CN112010427A - Construction method and application of two-step decay model of nitrite oxidizing bacteria - Google Patents

Construction method and application of two-step decay model of nitrite oxidizing bacteria Download PDF

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CN112010427A
CN112010427A CN202010932407.3A CN202010932407A CN112010427A CN 112010427 A CN112010427 A CN 112010427A CN 202010932407 A CN202010932407 A CN 202010932407A CN 112010427 A CN112010427 A CN 112010427A
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刘兵
陈飞勇
邵媛媛
宋扬
王静
吴金柱
李一凡
徐保建
杨寓超
徐成远
刘华
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Abstract

The invention relates to a construction method and application of a two-step decay model of nitrite oxidation bacteria, and solves the problem that the traditional bacterial decay model is only suitable for the primary stage (live cells → dead cells) of an NOB decay stage and cannot evaluate the conversion of the dead cells into soluble COD. The invention scientifically simulates the NOB apoptosis process under high-concentration nitrite, establishes a two-step apoptosis model (living cells → dead cells → soluble COD), and determines the model parameters of NOB apoptosis under nitrite inhibition. The invention establishes a two-step decline mathematical model of nitrite oxidizing bacteria for the first time, and the model can ensure the denitrification effect of short-cut nitrification and denitrification by controlling the concentration of nitrite. Improving the sludge treatment effect. Also provides technical support for further understanding the phenomena of microorganism inhibition and activation recovery.

Description

Construction method and application of two-step decay model of nitrite oxidizing bacteria
Technical Field
The invention belongs to the field of sewage treatment, relates to a construction method and application of a two-step decay model of nitrite oxidizing bacteria, and particularly relates to a two-step decay model of nitrite oxidizing bacteria under the inhibition of high nitrite concentration.
Background
The conventional biological denitrification process is a process in which ammonia nitrogen generates Nitrite under the action of ammonia oxidizing Bacteria, then the Nitrite is oxidized into nitrate by Nitrite-oxidizing Bacteria (NOB), and finally the nitrate generates nitrogen under the action of denitrifying Bacteria. The short-cut nitrification and denitrification process is to control the generation of nitrite so that denitrifying bacteria can directly complete the denitrification process by taking the nitrite as an electron acceptor. Compared with the traditional biological denitrification process, the method has the obvious advantages of less oxygen demand in the nitrification stage and less carbon source consumption in the denitrification stage. How to inhibit the generation of nitrate is a key factor for ensuring the efficiency of short-cut nitrification and denitrification. The nitrite concentration with high concentration can inhibit nitrite oxidizing bacteria and can be used as a control condition for developing short-cut nitrification and denitrification, so that the activity of Nitrite Oxidizing Bacteria (NOB) with high concentration is evaluated to be beneficial to the control of short-cut nitrification and denitrification. However, the traditional bacterial apoptosis model is only suitable for the primary stage (live cells → dead cells) of the nitrite oxidation bacterial apoptosis stage, and the conversion of the dead cells into soluble COD cannot be evaluated.
Therefore, in order to evaluate the NOB activity under high-concentration nitrite and improve the control effect on short-cut nitrification and denitrification, a new nitrite oxidizing bacteria decay model is urgently needed. The present invention is based on the premise that no model of this type is currently available in the art.
Disclosure of Invention
The invention mainly aims to scientifically simulate the decay process of Nitrite Oxidizing Bacteria (NOB) under high-concentration nitrite and establish a two-step decay model (living cells → dead cells → soluble COD). According to the invention, by measuring model parameters of NOB decay under the inhibition of nitrite, a complete two-step decay mathematical model is established, so that the shortcut nitrification and denitrification nitrogen removal effect can be ensured by controlling the concentration of nitrite.
In order to achieve the purpose, the invention adopts the technical scheme that: a method for constructing a two-step decline model of nitrite oxidizing bacteria comprises the following steps:
(1) the net specific growth rate of the simulated nitrous acid oxidizing bacteria was calculated and obtained using eq.1:
r＝u-btot (Eq.1)
wherein, r: net specific rate of NOB increase (d)-1) Specific growth rate of u, NOB (d)-1)，btot: total NOB specific decay Rate (d)-1)。
(2) Considering the inhibition of NOB by nitrite at high concentrations, the results were obtained using Eq.2:
Figure BDA0002670679780000021
wherein,
Figure BDA0002670679780000022
maximum specific growth rate (d) of NOB-1),
Figure BDA0002670679780000023
Nitrite concentration (mg-N/L),
Figure BDA0002670679780000024
the half saturation factor (mg-N/L) for NOB growth,
Figure BDA0002670679780000025
half saturation factor (mg-N/L) for NOB growth inhibition.
(3) Simulating and calculating the total ratio decay rate of NOB, introducing the toxicity influence of nitrite, wherein the toxicity of the nitrite is expressed by an S-type formula with power n being more than 1; obtained using eq.3:
Figure BDA0002670679780000026
wherein, b is NOB intrinsic specific decay Rate (d)-1)，
Figure BDA0002670679780000027
NOB nitrite toxic decay Rate (d)-1)，
Figure BDA0002670679780000028
Half-saturation factor (mg-N/L) for NOB nitrite toxicity decay, N: the power of nitrite concentration.
(4) Simulating the conversion of dead NOB to soluble COD, the total specific degradation of NOB was introduced and obtained using Eq.4:
Figure BDA0002670679780000029
wherein: bl,tot: NOB Total specific degradation Rate (d)-1)，bl: NOB intrinsic specific degradation rate (d)-1)，
Figure BDA00026706797800000210
NOB nitrite inhibited degradation rate (d)-1)，
Figure BDA00026706797800000211
NOB nitrite inhibited degradation half saturation factor (mg-N/L), m: the power of nitrite concentration.
(5) Modeling
TABLE 1 two-step decay model of nitrite-oxidizing bacteria
Figure BDA0002670679780000031
The two-step decay model can be used for sewage treatment, and the sewage treatment effect is improved. Furthermore, GPS-X software is used for simulating aerobic rate OUR and COD of nitrifying bacteria of the activated sludge based on the two-step decay model, the shortcut nitrification and denitrification effect is ensured by controlling the concentration of nitrite, and the sewage treatment effect is improved.
The invention provides a NOB intensified decay and death (X) for the first time based on an IWA activated sludge model NO.1(ASM1) commonly used in the fieldB，L→XB，D) And enhanced degradation (X)B,D→SB) The two-step decay model of (1). The invention scientifically simulates the NOB decay process under high-concentration nitrite, establishes a two-step decay model (living cells → dead cells → soluble COD), determines the model parameters of NOB decay under the inhibition of nitrite, and perfects the two-step decay mathematical model, thereby realizing the purpose of ensuring the denitrification effect by short-cut nitrification by controlling the concentration of nitrite. The invention provides important support for further understanding the phenomena of microorganism inhibition and activation recovery while establishing and perfecting a two-step decline mathematical model.
Compared with the prior art, the invention has the following advantages:
(1) the invention uses a mathematical model method to determine the influence of nitrite on NOB, and establishes a two-step apoptosis model (live cells → dead cells → soluble COD) for the first time. Compared with a mathematical model which cannot evaluate the conversion of dead cells into soluble COD by the traditional method, the model of the invention is obviously more perfect, precise and accurate.
(2) In the establishing process of the model, the nitrite concentration factor parameter of the NOB is determined, and the toxicity influence of the nitrite is introduced in the simulation calculation of the total NOB ratio decay rate for the first time. And in the second stage, the invention introduces the total specific degradation rate of NOB in order to simulate the conversion process of dead NOB into soluble COD. The invention integrates the influencing factors of all aspects and realizes the purpose of perfecting and enriching the NOB mathematical model in the field.
(3) The NOB two-step apoptosis model established by the invention solves the problem that the existing model in the field can not evaluate the conversion of dead cells into soluble COD. The model of the invention can be used for seeking an optimized environmental condition, thereby promoting the formation of the short-cut nitrification and denitrification by controlling the activity of NOB and finally improving the sewage treatment effect.
(4) The determination method is simple and easy to operate, has high repeatability, is convenient to popularize and use, and can promote the development and innovation of mathematical model methods in the field of sewage treatment. Also provides important support for further understanding the phenomena of microorganism inhibition and activation recovery.
Drawings
FIG. 1 is a schematic diagram of an OUR experimental apparatus of the present invention; in fig. 1: 1. a water bath magnetic stirrer; 2. a magnetic rotor; 3. BOD bottle; 4. a Dissolved Oxygen (DO) measurement probe; 5. dissolved Oxygen (DO) meter.
FIG. 2 is a graph of the simulation of OUR and COD of nitrite-oxidized bacteria NOB in example 2 using the two-step decay model of the present invention at a nitrite concentration of 50 mg-N/L.
FIG. 3 is a graph of the simulation of OUR and COD of nitrite-oxidized bacteria NOB in example 3 using the two-step decay model of the present invention at a nitrous acid concentration of 2000 mg-N/L.
Detailed Description
The invention is further described with reference to the following figures and specific embodiments. In the present invention, reagents used are those conventional in the art, and methods used are those conventional in the art, unless otherwise specified.
Example 1 construction of two-step decay model of nitrite-oxidizing bacteria of the present invention
1. With NaNO-containing gas2(500mg-N/L)、NH4CI(1.0mg-N/L)、KH2PO4(0.13mg-P/L) and Na2PO4(1.67-P/L) synthetic wastewater enriched with sludge (NOB) in a 5-L Sequencing Batch Reactor (SBR)The purpose of enrichment is to eliminate interference from other species). The pH, temperature and DO of the reactor were controlled at 7.3, 35 + -0.5 deg.C and above 5mg/L, respectively. The SRT of the SBR reactor was maintained for 50 days, and after 240 days of culture, experiments were carried out using concentrated NOB sludge.
2. NH for concentrated NOB sludge4CI(1.0mg-N/L)、KH2PO4(0.13mg-P/L) and Na2The synthetic wastewater of PO4(1.67-P/L) was centrifuged three times (5000rpm) for three minutes each. Before NOB sludge culture, three centrifugal washes of sludge samples were performed using inorganic synthetic wastewater without nitrite to ensure that the samples were nitrite free. Obtaining NOB concentrated sludge without nitrite for later use.
3. NOB-enriched sludge containing no nitrite was used for breath test, NOB culture was carried out under experimental conditions in which nitrite concentration was maintained at 0mg-N/L, 50mg-N/L, 500mg-N/L, 2000mg-N/L, and cultured in a constant temperature incubator at 35 + -0.2 ℃ for 7 days while dissolved oxygen in the culture vessel was maintained at 6mg-O2More than/L.
4. And (3) distinguishing the change of the number of live bacteria and dead bacteria in the culture boxes with different nitrite concentrations by using bacterial staining, and solving model parameters according to the measured data. See table 2 below:
TABLE 2 kinetic and stoichiometric parameters of NOB
Figure BDA0002670679780000061
5. Establishing a model equation of two-step decay of nitrite oxidizing bacteria NOB:
(1) the net specific growth rate of the simulated nitrous acid oxidizing bacteria was calculated and obtained using eq.1:
r＝u-btot (Eq.1)
wherein, r: net specific rate of NOB increase (d)-1) Specific growth rate of u, NOB (d)-1)，btot: total NOB specific decay Rate (d)-1)；
(2) Considering the inhibition of NOB by nitrite at high concentrations, the results were obtained using Eq.2:
Figure BDA0002670679780000071
wherein,
Figure BDA0002670679780000072
maximum specific growth rate (d) of NOB-1),
Figure BDA0002670679780000073
Nitrite concentration (mg-N/L),
Figure BDA0002670679780000074
the half saturation factor (mg-N/L) for NOB growth,
Figure BDA0002670679780000075
the half saturation factor (mg-N/L) for NOB growth inhibition;
(3) introducing the toxicity influence of nitrite in the simulation calculation of the NOB total ratio decay rate, wherein the toxicity of the nitrite is expressed by an S-type formula with power n being more than 1; obtained using eq.3:
Figure BDA0002670679780000076
wherein, b is NOB intrinsic specific decay Rate (d)-1)，
Figure BDA0002670679780000077
NOB nitrite toxic decay Rate (d)-1)，
Figure BDA0002670679780000078
Half-saturation factor (mg-N/L) for NOB nitrite toxicity decay, N: the power of nitrite concentration.
(4) To simulate the process of conversion of dead NOB to soluble COD, the overall specific degradation rate of NOB was introduced and obtained using Eq.4:
Figure BDA0002670679780000079
wherein: bl,tot: NOB Total specific degradation Rate (d)-1)，bl: NOB intrinsic specific degradation rate (d)-1)，
Figure BDA00026706797800000710
NOB nitrite inhibited degradation rate (d)-1)，
Figure BDA00026706797800000711
NOB nitrite inhibited degradation half saturation factor (mg-N/L), m: the power of nitrite concentration.
(5) Modeled in Table 1 below
TABLE 1 two-step decay model of nitrite-oxidizing bacteria
Figure BDA0002670679780000081
Example 2 OUR and COD of NOB were measured and simulated using the model equations set forth in example 1 with a nitrous acid concentration of 50 mg-N/L.
(1) With NaNO-containing gas2(500mg-N/L)、NH4CI(1.0mg-N/L)、KH2PO4(0.13mg-P/L) and Na2PO4(1.67-P/L) synthetic wastewater sludge was enriched in a 5-L Sequencing Batch Reactor (SBR) (NOB enrichment was aimed at eliminating interference from other species). The pH, temperature and DO of the reactor were controlled at 7.3, 35 + -0.5 deg.C and above 5mg/L, respectively. The SRT of the SBR reactor was maintained for 50 days, and after 240 days of culture, experiments were carried out using concentrated NOB sludge.
(2) NH for concentrated NOB sludge4CI(1.0mg-N/L)、KH2PO4(0.13mg-P/L) and Na2PO4The synthetic wastewater (1.67-P/L) was washed three times (5000rpm) by centrifugation for three minutes each. Before NOB sludge culture, three centrifugal washes of sludge samples were performed using inorganic synthetic wastewater without nitrite to ensure that the samples were nitrite free.
(3) Use of nitrite-free NOB enriched sludge forBreath test, NOB culture under the experimental condition that nitrite concentration is kept at 50mg-N/L, culturing at 35 + -0.2 deg.C for 7 days in a constant temperature incubator while dissolved oxygen in the culture vessel is required to be kept at 6mg-O2More than/L.
(4) In order to maintain the nitrite concentration during the test, a computer programmed syringe pump was installed to continuously feed a high concentration sodium nitrite solution (20000mg-N/L) into the culture vessel to control the nitrite concentration to within + -5% of the target.
(5) The activated sludge in the culture tanks was sampled manually at intervals of 6 to 8 hours, and placed in 100mL BOD bottles after adjusting the nitrite concentration to the same predetermined concentration for each culture tank. Using O2The gas sets the initial dissolved oxygen concentration of the sample to be in excess of 10mg-O2L, and the consumption of dissolved oxygen in the BOD bottle was recorded every 1 minute using a portable dissolved oxygen meter. OUR of each culture vessel was obtained from the decrease in the dissolved oxygen concentration with time. The concentration of dissolved oxygen is taken as a vertical axis, time is taken as a horizontal axis to be plotted, and the coefficient R is determined by the plotted straight line2And the slope of the straight line is more than or equal to 99, and the oxygen consumption rate of the NOB is obtained.
(6) 5ml of NOB sludge was taken out from the culture tank and washed 3 times by centrifugation (10000rpm, three minutes of washing) with 0.85% NaCl solution to remove the interference of impurities.
(7) COD of the NOB sludge washed in the culture tank was measured using a COD analysis kit (TNT Plus 821(COD range 3-150 ppm, U.S. Hash).
(8) A scatter diagram of nitrite concentration, NOB oxygen consumption rate (OUR) and sludge COD is made, the scatter diagram is fitted by using GPS-X software based on the two-step decay model equation of the invention, measured data and simulation data are respectively shown in tables 3 and 4, and experimental results and fitting effects are shown in figure 2.
TABLE 3
t 50mg-N/L t 50mg-N/L
OUR COD
d Measured value d Measured value
0.034722222 4.23 0.010416667 74.29447853
0.25 5.71956 0.96875 78.52760736
0.415277778 5.46 1.948611111 79.95008841
0.96875 5.54046 2.943055556 82.68033429
1.208333333 6.4506 3.932638889 85.27607362
1.44375 5.69322 4.929861111 92.02453988
1.975 5.6592 5.932638889 97.98347717
2.208333333 5.7528 6.936111111 104.2944785
2.460416667 5.811 7.93125 117.8064389
2.919444444 5.50668
3.20625 6.5637
3.413888889 6.1425
3.943055556 6.75
4.2125 6.87
4.414583333 7.68
4.934722222 8.9
5.202083333 8.52
5.427777778 7.25724
5.944444444 8.814
6.207638889 9.4776
6.413888889 9.3636
6.941666667 10.8739726
7.211805556 10.67664
7.475694444 12.08064
8.099305556 11.8734
TABLE 4
Figure BDA0002670679780000101
Figure BDA0002670679780000111
Example 3 OUR and COD of NOB were detected and simulated using the model equations set forth in example 1 with a nitrous acid concentration of 2000 mg-N/L.
(1) With NaNO-containing gas2(500mg-N/L)、NH4CI(1.0mg-N/L)、KH2PO4(0.13mg-P/L) and Na2PO4(1.67-P/L) of the synthetic wastewater was enriched with sludge in a 5-L Sequencing Batch Reactor (SBR). The pH, temperature and DO of the reactor were controlled at 7.3, 35 + -0.5 deg.C and above 5mg/L, respectively. The SRT of the SBR reactor was maintained for 50 days, and after 240 days of culture, experiments were carried out using concentrated NOB sludge.
(2) For concentrated NOB sludgeNH4CI(1.0mg-N/L)、KH2PO4(0.13mg-P/L) and Na2PO4The synthetic wastewater (1.67-P/L) was washed three times (5000rpm) by centrifugation for three minutes each.
(3) NOB-enriched sludge containing no nitrite was used for breath test, NOB culture was carried out under the experimental conditions that the nitrite concentration was maintained at 2000mg-N/L, and cultured in a constant temperature incubator at 35 + -0.2 ℃ for 7 days while the dissolved oxygen in the culture vessel was maintained at 6mg-O2More than/L.
(4) In order to maintain the nitrite concentration during the test, a computer programmed syringe pump was installed to continuously feed a high concentration sodium nitrite solution (20000mg-N/L) into the culture vessel to control the nitrite concentration to within + -5% of the target.
(5) The activated sludge in the culture tanks was sampled manually at intervals of 6 to 8 hours, and placed in 100mL BOD bottles after adjusting the nitrite concentration to the same predetermined concentration for each culture tank. Using O2The gas sets the initial dissolved oxygen concentration of the sample to be in excess of 10mg-O2L, and the consumption of dissolved oxygen in the BOD bottle was recorded every 1 minute using a portable dissolved oxygen meter. OUR of each culture vessel was obtained from the decrease in the dissolved oxygen concentration with time.
(6) 5ml of NOB sludge was taken out from the culture tank and washed 3 times by centrifugation (10000rpm, three minutes of washing) with 0.85% NaCl solution to remove the interference of impurities.
(7) COD of the NOB sludge washed in the culture tank was measured using a COD analysis kit (TNT Plus 821(COD range 3-150 ppm, U.S. Hash).
(8) A scatter diagram of nitrite concentration, NOB oxygen consumption rate (OUR) and NOB sludge COD is made, the scatter diagram is fitted by using GPS-X software based on the two-step decay model equation, experimental data are shown in tables 5 and 6, and experimental results and fitting effects are shown in figure 3.
TABLE 5
Figure BDA0002670679780000121
Figure BDA0002670679780000131
TABLE 6
t 50mg-N/L 50mg-N/L
OUR COD
d Simulation of Simulation of
0 4.351442 74
0.25 5.455606 74.70636
0.5 5.604481 75.47575
0.750001 5.729357 76.29351
1.000001 5.848524 77.14692
1.250001 5.966282 78.02398
1.500001 6.084471 78.91807
1.750001 6.203997 79.82719
2.000002 6.3253 80.75166
2.250002 6.448603 81.69247
2.500002 6.574032 82.6506
2.750002 6.701681 83.62686
3.000002 6.831623 84.62195
3.250003 6.963927 85.63643
3.500003 7.098653 86.67084
3.750003 7.235861 87.72568
4.000003 7.375605 88.80142
4.250003 7.517942 89.89853
4.500004 7.662924 91.01748
4.750004 7.810604 92.15872
5.000004 7.961036 93.32271
5.250004 8.114273 94.50991
5.500004 8.270367 95.72079
5.750005 8.429372 96.95581
6.000005 8.591342 98.21544
6.250005 8.756331 99.50016
6.500005 8.924395 100.8104
6.750005 9.095589 102.1468
6.958339 9.240683 103.2807
The data and simulations obtained in examples 2 and 3 above fully demonstrate the validity of the two-step decay model of nitrite-oxidizing bacteria of the present invention. Therefore, the nitrite concentration required for inhibiting the activity of NOB can be calculated through a two-step decay model, and the control of the short-cut nitrification and denitrification process is realized. Namely, the invention realizes that the short-cut nitrification and denitrification effect is ensured by controlling the concentration of the nitrite.

Claims (4)

1. A method for constructing a two-step decay model of nitrite oxidizing bacteria is characterized by comprising the following steps of:
(1) the net specific growth rate of the simulated nitrous acid oxidizing bacteria was calculated and obtained using eq.1:
r＝u-btot (Eq.1)
wherein, r: net specific rate of NOB increase (d)-1) Specific growth rate of u, NOB (d)-1)，btot: total NOB specific decay Rate (d)-1)；
(2) Considering the inhibition of NOB by nitrite at high concentrations, the results were obtained using Eq.2:
Figure FDA0002670679770000011
wherein,
Figure FDA0002670679770000012
maximum specific growth rate (d) of NOB-1)，
Figure FDA0002670679770000013
Nitrite concentration (mg-N/L),
Figure FDA0002670679770000014
the half saturation factor (mg-N/L) for NOB growth,
Figure FDA0002670679770000015
the half saturation factor (mg-N/L) for NOB growth inhibition;
(3) simulating and calculating the total ratio decay rate of NOB, introducing the toxicity influence of nitrite, wherein the toxicity of the nitrite is expressed by an S-type formula with power n being more than 1; obtained using eq.3:
Figure FDA0002670679770000016
wherein, b: NOB intrinsic specific decay Rate (d)-1)，
Figure FDA0002670679770000017
NOB nitrite toxic decay Rate (d)-1)，
Figure FDA0002670679770000018
Half-saturation factor (mg-N/L) for NOB nitrite toxicity decay, N: the power of nitrite concentration;
(4) simulating the conversion of dead NOB to soluble COD, the overall specific degradation rate of the introduced NOB was obtained using Eq.4:
Figure FDA0002670679770000019
wherein: bl，tot: NOB Total specific degradation Rate (d)-1)，bl: NOB intrinsic specific degradation rate (d)-1)，
Figure FDA00026706797700000110
NOB nitrite inhibited degradation rate (d)-1)，
Figure FDA00026706797700000111
NOB nitrite inhibited degradation half saturation factor (mg-N/L), m: the power of nitrite concentration;
(5) modeling
Figure FDA0002670679770000021
2. A two-step decay model of nitrite-oxidizing bacteria constructed by the method of claim 1.
3. The two-step decay model of nitrite oxidizing bacteria of claim 2, wherein the model is a two-step decay model of nitrite oxidizing bacteria under high nitrite concentration inhibition.
4. Use of the two-step decay model of claim 2 in wastewater treatment, wherein: calculating the nitrite concentration required to inhibit the activity of NOB by the two-step decay model; namely, the short-cut nitrification and denitrification nitrogen removal effect is ensured by controlling the concentration of the nitrite, and the sewage treatment effect is improved.
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