Study on internal electrolysis combine with aao - Mbbr to treat tnt wastewater

erimental 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 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 A2O- MBBR system, the software control the internal electrolytic system combined with A2O-MBBR to treat wastewater containing TNT. 5 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%. a b 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: 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. 6 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. 20 40 60 80 100 120 4.0E-6 6.0E-6 8.0E-6 1.0E-5 1.2E-5 1.4E-5 1.6E-5 Thời gian (phút) D o n g a n m o n i r (A ) Fe/Cu Fe/C 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 E 0 = 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/cm 2 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: 2 3 4 5 6 0 20 40 60 80 100 T N T (m g /L ) pH 0 20 40 60 80 100 120 140 160 180 0 20 40 60 80 100 Thời gian (phút) 2 2.5 3 3.5 4 4.5 5 5.5 6 T N T ( m g /L ) Figure 3.4: Treatment efficiency of TNT in different initial pH conditions at the time of 90 minutes Figure 3.5: Dependence treatment efficiency on initial pH over time 7 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. 10 20 30 40 50 60 0 4 8 12 16 20 24 28 32 T N T (m g /L ) Hàm lượng Fe/Cu (g/L) 0 30 60 90 120 150 180 -10 0 10 20 30 40 50 60 70 80 90 100 Thời gian (phút) T N T (m g /L ) 10 g/L 20 g/L 30 g/L 40 g/L 50 g/L 60 g/L Figure 3.6: Dependence of TNT treatment efficiency at 90th minutes on the content of Fe / Cu Figure 3.7: Change of TNT concentration over time 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. 8 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. 20 25 30 35 40 45 0 1 2 3 4 5 6 Nhiệt độ (o C) T N T (m g /L ) 0 20 40 60 80 100 120 140 160 180 0 20 40 60 80 100 Thời gian (phút) T N T ( m g /L ) 20 25 30 35 40 45 80 120 160 0 4 8 Figure 3.8: Dependence of TNT treatment efficiency on temperature at first 90 minutes Figure 3.9: The change in TNT concentration is treated by internal 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: 9 40 50 60 70 80 90 100 1.4 1.5 1.6 1.7 T N T (m g /L ) Nồng độ TNT ban đầu (mg/L) 20 40 60 80 100 120 140 160 180 -10 0 10 20 30 40 50 60 70 80 90 100 110 Thời gian (phút) T N T ( m g /L ) 40 60 80 100 30 40 50 60 70 80 90 100 0 20 40 Figure 3.10: Dependence of TNT concentration remaining after treatment on the initial concentration Figure 3.11: The change of TNT concentration after treatment over time with different initial TNT concentrations 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 10 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. . 0 20 40 60 80 100 120 3 4 5 6 7 8 9 10 11 Thời gian (phút) C io n F e (m g /L ) 0 50 100 150 200 250 300 350 0.0 0.2 0.4 0.6 0.8 1.0 Thời gian (phút) C t/ C o Figure 3.13: Dependence of dissolved Fe content on reaction time of internal electrolysis process Figure 3.14: Dependence of TNT concentration on the internal electrolytic reaction time of Fe / Cu materials 11 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 0 20 40 60 80 -3.5 -3.0 -2.5 -2.0 -1.5 -1.0 -0.5 0.0 Thời gian (phút) 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 ln (C t/C o ) 0 20 40 60 80 -4.5 -4.0 -3.5 -3.0 -2.5 -2.0 -1.5 -1.0 -0.5 0.0 0.5 10 g/L k=0.0126 20 g/L k=0.0205 30 g/L k=0.0339 40 g/L k=0.0452 50 g/L k=0.0459 60 g/L k=0.0459 Thời gian (phút) ln (C t/ C o ) Figure 3.16: Effect of initial pH on the rate of TNT decomposition Figure 3.17: Effect of Fe / Cu content on the rate of TNT decomposition 3.3.3. Effect of shaking speed and temperature 0 20 40 60 80 100 120 140 160 180 -6 -5 -4 -3 -2 -1 0 60 rpm k=0.013 90 rpm k=0.025 120rpm k=0.044 Thời gian (phút) ln ( C t/C o ) 0 20 40 60 80 -7 -6 -5 -4 -3 -2 -1 0 ln (C t/C o ) Thời gian (phút) 20 o C k=0.0325 25 o C k=0.0382 30 o C k=0.0462 35 o C k=0.0543 40 o C k=0.0691 45 o C k=0.0746 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 12 Thus the activation energy Ea is calculated based on the graph of the relationship between Ln k and 1 / T (Figure 3.20). 0.00315 0.00320 0.00325 0.00330 0.00335 0.00340 -3.4 -3.2 -3.0 -2.8 -2.6 ln k 1/T Equation y = a + b*x Weight No Weighting Residual Sum of Squares 0.00467 Pearson's r -0.99563 Adj. R-Square 0.98911 Value Standard Error lnk Intercept 7.64344 0.49879 Slope -3246.34703 152.20171 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 NO 2- radicals. Thereby it is possible to assess the existence of 3 NO 2- radicals on the TNT molecule. In other words, it is possible to evaluate the reduction of 3 NO 2- radicals of TNT molecule into NH2 amine. The result is shown in Figure 3.21 as follows: a b TNT TNT 0.10 0 -0.10 -0.20 -0.30 -0.40 -0.50 U (V) -40.0n -60.0n -80.0n -100n -120n -140n I ( A) TNT3 TNT2TNT1 TNT TNT 0.10 0 -0.10 -0.20 -0.30 -0.40 -0.50 U (V) -60.0n -80.0n -100n -120n -140n -160n I ( A) TNT1 TNT3 TNT2 13 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 NO 2- 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 NO 2- radical are nearly flat. In other words, the NO 2- 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, 2 - 61,3 TNT (mg/L) 95 –106,4 0 100 BOD5/COD 0,18 –0,2 0, 55 – 0,56 - pH 5 6,5 – 6,6 - 3.3.5.1. Treatment efficiency of TNT 0 2 4 6 8 10 12 14 16 0 20 40 60 80 100 120 In En T N T (m g /l) Times(day) Figure 3.21: Treatment efficiency of TNT TNT TNT 0.10 0 -0.10 -0.20 -0.30 -0.40 -0.50 U (V) -50.0n -75.0n -100n -125n -150n -175n -200n I ( A) TNT1 TNT TNT 0.10 0 -0.10 -0.20 -0.30 -0.40 -0.50 U (V) 0 -20.0n -40.0n -60.0n -80.0n -100n I ( A) TNT3 TNT1TNT2 14 a b Figure 3.24: HPLC spectrum of pre-treatment (a) and post-treatment (b) 3.3.5.2. COD removal efficiency 0 2 4 6 8 10 12 14 16 100 120 140 160 180 200 220 240 260 280 IN EN C O D (m g /l) Time(day) 3 4 5 6 7 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 B O D 5/ C O D pH 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 Mixed Liquor Suspended Solids MLSS (mg/L) Characteristics Aerobic 2120 ± 50 yellowish brown, mud suspended, the suspension Anoxic 1596 ± 50 Dark brown, big mud cotton, rapid sedimentation Anaerobic 1103 ± 50 black, heavy mud, very rapid sedimentation 15 3.4.1.2. Evaluation of activated sludge particle size Time (days) Anaerobic Anoxic Aerobic 30 12,11329 µm 13,57996 µm 20,44160 µm 90 14,13µ𝑚 82,88 µm 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: T1 T2 T3 T4 T5 T6 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 S E P S ( m g /g ) Thoi gian Proteins Pollysaccharides Total a T1 T2 T3 T4 T5 T6 0.0 0.2 0.4 0.6 0.8 1.0 B E P S ( m g /g ) Thoi gian Proteins Pollysaccharides Total b Figure 3.28: Polymer content of anaerobic tanks: SEPS (a) and BEPS (b) 16 T1 T2 T3 T4 T5 T6 0.0 0.1 0.2 0.3 0.4 0.5 0.6 S E P S ( m g /g ) Thoi gian Proteins Pollysaccharides Total T1 T2 T3 T4 T5 T6 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 B E P S ( m g /g ) Thoi gian Proteins Pollysaccharides Total a b Figure 3.29: Polymer content in anoxic tanks: SEPS (a) and BEPS (b) T1 T2 T3 T4 T5 T6 0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 S E P S ( m g /g ) Thoi gian Proteins Pollysaccharides Total T1 T2 T3 T4 T5 T6 0.0 0.1 0.2 0.3 0.4 0.5 0.6 B E P S ( m g /g ) Thoi gian Proteins Pollysaccharides Total 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. 0 5 10 15 20 25 30 5 6 7 8 %(3) %(4) pH influence pH Ky Khi p H 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: 17 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 + 0 2 4 6 8 10 12 14 16 50 100 150 200 250 300 IN EN C O D (m g /l) Times Figure 3.34: COD removal efficiency 0 2 4 6 8 10 12 14 16 10 15 20 25 30 35 40 45 50 B C NH 4 -N (m g/ l) Times 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: 0 10 20 30 40 50 60 70 80 90 0 10 20 30 40 50 60 70 80 90 100 110 120 C O D m g /L Time (day) Figure 3.36: COD removal efficiency on A2O-MBBR system 0 5 10 15 20 25 30 0 5 10 15 20 25 T N T r e m o v a l (% ) T N T c o n c e n tr a ti o n ( m g /L ) Time (day) Remove Vao Ra 100 80 60 40 20 0 200 250 300 350 400 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 A b s Wave Ky Khi Thieu khi Hieu Khi T N T r e m o v a l (% ) Before After 18 3.4.3.2. Efficient treatment of NH4 NH4 treatment results are presented in Figure 3.37: 0 20 40 60 80 0 5 10 15 20 25 30 35 N H 4 (m g /l ) Time(day) 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: a b c Figure 3.38: HPLC spectrum of TNT in anaerobic tanks (a); anoxic (b); aerobic (c) Table 3.3: Efficiency before and after electrolysis treatment Pre-treat Internal electrolytic A2O- MBBR Post-treat COD (mg/l) 220 - 270 85 - 110 33 -38 86 – 89 % TNT (mg/l) 95 – 106,4 0 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 5 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%. Pre-treat Post-treat 19 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. 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 B. alpina_PO-04-17-38 T _JF763852 B. oklahomensis_C6786 T _ABBG010005 B. pseudomallei_ATCC 23343 T _CWJA01000021 B. mallei_ATCC 23344 T _CP000011 B. thailandensis_E264 T _CP000086 99 B. singularis_LMG 28154T_FXAN01000134 50 B. plantarii_ATCC 43733T_CP007212 B. gladioli_NBRC 13700 T _BBJG01000151 B. glumae_LMG 2196T_AMRF01000003 73 B. pseudomultivorans_LMG 26883 T _HE962386 B. rinojensis_A396 T _KF650996 65 B. humptydooensis_MSMB43 T _CP01338 B. pyrrocinia_DSM 10685 T _CP011503 B. stabilis_ATCC BAA-67 T _CP016444 B. stagnalis_LMG 28156 T _LK023502 95 B. ubonensis_CIP 107078 T _EU024179 B. mesoacidophila_ATCC 31433 T _CP020739 B.latens_R-5630 T _AM747628 B.dolosa_LMG 18943 T _JX986970 B.multivorans_ATCC BAA- 247 T _ALIW01000278 51 B.vietnamiensis_LMG 10929 T _CP009631 88 B.territorii_LMG28158 T _LK023503 B.cepacia_ATCC 25416T_AXBO01000009 seminalis_R-24196 T _AM747631 50 B. anthina_R-4183 T _AJ420880 B. metallica_R-16017 T _AM747632 KK2-III TK3-III TK1-III KK1-II_ TK3-II 58 B. contaminans LMG 23361 T _LASD01000006 74 B. arboris_R-24201 T _AM747630 B. lata_383T_CP000150 80 87 B. cenocepacia_LMG T 53 B. ambifaria_AMMD T _CP000442 B. diffusa_R-15930 T _AM747629 B. puraquae_CAMPA T 67 51 63 61 0.005 20 Figure 3.39: Phylogeny of HK5-II, TK1-II và KK1-III, that close relative in Species of Bacillus genus. Ornithinibacillus contaminans CCUG 53201 T FN597064 is extrinsic group, bootstrap values> 50% are shown on the tree, bar 0.01 Figure 3.40: Phylogeny of HK2-III, TK2-II, that close relative in Species of Pseudomonas genus. Azotobacter_beijerinckii ATCC T 19360_AJ308319 is extrinsic group, bootstrap values> 50% are shown on the tree, bar 0.005. Ornithinibacillus contaminans CCUG 53201 T FN597064 B. drentensis_LMG 2183 T _AJ542506 B. endozanthoxylicus_1404 T _KX8651 B. oryzisoli_1DS3-10 T _KT886063 B. circulans_ATCC 4513 T _AY724690 95 B. dakarensis_ P3515T_LT707409 B. korlensis_ZLC-26 T _EU603328 88 70 B. purgationiresistens_DS22 T _FR66 B. depressus_BZ1 T _KP259553 B. herbersteinensis_D-1-5a T _AJ781 B. halosaccharovorans_E33 T _HQ4334 B. seohaeanensis_BH724 T _AY667495 B. carboniphilus_JCM 9731 T _AB021182 B. kexueae_Ma50-5 T _MF582327 B. manusensis_Ma50-5 T _MF582328 60 B. filamentosus_SGD-14 T _KF265351 B. endophyticus_2DT T _AF295302 B. humi_LMG 22167 T _AJ627210 B. sinesaloumensis_ P3516T_LT732529 B. timonensis_10403023 T _CAET01000 84 B. onubensis_0911MAR22V3 T _NSEB010 B. salidurans_KNUC7312 T _KX904715 96 100 B. taiwanensis_FJAT-14571 T _KF0405 72 100 53 50 KK1-III B. subtilis D7XPN1 T _JHCA01000027 TK1-II 99 HK5-II 99 B. shackletonii_LMG B. dabaoshanensis_GSS04 T _KJ818278 100 74 60 0.01 Azotobacter_beijerinckii ATCC T 19360_AJ308319 P. plecoglossicida_ NBRC 103162T_BBIV01000080 P. guariconensis_ LMG 27394T_FMYX01000029 P. glareae_KMM 9500 T _LC011944 99 P. fluvialis_ASS-1T_NMQV01000040 pharmacofabricae_ZYSR67-Z_KX91 P. linyingensis_LYBRD3-7T_HM24614 P. sagittaria_ JCM 18195 T _FOXM01000044 18195T_FOXM01000044 P. guangdongensis_CCTCC AB 2012022T_LT629780 100 P. oryzae_ KCTC 32247T_LT629751 93 P. resinovorans_LMG 2274T_Z76668 P. otitidis_MCC10330 T _AY953147 P. furukawaii_KF77 T _AJMR01000229 P. indica _ NBRC 103045T_BDAC01000046 KK2_II HK2-III-5 P.aeruginosa_JCM 5962 T _BAMA01000316 88 100 73 100 68 21 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. 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. Chryseobacterium piscium_LMG 23089T_AM040439 C. sediminis_IMT-174T_KR349467 C. viscerum_687B-08T_FR871426 C. rhizoplanae_JM-534T_KP033261 C. contaminans_DSM 23361T_LASD01000006 C. gallinarum_DSM 27622T_CP009928 C. joostei_DSM 16927T_jgi.1096615 85 61 C. indologenes_ NBRC 14944T_BAVL01000024 TK5-III TK5-II Chryseobacteriu