CN108178337B - Activation promoter for improving microbial treatment effect of daily chemical wastewater - Google Patents
Activation promoter for improving microbial treatment effect of daily chemical wastewater Download PDFInfo
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Abstract
The invention belongs to the field of biological enhanced treatment of wastewater, and particularly relates to an activation promoter for improving the microbial treatment effect of daily chemical wastewater. The daily chemical wastewater has the characteristics of high organic matter, high salt, high biological toxicity and the like, and is difficult to reach the standard by using a conventional biological method for treatment. The invention provides an activation promoter by using chitosan, betaine, ferrous sulfate, trehalose, citric acid, proline and compound vitamins, can effectively improve the survival reproductive capacity and metabolic activity of microorganisms under the conditions of high organic matters and high salt, and effectively improves the sewage treatment effect. The invention has low cost and convenient manufacture, and can be practically applied to the biological treatment process of daily chemical wastewater and other high-salt surfactant wastewater.
Description
Technical Field
The invention belongs to the field of biological enhanced treatment of wastewater, and particularly relates to an activation promoter for improving the microbial treatment effect of wastewater in the daily chemical industry.
Background
The daily chemical wastewater refers to wastewater generated in the production process of daily chemical products (such as shampoos, hand washing solutions, laundry detergents and the like). The wastewater is mainly characterized by large discharge amount, high concentration of organic matters (mainly containing surfactants, including anionic surfactants, namely Sodium Dodecyl Sulfate (SDS) and nonionic surfactants, namely peregal) and large salt content. Without effective control and treatment, surfactants in the wastewater slowly accumulate in the environment and pose a hazard to the ecosystem.
The current reports on the treatment of high-salt-content daily chemical wastewater are rare, and the reported treatment methods mainly comprise a Fenton method (the Fenton method is used for treating the daily chemical wastewater with high-concentration sulfate) and Fenton method-contact oxidation method combined treatment (Fenton oxidation-aerobic contact process is used for treating LAS wastewater with high-concentration sulfate; and the high-concentration daily chemical wastewater treatment research). Currently, the treatment of high-salinity wastewater is mostly performed by a physical and chemical method. For example, the high-salinity wastewater can be effectively treated by means of evaporative crystallization (a high-salinity desulfurization wastewater advanced treatment method and system; a high-salinity wastewater low-temperature multi-effect evaporation-MVR evaporative crystallization combined process; a high-salinity wastewater treatment method), adsorption (a high-salinity organic wastewater treatment device) and membrane interception (an organic silicon composite membrane for high-salinity wastewater treatment). It should be noted that although these methods are better at removing high concentrations of salts from wastewater, these equipment or materials are costly to produce and use; in addition, most of the high-salinity wastewater contains other pollutants at the same time, and the other pollutants in the high-salinity wastewater cannot be effectively removed only by the method. In the aspect of biological treatment, Zhanyonghai et al report a method for treating high-salinity wastewater by combining a biological rotating disc and a low-temperature MVR drying device (a zero-emission treatment device and a treatment method for saline wastewater), but the method does not explain the adaptability and treatment effect of microorganisms on saline wastewater; li jin et al reported a reinforcer and method for high-salt low-temperature wastewater treatment, whose action principle is to adjust the osmotic pressure of microorganism cells by adding the reinforcer, so that the microorganism can play a role in the biological treatment of high-salt wastewater (a reinforcer and method for high-salt low-temperature wastewater treatment). In the application process of the technical method, the main effect is to adjust the osmotic pressure of microbial cells, the adding amount of the enhancer is 5-25 kg/ton of water, and the use cost is high. The application cost and the operation complexity of various methods are comprehensively compared, and the microorganism treatment of the wastewater has certain technical and cost advantages. The key to efficient treatment of such wastewater is the provision of measures to increase the microbial cell tolerance to high salt conditions and surfactant degrading activity.
Therefore, the microbial activation accelerant for improving the degradation activity of the microorganisms in the high-salt daily chemical wastewater is provided, can obviously reduce the wastewater treatment cost, is environment-friendly and has important practical significance.
Disclosure of Invention
The invention aims to provide an activation promoter for improving the microbial treatment effect of daily chemical wastewater.
In order to achieve the purpose of the invention, the technical scheme adopted by the invention is as follows: a microbial activation promoter comprises 1L of water as a solvent, and comprises the following components: 5-35 g of betaine, 25-175 g of citric acid, 45-100 g of ferrous sulfate, 5-35 g of proline, 120-200 g of chitosan, 2-10 g of trehalose and 0.10-0.50 g of vitamin complex, wherein the mass of the citric acid is 4-5 times that of the proline.
Preferably, the preparation method of the compound vitamin comprises the following steps: adding thiamine, folic acid and biotin into 1L of distilled water according to the mass ratio of 1:1:1, and fully and uniformly mixing to obtain the composition.
Preferably, the chitosan accounts for 140-180 g.
Preferably, the feed additive comprises 10-25 g of betaine, 50-100 g of citric acid, 50-80 g of ferrous sulfate, 10-20 g of proline, 2-6 g of trehalose and 0.20-0.35 g of vitamin complex.
Correspondingly, the application method of the microbial activation accelerant comprises the following specific steps:
(1) inoculating a compound microbial agent into the wastewater to be treated according to the inoculation amount of 10% and the OD600 of 0.1-0.2;
(2) the microbial activation accelerant is added into the wastewater inoculated with the compound microbial inoculant, the addition amount of the microbial activation accelerant in every kiloliter of wastewater is one liter, and the working temperature is as follows: 15 to 30 ℃.
Correspondingly, the preparation method of the compound microbial agent comprises the following steps:
(1) preparing a wastewater sample: adding SDS and Brij35 in a mass ratio of 1:1, fully dissolving in water to obtain wastewater with an initial concentration of 1500mg COD/L, and adding 20-30 g/L NaCl to obtain wastewater;
(2) inoculating the Pseudomonas sp.SDS-N2 bacterial liquid and the Pseudomonas pseudoalcaligenes sp.PG-1 bacterial liquid into the wastewater in the step (1) according to the volume ratio of 1:1, and performing mixed culture, wherein the total inoculation amount of the mixed bacteria is 20%;
(3) n, P is added into the wastewater in the step (1) according to the mass ratio of COD to N to P being 200 to 5 to 1, 5mL/L of trace elements is added, and the mixture is continuously cultured for 24 hours under the environment that the temperature is 28 ℃, the pH value is 8.0 +/-0.5 and the dissolved oxygen is sufficient, so as to obtain the compound microbial inoculum.
Preferably, the preparation method of the trace elements comprises the following steps: 1L of water is used as a solvent, and the following components are fully dissolved: EDTA 0.5g/L, ZnSO4·7H2O 0.22g/L,CaCl20.055g/L,MnCl2·4H2O0.051g/L,FeSO4·7H2O 0.049g/L,(NH4)2Mo7O24·4H2O 0.011g/L,CuSO4·5H2O0.0157g/L,CoCl2·6H2O0.016 g/L, pH adjusted to 6.0 using KOH.
Correspondingly, the compound microbial agent is applied to daily chemical wastewater.
The invention has the following beneficial effects:
1. the invention provides a preparation method and application effects of an activation promoter suitable for microbial treatment of high-salt daily chemical wastewater (mainly comprising an anionic surfactant SDS and a nonionic surfactant Brij35), aiming at the problems that microbial cell osmotic pressure and degradation activity are influenced by a high-salt environment in the treatment process of the daily chemical wastewater, so that the degradation rate of the surfactant is slow, the treatment effect is poor and the like.
2. The invention comprehensively promotes the activity of microorganisms from various aspects. Wherein: the proline and the trehalose can maintain the osmotic pressure of microbial cells in a high-salt environment, maintain the stability and the fluidity of the microbial cells, protect biological macromolecules and organisms in a non-specific manner, and improve the high-salt resistance of the microbes. The citric acid is involved in tricarboxylic acid cycle in the microorganism body, can promote the generation of ATP and improve the metabolic activity of the microorganism. Ferrous ions are active central sites of a plurality of enzymes, and the proper addition of ferrous ions can promote the activity of the enzymes, thereby improving the metabolic activity of microorganisms. Betaine can regulate osmotic pressure of microbial cells to maintain osmotic balance inside and outside the cells; in addition, betaine plays an important role in the processes of resisting oxidative stress caused by high salt, protecting the respiration of microorganisms, and metabolizing enzyme energy. The compound vitamin as a growth factor can effectively promote the growth of microorganisms; the chitosan has a certain adsorption flocculation effect and can agglomerate microorganisms into clusters, so that the interference of external adverse factors is reduced.
3. By using the microbial activation accelerant prepared by the scheme of the invention, unknown combined action occurs among various added substances.
(1) When the citric acid is 15 times or more of the proline dosage, the combined action of oxidation resistance, growth promotion and microorganism osmotic pressure maintenance can be generated, the citric acid dosage of the invention is only 5 times of the proline dosage, and the microorganism growth achieves the effect equivalent to or even better than the conventional dosage, which is probably because the unknown combined action is generated among various substances added by the invention.
(2) Ferrous ions can promote the enzyme activity of microorganisms, but the effect is better when the concentration is 5-45 mg/L (for example, the effect of ferrous on denitrification of the horizontal subsurface flow constructed wetland; Effects of Fe2+The concentration of the ferrous ion is 50-80 mg/L, which is obviously higher than the conventional dosage, and the growth of the microorganism achieves the effect equivalent to or even better than the conventional dosage, probably because the unknown combined action occurs among various substances added in the invention.
(3) The chitosan has the functions of adsorption and flocculation, can agglomerate microorganisms to reduce the influence of external adverse factors on microbial cells, and the excessive chitosan can inhibit the activity of the microbial cells. Generally, the amount of chitosan added is preferably 120mg/L or less. In the research, the addition amount of the chitosan reaches 140-180 mg/L, which is obviously higher than the conventional amount, and the growth of microorganisms reaches the effect equivalent to or even better than the conventional amount, probably because unknown combined action occurs among various substances added in the invention.
4. Experiments prove that the microbial activation accelerant can effectively promote the growth and metabolic activity of microbes under the high-salt condition, improve the pollutant removal efficiency, obviously reduce the operation energy consumption and has obvious economic benefit. Meanwhile, the microbial activation accelerant is simple to prepare, low in cost and convenient to use.
Detailed Description
1. Preparing a wastewater sample:
SDS and Brij35 are added according to the proportion of 1:1(w/w, mass ratio), the concentration of salt (NaCl) is 20-30 g/L, and the salt (NaCl) is fully dissolved in water to obtain high-salt daily chemical wastewater with the initial concentration of 1500mg COD/L, which is used as a full-text wastewater sample to be treated.
2. Preparing a compound microbial agent:
(1) SDS degrading bacteria are selected from soil in Deyang city of Sichuan province and are subjected to plate-line drawing. The method comprises the following specific steps:
1) 20mL of soil was taken and acclimatized and cultured in a liquid medium containing 1g/L SDS.
The liquid culture medium is as follows: na (Na)2HPO4·12H2O 1.0g/L，KH2PO41.0g/L，MgSO4·7H2O 0.1g/L，(NH4)SO40.3g/L, 5mL/L of trace elements, 1.0g/L of SDS, and 8.0 of pH;
the trace elements are: 1L of water is taken as a solvent, and the components comprise: EDTA 0.5g/L, ZnSO4·7H2O 0.22g/L，CaCl20.055g/L，MnCl2·4H2O 0.051g/L，FeSO4·7H2O 0.049g/L，(NH4)2Mo7O24·4H2O0.011g/L，CuSO4·5H2O0.0157g/L，CoCl2·6H2O 0.016g/L，pH＝6.0(KOH)。
2) When the SDS removal rate of the soil sample exceeds 90% within 24 hours, 1mL of the sample was subjected to gradient dilution with distilled water (10)-1、10-2、10-3、10-4、10-5、10-6、10-7) Respectively taking 0.2mL of diluent, coating the diluent in a 1g/L SDS solid culture medium, placing the mixture in an incubator, culturing for 24-72 h at a constant temperature of 30 ℃, and culturing until macroscopic colonies grow out;
the preparation method of the SDS solid medium comprises the following steps: adding 0.2% agar powder into the liquid culture medium in the step 1).
3) And (3) selecting a single colony growing quickly in a 1g/LSDS solid culture medium, purifying the colony by a streaking separation method, and repeating the operation for 3-4 times so as to obtain a pure strain. Picking single colony, inoculating the strain into SDS solid culture medium, and preserving in refrigerator at 4 deg.c.
(2) The Birj35 degrading bacteria are selected from soil in Deyang city of Sichuan province and are subjected to plate marking. The method comprises the following specific steps:
1) 20mL of soil was taken and acclimatized in a liquid medium containing 0.5g/L of Birj 35.
The liquid culture medium is as follows: na (Na)2HPO4·12H2O 1.0g/L，KH2PO41.0g/L，MgSO4·7H2O 0.1g/L，(NH4)SO40.3g/L, 5ml/L of trace elements, Brij350.5g/L and 8.0 of pH;
the trace elements are: 1L of water is taken as a solvent, and the components comprise: EDTA 0.5g/L, ZnSO4·7H2O 0.22g/L，CaCl20.055g/L，MnCl2·4H2O 0.051g/L，FeSO4·7H2O 0.049g/L，(NH4)2Mo7O24·4H2O0.011g/L，CuSO4·5H2O0.0157g/L，CoCl2·6H2O 0.016g/L，pH＝6.0(KOH)。
2) When the removal rate of the soil sample to the Birj35 exceeds 90 percent within 24h, 1mL of the sample is subjected to gradient dilution with distilled water (10)-1、10-2、10-3、10-4、10-5、10-6、10-7) Respectively taking 0.2mL of diluent, coating the diluent in a 0.5g/L Birj35 solid culture medium, placing the mixture in an incubator, culturing for 24-72 h at a constant temperature of 30 ℃, and culturing until macroscopic colonies grow out.
The preparation method of the Birj35 solid culture medium comprises the following steps: adding 0.2% agar powder into the liquid culture medium in the step 1).
3) And (3) selecting a single colony growing quickly in a 0.5g/L Birj35 solid culture medium, purifying the colony by a streaking separation method, and repeating the operation for 3-4 times to ensure that a pure strain is obtained. A single colony was picked, and the strain was inoculated into Birj35 solid medium and stored in a refrigerator at 4 ℃.
(2) Selecting a Pseudomonas sp.SDS-N2 strain (Genbank No. MF457589, DNA sequence is shown as SEQ ID NO 1) preserved in 4 ℃ SDS solid medium, placing the strain in sterilized LB medium containing 1000mg/L SDS, and culturing at 28 ℃ for 12h, wherein the OD value of the bacterial liquid reaches 1.0-1.5.
(3) Selecting a Pseudomonas pseudoalcaligenss strain PG-1 (Genbank No. MG255175, the DNA sequence is shown in SEQ ID NO 2) stored on the inclined plane of a solid culture medium of Birj35 at 4 ℃, placing the strain into a sterilized LB culture medium containing 500mg/LBrij35, and culturing at the constant temperature of 28 ℃ for 12 hours until the OD value of a bacterial liquid reaches 0.6-1.0.
(4) Inoculating the Pseudomonas sp.SDS-N2 bacterial liquid and the Pseudomonas pseudoalcaligenes p.PG-1 bacterial liquid according to the proportion of 1:1(v/v, volume ratio) and carrying out mixed culture, wherein the total inoculation amount of the mixed bacteria is 20%, adding SDS: Brij35 according to the mass ratio of 1:1 (the final concentration is about 1500mg COD/L), adding enough nitrogen and phosphorus elements (the mass ratio, COD: N: P is 200:20:1) and 5mL/L of trace elements, carrying out culture at the temperature of 28 ℃, the pH value of 8.0 +/-0.5, and carrying out continuous culture for 24 hours to obtain the composite microbial inoculum.
(5) The preparation method of the trace elements comprises the following steps: 1L of water is used as a solvent, and the following components are fully dissolved: EDTA 0.5g/L, ZnSO4·7H2O 0.22g/L，CaCl20.055g/L，MnCl2·4H2O0.051g/L，FeSO4·7H2O 0.049g/L，(NH4)2Mo7O24·4H2O 0.011g/L，CuSO4·5H2O 0.0157g/L，CoCl2·6H2O 0.016g/L，pH＝6.0(KOH)。
3. Preparing the compound vitamin:
adding thiamine, folic acid and biotin into 1L of distilled water according to the mass ratio of 1:1:1, and fully and uniformly mixing to obtain the compound vitamin with the concentration of 0.1-0.5 g/L.
4. Data determination
(1) Protein content determination
The protein content is measured by using BCA protein concentration measuring kit of Biyuntian (Beyotime) company, and the protein concentration measuring instrument is a multifunctional enzyme-linked immunosorbent assay (EnVision, PerkinElmer company, USA). The specific detection steps are as follows:
1) firstly, reagent A (reagent in the kit, product number: p0012-1) and uniformly mixing by shaking, adding 50 volumes of reagent A and 1 volume of reagent B (reagent in the kit, product number: p0012-2) reagent to prepare a BCA working solution, and fully and uniformly mixing. The BCA working solution was stable within 24 hours.
2) Diluting the standard substance: a10. mu.l aliquot of a completely solubilized protein standard (5mg/mL BSA, stored at-20 ℃) was diluted to 100. mu.L with PBS to a final concentration of 0.5 mg/mL.
The diluted standard (0.5mg/mL) was added to the protein standard wells of a 96-well plate in 0. mu.L, 2. mu.L, 4. mu.L, 6. mu.L, 8. mu.L, 12. mu.L, 16. mu.L, and 20. mu.L, respectively, and the standard dilution was added to make up to 20. mu.L.
3) Add the appropriate volume of sample to the sample wells of a 96 well plate and make up to 20. mu.L with standard dilution.
4) And adding 200 mu L of BCA working solution into each hole, gently blowing and beating by using a sample adding gun, uniformly mixing the liquid, and then placing in a room at 37 ℃ for 30-60 min. The liquid was cooled to room temperature, the absorbance at 562nm was measured on a microplate reader, and finally the protein concentration (mg/L) was calculated from the standard curve.
(2) Electron Transport System (ETS) Activity assay
ETS activity was determined using the INT-ETS method. The principle of the method is as follows: iodine nitro tetrazole [2- (rho-iodophenyl) -3- (rho-nitrophenyl) -5-phenyl Tetrazolium Chloride, INT ] is used as an artificial electron acceptor, and the INT generates a mauve reduction product triphenylmethyl ester after receiving electrons, so that the activity of the microorganism can be indirectly reflected. The specific detection steps of ETS are as follows:
taking 1mL of the bacterial liquid, centrifuging at 8000rpm for 5min, discarding the supernatant, washing the sample with PBS buffer solution (pH 8.4), centrifuging for 3 times, finally supplementing the sample volume with PBS buffer solution (pH 8.4) and mixing uniformly. 0.5mL of 0.2% INT solution and 2mL of 0.1mol/L glucose solution were added, and the mixture was subjected to shake culture at 37 ℃ for 1 hour under dark conditions. Adding formaldehyde (the addition amount is 2% of the sample amount) to terminate the reaction, centrifuging for 3min (10000rpm), discarding the supernatant, adding 1mL ethanol and 1mL acetone, and extracting under shaking at 37 + -1 deg.C in dark for 15 min. Centrifuging at 10000rpm for 3min, collecting the upper layer solution, measuring absorbance at 485nm, and converting into ETS activity.
(3) Determination of Chemical Oxygen Demand (COD)
COD refers to the amount of oxidant consumed by oxidizing reducing substances in 1L of water sample under a certain condition, and is expressed by mg/L of oxygen; the determination adopts the bichromate method (GB 11914-89) which is the determination of the chemical oxygen demand of water quality. The method comprises the following specific steps:
1) putting 20.00mL of uniformly mixed water sample (or diluting a proper amount of water sample to 20.00mL) into a 250mL backflow conical flask with a ground opening, accurately adding 10.00mL of potassium dichromate standard solution and a plurality of anti-bumping glass beads, connecting a backflow condensation pipe with the ground opening, slowly adding 30mL of sulfuric acid-silver sulfate solution from the upper end of the condensation pipe, and continuously rotating the conical flask to uniformly mix the solution. The solution was heated to reflux for 2h since it started boiling.
2) After cooling, the wall of the condensation tube is washed by 20-30 mL of water, and the conical flask is taken down. The total volume of the solution should not be less than 140 mL.
3) After the solution is cooled to room temperature, 3 drops of ferron indicator solution are added, ammonium ferrous sulfate standard solution is used for titration, the end point is that the color of the solution is changed from yellow to reddish brown from blue green, and the using amount of the ammonium ferrous sulfate standard solution is recorded.
4) While measuring the water sample, 20.00mL of redistilled water was taken, and a blank experiment was performed in the same manner. And recording the dosage of the standard solution of the ammonium ferrous sulfate when the blank is titrated.
Calculating the formula:
in the formula, c is the concentration (mol/L) of the ammonium ferrous sulfate standard solution;
V1consumption of standard solution of ferrous ammonium sulfate in titration of blankAmount (mL);
V2consumption (mL) of standard solution of ferrous ammonium sulfate when titrating a water sample;
v — water sample volume (mL);
8-oxygen (1/2O)2) Molar mass (g/mol).
Example 1: influence of various additives on sewage treatment effect
1. The accelerator components of each set are shown in table 1. An equal amount of phosphate buffer was also set as a control.
TABLE 1 ingredient table of each component of accelerator
2. Respectively taking the same amount of the accelerant in each group in the step 1, wherein the addition amount of the microbial activation accelerant in each kiloliter of wastewater is one liter, the initial surfactant concentration is about 1500mg COD/L of wastewater, the salt concentration (NaCl) in the wastewater is 20-30 g/L, and then inoculating the composite microbial inoculum with the inoculation amount of 20% (OD)6000.1-0.2) and the working temperature is 20 ℃.
The microbial activation accelerant prepared according to the technical scheme of the invention can work well at 15-30 ℃, and is convenient to operate, and the microbial activation accelerant uniformly adopts 20 ℃ throughout the text.
3. Through shake flask experiments, COD values, biomass (expressed in protein concentration), metabolic activity (ETS) were observed throughout the degradation process. The results are shown in Table 2.
TABLE 2 effect of wastewater treatment
4. As shown in the table: the data of groups 1-7 demonstrate that, in terms of the importance of the components in the accelerator, betaine > citric acid > ferrous sulfate > proline > chitosan > trehalose > multivitamins; the complete accelerator prepared according to the technical scheme of the invention has obvious effect which is extremely superior to the accelerator and a blank control group which lack a certain substance.
Example 2: influence of proline and citric acid on microbial wastewater treatment in high-salt environment
1. Preparing an accelerant:
(1) positive control group: 18g of proline, 63g of ferrous sulfate, 270g of citric acid, 170g of chitosan, 21g of betaine, 4g of trehalose and 0.24g of compound vitamin are dissolved in 1L of water, and are fully and uniformly mixed to prepare the microbial activation promoter, and the microbial activation promoter is sealed and stored.
(2) Blank control group: equal amount of phosphate buffer.
(3) The test groups are shown in Table 3 for each component.
TABLE 3 Accelerator test groups Components
2. Respectively taking the same amount of the accelerators in each group in the step 1, adding one liter of the microbial activation accelerator into wastewater with the initial concentration of 1500mgCOD/L and the salt concentration of 20-30 g/L at 20 ℃, and then inoculating the composite microbial agent with the inoculation amount of 10% (OD)600＝0.1～0.2)。
3. Through shake flask experiments, COD values, biomass (expressed in protein concentration), metabolic activity (ETS) were observed throughout the degradation process. The results are shown in Table 4.
TABLE 4 Effect of proline and citric acid on microbial growth and wastewater treatment
4. The results show that: when the addition amount of the citric acid is only about 5 times of that of the proline, the effect is equivalent to or even slightly better than that when the addition amount reaches 15 times, and the treatment effect is extremely obviously better than that of a blank control group. This is probably due to the unknown association between the additives using the solution of the invention. Meanwhile, in a certain range, better effect can be achieved no matter how much citric acid and proline are added as long as the addition ratio of the citric acid to the proline is about 4-5 times.
Example 3: influence of ferrous ions on microbial wastewater treatment in high-salt environment
1. Preparing an accelerant:
(1) positive control group: dissolving proline 18g, ferrous sulfate 25g, citric acid 80g, chitosan 170g, betaine 21g, trehalose 4g, and vitamin complex 0.24g in 1L water, mixing, making into microbial activation promoter, and sealing for storage.
(2) Blank control group: equal amount of phosphate buffer.
(3) Test groups: the components are shown in table 5. Dissolving the components in 1L water, mixing well to obtain microbial activation promoter, and sealing for storage.
TABLE 5 test groups of Components
| Group of | Betaine | Citric acid | Ferrous sulfate | Proline | Chitosan | Trehalose | Compound vitamin |
| Group 1 | 21g | 80g | 50g | 18g | 170g | 4g | 0.24g |
| Group 2 | 21g | 80g | 58g | 18g | 170g | 4g | 0.24g |
| Group 3 | 21g | 80g | 70g | 18g | 170g | 4g | 0.24g |
2. Respectively taking the same amount of the accelerant in each group in the step 1, adding one liter of the microbial activation accelerant into one kiloliter of wastewater, adding the accelerant into the wastewater with the temperature of 20 ℃ and the initial concentration of 1500mgCOD/L, and then inoculating the compound microbial inoculum with the inoculation amount of 10% (OD)600＝0.1～0.2)。
3. Through shake flask experiments, COD values, biomass (expressed in protein concentration), metabolic activity (ETS) were observed throughout the degradation process. The results are shown in Table 6.
TABLE 6Fe2+Influence on microbial growth and wastewater treatment
4. The results show that: the enzyme activity of the microorganism can be effectively improved only when the ferrous ion is generally used at a concentration of 5-45 mg/L, but the dosage of the ferrous ion is 50-80 mg/L, which is obviously higher than the conventional dosage, and the microorganism growth achieves the effect equivalent to or slightly better than the conventional dosage, probably because unknown combined action occurs among various substances added in the invention.
Example 4: influence of chitosan on microbial wastewater treatment in high-salt environment
1. Preparing an accelerant:
(1) positive control group: 18g of proline, 63g of ferrous sulfate, 80g of citric acid, 100g of chitosan, 21g of betaine, 4g of trehalose and 0.24g of compound vitamin are dissolved in 1L of water, and are fully and uniformly mixed to prepare the microbial activation promoter which is sealed and stored.
(2) Blank control group: equal amount of phosphate buffer.
(3) The test groups were prepared by dissolving the components in 1L of water, mixing them, and sealing for storage, as shown in Table 7.
TABLE 7 test groups Components
2. Respectively adding one liter of the same amount of the accelerant in each group in the step 1 into one thousand liters of the wastewater, wherein the addition amount of the microbial activation accelerant is one literInoculating 10% (OD) of compound microorganism bacterium agent into wastewater with initial concentration of 1500mgCOD/L at 20 deg.C600＝0.1～0.2)。
3. Through shake flask experiments, COD values, biomass (expressed in protein concentration), metabolic activity (ETS) were observed throughout the degradation process. The results are shown in Table 8.
TABLE 8 Effect of Chitosan on microbial growth and wastewater treatment
4. The addition amount of the chitosan is preferably below 120mg/L, and the enzyme activity of the microorganism can be inhibited too much; but the table above shows: the addition amount of the chitosan reaches 140-180 mg/L, which is obviously higher than the conventional dosage, and the growth of microorganisms reaches the effect equivalent to or even better than the conventional dosage, probably because unknown combined action occurs among various substances added in the invention.
Example 5: effect of microorganism activation promoter on treating sewage
1. The accelerator test components are shown in table 9.
TABLE 9 Accelerator ingredient Table
2. Dissolving the above materials in 1L water respectively, mixing well to obtain microbial activation promoter, sealing and storing; an equal amount of physiological saline was used as a blank control. A positive control group was also prepared, mannitol: tetrahydropyrimidine: trehalose: amino acids: potassium salt: inositol methyl ester: and (3) a first glycan: n dimethyl sulfoxide: methanol: acetamide (18: 5:8:6:25:5:7:6:22:1 (molar ratio) was mixed and prepared in an amount of 1 liter per kiloliter of wastewater, and used as a positive control.
2. Respectively adding equal amount of the accelerant in each group in the step 1 into wastewater with the temperature of 20 ℃ and the concentration of 1500mg COD/L, wherein the addition amount of the microbial activation accelerant in each kiloliter of wastewater is one liter, and then inoculating the compound microbial inoculum with the inoculation amount of 10% (the inoculation amount is 10%)OD600＝0.1～0.2)。
3. Through shake flask experiments, COD values, biomass (expressed in protein concentration), metabolic activity (ETS) were observed throughout the degradation process. The results are shown in Table 10.
TABLE 10 effects of wastewater treatment
4. The results show that: the microbial activation accelerant has obvious promotion effect on the growth of microbes in a high-salt environment and sewage treatment. When the components are as follows: the effect is good when the content of betaine is 5-35 g, the content of citric acid is 25-175 g, the content of ferrous sulfate is 45-100 g, the content of proline is 5-35 g, the content of chitosan is 120-200 g, the content of trehalose is 2-10 g, and the content of vitamin complex is 0.10-0.50 g; but the components are as follows: 140-180 g of chitosan, 10-25 g of betaine, 50-100 g of citric acid, 50-80 g of ferrous sulfate, 10-20 g of proline, 2-6 g of trehalose and 0.20-0.35 g of vitamin complex, and the effect is particularly good.
Sequence listing
<110> institute of biological research of Chengdu of Chinese academy of sciences
<120> an activation promoter for improving the effect of microbial treatment of daily chemical wastewater
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gtgccagcag ccgcggtaat acgaagggtg caagcgttaa tcggaattac tgggcgtaaa 480
gcgcgcgtag gtggtttggt aagatggatg tgaaatcccc gggctcaacc tgggaactgc 540
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acaagcggtg gagcatgtgg tttaattcga agcaacgcga agaaccttac ctggccttga 900
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Claims (7)
1. A microbial activation promoter characterized by: 1L of water is taken as a solvent, and the components comprise: 5-35 g of betaine, 25-175 g of citric acid, 50-80 g of ferrous sulfate, 5-35 g of proline, 140-180 g of chitosan, 2-10 g of trehalose and 0.10-0.50 g of vitamin complex; the mass of the citric acid is 4-5 times of that of the proline.
2. A microbial activation accelerator according to claim 1, wherein: the preparation method of the compound vitamin comprises the following steps: adding thiamine, folic acid and biotin into 1L of distilled water according to the mass ratio of 1:1:1, and fully and uniformly mixing to obtain the composition.
3. A microbial activation accelerator according to claim 1, wherein: 1L of water is taken as a solvent, and the components comprise: 10-25 g of betaine, 50-100 g of citric acid, 50-80 g of ferrous sulfate, 10-20 g of proline, 2-6 g of trehalose and 0.20-0.35 g of vitamin complex.
4. A method of using the microbial activation accelerator according to claim 1, wherein: the method comprises the following specific steps:
(1) inoculating 10% of compound microbial agent in the wastewater to be treated, wherein the inoculation amount is OD600=0.1～0.2；
(2) The microorganism activation accelerant is added into the wastewater inoculated with the compound microorganism bacterium agent in the following proportion: adding one liter of microbial activation accelerant into every kiloliter of wastewater, wherein the working temperature is as follows: 15 to 30 ℃.
5. The use of the microbial activation accelerator according to claim 4, wherein: the preparation method of the compound microbial agent comprises the following steps:
1) preparing wastewater: fully dissolving SDS and Brij35 in water according to the mass ratio of 1:1, wherein the initial concentration is 1500mg COD/L, and adding 20-30 g/L NaCl to obtain wastewater;
2) will be provided withPseudomonassp, SDS-N2 bacterial solution andPseudomonas pseudoalcaligenesinoculating sp, PG-1 bacterial liquid into the wastewater in the step 1) according to the volume ratio of 1:1, and performing mixed culture, wherein the total inoculation amount of mixed bacteria is 20%;
3) the weight ratio is as follows: n, P is added into the wastewater in the step 1) according to the ratio of COD to N to P =200 to 5 to 1, 5mL/L of trace elements is added, and the mixture is continuously cultured for 24h under the environment with the temperature of 28 ℃, the pH =8.0 +/-0.5 and sufficient dissolved oxygen, so that the compound microbial agent is obtained.
6. The use of the microbial activation accelerator according to claim 5, wherein: the preparation method of the trace elements in the step 3) comprises the following steps: 1L of water is used as a solvent, 0.5g/L of EDTA, ZnSO4·7H2O 0.22g/L，CaCl20.055g/L，MnCl2·4H2O 0.051g/L，FeSO4·7H2O 0.049g/L，(NH4)2Mo7O24·4H2O 0.011g/L，CuSO4·5H2O0.0157g/L， CoCl2·6H2O0.016 g/L, adjusted pH =6.0 using KOH.
7. Use of the microbial activation accelerator according to any one of claims 1 to 3 in chemical wastewater.
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