CN111847663B - Domestication and enrichment reactor and domestication and enrichment method for sulfur autotrophic denitrifying bacteria in seawater substrate - Google Patents
Domestication and enrichment reactor and domestication and enrichment method for sulfur autotrophic denitrifying bacteria in seawater substrate Download PDFInfo
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Abstract
The invention discloses a reactor for domesticating and enriching sulfur autotrophic denitrifying bacteria, which comprises: the device comprises a reaction tank body, a mixing device, a temperature control device, a liquid guide pipe, an aeration pipe and a sealing cover; the reaction tank body is provided with an opening, the opening is sealed by a sealing cover, and the mixing device, the temperature control device, the liquid guide pipe and the aeration pipe respectively extend into the reaction tank body through an opening on the reaction tank body or the sealing cover; the aeration pipe comprises a protective air source, a flow meter and an aerator which are sequentially communicated through pipelines, the aerator is fixedly communicated with the tail end of the aeration pipe in the reaction tank body, and the tail end of the aeration pipe in the reaction tank body is communicated with the protective air source. The invention also discloses a method for domesticating the sulfur autotrophic denitrifying bacteria in the enriched seawater substrate. The method has the advantages that the rapid domestication and enrichment of the sulfur autotrophic denitrifying bacteria are realized in the seawater matrix, high-efficiency strains are provided for the biofilm reactor for treating the low-carbon-nitrogen ratio high-salinity wastewater, and the biofilm formation starting efficiency is improved.
Description
The technical field is as follows:
the invention relates to the field of biological sewage treatment, in particular to a domestication and enrichment reactor for sulfur autotrophic denitrifying bacteria in a seawater substrate and a method for domesticating and enriching the sulfur autotrophic denitrifying bacteria in a marine environment by using the reactor.
Background art:
at present, the heterotrophic denitrification process is widely applied to biological denitrification of sewage. The heterotrophic denitrifying bacteria utilize biodegradable organic matters in the water body as an electron donor and a carbon source, and nitrate as an electron acceptor to gradually convert the nitrate into gaseous nitrogen, thereby finally realizing the removal of total nitrogen in the water body. However, the demand for organic carbon sources limits the denitrification efficiency of heterotrophic denitrification in low carbon-to-nitrogen ratio wastewater (e.g., secondary sediment effluent, landfill leachate, mariculture wastewater, etc.). Therefore, in engineering practice, water-soluble organic matters such as methanol, ethanol, sodium acetate and the like are usually added into a reactor to supplement an organic carbon source for the heterotrophic denitrifying bacteria. However, the addition of water-soluble organic carbon sources presents two problems: (1) the price of the water-soluble organic carbon source is higher, so that the cost of sewage treatment is increased; (2) the denitrification effect of the heterotrophic denitrification is influenced by insufficient addition of the carbon source, and the secondary pollution of the water body is caused by excessive addition, so that the addition of the carbon source needs to be accurately controlled according to the content of nitrate nitrogen in the water body, and the process control of the heterotrophic denitrification becomes more complex. Therefore, the autotrophic denitrification process is widely concerned by researchers at home and abroad.
The autotrophic denitrifying bacteria can use reduced inorganic substance as electron donor and inorganic Carbon (CO)2、HCO3 -、CO3 2-) As a carbon source, nitrate nitrogen is reduced to gaseous nitrogen, thereby removing total nitrogen in the water body. It has been found that a wide variety of inorganic substances (e.g., reduced sulfur, hydrogen, reduced iron, etc.) can be used as electron donors for autotrophic denitrification. Among them, studies on an autotrophic denitrification process using reduced sulfur (e.g., elemental sulfur, sulfide, sulfite, pyrite, etc.) as an electron donor have been conducted. Moreover, sulfur-containing minerals such as sulfur, pyrite, pyrrhotite, etc. have begun to be used as carriers for microbial growth and denitrification electron donors in biofilm reactors (e.g., fixed bed reactors, fluidized bed reactors, etc.) for biological denitrification of low carbon-nitrogen ratio wastewater. Although the sulfur autotrophic denitrification is free from dependence on organic carbon sources, the method still has some defects, such as low denitrification efficiency, harsh operating conditions, weak impact load resistance and the like.
Compared with heterotrophic denitrifying bacteria, the growth rate of sulfur autotrophic denitrifying bacteria is low, so that the biofilm process based on sulfur autotrophic denitrification usually needs a longer start-up time, and the rapid popularization and application of the sulfur autotrophic denitrification process are limited to a certain extent, especially the application in the treatment of high-salinity wastewater (such as mariculture wastewater, seafood processing wastewater and the like). The activity of sulfur autotrophic denitrifying bacteria can be inhibited by excessively high salinity in the water body, so that the denitrification performance of the biological membrane is greatly reduced. In current research and engineering practice, activated sludge of a sewage treatment plant is usually used as seed sludge to start biofilm formation of a biofilm reactor, and the method is difficult to be applied to high-salinity wastewater. Therefore, it is of great significance to find a quick starting method of the sulfur autotrophic denitrification biomembrane process suitable for high-salinity wastewater. Numerous studies have shown that in marine environments certain species of microorganisms can utilize reduced sulfur as an electron donor for autotrophic denitrification to sustain their own growth and reproduction. If the acclimation and enrichment of the sulfur autotrophic denitrifying bacteria in the marine environment can be carried out under the condition of artificial control, and the acclimated and enriched denitrifying bacteria are used as strains for the biofilm formation starting of the subsequent biofilm process, the starting time of the biofilm can be greatly shortened, and the adaptability of the biofilm to the overhigh salinity of the water body is improved.
The invention content is as follows:
in order to solve the problems that the start of the sulfur autotrophic denitrification biomembrane process in the high-salinity wastewater is difficult and the denitrification performance is inhibited, the invention provides a method for domesticating and enriching sulfur autotrophic denitrifying bacteria in a seawater substrate, which domesticates and enriches the sulfur autotrophic denitrifying bacteria in the marine environment under the condition of artificial control so as to be used as strains of the sulfur autotrophic denitrification biomembrane process.
The invention provides a reactor for domesticating and enriching sulfur autotrophic denitrifying bacteria, which comprises:
the device comprises a reaction tank body, a mixing device, a temperature control device, a liquid guide pipe, an aeration pipe and a sealing cover;
the reaction tank body is provided with an opening, the opening is sealed by a sealing cover, and the mixing device, the temperature control device, the liquid guide pipe and the aeration pipe respectively extend into the reaction tank body through an opening on the reaction tank body or the sealing cover; the aeration pipe comprises a protective air source, a flow meter and an aerator which are sequentially communicated through pipelines, the aerator is fixedly communicated with the tail end of the aeration pipe in the reaction tank body, and the tail end of the aeration pipe in the reaction tank body is communicated with the protective air source.
In one embodiment of the invention, the sealing cover is provided with a plurality of openings, and the mixing device, the temperature control device, the liquid guide pipe and the aeration pipe respectively extend into the reaction tank body through the openings on the sealing cover.
In one embodiment according to the invention, the reactor further comprises a thermostatic control device.
In one embodiment of the invention, the thermostatic control device comprises a temperature-controlled heater and a water bath device, the water bath device is arranged in the wall of the reaction tank body, one end of the temperature-controlled heater is connected with a power supply, and the other end of the temperature-controlled heater is arranged in the water bath device.
In another aspect of the present invention, a method for acclimatizing and enriching autotrophic denitrifying bacteria in seawater substrate is provided, which comprises:
1) adding a certain amount of seed sludge, culture solution and mineral powder containing reduced sulfur into a sulfur autotrophic denitrifying bacteria domestication enrichment reactor, wherein the mixed solution is in a completely mixed state;
2) keeping an anoxic and constant-temperature environment in the reactor, operating in a sequencing batch reactor mode, and performing three stages of adaptation, domestication and enrichment to obtain a high and stable nitrate removal effect and finish domestication and enrichment of sulfur autotrophic denitrifying bacteria;
wherein the seed sludge is selected from the group consisting of marine sediments, mariculture solid waste, and sludge from anaerobic sections of sewage plants;
the culture solution is prepared from natural seawater or artificial simulated seawater.
In one embodiment according to the invention, step 1) further comprises seed-sludge elutriation, said seed-sludge elutriation being effected by a process comprising the steps of:
when the seed sludge is marine sediment or mariculture solid waste, fully stirring and mixing the seed sludge and sufficient natural seawater or artificial simulated seawater, sieving by using a 50-100-mesh sieve, standing the sieved mixed solution for a period of time, removing supernatant, and keeping bottom precipitate; when the seed sludge is anaerobic section sludge of a sewage plant, standing for a period of time for precipitation, discarding the supernatant, uniformly mixing the precipitate with sufficient culture solution, standing for precipitation again for a period of time, discarding the supernatant, repeating the steps for several times, and then retaining the bottom precipitate.
In one embodiment according to the present invention, step 2) further comprises:
the HRT is controlled to be 36-60 hours in the adaptation stage, and the residual sludge is not discharged;
entering an acclimation stage when the nitrate removal rate exceeds 40%;
the HRT of the acclimation stage is controlled to be 18-30 hours, no excess sludge is discharged, and the enrichment stage is carried out when the nitrate removal rate of two continuous operation periods exceeds 60%;
and (3) controlling the HRT in the enrichment stage to be about 12 hours, discharging a certain amount of residual sludge every day, keeping the SRT for 20-40 days, and completing the acclimation and enrichment of the sulfur autotrophic denitrifying bacteria when the nitrate removal rate of three continuous operation periods exceeds 60% and reaches a steady state.
In one embodiment according to the invention, the initial seed sludge concentration in the reactor is 3-4 g/L; the mineral is selected from one or more of sulfur, pyrite or pyrrhotite, and the dosage of the mineral powder is 1-2 g/L.
In one embodiment according to the present invention, the culture liquid contains 3.0g/L of Na as a main component2S2O3·5H2O、2.0g/L KNO3、0.5g/L NaHCO3、0.1g/L NH4Cl、0.1g/L KH2PO4。
In one embodiment according to the present invention, the concentration of dissolved oxygen during said acclimatization and enrichment process is controlled below 1mg/L and the temperature is controlled at 30 + -1 deg.C.
The invention has the beneficial effects that:
1) the domestication and enrichment of the sulfur autotrophic denitrifying bacteria are realized in the seawater substrate, the operation is simple, the speed is high, and the cost is low;
2) the domesticated and enriched sulfur autotrophic denitrifying bacteria can perform denitrification in a low-organic-carbon and high-salt environment, and have strong impact load resistance;
3) the domesticated and enriched sulfur autotrophic denitrifying bacteria can be used as strains of an autotrophic or mixotrophic biofilm reactor for treating high-salt wastewater, so that the biofilm formation starting efficiency is improved;
4) the method can also be used as a seed conservation means for sulfur autotrophic denitrifying bacteria in high-salt environment.
Drawings
FIG. 1 is a sulfur autotrophic denitrifying bacteria domestication enrichment reactor.
FIG. 2 shows the effect of acclimatization and enrichment of sulfur autotrophic denitrifying bacteria by using elemental sulfur.
FIG. 3 shows the enrichment effect of sulfur autotrophic denitrifying bacteria by pyrite.
FIG. 4 shows the variation of nitrate nitrogen concentration in inlet and outlet water during biofilm reactor biofilm formation start-up with different strains.
Description of reference numerals:
1 protective gas, 2 gas flow meters, 3 sealing covers, 3a stirring rod holes, 3b protective gas guide pipe holes, 3c liquid feeding/discharging pipe holes, 3d thermometer holes, 3e water quality parameter detection holes, 3f exhaust guide pipe holes, 4 mixing devices, 5 glass guide pipes, 6 reaction tank bodies, 7 temperature control heaters, 8 aerators and 9 water baths.
The specific implementation mode is as follows:
the following detailed description of the preferred embodiments of the present invention, taken in conjunction with the accompanying drawings, will make the advantages and features of the invention more readily understood by those skilled in the art, and thus will more clearly and distinctly define the scope of the invention.
Example 1 bench scale acclimatization and enrichment reactor for sulfur autotrophic denitrifying bacteria
The invention relates to a sulfur autotrophic denitrifying bacteria domestication enrichment reactor which mainly comprises a reaction tank body, a mixing device, a temperature control device, a liquid feeding/discharging device, a protective gas aeration device and a water quality parameter detection device. Thus, a set of small-scale sulfur autotrophic denitrifying bacteria domestication enrichment reactor is constructed, and the structural schematic diagram of the reactor is shown in figure 1. A brown glass jar is used as a reaction tank body 6, and the effective volume is 5L; a rubber plug is used as a sealing cover 3, and holes are punched on the cover according to the position shown in figure 1; a mechanical stirring rod is inserted into the stirring rod hole 3a, a glass guide pipe 5 is inserted into the protective gas guide pipe hole 3b, the liquid feeding/discharging pipe hole 3c, the water quality parameter detection hole 3e and the exhaust guide pipe hole 3f, and a thermometer is inserted into the thermometer hole 3 d; the mixing device 4 adopts a speed-adjustable mechanical stirrer (not shown) as a power device; nitrogen is used as protective gas 1, a gas flow meter 2 is used for controlling the gas flow, and the gas is uniformly introduced into a reaction tank body 6 through an aerator 8 to create an anoxic environment for the reaction tank body 6; placing a temperature control heater 7 in a water bath 9 to provide constant temperature conditions for the reaction tank body 6; adding fresh culture solution and discharging waste liquid through a siphon reaction tank body; the tail end of the exhaust conduit is sealed by water.
Of course, the self-made small-scale reactor can also be used for modifying equipment such as a fermentation tank and the like based on the principle.
Example 2 acclimatization and enrichment of Sulfur autotrophic denitrifying bacteria Using elemental Sulfur
The seawater is adopted to prepare artificial simulated seawater, and the salinity is about 3.2%. Fully mixing the marine sediments and the mariculture solid wastes with artificial simulated seawater respectively, then sieving the mixture by a sieve with 50-100 meshes, standing the sieved mixture for 10 minutes, removing the supernatant, and keeping the bottom sediments. Standing and precipitating the sludge in the anaerobic section of the sewage plant for 30 minutes, discarding the supernatant, then uniformly mixing the precipitate with the artificial simulated seawater again, standing and precipitating for 30 minutes again, discarding the supernatant, repeating the steps for 2 times, and finally retaining the bottom precipitate. Adding analytically pure Na into artificial simulated seawater2S2O3·5H2O(3.0g/L)、KNO3(2.0g/L)、NaHCO3(0.5g/L)、NH4Cl(0.1g/L)、KH2PO4(0.1g/L) to prepare a culture solution of the sulfur autotrophic denitrifying bacteria. Adding the elutriated seed sludge into the sulfur autotrophic denitrifying bacteria domestication enrichment reactor in the embodiment 1, then injecting a culture solution into the reactor, and starting a mechanical stirrer to uniformly mix the sludge; the initial concentration of the seed sludge was determined to be 3.3 g/L. Finally, the process is carried out in a batch,sulfur powder (1.5g/L) was added to the reactor. Introducing protective gas into the reactor for 15 minutes to drive air at the top of the reactor and in the sludge mixed liquor, so as to create an anoxic environment; starting a temperature control heater to heat the water bath, and setting the temperature to be 30 ℃.
The acclimatization and enrichment process of the sulfur autotrophic denitrifying bacteria is divided into three stages of adaptation, acclimatization and enrichment. HRT in the adaptation stage is 48 hours, no excess sludge is discharged, and when the nitrate removal rate exceeds 40%, the acclimation stage is started; HRT of the acclimation stage is 24 hours, residual sludge is not discharged, and the enrichment stage is carried out when the nitrate removal rate of two continuous operation periods exceeds 60 percent; HRT in the enrichment stage is 12 hours, 160mL of sludge mixed liquor is discharged every day, SRT is kept for about 30 days, and when the nitrate removal rate in three continuous operation periods exceeds 60% and reaches a steady state, the domestication and enrichment of the sulfur autotrophic denitrifying bacteria are completed. During the reactor run, the nitrate nitrogen concentration in the effluent was measured at the end of each cycle. As shown in fig. 2, after the reactor was operated for 4 days, the nitrate removal rate reached 40.2%; from day 5, the reactor was operated into the acclimation phase, with nitrate removal rates higher than 60% on days 8 and 9; from day 10 on, the reactor was run into the enrichment phase and the nitrate removal rate after day 11 was over 60% and remained relatively stable. Therefore, after the reactor is operated for 11 days, the acclimatization and enrichment of the sulfur autotrophic denitrifying bacteria are completed.
Example 3 acclimatization and enrichment of autotrophic denitrifying bacteria on pyrite
The same procedure as in example 2 was repeated except that pyrite powder was used instead of the sulfur powder in example 2, and the effect of acclimatization and enrichment was as shown in FIG. 3. It can be seen that the nitrate removal rate reaches 41.1% after the reactor is operated for 4 days; from day 5, the reactor was run into the acclimation phase with nitrate removal rates of 65.4% and 61.6% on days 8 and 9, respectively; from day 10 on, the reactor was run into the enrichment phase and the nitrate removal rate after day 11 was over 60% and remained relatively stable. Thus, after 11 days of operation, the acclimatization and enrichment of the sulfur autotrophic denitrifying bacteria are completed.
Example 4: influence of different strains on biofilm reactor biofilm formation start
The sulfur autotrophic denitrifying bacteria and the sludge in the anaerobic section of the sewage plant in the example 2 are respectively inoculated into two same biofilm reactors, the initial concentrations of the sludge are respectively 1.3 and 1.4g/L, and the sludge is subjected to biofilm formation starting. The biomembrane reactor adopts a packed column form, takes sulfur and limestone particles as fillers, and the particle size of the sulfur and the limestone particles is about 5 mm. Analytically pure drugs such as potassium nitrate, monopotassium phosphate and the like are added into the artificial simulated seawater described in example 1, artificial simulated mariculture wastewater is prepared and used as inlet water of the two reactors. The change of nitrate nitrogen concentration in the inlet water and the outlet water during the start-up of biofilm formation in the two reactors is shown in figure 4. It can be seen that for the biofilm reactor inoculated with the sulfur autotrophic denitrifying bacteria after domestication and enrichment, the concentration of nitrate nitrogen in effluent water rapidly decreases along with the prolonging of the starting time, the nitrate nitrogen concentration in effluent water reaches relatively stable after 12 days, the removal rate is kept above 80%, and the reactor can be considered to be successfully started by biofilm formation. For the biofilm reactor for inoculating the sludge in the anaerobic section of the sewage plant, the reduction speed of the nitrate nitrogen concentration in the effluent along with the starting time is obviously smaller than that of the former, the nitrate nitrogen removal rate is 65.4 percent after the effluent is operated for 30 days, but the relative stable state is not reached yet, and the biofilm formation starting is required to be continued. Therefore, the membrane hanging starting efficiency of the biofilm reactor can be greatly improved by taking the domesticated and enriched sulfur autotrophic denitrifying bacteria as strains.
The above summary and the detailed description are intended to demonstrate the practical application of the technical solutions provided by the present invention, and should not be construed as limiting the scope of the present invention. Various modifications, equivalent substitutions, or improvements may be made by those skilled in the art within the spirit and principles of the invention. The scope of the invention is to be determined by the appended claims.
Claims (2)
1. A method for domesticating sulfur autotrophic denitrifying bacteria in enriched seawater substrates is characterized by comprising the following steps:
1) adding a certain amount of seed sludge, culture solution and mineral powder containing reduced sulfur into a sulfur autotrophic denitrifying bacteria domestication enrichment reactor, wherein the mixed solution is in a completely mixed state;
2) keeping an anoxic and constant-temperature environment in the reactor, operating in a sequencing batch reactor mode, and performing three stages of adaptation, domestication and enrichment to obtain a high and stable nitrate removal effect and finish domestication and enrichment of sulfur autotrophic denitrifying bacteria;
wherein the seed sludge is selected from the group consisting of marine sediments, mariculture solid waste, and sludge from anaerobic sections of sewage plants;
the culture solution is prepared from natural seawater or artificial simulated seawater;
the method also comprises seed sludge elutriation in the step 1), and the seed sludge elutriation is realized by a method comprising the following steps:
when the seed sludge is marine sediment or mariculture solid waste, fully stirring and mixing the seed sludge and sufficient natural seawater or artificial simulated seawater, sieving by using a 50-100-mesh sieve, standing the sieved mixed solution for a period of time, removing supernatant, and keeping bottom precipitate; when the seed sludge is the anaerobic section sludge of the sewage plant, standing for a period of time, discarding the supernatant, uniformly mixing the precipitate with sufficient natural seawater or artificial simulated seawater, standing for a period of time again, discarding the supernatant, repeating the steps for several times, and then retaining the bottom precipitate;
the step 2) also comprises the following steps:
the HRT is controlled to be 36-60 hours in the adaptation stage, and the residual sludge is not discharged;
entering an acclimation stage when the nitrate removal rate exceeds 40%;
the HRT of the acclimation stage is controlled to be 18-30 hours, no excess sludge is discharged, and the enrichment stage is carried out when the nitrate removal rate of two continuous operation periods exceeds 60%;
the HRT in the enrichment stage is controlled to be 12 hours, a certain amount of residual sludge is discharged every day, the SRT is kept for 20-40 days, and when the nitrate removal rate of three continuous operation periods exceeds 60% and reaches a steady state, the acclimation and enrichment of the sulfur autotrophic denitrifying bacteria are completed;
the initial seed sludge concentration in the reactor is 3-4 g/L; the mineral is selected from one or two of pyrite or pyrrhotite, and the dosage of the mineral powder is 1-2 g/L; the concentration of dissolved oxygen in the domestication and enrichment process is controlled to be below 1mg/L, and the temperature is controlled to be 30 +/-1 ℃.
2. The method according to claim 1, wherein the culture medium contains 3.0g/L Na as a main component2S2O3·5H2O、2.0 g/L KNO3、0.5 g/L NaHCO3、0.1 g/L NH4Cl、0.1 g/L KH2PO4。
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