CN109055285B - High-density enrichment method of autotrophic nitrifying bacteria consortium - Google Patents
High-density enrichment method of autotrophic nitrifying bacteria consortium Download PDFInfo
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Abstract
The invention belongs to the field of microbial fermentation, and particularly relates to a high-density enrichment method of an autotrophic nitrifying bacteria consortium and application of the obtained high-density autotrophic nitrifying bacteria consortium in sewage treatment. Inoculating thallus in autotrophic nitrifying bacteria initial culture medium, culturing at 28-32 deg.C, pH of 7.5-8.0 and Dissolved Oxygen (DO) of 20-40% under stirring condition, and culturing with NO in the culture process3And (4) monitoring the change of the pH value in the culture process through a pH electrode to realize the supplement of an inorganic nitrogen source and an inorganic carbon source in the culture system, controlling the ammonia nitrogen concentration in the culture process to be 150-300mg/L, and controlling the pH value of the system to be 7.5-8.0 through the supplemented carbon source so as to realize the high-density automatic propagation of the autotrophic nitrifying bacteria consortium. The invention effectively solves the problem of poor ammonia nitrogen treatment effect caused by the reasons that the consortium has poor adaptability to strains, loss of the added strains, low cost of the added strains and the like in the ammonia nitrogen wastewater treatment application process.
Description
Technical Field
The invention belongs to the field of microbial fermentation, and particularly relates to a high-density enrichment method of an autotrophic nitrifying bacteria consortium and application of the obtained high-density autotrophic nitrifying bacteria consortium in sewage treatment.
Background
Autotrophic nitrifying bacteria are a general name of nitrosobacteria and nitrifying bacteria consortia, and the microorganisms can convert ammonia nitrogen into nitrite nitrogen and convert the nitrite nitrogen into nitrate nitrogen, and play an important role in the N element circulation in the nature. With the development of microbial isolation technology, such microorganisms are used in various fields, for example, patent CN101039692A discloses the application of a consortium consisting of nitromonas eutropea and nitrobacter winogradskyi in aquaculture. The subsequent extensive application of this kind of synbiotics in ammonia nitrogen wastewater treatment, but the application effect of this kind of microorganism in this sewage treatment field is not very ideal, the main reason lies in the sewage treatment process, the water quality situation of various waste waters produces certain influence to this kind of microorganism; compared with aquaculture, sewage treatment belongs to a fluid ecosystem, so that the problem of loss after adding strains also influences the treatment effect; in addition, the factors such as dissolved oxygen, temperature and pH fluctuate in the sewage treatment process can cause the death of the microorganisms. The current patents in the related art are mainly divided into three aspects. Firstly, in the aspect of strain enrichment and screening, for example, patents CN106350471A, CN106396132A, CN108102952A and the like all disclose enrichment or screening methods of nitrifying bacteria, which hope to screen out a microorganism that can adapt to the actual environment of wastewater so as to improve the treatment effect, but the types of wastewater are different in the field of wastewater treatment, and it is not practical to obtain a broad-spectrum microorganism; secondly, the expansion culture method of nitrifying bacteria, such as patents CN106520610A, CN102070251A, CN106396132A, etc., disclose how to expand autotrophic nitrifying bacteria. CN106396132A discloses an intermittent, ammonia nitrogen fed-batch operation alternate nitrobacteria culture method, which is the closest scheme to the technology of the invention in the prior art, the patent utilizes a bacterial fermentation tank to inoculate activated sludge containing nitrobacteria, ammonia nitrogen is intermittently supplemented in daytime, the ammonia nitrogen fed-batch speed at night is calculated according to the ammonia nitrogen consumption speed in daytime, and the patents hope to improve the treatment effect by improving the fermentation efficiency of autotrophic nitrobacteria so as to increase the added bacteria quantity in the sewage treatment process; thirdly, nitrobacteria propagation devices such as CN207596864U, CN207452089U, CN206467042U, CN206173357U and the like disclose nitrobacteria propagation devices, and such patents also aim to improve the fermentation efficiency of nitrobacteria.
In the prior art, the bacteria can be divided into autotrophic nitrifying bacteria and heterotrophic nitrifying bacteria from the aspect of genus, heterotrophic microorganisms are easy to separate and expand, but most of the degradation of ammonia nitrogen is assimilation in the culture process, and the ammonia nitrogen is not converted into nitrate nitrogen, so that the total nitrogen is not favorably removed. In addition, the ammonia nitrogen of the heterotrophic nitrifying bacteria is not the only nitrogen source for the metabolism of the heterotrophic bacteria, and the selection of the nitrogen source in the actual environment has diversity. The autotrophic nitrifying bacteria consortium cannot be influenced by organic carbon sources and other heterotrophic microorganisms in an actual environment, ammonia nitrogen is also the only nitrogen source of the microorganisms, the microorganisms are the dominant flora in the biological denitrification process, the number of the flora determines the denitrification capacity of the system, and the culture expanding efficiency of the autotrophic nitrifying bacteria consortium in the prior art is too low; autotrophic nitrifying bacteria are directly enriched from the sludge, and the number of the nitrifying bacteria in the enriched product cannot meet the required adding amount of sewage treatment. Meanwhile, the autotrophic nitrifying bacteria consortium has strict requirements on the growth environment, so that in an actual environment, the autotrophic nitrifying bacteria are inoculated with high-density autotrophic nitrifying bacteria for inoculation and are continuously inoculated for multiple times to enable the system to have stable autotrophic nitrifying bacteria flora, and the premise of continuous inoculation is a high-efficiency and automatic culture expanding process. However, the prior art does not well solve the problems, particularly the influence of the supplemented carbon source on the pH of the system in the inorganic carbon source supplementing process, and the automatic supplementing is not realized in the inorganic nitrogen source supplementing process. The expanding culture of the autotrophic nitrifying bacteria is different from other microorganisms, the inorganic carbon source and the inorganic nitrogen source which need to be supplemented can cause the change of the pH value of the system, the pH value of the system influences the growth speed of thalli, and the key for improving the expanding culture efficiency of the autotrophic nitrifying bacteria is how to adjust the relationship of the inorganic carbon source and the inorganic nitrogen source. How to utilize the existing signal or the additional signal of the fermentation tank to control the peristaltic pump to realize the automatic replenishment of the inorganic carbon source and the inorganic nitrogen source is a key factor for realizing the automatic culture of the autotrophic nitrobacteria.
Disclosure of Invention
The invention aims to provide a high-density enrichment method of autotrophic nitrifying bacteria consortium.
In order to achieve the purpose, the invention adopts the technical scheme that:
a high-density enrichment method of autotrophic nitrifying bacteria consortium comprises inoculating thallus into autotrophic nitrifying bacteria initial culture medium, culturing at 28-32 deg.C, pH 7.5-8.0, Dissolved Oxygen (DO) controlled at 20-40%, stirring, and culturing with NO in the culture process3And (4) monitoring the change of the pH value in the culture process through a pH electrode to realize the supplement of the inorganic carbon source and the inorganic nitrogen source in the culture system, controlling the ammonia nitrogen concentration in the culture process to be 150-300mg/L, and controlling the pH value of the system to be 7.5-8.0 through the supplemented inorganic carbon source so as to realize the high-density automatic propagation of the autotrophic nitrifying bacteria consortium.
By adopting the method, autotrophic nitrifying bacteria are rapidly propagated under the optimal growth condition all the time to finally obtain high-density bacterial liquid, and the automatic culture method realizes online culture and continuous feeding of the autotrophic nitrifying bacteria in the field of sewage treatment, effectively improves the nitrification capacity of a biochemical system, and solves the problem of ammonia nitrogen in various waste water.
And controlling the consumption of the inorganic carbon source according to the detected pH value, further supplementing the carbon source, and calculating the supplementing amount of the inorganic nitrogen source in the corresponding time point according to the consumption of the inorganic carbon source in different time in the culture process.
The culture is started after inoculation:
the addition amount of the inorganic nitrogen source in 1h-2h is as follows: { (X)1-X0) X 0.1745 x 80 divided by V +68.826 }. times V divided by 3120, in mL/min;
the addition amount of the inorganic nitrogen source within 2h-3h is as follows: { (X)2-X1) X 0.1745 x 80 divided by V +68.826 }. times V divided by 3120, in mL/min;
……
the addition amount of the inorganic nitrogen source in nh- (n +1) h is as follows: { (X)n-X(n-1)) X 0.1745 x 80 divided by V +68.826 }. times V divided by 3120, in mL/min;
wherein the culture time is every other hour from 0hThe consumption of the obtained inorganic carbon source is X0、X1、X2、X3、X4……Xn、Xn+1… … in mL.
When the cumulative amount X value of the consumption of the inorganic carbon source reaches V/4(L), the nitrification rate of the culture solution for propagation is more than 160mg (NO)3-N)/L/h, after the culture expansion is finished, stopping stirring and standing for 1h, collecting the nitrosobacteria and nitrifying bacteria consortium, wherein the nitrification rate of the harvested consortium per unit mass is more than 600mg (NO)3-N)/g/h。
The inorganic carbon source supplemented in the culture expanding process is 5-10 wt% of Na2CO3A solution; the supplementary inorganic nitrogen source is 10 to 20 weight percent of NH4And (4) Cl solution.
The autotrophic nitrifying bacteria initial culture medium contains Na in each liter of water2CO3 2g-4g、MgSO40.2-0.4 g, KCl 0.2-0.4 g, pH adjusted to 7.5-8.0 with phosphoric acid, NH4Cl 0.5g-1.0g, initial NH4-N concentration 150mg/L-300mg/L, NO in the initial medium3The concentration of-N is controlled to be 0mg/L-10 mg/L.
The thallus is autotrophic nitrifying bacteria consortium or activated sludge containing autotrophic nitrifying bacteria; the inoculation amount is controlled to be 0.5wt% -1 wt%.
The stirring speed is controlled to be constant between 150r/min and 300r/min in the expanding culture process; the Dissolved Oxygen (DO) in the system is controlled at 20% -40%, and the electromagnetic ventilation valve is controlled by the dissolved oxygen electrode signal to realize automatic control; the temperature in the system is controlled by a temperature probe to control the temperature of jacket water, so that the temperature in the reaction process is between 28 and 32 ℃.
The application of the high-density nitrosobacteria and nitrobacteria consortium collected by propagation in sewage treatment.
The invention has the advantages that: compared with the prior art, the automatic culture method of the autotrophic nitrobacteria has the advantages that the inorganic carbon source can be accurately and automatically supplemented through the change signal of the pH value in the system, meanwhile, the amount of the inorganic nitrogen source needed in the system is judged according to the amount of the inorganic carbon source supplemented in the system and is automatically supplemented, and the whole cultured autotrophic nitrobacteria is ensured to be completely culturedIn the process, the inorganic carbon source, the inorganic nitrogen source and the pH are controlled within the most suitable range of the autotrophic nitrifying bacteria consortium, and the high-density automatic culture process is realized. The inorganic carbon source is controlled and supplemented by using the pH electrode, and the inorganic nitrogen source is controlled and supplemented by using the cumulant signal of the inorganic carbon source. The method can realize the automatic culture of the autotrophic nitrifying bacteria consortium or the activated sludge containing autotrophic nitrifying bacteria, and the whole-process automatic culture of the ammonia nitrogen concentration in a detection system is not needed in the culture process. The nitrifying rate of the cultured nitrifying bacteria culture solution can reach 200mg (NO)3about-N)/L/h, the nitrification rate of the thallus per unit mass can reach 600mg (NO)3-N)/g/h or more; meanwhile, the nitrification rate of the nitrifying bacteria enrichment liquid in the enriched activated sludge by using the method can reach 158mg (NO)3-N)/L/h, the nitrification rate of the flocculating constituent unit mass can reach 465mg (NO)3-N)/g/h. The success rate of automatic culture reaches 100%, which is the premise of on-line automatic culture and continuous addition, and realizes on-line culture and continuous addition, thereby improving the nitrification rate in various wastewater treatment systems and reducing the concentration of ammonia nitrogen in effluent.
Drawings
Fig. 1 is a reaction apparatus provided in an embodiment of the present invention, in which 1, a fermentation tank, 2, a carbon source storage tank, 3, a peristaltic pump a, 4, a pH electrode, 5, a control system, 6, a nitrogen source storage tank, 7, a peristaltic pump B, 8, a dissolved oxygen electrode, 9, a temperature probe, 10, a stirring motor, 11, an air compressor, 12, and a heater.
FIG. 2 shows the nitrification amount of sodium carbonate solution and NO in the 100L scale culture process of autotrophic nitrifying bacteria consortium according to the present invention3-a linear plot of the cumulative amount of N.
FIG. 3 shows the high throughput sequencing result before enrichment of activated sludge in the biochemical pool for pesticide wastewater according to the embodiment of the present invention.
FIG. 4 is a high throughput sequencing result of the method for enriching the activated sludge in the pesticide wastewater biochemical pool.
FIG. 5 is a data diagram of ammonia nitrogen in the effluent of an aerobic biochemical tank of a pesticide enterprise after autotrophic nitrifying bacteria consortium are continuously added according to an embodiment of the present invention.
Detailed Description
The invention is further explained below with reference to the figures and examples.
The invention uses NO in the process of autotrophic nitrifying bacteria culture3-N accumulation with Na2CO3An automatic nitrogen supplementing scheme is formulated according to the relation of solution consumption, the synergistic relation among inorganic carbon source supplement, inorganic nitrogen source supplement and system pH of the autotrophic nitrifying bacteria consortium in the culture process is effectively solved, high-density automatic propagation of the autotrophic nitrifying bacteria consortium is realized, high-density continuous feeding of the consortium in sewage treatment is further realized, and the problem of poor ammonia nitrogen treatment effect caused by the reason that the consortium is poor in ammonia nitrogen wastewater treatment application process, fed strains run off, the number of fed strains is not high enough, and the like is effectively solved.
Example 1:
the reaction device comprises a fermentation tank, a carbon source storage tank, a peristaltic pump A, a pH electrode, a control system, a nitrogen source storage tank, a peristaltic pump B, a dissolved oxygen electrode, a temperature probe, a stirring motor, an air compressor and a heater. The fermentation tank is internally inserted with a pH electrode, a dissolved oxygen electrode and a temperature probe which are respectively connected with a control system through leads, the fermentation tank is respectively connected with an external stirrer, a peristaltic pump A, a peristaltic pump B, an air compressor and a heater through leads, the peristaltic pump A, the peristaltic pump B, the air compressor and the heater are respectively connected with the control system through leads, and in addition, the peristaltic pump A is connected with a carbon source storage tank and the peristaltic pump B through pipelines and a nitrogen source storage tank.
The reaction process is as follows:
setting a fixed stirring rotating speed through a stirring motor in the culture process; the temperature in the fermentation tank can transmit signals to a control system through a temperature probe, and the temperature is controlled by controlling a heater; the oxygen content in the fermentation tank transmits a signal to a control system through a dissolved oxygen electrode, and the dissolved oxygen is controlled by controlling an air compressor; when the pH value of the fermentation tank is set to 7.5-8.0 and the pH value of the liquid in the fermentation tank is reduced to 7.47 in the culture process, the pH electrode transmits a signalSending to a control system to supplement inorganic carbon source (alkali/10% NaCO) stored in a carbon source storage tank by controlling a peristaltic pump A3) The peristaltic pump A rotates for 1s every 10s until the pH value is restored to 7.50, the rotation is stopped, at the moment, the control system can automatically calculate the amount of the added inorganic carbon source according to the rotation time of the peristaltic pump A and the pump speed of the peristaltic pump A, and the control system generates a signal through the difference value of the added amounts of the inorganic carbon source at different time periods to control the pump speed calculated by the peristaltic pump B to carry out inorganic nitrogen source (20% NH)4Cl).
The cumulative amount of inorganic carbon source consumption in the culture process is X, the value is automatically generated according to the time of controlling the rotation of the peristaltic pump A by the pH electrode and the input corrected peristaltic pump speed, the cumulative amount is displayed on a control screen of the culture expanding system, the culture is started after inoculation, the culture time is 0h, and Na is added every other hour from 0h2CO3Cumulative amounts of solution consumed were X, respectively0、X1、X2、X3、X4……Xn、Xn+1… … in mL.
And controlling a peristaltic pump B to realize automatic supplement of the electrodeless nitrogen source through signal change of the cumulant X consumed by the inorganic carbon source in the expanding culture process, wherein the peristaltic pump B automatically supplements the electrodeless nitrogen source from the reaction culture time of 1h, and the specific supplement scheme is as follows.
The pump speed of the peristaltic pump B of 1h-2h is as follows: { (X)1-X0) X0.1745X 80/V +68.826 }. times.V/3120 in mL/min.
The pump speed of the peristaltic pump B for 2-3 h is as follows: { (X)2-X1) X0.1745X 80/V +68.826 }. times.V/3120 in mL/min.
The pump speed of the peristaltic pump B of nh to (n +1) h is as follows: { (X)n-X(n-1)) X0.1745X 80/V +68.826 }. times.V/3120 in mL/min.
When Na is present2CO3When the cumulative X value of the solution consumption reaches V/4(L), the propagation is finished, and the nitrification rate of the propagation liquid is more than 160mg (NO)3-N)/L/h. The autotrophic nitrifying bacteria consortium was collected after the stirring was stopped and the rest for 1h, and the nitrification rate per unit mass of the harvested consortium was > 600mg (NO)3-N)/g/h. Example 2100L Scale nitrosationAutomated propagation of bacterial and nitrifying bacterial interpolymers
The volume of the fermentation tank is 100L, the consortium of nitrosobacteria and nitrobacteria is used as inoculum, the inoculum size is 1%, and the initial volume V of the culture medium is 80L. The stirring speed is 220r/min, the culture is kept unchanged in the culture expanding process, and the inorganic carbon source is 10 percent of Na2CO3Solution with inorganic nitrogen source of 20% NH4And (4) Cl solution. The dissolved oxygen DO is controlled at 20% and the pH is controlled at 7.50 during the propagation process. Wherein, the autotrophic nitrifying bacteria initial culture medium contains Na in each liter of water2CO3 2g-4g、MgSO40.2-0.4 g, KCl 0.2-0.4 g, pH adjusted to 7.5-8.0 with phosphoric acid, NH4Cl 0.5g-1.0g, initial NH4-N concentration 150mg/L-300mg/L, NO in the initial medium3The concentration of-N is controlled to be 0mg/L-10 mg/L.
During the cultivation process, NO is added during the cultivation process3Continuous accumulation of-N, continuous consumption of nitrogen and carbon sources, and utilization of NO3The relation between the accumulation amount of-N and the consumption amount of the inorganic carbon source (see figure 1), the pH electrode is used for monitoring the change of the pH value in the culture process to realize the supplement of the inorganic nitrogen source and the inorganic carbon source in the culture system, so that the ammonia nitrogen concentration in the culture process is controlled to be 150-300mg/L, the pH value is controlled to be 7.50, when the pH value is reduced to 7.47, the pH electrode is used for transmitting a signal to the control system to supplement the inorganic carbon source (alkali/10 percent NaCO) by controlling the peristaltic pump A3) The peristaltic pump A rotates for 1s every 10s until the pH value is restored to 7.50 and then stops rotating, at the moment, the control system can automatically calculate the amount of the added inorganic carbon source according to the rotation time of the peristaltic pump A and the pump speed of the peristaltic pump A, and the control system generates a signal through the difference value of the added amounts of the inorganic carbon source at different time periods to control the peristaltic pump B to carry out the inorganic nitrogen source (20% NH) according to the pump speed calculated by the formula related to the patent4Cl). Wherein the inorganic nitrogen source utilizes Na in an expanding culture system2CO3Cumulative volume consumed X for solution (carbon source)n(mL) according to the formula { (X)n-X(n-1)) The unit of multiplied by 0.1745 multiplied by 80 divided by V +68.826 multiplied by V divided by 3120 is mL/min, and the nitrogen supplement speed is changed every 1 h; wherein, the speed formula for automatically replenishing the electrodeless nitrogen source is based on the consumption of Na in the process of expanding culture2CO3Cumulative amount of solution and systemIn NO3-N cumulative amount is calculated as a linear relationship.
When Na is present2CO3When the cumulative amount of solution consumed X reached V/4 ═ 20(L), the amplification was terminated, and the results of 3 continuous cultures are shown in table 1.
Table 1: automatic culture result of 100L scale nitrosation bacteria nitrobacteria interpolymer
| Expanding culture batch | Nitrification rate of propagation liquid | Nitration Rate per Mass of harvested interpolymer |
| Batch 1 | 205mg(NO3-N)/L/h | 635mg(NO3-N)/g/h |
| Batch 2 | 216mg(NO3-N)/L/h | 646mg(NO3-N)/g/h |
| Batch 3 | 198mg(NO3-N)/L/h | 615mg(NO3-N)/g/h |
The linear relationship between the two in the 3-batch culture process in Table 1 is shown in FIG. 2, and the values of the carbon source supplementation amount at different time points in the 3-batch culture process in Table 1 are shown in Table 2.
Table 2: values of carbon source addition amounts of different batches at different time points
Example 3: automatic enrichment and propagation of 100L scale activated sludge autotrophic nitrifying bacteria flora
The volume of the fermentation tank is 100L, the activated sludge of the pesticide wastewater biochemical pool is used as inoculation liquid, the inoculation amount is 10%, and the initial volume V of the culture medium is 80L. The stirring speed is 220r/min, the culture is kept unchanged in the culture expanding process, and the inorganic carbon source is 10 percent of Na2CO3Solution with inorganic nitrogen source of 20% NH4And (4) Cl solution. The dissolved oxygen DO is controlled at 20% and the pH is controlled at 7.50 during the propagation process. Wherein, the autotrophic nitrifying bacteria initial culture medium contains Na in each liter of water2CO3 2g-4g、MgSO40.2-0.4 g, KCl 0.2-0.4 g, pH adjusted to 7.5-8.0 with phosphoric acid, NH4Cl 0.5g-1.0g, initial NH4-N concentration 150mg/L-300mg/L, NO in the initial medium3The concentration of-N is controlled to be 0mg/L-10 mg/L.
In the expanding culture process, NO is added in the culture process3Continuous accumulation of-N, continuous consumption of nitrogen and carbon sources, and utilization of NO3The relation between the-N accumulation amount and the consumption amount of the inorganic carbon source, the inorganic carbon source and the inorganic nitrogen source are supplemented in the culture system by monitoring the change of the pH value in the culture process through the pH electrode, the ammonia nitrogen concentration in the culture process is controlled to be 150-300mg/L, the pH is controlled to be 7.5, when the pH is reduced to 7.47, the pH electrode transmits a signal to a control system to supplement the inorganic carbon source (alkali/10 percent NaCO) through controlling a peristaltic pump A3) The peristaltic pump A rotates for 1s every 10s until the pH value is restored to 7.50 and then stops rotating, at the moment, the control system can automatically calculate the amount of the added inorganic carbon source according to the rotation time of the peristaltic pump A and the pump speed of the peristaltic pump A, and the control system generates a signal through the difference value of the added amounts of the inorganic carbon source at different time periods to control the peristaltic pump B to carry out the inorganic nitrogen source (20% NH) according to the pump speed calculated by the formula related to the patent4Cl). Na in inorganic nitrogen source utilization expanding culture system2CO3Cumulative volume consumed of solution Xn(mL) according to the formula { (X)n-X(n-1))×The nitrogen supplement rate was changed every 1h at 0.1745X 80/V +68.826 }. times V/3120 units of mL/min. When Na is present2CO3And finishing the propagation when the cumulative N value of the solution consumption reaches V/4-20 (L).
The supplement speed of the inorganic nitrogen source in the expanding culture process is changed every 1h according to a formula { (Xn-X (n-1)) × 0.1745 × 80 ÷ V +68.826} × V ÷ 3120 unit of mL/min. Sampling every 4h in the re-expansion process to carry out NH4The results of measurement of the-N concentration are shown in Table 3.
The activated sludge sample was subjected to high throughput sequencing, and the sequencing result is shown in fig. 3, wherein the activated sludge contains nitrosolomas (nitrosobacteria) capable of autotrophic degradation of ammonia nitrogen, and the proportion of the genus is 0.12%. Furthermore, it was found that autotrophic nitrifying bacteria must be contained in the inoculated activated sludge.
Table 3: NH of different time point systems in the process of expanding culture4Concentration of-N
| Time of expanding culture | NH4Concentration of-N |
| 0h | 180.04mg/L |
| 4h | 181.12mg/L |
| 8h | 177.15mg/L |
| 12h | 147.86mg/L |
| 16h | 160.57mg/L |
| 20h | 176.12mg/L |
| 24h | 180.51mg/L |
| 28h | 174.34mg/L |
| 32h | 174.96mg/L |
| 36h | 182.27mg/L |
According to the formula of the above example and table 3, the nitrogen source non-addition amount at different time points is calculated and obtained, and the nitrification rate of the culture solution after the enrichment is finished can reach 158mg (NO)3-N)/L/h, the nitrification rate of per unit mass of the flocculating constituent collected after the culture is finished can reach 465mg (NO)3-N)/g/h, most heterotrophic microorganisms in the expanded floccule die, the number of autotrophic nitrifying bacteria is remarkably increased, and the high-throughput sequencing result of the expanded floccule is shown in figure 4. The proportion of Nitrosomonas is 63.55%.
Example 4: application of autotrophic nitrifying bacteria consortium in automatic continuous culture of pesticide wastewater
The effective volume of an aerobic biochemical tank of a pesticide enterprise is 6800 cubic, the daily treatment capacity of wastewater is 420 vertical/day, and the retention time is 16 hours. By continuously culturing autotrophic nitrifying bacteria copolymer with 2-cube-scale automatic fermentation culture system, 80-100L of concentrated solution of autotrophic nitrifying bacteria consortium with nitrification rate > 3000mg (NO) can be obtained every 36-48h according to the enrichment and propagation process of the above embodiment3-N)/L/h, reserving 5L of inoculating liquid for the next culture, and putting the rest into an aerobic tankAfter the biochemical tank is repeatedly circulated and continuously added for 1 month, the average concentration of the ammonia nitrogen in the inlet water of the aerobic tank reaches about 30mg/L, the concentration of the ammonia nitrogen in the outlet water is about 1mg/L, and the treatment effect is obviously improved (see figure 5). The process data is shown in figure 5.
Claims (4)
1. A high-density enrichment method of autotrophic nitrifying bacteria consortium is characterized in that: inoculating thallus in autotrophic nitrifying bacteria initial culture medium, culturing at 28-32 deg.C, pH of 7.5-8.0 and Dissolved Oxygen (DO) of 20-40% under stirring condition, and culturing with NO in the culture process3The relation between the accumulation amount of N and the consumption amount of the inorganic carbon source, the pH value change in the culture process is monitored through a pH electrode, the inorganic nitrogen source and the inorganic carbon source are supplemented into the culture system, the ammonia nitrogen concentration in the culture process is controlled to be 150-300mg/L, the pH value of the system is controlled to be 7.5-8.0 through the supplemented carbon source, and the high-density automatic propagation of the autotrophic nitrifying bacteria consortium is further realized;
the autotrophic nitrifying bacteria initial culture medium contains Na in each liter of water2CO3 2g-4g、MgSO40.2-0.4 g, KCl 0.2-0.4 g, pH adjusted to 7.5-8.0 with phosphoric acid, NH4Cl 0.5g-1.0g, initial NH4-N concentration 150mg/L-300mg/L, NO in the initial medium3The concentration of N is controlled between 0mg/L and 10 mg/L;
controlling the consumption of the inorganic carbon source according to the detected pH value, further supplementing the carbon source, and calculating the supplementing amount of the inorganic nitrogen source in the corresponding time point according to the consumption of the inorganic carbon source in different time in the culture process;
culture was started after inoculation:
the addition amount of the inorganic nitrogen source in 1h-2h is as follows: { (X)1- X 0) X 0.1745 x 80 divided by V +68.826 }. times V divided by 3120, in mL/min;
the addition amount of the inorganic nitrogen source within 2h-3h is as follows: { (X) 2- X 1) X 0.1745 x 80 divided by V +68.826 }. times V divided by 3120, in mL/min;
……
the addition amount of the inorganic nitrogen source in nh- (n +1) h is as follows: { (X) n- X（n-1）) X 0.1745 x 80V +68.826 }. times.v 3120, unitIs mL/min;
wherein the consumption of the carbon source obtained every hour from 0h of the culture time is X0、X1、X2、X3、X4……Xn、Xn+1… … in mL, V is the initial volume of medium in L;
the inorganic carbon source supplemented in the culture expanding process is 5-10 wt% of Na2CO3A solution; the supplementary inorganic nitrogen source is 10 to 20 weight percent of NH4And (4) Cl solution.
2. The method for high density enrichment of an autotrophic nitrifying bacteria consortium according to claim 1, wherein: when the cumulative X value of the consumption of the inorganic carbon source reaches V/4, the nitrification rate of the culture solution for propagation is more than 160mg (NO)3-N)/L/h, after the culture expansion is finished, stopping stirring and standing for 1h, collecting the nitrosobacteria and nitrifying bacteria consortium, wherein the nitrification rate of the harvested consortium per unit mass is more than 600mg (NO)3-N）/g/h。
3. The method for high density enrichment of an autotrophic nitrifying bacteria consortium according to claim 1, wherein: the thallus is autotrophic nitrifying bacteria consortium or activated sludge containing autotrophic nitrifying bacteria; the inoculation amount is controlled to be 0.5wt% -1 wt%.
4. Use of the high-density nitrifying bacteria and nitrifying bacteria consortium harvested by the high-density enrichment method of an autotrophic nitrifying bacteria consortium according to any of claims 1-3 in sewage treatment.
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| CN110499265B (en) * | 2019-07-29 | 2022-12-09 | 辽宁格瑞凯特科技有限公司 | Autotrophic nitrifying bacteria consortium and culture expanding method thereof |
| CN112746031B (en) * | 2019-10-30 | 2023-02-03 | 中国石油化工股份有限公司 | Mixed culture method of nitrifying bacteria |
| CN112746032B (en) * | 2019-10-30 | 2023-02-03 | 中国石油化工股份有限公司 | Enrichment culture method of sulfur bacteria |
| CN114438069B (en) * | 2020-10-31 | 2023-07-28 | 中国石油化工股份有限公司 | Double microcarriers for culturing nitrifying bacteria and preparation method and application thereof |
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