Summary of Doctoral thesis Chemical engineering: Study on internal electrolysis combine with AAO-MPPR to treat TNT wastewater

MINISTRY OF EDUCATION AND TRAINING

VIETNAM ACADEMY OF 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: 9 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:

1. Assoc. Prof. Le Thi Mai Huong

2. 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

INTRODUCTION

I. 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.

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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 8. 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.

link and the

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 are: corresponding https://www.ncbi.nlm.nih.gov/nuccore/LC483151; https://www.ncbi.nlm.nih.gov/nuccore/Lc483155

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1.4. The layout of dissertation

The dissertation consists of 191 pages with 24 tables, 101 pictures, 139 references and 2 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 -1.00V- 0.0V, scanning speed 10 mV/s, Electrodes compare Ag/AgCl

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(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

1. Fabrication of Nano Fe / Cu materials: by CuSO4 plating method on powder Fe average size of 100 nm on magnetic stirrer. 2. 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. 3. Experimental planning method: Follow the quadratic planning Box- Behnken and Design-Expert optimization software version 11. 4. 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. 5. 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

includes: establishing conditions

for The chapter’s content 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 A2O- MBBR system, the software control the internal electrolytic system combined with A2O-MBBR to treat wastewater containing TNT.

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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 2 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 2 types of bimetal nanomaterials Fe/C and Fe/Cu are shown in Figure 3.2:

b a

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.

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Figure 3.3 shows that the corrosion speed of Fe / Cu material is 8,187.10-2 mm/year, which is nearly 2 times higher than that of Fe / C material, only 4,811.10-2 mm/year.

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:

in different

Figure 3.4: Treatment efficiency of TNT initial pH conditions at the time of 90 minutes Figure 3.5: Dependence treatment efficiency on initial pH over time

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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 6 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.

over time

Figure 3.6: Dependence of TNT treatment at 90th efficiency minutes on the content of Fe / Cu Figure 3.7: Change of TNT concentration at 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.

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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.

Figure 3.8: Dependence of TNT treatment efficiency on temperature at first 90 minutes according

Figure 3.9: The change in TNT concentration is treated by internal electrolyte material 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:

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after

Figure 3.11: The change of TNT concentration treatment over time with different initial TNT concentrations

Figure 3.10: Dependence of remaining TNT concentration initial the treatment on after concentration Figure 3.10; 3.11 shows that the lower the concentration of TNT, the higher the processing efficiency and conversely. After 90 minutes, the remaining TNT concentration was 1.35; 1.42; 1.51; 1.68 mg/L corresponds to the initial TNT concentrations of 40; 60; 80; 100 mg / L. In the next phase, from 90 to 180 minutes, the effect of the initial TNT concentration on the processing speed and efficiency is almost no difference. At 180 minutes, the remaining TNT concentration was corresponding to 0.15; 0.19; 0.21 and 0.23 mg/L. 3.2.5. Optimize the process of treating TNT wastewater Applying Box-Behnken method for pH, temperature, shaking speed, reaction time for regression equations: Y = 93.16 + 1.05B + 3.02C + 8.62D - 0.265BC - 4.73CD + 1.12A2 - 1.11C2 - 3D2. Optimal conditions are determined from the regression equation corresponding to: pH = 3.24, temperature at 32.6 ℃, shaking speed of 91 rpm for 140 minutes and get TNT treatment efficiency of 98.29%. Among the factors that affect TNT's processing performance, the time is greatest affect, follow is the temperature, but to a lesser, the shaking speed and pH have little effect.

a b

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c d

e f

Figure 3.12: Relationship between factors on efficiency of TNT treatment. (a): pH and time; (b) pH and temperature; (c) pH and shaking speed; (d) temperature and time; (e) temperature and shaking speed; (f) shaking speed and time. 3.3. Some kinetic characteristics of the internal electrolysis process TNT 3.3.1. Iron corrosion rate and TNT decomposition kinetics This section presents the results of the iron corrosion rate and the correlation between the rate of TNT decomposition.

3.13: Dependence

. of Figure dissolved Fe content on reaction time of internal electrolysis process

Figure 3.14: Dependence of concentration on TNT the reaction electrolytic internal time of Fe / Cu materials

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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 1 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

Figure 3.17: Effect of Fe / Cu content on the rate of TNT decomposition

Figure 3.16: Effect of initial pH on the rate of TNT decomposition 3.3.3. Effect of shaking speed and temperature

Figure 3.18: Effect of shaking speed on the rate of TNT decomposition Figure 3.19: Effect of temperature on the rate of TNT decomposition

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Thus the activation energy Ea is calculated based on the graph of the relationship between Ln k and 1 / T (Figure 3.20).

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 6 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 NO2- radicals. Thereby it is possible to assess the existence of 3 NO2- radicals on the TNT molecule. In other words, it is possible to evaluate the reduction of 3 NO2- radicals of TNT molecule into NH2 amine. The result is shown in Figure 3.21 as follows:

a b

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c d

Figure 3.21: Von - Amper spectrum of TNT decomposition process at time 0 minutes (a); 15 minutes (b); 90 minutes (c); 330 minutes (d) It can be seen that, at the 0 minutes, there were still 3 spectral peaks equivalent to 3 NO2- radicals, after 15 minutes response the spectral peaks was lower and to 90 minutes, there was only 1 spectral peak but it was lower so many. At 330 minutes, the spectral peaks of the NO2- radical 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 After treat 85 - 110 0 0, 55 – 0,56 6,5 – 6,6 Eficiency (%) 59, 2 - 61,3 100 - - COD (mg/L) TNT (mg/L) BOD5/COD pH Initial 220 - 270 95 –106,4 0,18 –0,2 5

3.3.5.1. Treatment efficiency of TNT

Figure 3.21: Treatment efficiency of TNT

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a b

Figure 3.24: HPLC spectrum of pre-treatment (a) and post-treatment (b) 3.3.5.2. COD removal efficiency

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

Condition Characteristics

Mixed Liquor Suspended Solids MLSS (mg/L)

Aerobic 2120 ± 50

Anoxic 1596 ± 50

rapid Anaerobic 1103 ± 50 yellowish brown, mud suspended, the suspension Dark brown, big mud cotton, rapid sedimentation black, heavy mud, very sedimentation

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Anoxic Aerobic 3.4.1.2. Evaluation of activated sludge particle size Time (days) Anaerobic

20,44160 µm 13,57996 µm 12,11329 µm

82,88 µm 14,13µ𝑚 163,55µ𝑚

180

14,12941 µm 14,32089 µm 67,01550 µm

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 6 months and give results shown in Figure 3.28; 3.29; 3.30:

b a

Figure 3.28: Polymer content of anaerobic tanks: SEPS (a) and BEPS (b)

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a b

Figure 3.29: Polymer content in anoxic tanks: SEPS (a) and BEPS (b)

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.

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:

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)

%

( l a v o m e r T N T

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

Before After

Figure 3.34: COD removal efficiency Figure 3.35: Ammonium removal efficiency

3.4.3. Combining the method of internal electrolysis and A2O- MBBR 3.4.3.1. COD removal efficiency COD treatment results of the reaction system are presented in Figure 3.36:

Figure 3.36: COD removal efficiency on A2O-MBBR system

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3.4.3.2. Efficient treatment of NH4 NH4 treatment results are presented in Figure 3.37:

Pre-treat

Post-treat

Figure 3. 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 A2O- MBBR 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

Post-treat Pre-treat

Internal electrolytic 85 - 110 0 0, 55 – 0,56 18 - 32 6,5 – 6,6 A2O- MBBR 33 -38 0 0,29 -0,5 5,8 -7,9 6,5-7,2 220 - 270 95 – 106,4 0,18 – 0,2 23 - 45 5 86 – 89 % 100 - 73- 82 -

COD (mg/l) TNT (mg/l) BOD5/COD +(mg/l) NH4 pH 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%.

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0.005

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 7 genera: Candida, Bacillus, Burkholderia, Chryseobacterium, Novosphingobium, Pseudomonas and Trichosporon, 8 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.

B. puraquae_CAMPAT

B. diffusa_R-15930T_AM747629 B. ambifaria_AMMDT_CP000442

B. cenocepacia_LMGT 87

B. lata_383T_CP000150 B. arboris_R-24201T_AM747630

B. contaminans LMG 23361T_LASD01000006

TK3-II

53 80

KK1-II_ TK1-III

TK3-III

KK2-III

B. metallica_R-16017T_AM747632 B. anthina_R-4183T_AJ420880 seminalis_R-24196T_AM747631 B.cepacia_ATCC 25416T_AXBO01000009 B.territorii_LMG28158 T_LK023503

B.vietnamiensis_LMG 10929T_CP009631

B.multivorans_ATCC BAA- B.dolosa_LMG 18943T_JX986970 247T_ALIW01000278 B.latens_R-5630T_AM747628

51 B. mesoacidophila_ATCC 31433T_CP020739

B. ubonensis_CIP 107078T_EU024179

B. stagnalis_LMG 28156T_LK023502

B. stabilis_ATCC BAA-67T_CP016444 B. pyrrocinia_DSM 10685T_CP011503

B. humptydooensis_MSMB43T_CP01338

B. rinojensis_A396T_KF650996 B. pseudomultivorans_LMG 26883T_HE962386 B. glumae_LMG 2196T_AMRF01000003

B. gladioli_NBRC 13700T_BBJG01000151 B. plantarii_ATCC 43733T_CP007212 B. singularis_LMG 28154T_FXAN01000134

B. thailandensis_E264T_CP000086

B. mallei_ATCC 23344T_CP000011

B. pseudomallei_ATCC 23343T_CWJA01000021

B. oklahomensis_C6786T_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

100

B. dabaoshanensis_GSS04T_KJ818278 B. shackletonii_LMG HK5-II TK1-II B. subtilis D7XPN1T_JHCA01000027

99 99

100

96

84 100 60

KK1-III B. taiwanensis_FJAT-14571T_KF0405 B. salidurans_KNUC7312T_KX904715 B. onubensis_0911MAR22V3T_NSEB010 B. timonensis_10403023T_CAET01000 B. sinesaloumensis_ P3516T_LT732529 B. humi_LMG 22167T_AJ627210 B. endophyticus_2DTT_AF295302 B. filamentosus_SGD-14 T_KF265351 B. manusensis_Ma50-5T_MF582328 B. kexueae_Ma50-5T_MF582327 B. carboniphilus_JCM 9731T_AB021182 B. seohaeanensis_BH724T_AY667495 B. halosaccharovorans_E33T_HQ4334

B. herbersteinensis_D-1-5aT_AJ781

B. depressus_BZ1T_KP259553

B. endozanthoxylicus_1404T_KX8651

B. purgationiresistens_DS22T_FR66 B. korlensis_ZLC-26T_EU603328 B. dakarensis_ P3515T_LT707409 B. circulans_ATCC 4513T_AY724690 B. oryzisoli_1DS3-10T_KT886063 B. drentensis_LMG 2183T_AJ542506

Ornithinibacillus contaminans CCUG 53201TFN597064

Figure 3.39: Phylogeny of HK5-II, TK1-II và 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

100

88 P.aeruginosa_JCM 5962T_BAMA01000316 HK2-III-5 KK2_II

P. indica_ NBRC 103045T_BDAC01000046

P. furukawaii_KF77T_AJMR01000229

P. otitidis_MCC10330T_AY953147

P. resinovorans_LMG 2274T_Z76668

P. oryzae_ KCTC 32247T_LT629751

73

93

100 P. guangdongensis_CCTCC AB 2012022T_LT629780 P. sagittaria_ JCM 18195T_FOXM01000044 P. linyingensis_LYBRD3-7T_HM24614 18195T_FOXM01000044 pharmacofabricae_ZYSR67-Z_KX91

100 P. fluvialis_ASS-1T_NMQV01000040

68 P. glareae_KMM 9500T_LC011944 P. guariconensis_ LMG 27394T_FMYX01000029

99 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

79

59 C. vietnamense_GIMN1.005T_HM21241 C. aquifrigidense_CW9T_EF644913 C. flavum_CW-E_EF154516

C. arthrosphaerae_CC-VM-7T_MAYG01

Chryseobacterium_gleum_ATCC 35910T_ACKQ01000057

68

99 TK5-II TK5-III

100 C. indologenes_ NBRC 14944T_BAVL01000024

C. joostei_DSM 16927T_jgi.1096615

61

85 C. gallinarum_DSM 27622T_CP009928 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

N._oryzae NR_147755_T N. humi R1-4TKY658458

100

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. sediminicola AH51FJ177534T HK4-II HK4-III

100

N. subterraneum_DSM12447_T__JRVC0 N. aromaticivorans_CP000248_DSM12

N. fontis LN890293T

N. naphthalenivorans NBRC_02051T

N. barchaimii_KQ130454T N. gossypii KP657488T N._guangzhouense KX215153T HK1-II 100 HK1-III N. arvoryzae HF548596T

Blastomonas_natatoria_AB024288

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

Candida tropicalis KF281607 Candida dubliniensis MH591468

100

HK3-III HK3_II TK2-III 56 100 67

100

Trichosporon cutaneum Trichosporon mucoides AB305103 Trichosporon dermatis AB305104 Trichosporon terricola HM802130 Trichosporon middelhovenii AB086382 HK2-III AB180198 TK2-II HK2-II

Saccharomyces cerevisiae

Figure 3.43: Phylogeny of HK3-II, HK3-III, TK2-III, HK2-III, TK2-II và HK2-II, that close relative in Species of Candida và 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

No. Name TCVN/QS 658:2012

TNT (mg/l) 96

0,5

Untreated waste water Wastewater after internal electrolysis treatment KPH KPH Waste water after A2O-MBBR treatment 115 Untreated waste water Wastewater after internal electrolysis treatment KPH KPH Waste water after A2O-MBBR treatment 36 Untreated waste water Wastewater after internal electrolysis treatment KPH KPH Waste water after A2O-MBBR treatment 85 Untreated waste water Wastewater after internal electrolysis treatment KPH KPH Waste water after A2O-MBBR treatment

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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 8. 87*10-2 mm year. (2) Some kinetic characteristics of internal electrolytic reactions on / Cu nanomaterials. The reaction rate of TNT bimetal Fe 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

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Bacillus, Burkholderia,

internal electrolytic

the A2O-MBBR system treating TNT mainly consists of 7 genera: Candida, Chryseobacterium, Novosphingobium, Pseudomonas and Trichosporon, 8 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 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.

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PUBLISHED WORKS

Technology.2018. V.61. N

1. Treatment of wastewater containing aromatic nitro compounds using the A2O-MBBR method. Rusian journal of Chemistry and 9-10. DOI: Chemical 10.6060/ivkkt.20186109-10.5541.

2. 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.

3. 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.

4. Treating wastewater contaminated with TNT, NH4NO3 by internal electrolysis and A20-MBBR method.

combining Journal of Chemistry. 10/2015. 53 (5el) 212-217.

treatment process by

5. Treatment of TNT wastewater by internal electrolysis method). Journal of Science and Technology 51 (3A) (2013) 294-302. 6. Characteristics of corrosive line and optimization of TNT- internal contaminated wastewater electrolysis method with Fe/Cu bimetal nanomaterials. Journal of Chemistry. National Chemistry Conference 2019.

7. 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