g and water 1L.
Further, the inoculation amounts of the bacillus licheniformis, the bacillus subtilis, the yeast, the lactobacillus and the pseudomonas stutzeri are all 1.2-3.3% of the volume of the LB liquid culture medium.
The invention has the following advantages: no protective agent is added, and active ingredients in the microbial inoculum are not damaged in a high-salt environment, so that the high-salt wastewater treatment efficiency is reduced; meanwhile, the TOC treatment efficiency of the high-salinity wastewater is improved under the synergistic effect of the bacillus licheniformis, the bacillus subtilis, the saccharomycetes, the lactobacillus and the pseudomonas stutzeri; and the application range is wide, and the method is suitable for both aerobic environment and anaerobic environment.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
Example 1
The preparation method of the microbial agent for treating the high-salinity wastewater comprises the following steps
S11: the bacillus licheniformis and the bacillus subtilis are mixedThe yeast, the lactobacillus and the pseudomonas stutzeri are respectively put into an LB liquid culture medium with the salinity of 6% (the formula of the LB liquid culture medium comprises 10g of tryptone, 50g of NaCl, 15g of agar, 20g of glucose and 1L of water), and the inoculation amounts of the bacillus licheniformis, the bacillus subtilis, the yeast, the lactobacillus and the pseudomonas stutzeri are respectively 1.2% of the volume of the LB liquid culture medium; at 25 deg.C and 150r min－1Cultured under the conditions of (1), and OD was measured every 2 hours600When OD of the strain600Respectively inoculating strains 1 with salt content of 8% in LB liquid culture medium at 25 deg.C and 150r min to obtain strains 1－1Cultured under the conditions of (1), and OD was measured every 2 hours600When OD of the strain600When the values respectively reach 1.2, obtaining the domesticated bacillus licheniformis, bacillus subtilis, saccharomycetes, lactobacillus and pseudomonas stutzeri;
s12: respectively culturing the domesticated bacillus licheniformis, bacillus subtilis, saccharomycetes, lactobacillus and pseudomonas stutzeri to a plateau stage;
s2: preparing the culture medium by mixing glucose 30g, tryptone 10g, sodium chloride 50g, agar 15g, 4gKH2PO4Uniformly mixing the raw materials of 1L of water for preparing the culture medium, adjusting the pH value to 6.0, sterilizing at 110 ℃ for 18min, and cooling to 37 ℃;
s3: and (3) culturing the bacillus licheniformis, the bacillus subtilis, the microzyme, the lactobacillus and the pseudomonas stutzeri which are cultured to a plateau stage according to the effective viable bacteria ratio of 8: 2: 3: 3: 1, uniformly mixing, and controlling the total effective viable count of the active ingredients to be 1 multiplied by 1010CFU/ml, then adding the mixture into the culture medium at 37 ℃, wherein the inoculation amount of the effective viable bacteria is 1 percent of the volume of the culture medium, and culturing for 68 hours after uniformly mixing.
Example 2
The preparation method of the microbial agent for treating the high-salinity wastewater comprises the following steps
S11: respectively putting the bacillus licheniformis, the bacillus subtilis, the microzyme, the lactobacillus and the pseudomonas stutzeri into an LB liquid culture medium with the salinity of 6 percent (the LBThe formula of the liquid culture medium comprises 10g of tryptone, 50g of NaCl50g, 15g of agar, 20g of glucose and 1L of water, wherein the inoculation amounts of the bacillus licheniformis, the bacillus subtilis, the yeast, the lactobacillus and the pseudomonas stutzeri are respectively 2.3 percent of the volume of the LB liquid culture medium; at 25 deg.C and 150r min－1Cultured under the conditions of (1), and OD was measured every 2 hours600When OD of the strain600When the values respectively reach 0.6, obtaining strains 1, respectively inoculating the strains 1 into LB liquid culture medium with salinity of 8%, at 25 deg.C and 150 r.min－1Cultured under the conditions of (1), and OD was measured every 2 hours600When OD of the strain600When the values respectively reach 1.5, obtaining the domesticated bacillus licheniformis, bacillus subtilis, saccharomycetes, lactobacillus and pseudomonas stutzeri;
s12: respectively culturing the domesticated bacillus licheniformis, bacillus subtilis, saccharomycetes, lactobacillus and pseudomonas stutzeri to a plateau stage;
s2: preparing the culture medium by mixing 37g of glucose, 13g of tryptone, 57g of sodium chloride, 16g of agar and 7gKH2PO4Uniformly mixing the raw materials of 1L of water for preparing the culture medium, adjusting the pH value to 6.8, sterilizing at 114 ℃ for 23min, and cooling to 37 ℃;
s3: and (3) culturing the bacillus licheniformis, the bacillus subtilis, the microzyme, the lactobacillus and the pseudomonas stutzeri which are cultured to a plateau stage according to the effective viable bacteria ratio of 8: 2: 3: 3: 1, uniformly mixing, and controlling the total effective viable count of the active ingredients to be 1 multiplied by 1011CFU/ml, then adding the mixture into the culture medium at 37 ℃, wherein the inoculation amount of the effective viable bacteria is 2% of the volume of the culture medium, and uniformly mixing and culturing for 72 hours.
Example 3
The preparation method of the microbial agent for treating the high-salinity wastewater comprises the following steps
S11: respectively putting the bacillus licheniformis, the bacillus subtilis, the microzyme, the lactobacillus and the pseudomonas stutzeri into an LB liquid culture medium with the salinity of 6% (the formula of the LB liquid culture medium comprises 10g of tryptone, 50g of NaCl, 50g, 15g of agar, 20g of glucose and 1L of water), and putting the bacillus licheniformis, the bacillus subtilis, the microzyme, the lactobacillus and the pseudomonas stutzeri into the LB liquid culture medium with the salinity of 6% (the formula of the LB liquid culture medium comprises 10g of tryptone, 50g of NaCl, g, 15g of agar, 20g of glucose and 1L of water)The inoculation amounts of bacillus, bacillus subtilis, yeast, lactobacillus and pseudomonas stutzeri are respectively 3.3 percent of the volume of the LB liquid culture medium; at 25 deg.C and 150r min－1Cultured under the conditions of (1), and OD was measured every 2 hours600When OD of the strain600When the values respectively reach 0.7, obtaining strains 1, respectively inoculating the strains 1 into LB liquid culture medium with salinity of 8%, at 25 deg.C and 150r min－1Cultured under the conditions of (1), and OD was measured every 2 hours600When OD of the strain600When the values respectively reach 1.6, obtaining the domesticated bacillus licheniformis, bacillus subtilis, saccharomycetes, lactobacillus and pseudomonas stutzeri;
s12: respectively culturing the domesticated bacillus licheniformis, bacillus subtilis, saccharomycetes, lactobacillus and pseudomonas stutzeri to a plateau stage;
s2: preparing the culture medium by mixing glucose 40g, tryptone 16g, sodium chloride 63g, agar 18g, 4-8gKH2PO4Uniformly mixing the raw materials of 1L of water for preparing the culture medium, adjusting the pH value to 7.2, sterilizing at 118 ℃ for 27min, and cooling to 37 ℃;
s3: and (3) culturing the bacillus licheniformis, the bacillus subtilis, the microzyme, the lactobacillus and the pseudomonas stutzeri which are cultured to a plateau stage according to the effective viable bacteria ratio of 8: 2: 3: 3: 1, uniformly mixing, and controlling the total effective viable count of the active ingredients to be 1 multiplied by 1012CFU/ml, then adding the mixture into the culture medium at 37 ℃, wherein the inoculation amount of the effective viable bacteria is 3% of the volume of the culture medium, and uniformly mixing and culturing for 75 h.
Comparative example 1:
the conventional microbial agent for treating high-salinity wastewater on the market is purchased, the same amount of high-salinity wastewater is treated by the microbial agent prepared in the comparative examples 1-3, the TOC removal rate of the treated high-salinity wastewater is detected, and the experimental data are shown in Table 1.
Comparative example 2:
after the existing microbial agent for treating high-salinity wastewater on the market, which is the same as that of the microbial agent for treating high-salinity wastewater in the comparative example 1, is purchased for acclimatization treatment, the same amount of high-salinity wastewater is treated by the microbial agent prepared in the comparative examples 1-3, the TOC removal rate of the treated high-salinity wastewater is detected, and the experimental data are shown in Table 1
Table 1: TOC value recording table
| Example 1 | Example 2 | Example 3 | Comparative example 1 | Comparative example 2 | |
| TOC removal (%) | 94.6 | 98.7 | 90.2 | 73.5 | 89.1 |
As shown in Table 1, the microbial agent for treating high-salinity wastewater has higher TOC removal efficiency of the high-salinity wastewater, which can reach 98.7% at most, compared with the existing microbial agent on the market.
The present invention is not limited to the above-described embodiments, and any variations, modifications, and substitutions which may occur to those skilled in the art may be made without departing from the spirit of the invention.
Claims (8)
1. The microbial agent for treating the high-salinity wastewater comprises active ingredients, and is characterized in that the active ingredients comprise: the bacillus licheniformis, the bacillus subtilis, the microzyme, the lactobacillus and the pseudomonas stutzeri have the effective viable count ratio of 8: 2: 3: 3: 1.
2. the microbial agent for treating high-salinity wastewater according to claim 1, wherein the total effective viable count of the active ingredients is 1 x 1010-1×1012CFU/ml。
3. The microbial inoculant for the treatment of high-salinity wastewater of claim 1, wherein: the microbial inoculum also comprises a culture medium mixed with the active ingredients, and the volume of the microbial inoculum is 1-3% of the volume of the culture medium.
4. The microbial inoculant for the treatment of high-salinity wastewater of claim 3, wherein: the formula of the culture medium comprises 30-40g of glucose, 10-16g of tryptone, 50-63g of sodium chloride, 15-18g of agar and 4-8gKH2PO4And 1L of water.
5. The method for preparing a microbial agent for treating high-salinity wastewater according to claim 1, characterized by comprising the following steps
S1: respectively culturing the domesticated bacillus licheniformis, bacillus subtilis, saccharomycetes, lactobacillus and pseudomonas stutzeri to a plateau stage;
s2: preparing the culture medium, uniformly mixing the raw materials for preparing the culture medium according to a proportion, adjusting the pH value to be 6.0-7.2, sterilizing at the temperature of 110-;
s3: uniformly mixing the bacillus licheniformis, the bacillus subtilis, the yeast, the lactobacillus and the pseudomonas stutzeri which are cultured to a plateau stage according to a proportion, then adding the mixture into the culture medium at 37 ℃, wherein the volume of the microbial inoculum is 1-3% of the volume of the culture medium, and culturing for 68-75h after uniform mixing.
6. The method of claim 1, wherein the acclimating step comprises respectively placing the Bacillus licheniformis, Bacillus subtilis, yeast, Lactobacillus, and Pseudomonas stutzeri in LB liquid medium with a salinity of 6%, and culturing at 25 deg.C and 150 r-min－1Cultured under the conditions of (1), and OD was measured every 2 hours600When OD of the strain600Respectively inoculating strains 1 to LB liquid culture medium with salinity of 8% at 25 deg.C and 150r min to obtain strains 1 respectively－1Cultured under the conditions of (1), and OD was measured every 2 hours600When OD of the strain600When the values respectively reach 1.2-1.6, the domesticated bacillus licheniformis, bacillus subtilis, saccharomycete, lactobacillus and pseudomonas stutzeri are respectively obtained.
7. The method for preparing a microbial inoculant for treating high-salinity wastewater according to claim 6, wherein the microbial inoculant comprises: the LB liquid medium comprises 10g of tryptone, 50g of NaCl50g, 15g of agar, 20g of glucose and 1L of water.
8. The method for preparing a microbial inoculant for treating high-salinity wastewater according to claim 6, wherein the microbial inoculant comprises: during acclimation, the inoculation amounts of the bacillus licheniformis, the bacillus subtilis, the saccharomycetes, the lactobacillus and the pseudomonas stutzeri are all 1.2-3.3% of the volume of the LB liquid culture medium.
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| CN117904011A (en) * | 2024-03-20 | 2024-04-19 | 江苏省环境工程技术有限公司 | Microbial agent for treating high-salt refractory industrial wastewater and preparation method and application thereof |
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| CN117904011A (en) * | 2024-03-20 | 2024-04-19 | 江苏省环境工程技术有限公司 | Microbial agent for treating high-salt refractory industrial wastewater and preparation method and application thereof |
| CN117904011B (en) * | 2024-03-20 | 2024-05-28 | 江苏省环境工程技术有限公司 | Microbial agent for treating high-salt refractory industrial wastewater and preparation method and application thereof |
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