MINISTRY OF EDUCATION VIETNAM ACADEMY OF AND TRAINING SCIENCE AND TECHNOLOGY GRADUATE UNIVERSITY OF SCIENCE AND TECHNOLOGY VU DUY NHAN STUDY ON INTERNAL ELECTROLYSIS COMBINE WITH AAO-MBBR TO TREAT TNT WASTEWATER Speciality: Chemical Engineering Code: 52 03 01 PhD DISSERTATION SUMMARY ON CHEMICAL ENGINEERING Ha Noi - 2020 The work was completed at: Vietnam Academy of Science and Technology Science instructor: Assoc Prof Le Thi Mai Huong Prof Le Mai Huong Reviewer 1: Reviewer 2: Reviewer 3: The dissertation will be presented in front of Dissertation Evaluation Council at Institute level at the Institute of Natural Products Chemistry - Vietnam Academy of Science and Technology, No 18 Hoang Quoc Viet, Cau Giay, Hanoi At , / / The dissertation can be found at: 1, National Library of Viet Nam 2, Library of Institute of Natural Products Chemistry Vietnam Academy of Science and Technology I INTRODUCTION 1.1 Background 2,4,6 trinitrotoluene (TNT) is a chemical widely used in defense and economy The explosive manufacturing industry discharges a large amount of wastewater containing toxic chemicals such as TNT In fact, about 50 years after World War II, in places where gunpowder factories were built, large amounts of TNT and their isomers were found in soil and water environments [1, 2, 21] This proves that TNT is capable of long-term survival in nature or in other words, TNT is difficult to biodegrade In our country, besides the factories is producing ammunition, explosives, and launchers in the defense industry, there are still a large amount of wastewater containing TNT which needs to be treated in warehouses for repairing and collecting ammunition The commonly methods are used to treat wastewater containing TNT including: physical method (adsorption by activated carbon, electrolysis); chemical method (Fenton, UV - Fenton, internal electrolysis), biological method (aerobic activated sludge, MBBR, UASB, MBR, plants, enzymes, white rot fungi) These measures may be used independent or combination with each other, depending on the nature of the wastewater and the material facilities and economic conditions of the manufacture establishment This dissertation focuses on establishing the process of manufacturing bimetallic Fe / Cu internal electrolysis nanomaterial, thereby studying some characteristics correlation between corrosive line and TNT decomposition kinetics and time Setting and optimizing the internal electrolysis process by bimetal Fe / Cu nanomaterials combined with biological method A2O-MBBR (moving bed Biological reactor) to treat TNT wastewater at laboratory scale and Pilot scale at the scene At the same time, the first step establishing the control automatic or semi-automatic operation software with the conditions of the treatment process are determined 1.2 Research objectives Bimetal Fe / Cu internal electrolytic nanomaterials Internal electrolysis method and biological method A2O - MBBR to treat wastewater containing TNT 1.3 New contributions 1.3.1 Successfully fabricated bimetallic Fe / Cu electrolytic internal materials with average size of 100 nm, potential (voltage) E0 = 0.777 V In electrolyte solution pH=3 with TNT concentration of 100 mg/L, corrosive current is reaching 14.85*10-6 A/cm2 and corrosion speed reach 87*10-2 mm / year Therefore has increased the reaction rate, processing efficiency is higher, faster Concurrently, It has been determined corrosion current and its relationship with LnCt / C0 depend on the duration of the TNT reduction process by the corrosion current measurement method There has not been any announcement using this method Some related publication determined the relationship between TNT reduction speed and reduction speed of H+ to H2 1.3.2 Establishing TNT treatment technology by combining the internal electrolysis method using bimetal Fe/Cu nanomaterials with biological method A2O-MBBR Nowadays, no announcement has been made which combined these two methods to treat wastewater The microorganisms in the A2O-MBBR system used to treat wastewater containing TNT has been identified, among them two strains can be new: Novosphingobium sp (HK1-II, HK1-III) have bootstrap value of 97.4-97.92% to Novosphingobium sediminicola sp and Trichosporon (HK2-II, TK2-II and HK2-III) have bootstrap value of 97.7% to middelhonenii sp These two species were published on the international gene bank with the code GenBank: LC483151.1; LC483155.1 and the corresponding link are: https://www.ncbi.nlm.nih.gov/nuccore/LC483151; https://www.ncbi.nlm.nih.gov/nuccore/Lc483155 1.4 The layout of dissertation The dissertation consists of 191 pages with 24 tables, 101 pictures, 139 references and appendices The layout of dissertation: Introduction (3 pages), Chapter 1: Literature review (44 pages), Chapter 2: Materials and methods (15 pages), Chapter 3: Results and discussion (79 pages) ), Conclusion (2 pages), Published works (1 page), References (15 pages), Appendix (17 pages) II CONTENTS INTRODUCTION The introduction refers to the scientific and practical significance of the dissertation CHAPTER 1: LITERATURE REVIEW Overview of international and domestic studies on issues such as: The studies on treatment methods wastewater containing TNT The studies on the internal electrolysis method to treat wastewater Studies on Fe / Cu bimetallic materials fabrication method for wastewater treatment Studies on combines biological method of A2O-MBBR to wastewater treatment The studies on software controls the wastewater treatment system CHAPTER 2: MATERIALS AND METHODS 2.1 Materials Pure TNT Wastewater containing TNT is collected from national defense production facilities 121 100 nm size iron powder 2.2 Methods  Analytical methods: Analytical methods to determine the structure, size, composition of Fe / Cu bimetal nano: SEM, ERD, EDX Methods of measuring corrosive lines: potential range -1.00V0.0V, scanning speed 10 mV/s, Electrodes compare Ag/AgCl (saturation) The corrosion line and the corrosion potential were measured using Autolab PG30 (Netherlands) TNT analytical methods: HPLC, Von – Amper Methods of determining Fe content Proceed to determine Fe ion content according to EPA 7000B method on Contraa 700 device Method of determining COD, T-N, T-P, NH4 +: According to TCVN or ISO  Experimental method Fabrication of Nano Fe / Cu materials: by CuSO4 plating method on powder Fe average size of 100 nm on magnetic stirrer Treatment of TNT wastewater: Prepare a 100 mg/L TNT solution into a 500 ml erlenmeyer flask, change the conditions reaction as pH, temperature, shaking speed, Fe/Cu content to each corresponding research Experimental planning method: Follow the quadratic planning BoxBehnken and Design-Expert optimization software version 11 Isolation of activated sludge: To activate, take activated sludge from wastewater containing TNT treatment stations of production facilities 121, 115 Then, activated sludge in anaerobic, anoxic and oxic activated condition of 30 days Then proceed to isolate microorganism system in the sludge is activated Microbiological classification method: Conduct DNA sequencing of selected strains, then compare with the DNA sequence of 16S are published species by the DDBJ, EMBL, GenBank CHAPTER 3: RESULTS AND DISCUSSIONS The chapter’s content includes: establishing conditions for manufacturing bimetal Fe/Cu nanomaterials, the effects of internal electrolytic factors, A2O-MBBR to treat wastewater containing TNT and optimize treatment conditions, Kinetic characteristics of internal electrolytic reaction, the diversity of microorganisms in the A2OMBBR system, the software control the internal electrolytic system combined with A2O-MBBR to treat wastewater containing TNT 3.1 Fabrication of internal electrolytic materials Nano bimetal Fe/Cu This section write details the results of the research to establish the reaction conditions for creating Fe / Cu materials: Fe powder of 100 nm size is plated by CuSO4 solution at a concentration of 6% in minutes Fe/Cu materials have Cu concentration on the surface of 68.44% and copper atomic mass reaches 79.58% b a Figure 3.1: SEM image (a) and EDS spectrum of Fe / Cu bimetallic nanomaterials Survey results and comparison of corrosion lines between types of bimetal nanomaterials Fe/C and Fe/Cu are shown in Figure 3.2: a b Figure 3.2: Tafel line of galvanic corrosion of Fe/C electrode system (a) and Fe/Cu after plating (b) at different time values From Figure 3.2, it can be seen that the corrosion potential (EĂM) of Fe materials has the descending rule towards the negative side However, the potential of Fe/Cu electrolytic internal materials reaches - 0.563 V÷-0.765 V with absolute value higher than the corrosion potential of Fe/C, only from - 0.263 V÷- 0.6693V Figure 3.3 shows that the corrosion speed of Fe / Cu material is 8,187.10-2 mm/year, which is nearly times higher than that of Fe / C material, only 4,811.10-2 mm/year 1.6E-5 Dong an mon ir (A) 1.4E-5 Fe/Cu Fe/C 1.2E-5 1.0E-5 8.0E-6 6.0E-6 4.0E-6 20 40 60 80 100 120 Thời gian (phút) Figure 3.3: The dependent on time of corrosion line of electrode material system: Fe/C before plating -- (a) and Fe/Cu after chemical plating -■- (b) Thus, bimetallic Fe/Cu electrolytic internal material has been synthesized with average size of 100 nm, potential voltage E0 = 0.777 V In electrolyte solution which have pH=3, concentration of TNT 100 mg/L, Fe/Cu materials have corrosion current density 14.85*10-6 A/cm2 and corrosion speed 8,187*10-2 mm/year 3.2 Effect of factors on the efficiency of TNT treatment 3.2.1 Effect of pH The effectiveness of TNT treatment depends on the initial pH value of the electrolyte solution The results are shown in the Figure 3.4: 100 100 80 60 TNT (mg/L) TNT(mg/L) 60 40 20 40 20 2 2.5 3.5 4.5 5.5 80 pH 20 40 60 80 100 120 140 160 180 Thời gian (phút) Figure 3.4: Treatment efficiency of Figure 3.5: Dependence TNT in different initial pH treatment efficiency on initial pH conditions at the time of 90 minutes over time Figures 3.4 and 3.5 show that during the first 90 minutes, the reaction speed was very fast, achieving high processing efficiency At 90 minutes, the TNT concentration reached 1.61; 1.62; 1.71 and 1.72 mg/L and treatment efficiency in turn 98.29; 98.22; 98.34 and 98.22% correspond to the initial pH values of 2.0; 2.5; 3.0; 3.5 For pH 4.0; 4,5; achieved a lower efficiency and the corresponding TNT concentration was 3.05; 13.09 mg/L Values pH 5.0; 5.5 and have the lowest treatment efficiency, with TNT concentrations respectively are 26.03; 56.36 and 89.03 mg/L From 90th to 180th minute, the processing efficiency slows down and does not change significantly 3.2.2 Effect of Fe/Cu material content Conducting survey on the influence of different Fe/Cu material content inTNT treatment efficiency The experiments have been conducted with 10; 20; 30; 40; 50; 60 g/L of Fe/Cu The result is shown in Figure 3.11; 3.12 and 3.13 32 100 90 28 80 24 20 60 TNT(mg/L) TNT(mg/L) 10 g/L 20 g/L 30 g/L 40 g/L 50 g/L 60 g/L 70 16 12 50 40 30 20 10 0 -100 10 20 30 40 50 30 60 90 120 150 180 60 Thời gian (phút) Hàm lượng Fe/Cu (g/L) Figure 3.6: Dependence of TNT Figure 3.7: Change of TNT treatment efficiency at 90th concentration over time at minutes on the content of Fe / Cu different Fe / Cu content The Figures 3.6 and 3.7 show that the content of materials has effectted on the efficiency of TNT treatment Thus, the effectiveness of TNT treatment depends on the content of Fe/Cu electrolytic internal materials into the reaction With material content Fe/Cu is 30; 40; 50; 60 g/L, after 180 minutes of reaction, reached the highest treatment efficiency of 99.99% and pH value increased to 5.5 3.2.3 Effect of temperature Temperature has an effect on the rate of internal electrolysis reaction, the higher the temperature, the faster the reaction speed and conversely 100 20 25 30 35 40 45 80 TNT (mg/L) TNT(mg/L) 60 40 80 20 120 160 020 25 30 35 40 45 Nhiệt độ (o C) 20 40 60 80 100 120 140 160 180 Thời gian (phút) Figure 3.8: Dependence of Figure 3.9: The change in TNT TNT treatment efficiency on concentration is treated by internal temperature at first 90 minutes electrolyte material according to reaction time at different temperatures Figures 3.8 and 3.9 show that the higher the temperature and the faster the reaction speed and conversely At the time of 90 minutes, the temperatures at 40℃ and 45℃ treated TNT were most effective, the concentration of TNT decreased to 0.57; 0.63 mg / L; next at 30℃, 35℃ is 1.76; 1.71 mg / L and finally at 20℃, 25℃ to 5.31; 3.60 mg / L Thus, it is clear that the higher the temperature and the faster the reaction speed, the highest processing efficiency is at 45℃ and the lowest is 20℃ The next phase, from 90 to 120 minutes, the reaction speed slows down 3.2.4 Effect of TNT concentration The initial concentration of TNT affects the reaction speed and the processing efficiency due to the following reasons: (1) contaminants and intermediate decomposition products will compete with each other on the surface of electrodes (2) Different concentrations of contaminants make the dispersion phase in contact between pollutants with Fe / Cu electrode surface different: Figure 3.13 and Figure 3.14 show the causal relationship between the rate of iron corrosion and the iron concentration in TNT treatment process depend on time Figure 3.15: Relationship between logarithms of concentration and time Figure 3.15 proves that TNT is reduced by Fe / Cu internal electrolysis reaction fit Level Kinetic assumptions model The reaction rate constant is calculated by the slope (angular coefficient) of the linear regression line 3.3.2 Effect of pH and Fe/Cu content 0.0 0.5 0.0 -0.5 -0.5 -1.0 -1.5 pH=2 k=0.0371 pH=2.5 k=0.0369 pH=3 k=0.0367 pH=3.5 k=0.0366 pH=4 k=0.0307 pH=4.5 k=0.0224 pH=5 k=0.0084 pH=5.5 k=0.0059 pH=6 k=0.0011 -2.0 -2.5 -3.0 ln(Ct/Co) ln(Ct/Co) -1.0 -1.5 -2.0 10 g/L 20 g/L 30 g/L 40 g/L 50 g/L 60 g/L -2.5 -3.0 -3.5 -3.5 k=0.0126 k=0.0205 k=0.0339 k=0.0452 k=0.0459 k=0.0459 -4.0 20 40 60 -4.5 80 Thời gian (phút) 20 40 60 80 Thời gian (phút) Figure 3.16: Effect of initial pH Figure 3.17: Effect of Fe / Cu content on the rate of TNT decomposition on the rate of TNT decomposition 3.3.3 Effect of shaking speed and temperature 0 -1 -1 -2 ln(Ct/Co) ln (Ct/Co) -2 -3 -4 20 oC 25 oC 30 oC 35 oC 40 oC 45 oC -4 -5 60 rpm k=0.013 90 rpm k=0.025 120rpm k=0.044 -5 -3 -6 -6 k=0.0325 k=0.0382 k=0.0462 k=0.0543 k=0.0691 k=0.0746 -7 20 40 60 80 100 120 140 160 180 20 40 60 80 Thời gian (phút) Thời gian (phút) Figure 3.18: Effect of shaking speed Figure 3.19: Effect of temperature on the rate of TNT decomposition on the rate of TNT decomposition 11 Thus the activation energy Ea is calculated based on the graph of the relationship between Ln k and / T (Figure 3.20) -2.6 Equation y = a + b*x Weight No Weighting Residual Sum of Squares -2.8 Adj R-Square 0.98911 Value Intercept lnk lnk 0.00467 -0.99563 Pearson's r Slope Standard Error 7.64344 0.49879 -3246.34703 152.20171 -3.0 -3.2 -3.4 0.00315 0.00320 0.00325 0.00330 0.00335 0.00340 1/T Figure 3.20: Relationship between Lnk and 1/T: y = - 3246x + 7.6434 R2 = 0.9891 In Figure 3.20, it can be seen that the correlation coefficients of these points on the regression line reach 0.9915, the Lnk and 1/T have a strong linear relationship The activation energy of the entire reaction has been calculated: Ea = 3246 * 8.314 = 26.99 KJ/mol and indicates that the TNT decomposition is in the diffusion domain, which in accordance with the above research results 3.3.4 Evaluate TNT molecular reduction process Extreme spectrum Von - Amper for analyzing the position of NO2radicals Thereby it is possible to assess the existence of NO2radicals on the TNT molecule In other words, it is possible to evaluate the reduction of NO2- radicals of TNT molecule into NH2 amine The result is shown in Figure 3.21 as follows: TNT TNT TNT TNT -160n -140n TNT3 TNT1 TNT2 -140n -120n -120n I (A) I (A) TNT1 -100n -100n TNT2 TNT3 -80.0n -80.0n -60.0n -60.0n -40.0n 0.10 -0.10 -0.20 -0.30 -0.40 0.10 -0.50 -0.10 -0.20 U (V) U (V) a b 12 -0.30 -0.40 -0.50 TNT TNT TNT TNT -100n -200n -80.0n -175n -60.0n I (A) I (A) -150n -125n -40.0n TNT1 TNT3 -100n -20.0n -75.0n 0.10 TNT1TNT2 -50.0n -0.10 -0.20 -0.30 -0.40 0.10 -0.50 -0.10 -0.20 -0.30 -0.40 -0.50 U (V) U (V) c d Figure 3.21: Von - Amper spectrum of TNT decomposition process at time minutes (a); 15 minutes (b); 90 minutes (c); 330 minutes (d) It can be seen that, at the minutes, there were still spectral peaks equivalent to NO2- radicals, after 15 minutes response the spectral peaks was lower and to 90 minutes, there was only spectral peak but it was lower so many At 330 minutes, the spectral peaks of the NO2radical are nearly flat In other words, the NO2- on the TNT molecule no longer exists 3.3.5 Operating TNT wastewater treatment at laboratory with Fe / Cu material This section presents the results of TNT wastewater treatment at laboratory using electrolytic internal material for 30 days Table 3.1: TNT wastewater treatment efficiency Initial After treat Eficiency (%) COD (mg/L) 220 - 270 85 - 110 59, - 61,3 TNT (mg/L) 95 –106,4 100 BOD5/COD 0,18 –0,2 0, 55 – 0,56 pH 6,5 – 6,6 3.3.5.1 Treatment efficiency of TNT 120 100 TNT(mg/l) 80 In 60 En 40 20 0 10 12 14 16 Times(day) Figure 3.21: Treatment efficiency of TNT 13 a b Figure 3.24: HPLC spectrum of pre-treatment (a) and post-treatment (b) 3.3.5.2 COD removal efficiency 280 0.7 260 0.6 240 0.5 200 IN 180 EN BOD5/COD COD(mg/l) 220 160 0.4 0.3 0.2 140 0.1 120 0.0 100 pH 10 12 14 16 Time(day) Figure 3.25: COD removal efficiency Figure 3.26: The change of BOD5 / COD ratio after treatment 3.4 Techniques A2O-MMBR treating TNT 3.4.1 Research isolated activated sludge 3.4.1.1 Isolation Table 3.2: Characteristic of domesticated activated sludge Mixed Liquor Condition Suspended Solids Characteristics MLSS (mg/L) yellowish brown, mud suspended, Aerobic 2120 ± 50 the suspension Dark brown, big mud cotton, rapid Anoxic 1596 ± 50 sedimentation black, heavy mud, very rapid Anaerobic 1103 ± 50 sedimentation 14 3.4.1.2 Evaluation of activated sludge particle size Time (days) Anaerobic Anoxic Aerobic 12,11329 µm 13,57996 µm 20,44160 µm 14,13µ𝑚 82,88 µm 163,55µ𝑚 14,12941 µm 14,32089 µm 67,01550 µm 30 90 180 Figure 3.27: Spectral size distribution of activated sludge 3.4.1.3 Survey of biological polymer content Conducting SEPS and BEPS content survey for months and give results shown in Figure 3.28; 3.29; 3.30: Proteins Pollysaccharides Total 0.8 Proteins Pollysaccharides Total 1.0 0.7 0.8 0.5 BEPS (mg/g) SEPS (mg/g) 0.6 0.4 0.3 0.2 0.6 0.4 0.2 0.1 0.0 0.0 T1 T2 T3 T4 T5 T1 T6 T2 T3 T4 T5 T6 Thoi gian Thoi gian b a Figure 3.28: Polymer content of anaerobic tanks: SEPS (a) and BEPS (b) 15 Proteins Pollysaccharides Total Proteins Pollysaccharides Total 0.7 0.6 0.6 0.5 BEPS (mg/g) SEPS (mg/g) 0.5 0.4 0.3 0.4 0.3 0.2 0.2 0.1 0.1 0.0 T1 0.0 T1 T2 T3 T4 T5 T2 T3 T6 T4 T5 T6 Thoi gian Thoi gian a b Figure 3.29: Polymer content in anoxic tanks: SEPS (a) and BEPS (b) Proteins Pollysaccharides Total Proteins Pollysaccharides Total 0.40 0.6 0.35 0.5 BEPS (mg/g) SEPS (mg/g) 0.30 0.25 0.20 0.15 0.4 0.3 0.2 0.10 0.1 0.05 0.0 T1 0.00 T1 T2 T3 T4 T5 T2 T3 T6 T4 T5 T6 Thoi gian Thoi gian a b Figure 3.30: Polymer content of aerobic tank: SEPS (a) and BEPS (b) 3.4.2 Treatment of TNT by A2O-MBBR method 3.4.2.1 Evaluate the processing efficiency of A2O-MBBR system The results of monitoring the change of pH in the reaction tanks are shown in Figure 3.31 pH 5 pH influence %(3) pH Ky Khi %(4) 10 15 20 25 30 Time (day) Figure 3.31: The change of pH at the reaction tank The efficiency of wastewater treatment containing TNT by the independent A2O-MBBR method is shown in Figure 3.32; 3.33 as follows: 16 25 20 20 4.5 10 60 Remove Vao Ra 3.5 3.0 Ky Khi Thieu Hieu Khi 2.5 Abs 40 TNT removal (%) 15 TNT removal (%) TNT concentration ( mg/L) 4.0 2.0 1.5 80 1.0 0 10 15 20 25 30 100 0.5 Time (day) 0.0 200 250 300 350 400 Wave Figure 3.32: TNT removal efficiency by A2O - MBBR system Figure 3:33: The transformation of substances in A2O-MBBR system Treatment efficiency of COD and NH4+ 300 50 45 B C 250 200 NH4-N(mg/l) COD(mg/l) 40 IN EN 150 Before After 35 30 25 20 100 15 50 10 12 14 10 16 Times 10 12 14 16 Times Figure 3.34: COD removal Figure 3.35: Ammonium efficiency removal efficiency 3.4.3 Combining the method of internal electrolysis and A2OMBBR 3.4.3.1 COD removal efficiency COD treatment results of the reaction system are presented in Figure 3.36: 120 110 100 90 COD mg/L 80 70 60 50 40 30 20 10 0 10 20 30 40 50 60 70 80 90 Time (day) Figure 3.36: COD removal efficiency on A2O-MBBR system 17 3.4.3.2 Efficient treatment of NH4 NH4 treatment results are presented in Figure 3.37: 35 30 NH4(mg/l) 25 20 15 Pre-treat 10 Post-treat 0 20 40 60 80 Time(day) Figure 37 NH4 treatment efficiency of A2O-MBBR 3.4.3.3 TNT treatment efficiency Through internal electrolysis process, TNT has been completely decomposed, however, we still tested the TNT content in A2OMBBR system by high-pressure liquid chromatography and the results shown in Figure 3.38: c b a Figure 3.38: HPLC spectrum of TNT in anaerobic tanks (a); anoxic (b); aerobic (c) Table 3.3: Efficiency before and after electrolysis treatment Internal A2OPre-treat Post-treat electrolytic MBBR COD (mg/l) 220 - 270 85 - 110 33 -38 86 – 89 % TNT (mg/l) 95 – 106,4 0 100 BOD5/COD 0,18 – 0,2 0, 55 – 0,56 0,29 -0,5 + NH4 (mg/l) 23 - 45 18 - 32 5,8 -7,9 73- 82 pH 6,5 – 6,6 6,5-7,2 Thus, the process of combining the internal electrolysis method and A2O-MBBR to treat TNT and NH4NO3 in the actual wastewater samples at the factory were successful, in which the efficiency of TNT, COD and NH4 removal, respectively were 100%, 86 - 89%, 73-85% 18 3.4.4 Microorganism diversity in A2O-MBBR system The results showed that the microorganism in the A2O-MBBR system treating TNT mainly consists of genera: Candida, Bacillus, Burkholderia, Chryseobacterium, Novosphingobium, Pseudomonas and Trichosporon, species In which there are 02 strains can be new, namely: Novosphingobium sp (HK1-II, HK1-III) have 97.4-97.92% similarity to Novosphingobium sediminicola Trichosporon sp (HK2II, TK2-II and HK2-III) have 97.7% similarity to middelhonenii 0.005 T B puraquae_CAMPA B diffusa_R-15930T_AM747629 T _CP000442 67 B ambifaria_AMMD T B cenocepacia_LMG 87 B lata_383T_CP000150 B arboris_R-24201T_AM747630 T 53 B contaminans LMG 23361 _LASD01000006 TK3-II 80 KK1-II_ 74 TK1-III 58 TK3-III KK2-III B metallica_R-16017T_AM747632 B anthina_R-4183T_AJ420880 seminalis_R-24196T_AM747631 50 B.cepacia_ATCC 25416T_AXBO01000009 B.territorii_LMG28158 T_LK023503 B.vietnamiensis_LMG 10929T_CP009631 B.multivorans_ATCC BAAT B.dolosa_LMG 18943T_JX986970 247 _ALIW01000278 T 51 B.latens_R-5630 _AM747628 B mesoacidophila_ATCC 31433T_CP020739 B ubonensis_CIP 107078T_EU024179 88 B stagnalis_LMG 28156T_LK023502 B stabilis_ATCC BAA-67T_CP016444 51 T 95 B pyrrocinia_DSM 10685 _CP011503 B humptydooensis_MSMB43T_CP01338 B rinojensis_A396T_KF650996 B pseudomultivorans_LMG 26883T_HE962386 61 B glumae_LMG 2196T_AMRF01000003 B gladioli_NBRC 13700TT_BBJG01000151 65 B plantarii_ATCC 43733 _CP007212 73 B singularis_LMG 28154T_FXAN01000134 B thailandensis_E264T_CP000086 63 B mallei_ATCC 23344T_CP000011 50 B pseudomallei_ATCC 23343T_CWJA01000021 99 T B oklahomensis_C6786 _ABBG010005 B alpina_PO-04-17-38T_JF763852 Figure 3.38: Phylogeny of TK3-II, KK1-II, TK1-II, TK1-III, TK3-III and KK2-III, that close relative in Species of in Burkholderia genus B alpine PO-04-17-38T_JF763852 is extrinsic group, bootstrap values> 50% are shown on the tree, bar 0.005 19 0.01 T B dabaoshanensis_GSS04 _KJ818278 B shackletonii_LMG HK5-II 100 99 TK1-II B subtilis D7XPN1T_JHCA01000027 99 KK1-III T B taiwanensis_FJAT-14571 _KF0405 T 53 100 60 B salidurans_KNUC7312 _KX904715 T B onubensis_0911MAR22V3 _NSEB010 B timonensis_10403023T_CAET01000 50 72 T B sinesaloumensis_T P3516 _LT732529 96 84 B humi_LMG 22167T _AJ627210 B endophyticus_2DT _AF295302 T B filamentosus_SGD-14 _KF265351 100 T B manusensis_Ma50-5 _MF582328 T 60 B kexueae_Ma50-5 _MF582327 T B carboniphilus_JCM 9731 _AB021182 B seohaeanensis_BH724T_AY667495 T B halosaccharovorans_E33 _HQ4334 T 74 B herbersteinensis_D-1-5a _AJ781 T B depressus_BZ1 _KP259553 T B purgationiresistens_DS22 _FR66 T 70 B korlensis_ZLC-26 _EU603328 T B dakarensis_ P3515 _LT707409 T B circulans_ATCC 4513 _AY724690 T B oryzisoli_1DS3-10 _KT886063 88 95 B endozanthoxylicus_1404T_KX8651 B drentensis_LMG 2183T_AJ542506 Ornithinibacillus contaminans CCUG 53201TFN597064 Figure 3.39: Phylogeny of HK5-II, TK1-II KK1-III, that close relative in Species of Bacillus genus Ornithinibacillus contaminans CCUG 53201TFN597064 is extrinsic group, bootstrap values> 50% are shown on the tree, bar 0.01 73 100 68 99 93 100 P.aeruginosa_JCM 5962T_BAMA01000316 HK2-III-5 88 KK2_II P indicaT_ NBRC 103045T_BDAC01000046 P furukawaii_KF77 _AJMR01000229 P otitidis_MCC10330T_AY953147T P resinovorans_LMG 2274 _Z76668 P oryzae_ KCTC 32247T_LT629751 P guangdongensis_CCTCC AB 2012022T_LT629780 P sagittaria_ JCM 18195T_FOXM01000044 100 P.18195T_FOXM01000044 linyingensis_LYBRD3-7T_HM24614 pharmacofabricae_ZYSR67-Z_KX91 P fluvialis_ASS-1T_NMQV01000040 P glareae_KMM 9500T_LC011944 P guariconensis_ LMG 27394T_FMYX01000029 P plecoglossicida_ NBRC 103162T_BBIV01000080 Azotobacter_beijerinckii ATCCT 19360_AJ308319 Figure 3.40: Phylogeny of HK2-III, TK2-II, that close relative in Species of Pseudomonas genus Azotobacter_beijerinckii ATCCT 19360_AJ308319 is extrinsic group, bootstrap values> 50% are shown on the tree, bar 0.005 20 0.01 C vietnamense_GIMN1.005T_HM21241 C aquifrigidense_CW9T_EF644913 59 C flavum_CW-E_EF154516 C arthrosphaerae_CC-VM-7T_MAYG01 Chryseobacterium_gleum_ATCC 35910T_ACKQ01000057 68 TK5-II 99 100 TK5-III C indologenes_ NBRC 14944T_BAVL01000024 C joostei_DSM 16927T_jgi.1096615 61 C gallinarum_DSM 27622T_CP009928 85 C contaminans_DSM 23361T_LASD01000006 58 C rhizoplanae_JM-534T_KP033261 C viscerum_687B-08T_FR871426 C sediminis_IMT-174T_KR349467 Chryseobacterium piscium_LMG 23089T_AM040439 79 Figure 3.41: Phylogeny of TK5-II TK5-III, that close relative in Species of Chryseobacterium genus Chryseobacterium piscium_LMG 23089T_AM040439 is extrinsic group, bootstrap values> 50% are shown on the tree, bar 0.01 N._oryzae NR_147755_T N humi R1-4TKY658458 N sediminicola AH51FJ177534T 79 HK4-II 100 100 HK4-III N subterraneum_DSM12447_T JRVC0 100 N aromaticivorans_CP000248_DSM12 N fontis LN890293T N naphthalenivorans NBRC_02051T 51 N barchaimii_KQ130454T 84 79 N gossypii KP657488T 67 N._guangzhouense KX215153T HK1-II 100 HK1-III N arvoryzae HF548596T Blastomonas_natatoria_AB024288 72 Figure 3.42: Phylogeny of HK4-II, HK4-III, HK1-II VÀ HK1-II, that close relative in Species of Novosphingobium genus Blastomonas_natatoria_AB024288 is extrinsic group, bootstrap values> 50% are shown on the tree, bar 0.01 21 0.05 100 100 100 Candida tropicalis KF281607 Candida dubliniensis MH591468 HK3-III 100 HK3_II 87 TK2-III 56 Trichosporon cutaneum 100 AB305103 Trichosporon mucoides 67 AB305104 Trichosporon dermatis Trichosporon terricola HM802130 99 Trichosporon AB086382 middelhovenii 100 HK2-III AB180198 TK2-II 100 HK2-II Saccharomyces cerevisiae Figure 3.43: Phylogeny of HK3-II, HK3-III, TK2-III, HK2-III, TK2-II HK2-II, that close relative in Species of Candida Trichosporon genus Saccharomyces cerevisiae DAOM216365 is extrinsic group, bootstrap values> 50% are shown on the tree, bar 0.05 3.5 Design and operate testing of TNT's pilot wastewater treatment system at Z121 Pilot system to treat wastewater contaminated with TNT, NH4NO3 is located at the wastewater treatment station of Factory 4, Enterprise 121 with a capacity of 250 liters/day and night This system has run the trial continuously for 40 days Table 3.4: Results of TNT analysis during the test TNT TCVN/QS No Name (mg/l) 658:2012 Untreated waste water 96 Wastewater after internal electrolysis treatment KPH Waste water after A2O-MBBR treatment KPH Untreated waste water 115 Wastewater after internal electrolysis treatment KPH Waste water after A2O-MBBR treatment KPH 0,5 Untreated waste water 36 Wastewater after internal electrolysis treatment KPH Waste water after A2O-MBBR treatment KPH Untreated waste water 85 Wastewater after internal electrolysis treatment KPH Waste water after A2O-MBBR treatment KPH 22 Thus, through the process of pilot testing practice shows that: The internal electrolytic treatment system combining A2O-MBBR system has high treatment efficiency, the TNT, COD, BOD5, NH4 + all meet QCVN 40: 2011 / BTNMT CONCLUSION (1) Successfully fabricated bimetallic Fe/Cu electrolytic internal materials with average size of 100 nm, potential E0 = 0.777 V to replace Fe/C materials In electrolyte solution pH=3 with the TNT concentration of 100 mg/L corrosive line reaches 14.85*10-6 A/cm2 and corrosion speed is 87*10-2 mm year (2) Some kinetic characteristics of internal electrolytic reactions on bimetal Fe / Cu nanomaterials The reaction rate of TNT decomposition over time follows the rules of first order reaction assumption in 90 minutes and has an activation energy of Ea = 26.99 kJ/mol This process is dominated by diffusion domain The mechanism of TNT decomposition has been shown that: TNT is reduced on the cathode surface by electrons received from Fe corrosion and is oxidized by Fenton reaction in the electrolyte solution The relationship between corrosion line, Fe ion generation rate and TNT treatment efficiency was determined based on reaction time Determined the K rate constants of the influencing factors in the electrolytic reaction (3) The specifications for TNT treatment are established by internal electrolysis method using bimetallic Fe/Cu nanomaterials The specifications are optimized by Experimental Box - Benken method and are selected as: pH 3; shaking speed of 120 rpm; time 180 minutes; Fe/Cu content of 50 g/L; at 30oC, with a concentration of TNT 100 mg/L, so the treatment efficiency reaches 98.29% The technical process is experimented by laboratory model and Pilot model in the factory (4) The technical parameters of A2O-MBBR method to treat wastewater TNT are established directly or indirectly through pretreatment by internal electrolysis method Technical process is tested by laboratory model and Pilot model in the field (5) Microorganism diversity and species variation of A2O-MBBR system are evaluated during treatment of TNT The microorganism in 23 the A2O-MBBR system treating TNT mainly consists of genera: Candida, Bacillus, Burkholderia, Chryseobacterium, Novosphingobium, Pseudomonas and Trichosporon, species In which there are 02 strains can be new, namely: Novosphingobium sp (HK1-II, HK1-III) have 97.4-97.92% similarity to Novosphingobium sediminicola Trichosporon sp (HK2-II, TK2-II and HK2-III) have 97.7% similarity to middelhonenii These two species were published on the international gene bank with the code GenBank: LC483151.1; LC483155.1 and the corresponding link are: https://www.ncbi.nlm.nih.gov/nuccore/LC483151; https://www.ncbi.nlm.nih.gov/nuccore/Lc483155 (6) Automatic and semi-automatic control software for treatment system are established according to the internal electrolysis process combined with A2O-MBBR method (7) The TNT wastewater treatment system at factory was designed, installed and operated using internal electrolytic technology combining A2O-MBBR Which according to the conditions is determined above and using nano bimetal Fe/Cu electrolytic internal materials The treatment efficiency reached column B, standard QCVN 40: 2011/BTNMT 24 PUBLISHED WORKS Treatment of wastewater containing aromatic nitro compounds using the A2O-MBBR method Rusian journal of Chemistry and Chemical Technology.2018 V.61 N 9-10 DOI: 10.6060/ivkkt.20186109-10.5541 Enhanced efficiency of treatment of TNT wastewater by internal electrolysis reaction use bimetallic materials Fe-Cu Journal of Science and Technology 54 (4B) (2016) 11-18 Enhancing the oxidation of the internal electrolysis to treat TNT wastewater by EDTA and H2O2 Journal of Science and Technology 53 (1B) (2015) 326-332 Treating wastewater contaminated with TNT, NH4NO3 by combining internal electrolysis and A20-MBBR method Journal of Chemistry 10/2015 53 (5el) 212-217 Treatment of TNT wastewater by internal electrolysis method) Journal of Science and Technology 51 (3A) (2013) 294-302 Characteristics of corrosive line and optimization of TNTcontaminated wastewater treatment process by internal electrolysis method with Fe/Cu bimetal nanomaterials Journal of Chemistry National Chemistry Conference 2019 Two species of microorganisms have been published on the international gene bank with the GenBank code: LC483155.1; LC483155.1 and the corresponding link are: - https://www.ncbi.nlm.nih.gov/nuccore/LC483151; - https://www.ncbi.nlm.nih.gov/nuccore/Lc483155 ... U (V) U (V) a b 12 -0.30 -0.40 -0.50 TNT TNT TNT TNT -100n -200n -80.0n -175n -60.0n I (A) I (A) -150n -125n -40.0n TNT1 TNT3 -100n -20.0n -75.0n 0.10 TNT1 TNT2 -50.0n -0.10 -0.20 -0.30 -0.40 0.10... efficiency of A2O- MBBR 3.4.3.3 TNT treatment efficiency Through internal electrolysis process, TNT has been completely decomposed, however, we still tested the TNT content in A2OMBBR system by... on the TNT molecule In other words, it is possible to evaluate the reduction of NO2- radicals of TNT molecule into NH2 amine The result is shown in Figure 3.21 as follows: TNT TNT TNT TNT -160n