CN110697887A - Method for domesticating humic acid reducing bacteria based on humic acid reduction-denitrification coupling - Google Patents
Method for domesticating humic acid reducing bacteria based on humic acid reduction-denitrification coupling Download PDFInfo
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Abstract
The invention relates to a method for domesticating humic acid reducing bacteria under the coupling condition of humic acid reduction-denitrification, which comprises the steps of firstly, respectively adding active sludge containing the humic acid reducing bacteria from a sewage treatment plant into the bottom ends of reaction chambers of an anaerobic baffle reactor through exhaust ports of the reaction chambers; pumping wastewater containing glucose and humic acid analogues into a first reaction chamber of the anaerobic baffle plate reactor as a first reaction solution through a water inlet; pumping the waste water containing nitrate as a second reaction liquid into a second reaction chamber and a fourth reaction chamber; and (5) domesticating reaction. In the reaction process, not only humic acid reducing bacteria are obtained, but also the effect of denitrification is achieved. In the technical scheme, the proportion of the bacteria with humic acid reducing capability in the four reaction chambers is increased from initial 0.31% to 21.12%, 19.89%, 25.78% and 20.03% respectively. Provides a new method for industrial application.
Description
Technical Field
The invention relates to the technical field of microbial culture or water body bioremediation, in particular to a method for domesticating humic acid reducing bacteria based on humic acid reduction-denitrification coupling.
Background
The humic acid reducing bacteria are anaerobic microorganisms with humus breathing capacity, organic pollutants can be used as electron donors, humus is used as electron acceptors, and coupling energy is used for cell growth, so that mineralization of the organic pollutants is accelerated. Therefore, the humic acid reducing bacteria have wide application prospects in the aspects of water self-purification, sediment remediation, sewage treatment and the like, and domesticating the high-efficiency humic acid reducing bacteria becomes a research hotspot at present.
At present, there are two ways for domesticating humic acid reducing bacteria: one is to add culture medium containing humic acid analogues into a serum bottle for culture; another method adopts an anaerobic culture device, and culture is carried out by pumping in a culture medium containing humic acid analogues (CN 106222091). However, in both culture modes, an aeration mode is adopted to realize the conversion of humic acid analogues from a reduction state to an oxidation state, and then oxygen in the humic acid analogues is discharged in a nitrogen blowing or chemical oxygen removal mode, so that the domesticated wastewater can be recycled. Taking patent CN106222091 as an example, an anaerobic culture device is adopted to acclimate humic acid reducing bacteria, the culture solution introduced by the device needs to further enter an aeration tank after being utilized, the culture solution enters a deoxygenation tank after being pumped by a centrifugal pump, sodium sulfite serving as a deoxygenating agent is added into the deoxygenation tank, and deoxygenation is further performed (see the attached drawing of patent CN106222091 for details). The device technology is comparatively loaded down with trivial details, and equipment is comparatively complicated, has not only increased the running cost, and the waste water after the domestication simultaneously is rich in multiple microorganism metabolite, and reuse leads to domesticating the effect not good, consequently has the domestication inefficiency, with high costs scheduling problem.
Disclosure of Invention
Aiming at the technical defects in the existing method for domesticating the humic acid reducing bacteria, a method for domesticating the humic acid reducing bacteria based on the coupling of humic acid reduction and denitrification is provided; nitrate can be used as a final electron acceptor to directly oxidize the reduced humic acid analogues, and simultaneously, the nitrate is reduced into nitrogen by microorganisms. By utilizing the anaerobic baffle plate reactor structure, the humic acid reduction reaction and the denitrification reaction are respectively carried out in different reaction chambers, so that the problems of low utilization rate and high cost of humic acid analogues are solved, the denitrification rate is accelerated, and a method is provided for industrial application.
The technical scheme of the invention is as follows:
a method for domesticating humic acid reducing bacteria based on humic acid reduction-denitrification coupling; the method utilizes an anaerobic baffle plate reactor to separate humic acid reduction and denitrification in different reaction chambers, and comprises the following steps:
1) firstly, adding activated sludge containing humic acid reducing bacteria from a sewage treatment plant into the bottom end of each reaction chamber of an anaerobic baffle reactor through an exhaust port of each reaction chamber;
2) pumping wastewater containing glucose and humic acid analogues into a first reaction chamber of the anaerobic baffle plate reactor as a first reaction solution through a water inlet; pumping the waste water containing nitrate as a second reaction liquid into a second reaction chamber and a fourth reaction chamber;
3) and (5) domesticating reaction.
The anaerobic baffle plate reactor comprises 4 reaction chambers, wherein a water inlet is formed in the side wall of a reactor body, and a water outlet is formed in the other side wall of the reactor body; the top of each reaction chamber is provided with a reaction chamber exhaust port; the lower end of the side wall of the second reaction chamber is provided with a first charging hole, and the lower end of the side wall of the fourth reaction chamber is provided with a second charging hole; a sampling port is arranged between every two reaction chambers; each reaction chamber of the anaerobic baffle reactor comprises a guide plate, a mudguard and an inclined plate; the inclined plate is used for preventing dead angles from being generated in the reaction chamber; an S-shaped channel for liquid to flow is formed between the mudguard of the front chamber and the guide plate of the rear chamber.
Preferred conditions are as follows:
the sludge concentration is 1000-2000 mg/L.
The humic acid analog refers to quinone-containing compounds such as lawsone, anthraquinone 1 sodium sulfonate, anthraquinone disulfonic acid sodium or anthraquinone 2,6 sodium sulfonate.
The concentration of the quinone-based compound is 0.05-0.10 mmol/L.
The COD of the wastewater containing glucose is 1-2 g/L.
The domestication process in the step 3) is divided into four stages for 28 days.
The first stage is day 1-8, the water inlet flow of the water inlet is 200-225mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 50-75mL/h, and the concentration of the nitrate nitrogen of the water inlet is 100 mg/L;
the second stage is 9-15 days, the water inlet flow of the water inlet is 250-300mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 75-100mL/h, and the concentration of the nitrate nitrogen of the water inlet is 100 mg/L;
the third stage is day 16-22, the water inlet flow of the water inlet is 300mL/h together with 250-100 mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same, and the concentration of the nitrate nitrogen in the inlet water is 150 mg/L;
the fourth stage is 23-28 days, the water inlet flow of the water inlet is 250-300mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 75-100mL/h, and the concentration of the nitrate nitrogen of the inlet water is 200 mg/L; the concentrations of the quinone-based compound and glucose were unchanged throughout the run.
In the technical scheme, the wastewater containing glucose and humic acid analogues enters the first reaction chamber from the water inlet and mainly undergoes a humic acid reduction reaction. The metabolite of the first reaction chamber enters the second reaction chamber along the channel formed by the first splash guard and the second guide plate. The nitrate wastewater enters the second reaction chamber from the first feed inlet, and the reduced humic acid and the nitrate react in the second reaction chamber to generate the oxidized humic acid. The effluent of the second reaction chamber enters a third reaction chamber along a channel formed by the second mud baffle and the third guide plate, and the humic acid reduction reaction continues to occur in the third reaction chamber. The effluent of the third reaction chamber enters a fourth reaction chamber along a channel formed by a third mud baffle and a fourth guide plate, the nitrate wastewater enters the fourth reaction chamber from a second feed inlet, and the reduction-state humic acid and the nitrate react in the fourth reaction chamber to generate oxidation-state humic acid.
In the reaction process, not only humic acid reducing bacteria are obtained, but also the effect of denitrification is achieved. In another aspect of the invention, based on the method for domesticating humic acid reducing bacteria under the coupling condition of humic acid reduction and denitrification, in the technical scheme, the ratio of the bacteria with humic acid reducing capability in the four reaction chambers is increased from the initial 0.31% to 21.12%, 19.89%, 25.78% and 20.03%, respectively (figure 2).
Compared with the prior art, the invention has the beneficial effects that:
1. the use of an aeration device and a deoxidant is reduced, and the operation cost is reduced.
2. The utilization rate of humus analogue lawsone is improved through the special structure of ABR;
3. the humic acid reducing bacteria are domesticated by a humic acid reduction-denitrification coupling method, so that the problems of low utilization rate and high cost of humic acid analogues are solved, and the denitrification rate is accelerated;
4. the quinone reduction reaction and the quinone oxidation reaction can be effectively separated by using the anaerobic baffle plate reactor, the process is simple, and the method is easy to be applied in practice; provides a new method for industrial application.
Drawings
FIG. 1 is a schematic view of an anaerobic baffled reactor.
FIG. 2 is a diagram showing the domestication ratio of humic acid reducing bacteria in the first embodiment.
FIG. 3 is a second example of the acclimatization ratio of humic acid-reducing bacteria.
Detailed Description
The invention is described in further detail below with reference to the figures and specific examples. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1:
the anaerobic baffled reactor is shown in figure 1: comprises a reactor body 1: a water inlet 2 is arranged on the side wall, a first feed inlet 5 is arranged on the side wall of the lower part of the second reaction chamber 1-2, a second feed inlet 6 is arranged on the side wall of the lower part of the fourth reaction chamber 1-4, a first reaction chamber exhaust port 11, a second reaction chamber exhaust port 12, a third reaction chamber exhaust port 13 and a fourth reaction chamber exhaust port 14 are arranged at the topmost end and are respectively positioned at the top of each reaction chamber, a sampling port 7 is arranged between the first reaction chamber 1-1 and the second reaction chamber 1-2, a sampling port 8 is arranged between the second reaction chamber 1-2 and the third reaction chamber 1-3, a sampling port 9 is arranged at the upper part between the third reaction chamber 1-3 and the fourth reaction chamber 1-4, and a water outlet 10 is arranged on the side wall;
sludge siltation prevention mechanism: the setting is inside reactor body 1, including first guide plate 3, second guide plate 16, third guide plate 18 and fourth guide plate 20, the guide plate bottom is 120 with guide plate vertical part contained angle, and first reaction chamber 1-1 is inside all to be equipped with swash plate 4, and swash plate 4 is 45 with 15 contained angles of fender, first fender 15, second fender 17 and third fender 19 respectively with second guide plate 16, the S type passageway that supplies the liquid circulation with the gap formation between third guide plate 18 and the fourth guide plate 20.
The exhaust port is respectively positioned right above each reaction chamber and has the functions of a detection port, a mud adding port and a mud taking port.
The operation steps are as follows:
and 2, pumping the wastewater containing glucose and humic acid analogues into a first reaction chamber of the anaerobic baffled reactor as a first reaction liquid through a water inlet 2, and pumping the wastewater containing nitrate into a second reaction chamber and a fourth reaction chamber as a second reaction liquid.
And 3, domesticating reaction.
The humic acid analog is selected from lawsone; the concentration of the lawsone is 0.05 mmol/L;
the COD of the wastewater containing glucose was 1 g/L.
The domestication process in the step 3) is divided into four stages for 28 days. The first stage is 1-8 days, the water inlet flow of the water inlet is 225mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 75mL/h, and the concentration of the nitrate and the nitrogen of the inlet water is 100 mg/L; the second stage is 9-15 days, the water inlet flow of the water inlet is 300mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 100mL/h, and the concentration of the nitrate and the nitrogen of the inlet water is 100 mg/L; the third stage is day 16-22, the water inlet flow of the water inlet is 300mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 100mL/h, and the concentration of the nitrate and the nitrogen of the inlet water is 150 mg/L; the fourth stage is 23-28 days, the water inlet flow of the water inlet is 300mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 100mL/h, and the concentration of the nitrate nitrogen of the inlet water is 200 mg/L; the concentrations of the quinone-based compound and glucose were unchanged throughout the run. The operating conditions of the anaerobic baffled reactor are shown in table 1:
TABLE 1 anaerobic baffled reactor operating conditions
In the technical scheme, the wastewater containing glucose and humic acid analogues enters the first reaction chamber from the water inlet 2 and mainly undergoes a humic acid reduction reaction. The first reaction chamber metabolite enters the second reaction chamber along the channel formed by the first splash guard 15 and the second guide plate 16. The nitrate wastewater enters the second reaction chamber from the first feed inlet 5, and the reduced humic acid and the nitrate react in the second reaction chamber to generate the oxidized humic acid. The effluent of the second reaction chamber enters a third reaction chamber along a channel formed by a second baffle 17 and a third guide plate 18, and the humic acid reduction reaction continues to occur in the third reaction chamber. Effluent of the third reaction chamber enters a fourth reaction chamber along a channel formed by a third mud baffle 19 and a fourth guide plate 20, nitrate wastewater enters the fourth reaction chamber from a second feeding hole 6, and reduced humic acid and nitrate react in the fourth reaction chamber to generate oxidized humic acid.
After the domestication by the humic acid reduction-denitrification coupling process, compared with the domestication before, the process can domesticate a large amount of humic acid reducing bacteria, wherein the flora structure is subjected to high-throughput sequencing, and finally, the colony structure analysis is carried out to obtain the proportion of the humic acid bacteria. According to the determination, the ratio of the bacteria with humic acid reducing capability in the four reaction chambers under the coupling condition of humic acid reduction and denitrification is increased to 21.12%, 19.89%, 25.78% and 20.03% from the initial 0.31% respectively; as shown in fig. 2.
Through determination, the reduction rate of the microbial quinone after disturbance load domestication is 80% -90%, and the reduction rate of the microbial quinone before domestication is 0.2%, wherein the reduction capability of the microbial quinone is determined through light absorption values, specifically, a standard curve is fitted through the light absorption values of solutions with different quinone concentrations, and then the concentration of the quinone in the solution under the corresponding light absorption value is calculated through a linear regression equation, so that the reduction rate is calculated.
In conclusion, the invention not only can domesticate the high-efficiency humic acid reducing bacteria, but also can improve the denitrification rate and the quinone utilization rate.
Example 2
The domestication method and the device are the same as the first embodiment, and the differences are as follows:
the concentration of the activated sludge in the first reaction chamber, the second reaction chamber, the third reaction chamber and the fourth reaction chamber is 2000 mg/L;
the concentration of lawsone in the first reaction liquid is 0.10mmol/L, and the COD is 2 g/L.
The first stage is 1-8 days, the water inlet flow of the water inlet is 200mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 50mL/h, and the concentration of the nitrate and the nitrogen of the inlet water is 100 mg/L; the second stage is 9-15 days, the water inlet flow of the water inlet is 250mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 75mL/h, and the concentration of the nitrate and the nitrogen of the inlet water is 100 mg/L; the third stage is 16-22 days, the water inlet flow of the water inlet is 250mL/h, the water inlet flow of the first feeding hole is 75mL/h as the water inlet flow of the second feeding hole, and the concentration of the nitrate and the nitrogen of the inlet water is 150 mg/L; the fourth stage is 23-28 days, the water inlet flow of the water inlet is 250mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 75mL/h, and the concentration of the nitrate nitrogen of the inlet water is 200 mg/L; the concentrations of the quinone-based compound and glucose were unchanged throughout the run. The operating conditions of the anaerobic baffled reactor are shown in the table 2:
TABLE 2 anaerobic baffled reactor operating conditions
After the humic acid reduction-denitrification coupling process is used for domestication, the process is compared with the process before domestication, and the process can be used
The humic acid reducing bacteria are domesticated by the humic acid reduction-denitrification coupling method, and the ratio of the bacteria with humic acid reducing capability in the four reaction chambers under the coupling condition of humic acid reduction and denitrification is respectively increased to 9.00%, 8.20%, 4.57% and 12.55% from the initial 0.31% (figure 3).
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (8)
1. A method for domesticating humic acid reducing bacteria based on humic acid reduction-denitrification coupling; the method is characterized in that an anaerobic baffle reactor is utilized to separate humic acid reduction and denitrification in different reaction chambers, and comprises the following steps:
1) firstly, adding activated sludge containing humic acid reducing bacteria from a sewage treatment plant into the bottom end of each reaction chamber of an anaerobic baffle reactor through an exhaust port of each reaction chamber;
2) pumping wastewater containing glucose and humic acid analogues into a first reaction chamber of the anaerobic baffle plate reactor as a first reaction solution through a water inlet; pumping the waste water containing nitrate as a second reaction liquid into a second reaction chamber and a fourth reaction chamber;
3) and (5) domesticating reaction.
2. The method as claimed in claim 1, wherein the anaerobic baffled reactor comprises 4 reaction chambers, a water inlet is formed on one side wall of the reactor body, and a water outlet is formed on the other side wall; the top of each reaction chamber is provided with a reaction chamber exhaust port; the lower end of the side wall of the second reaction chamber is provided with a first charging hole, and the lower end of the side wall of the fourth reaction chamber is provided with a second charging hole; a sampling port is arranged between every two reaction chambers; each reaction chamber of the anaerobic baffle reactor comprises a guide plate, a mudguard and an inclined plate; the inclined plate is used for preventing dead angles from being generated in the reaction chamber; an S-shaped channel for liquid to flow is formed between the mudguard of the front chamber and the guide plate of the rear chamber.
3. The method as set forth in claim 1, wherein the activated sludge concentration is 1000-2000 mg/L.
4. The method as set forth in claim 1, wherein the humic acid analog is a quinone group-containing compound selected from the group consisting of lawsone, anthraquinone 1 sodium sulfonate, anthraquinone disulfonate, and anthraquinone 2,6 sodium sulfonate.
5. The process as claimed in claim 1, wherein the concentration of the quinonyl-containing compound is from 0.05 to 0.10 mmol/L.
6. The method according to claim 1, wherein the COD of the wastewater containing glucose is 1 to 2 g/L.
7. The method as set forth in claim 1, wherein the acclimatization process of the step 3) is divided into four stages for 28 days.
8. The method as set forth in claim 7, wherein the first stage is day 1-8, the water inlet flow rate of the water inlet is 200-225mL/h, the water inlet flow rates of the first feed port and the second feed port are the same and are 50-75mL/h, and the concentration of the nitrate and the nitrogen in the water is 100 mg/L; the second stage is 9-15 days, the water inlet flow of the water inlet is 250-300mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 75-100mL/h, and the concentration of the nitrate nitrogen of the water inlet is 100 mg/L; the third stage is day 16-22, the water inlet flow of the water inlet is 300mL/h together with 250-100 mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same, and the concentration of the nitrate nitrogen in the inlet water is 150 mg/L; the fourth stage is 23-28 days, the water inlet flow of the water inlet is 250-300mL/h, the water inlet flow of the first feed inlet and the water inlet flow of the second feed inlet are the same and are 75-100mL/h, and the concentration of the nitrate nitrogen of the inlet water is 200 mg/L; the concentrations of the quinone-based compound and glucose were unchanged throughout the run.
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