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1 | (6) |
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2 Activated Sludge Activity Tests |
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7 | (126) |
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7 | (2) |
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2.2 Enhanced Biological Phosphorus Removal |
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9 | (45) |
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2.2.1 Process description |
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9 | (2) |
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2.2.2 Experimental set-up |
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11 | (1) |
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11 | (5) |
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2.2.2.2 Activated sludge sample collection |
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16 | (1) |
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2.2.2.3 Activated sludge sample preparation |
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16 | (1) |
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17 | (2) |
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2.2.2.5 Analytical procedures |
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19 | (3) |
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2.2.2.6 Parameters of interest |
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22 | (1) |
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2.2.3 EBPR batch activity tests: Preparation |
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22 | (1) |
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22 | (2) |
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24 | (1) |
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2.2.3.3 Media preparation |
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24 | (1) |
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2.2.3.4 Material preparation |
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25 | (3) |
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2.2.3.5 Activated sludge preparation |
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28 | (1) |
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2.2.4 Batch activity tests: Execution |
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29 | (1) |
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2.2.4.1 Anaerobic EBPR batch activity tests |
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30 | (3) |
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2.2.4.2 Anoxic EBPR batch tests |
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33 | (1) |
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2.2.4.3 Aerobic EBPR batch tests |
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34 | (2) |
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36 | (1) |
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2.2.5.1 Estimation of stoichiometric parameters |
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36 | (5) |
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2.2.5.2 Estimation of kinetic parameters |
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41 | (1) |
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2.2.6 Data discussion and interpretation |
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42 | (1) |
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2.2.6.1 Anaerobic batch activity tests |
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42 | (3) |
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2.2.6.2 Aerobic batch activity tests |
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45 | (1) |
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2.2.6.3 Anoxic batch activity tests |
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46 | (1) |
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47 | (1) |
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47 | (1) |
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47 | (4) |
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2.2.8 Additional considerations |
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51 | (1) |
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2.2.8.1 GAO occurrence in EBPR systems |
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51 | (1) |
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2.2.8.2 The effect of carbon source |
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51 | (1) |
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2.2.8.3 The effect of temperature |
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51 | (1) |
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52 | (1) |
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2.2.8.5 Denitrification by EBPR cultures |
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52 | (1) |
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2.2.8.6 Excess and shortage of intracellular compounds |
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52 | (1) |
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2.2.8.7 Excessive aeration |
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53 | (1) |
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2.2.8.8 Shortage of essential ions |
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53 | (1) |
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2.2.8.9 Toxicity/inhibition |
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53 | (1) |
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2.3 Biological Sulphate Reduction |
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54 | (19) |
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2.3.1 Process description |
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54 | (2) |
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2.3.2 Sulphide speciation |
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56 | (1) |
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2.3.3 Effects of environmental and operating conditions on SRB |
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57 | (1) |
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57 | (1) |
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2.3.3.2 COD to SO42- ratio |
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58 | (1) |
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58 | (1) |
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59 | (1) |
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59 | (1) |
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2.3.4 Experimental set-up |
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60 | (1) |
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2.3.4.1 Estimation of volumetric and specific rates |
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60 | (1) |
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60 | (1) |
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61 | (1) |
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61 | (1) |
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2.3.4.5 Temperature control |
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61 | (1) |
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2.3.4.6 Sampling and dosing ports |
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62 | (1) |
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2.3.4.7 Sample collection |
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62 | (1) |
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62 | (1) |
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2.3.5 Analytical procedures |
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63 | (1) |
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2.3.5.1 CODorganic and CODtotal |
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63 | (1) |
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64 | (1) |
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64 | (1) |
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2.3.6 SRB batch activity tests: preparation |
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65 | (1) |
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65 | (1) |
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65 | (1) |
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65 | (1) |
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2.3.6.4 Material preparation |
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66 | (1) |
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2.3.6.5 Mixed liquor preparation |
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67 | (1) |
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2.3.6.6 Sample collection and treatment |
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68 | (1) |
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2.3.7 Batch activity tests: execution |
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68 | (1) |
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69 | (1) |
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2.3.8.1 Mass balances and calculations |
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69 | (1) |
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2.3.8.2 Data discussion and interpretation |
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70 | (1) |
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70 | (2) |
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2.3.10 Practical recommendations |
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72 | (1) |
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2.4 Biological Nitrogen Removal |
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73 | (38) |
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2.4.1 Process description |
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73 | (1) |
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74 | (1) |
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75 | (1) |
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2.4.1.3 Anaerobic ammonium oxidation (Anammox) |
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76 | (1) |
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2.4.2 Process-tracking alternatives |
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76 | (1) |
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2.4.2.1 Chemical tracking |
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77 | (1) |
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2.4.2.2 Titrimetric tracking |
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77 | (1) |
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2.4.2.3 Manometric tracking |
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78 | (1) |
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2.4.3 Experimental set-up |
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79 | (1) |
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79 | (1) |
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2.4.3.2 Instrumentation for titrimetric tests |
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79 | (1) |
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2.4.3.3 Instrumentation for manometric tests |
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80 | (1) |
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2.4.3.4 Activated sludge sample collection |
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81 | (1) |
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2.4.3.5 Activated sludge sample preparation |
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82 | (1) |
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82 | (1) |
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2.4.3.7 Analytical procedures |
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83 | (1) |
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2.4.3.8 Parameters of interest |
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83 | (3) |
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2.4.3.9 Type of batch tests |
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86 | (1) |
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2.4.4 Nitrification batch activity tests: Preparation |
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86 | (1) |
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86 | (1) |
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86 | (1) |
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2.4.4.3 Media preparation |
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86 | (1) |
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2.4.5 Nitrification batch activity tests: Execution |
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87 | (5) |
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2.4.6 Denitrification batch activity tests: Preparation |
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92 | (1) |
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92 | (1) |
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93 | (1) |
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2.4.6.3 Working solutions |
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93 | (1) |
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2.4.6.4 Materials preparation |
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93 | (1) |
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2.4.7 Denitrification batch activity tests: Execution |
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93 | (6) |
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2.4.8 Anammox batch activity tests: Preparation |
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99 | (1) |
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99 | (1) |
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99 | (1) |
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2.4.8.3 Working solutions |
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99 | (1) |
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2.4.8.4 Materials preparation |
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100 | (1) |
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2.4.9 Anammox batch activity tests: Execution |
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100 | (3) |
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103 | (1) |
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2.4.10.1 Nitrification batch activity test |
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103 | (2) |
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2.4.10.2 Denitrification batch activity test |
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105 | (2) |
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2.4.10.3 Anammox batch activity test |
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107 | (2) |
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2.4.11 Additional considerations |
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109 | (1) |
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2.4.11.1 Presence of other organisms |
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109 | (1) |
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2.4.11.2 Shortage of essential micro- and macro-nutrients |
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109 | (1) |
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2.4.11.3 Toxicity or inhibition effects |
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110 | (1) |
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2.4.11.4 Effects of carbon source on denitrification |
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110 | (1) |
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2.5 Aerobic Organic Matter Removal |
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111 | (22) |
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2.5.1 Process description |
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111 | (1) |
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2.5.2 Experimental set-up |
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112 | (1) |
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112 | (1) |
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2.5.2.2 Activated sludge sample collection |
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112 | (1) |
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2.5.2.3 Activated sludge sample preparation |
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113 | (1) |
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113 | (1) |
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114 | (1) |
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2.5.2.6 Parameters of interest |
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114 | (1) |
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2.5.3 Aerobic organic matter batch activity tests: Preparation |
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115 | (1) |
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115 | (1) |
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115 | (1) |
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2.5.3.3 Working solutions |
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115 | (1) |
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2.5.3.4 Material preparation |
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116 | (1) |
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2.5.3.5 Activated sludge preparation |
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117 | (1) |
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2.5.4 Aerobic organic matter batch activity tests: Execution |
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117 | (1) |
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118 | (1) |
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119 | (1) |
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119 | (1) |
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119 | (2) |
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2.5.7 Additional considerations and recommendations |
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121 | (1) |
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2.5.7.1 Simultaneous storage and microbial growth |
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121 | (1) |
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2.5.7.2 Lack of nutrients |
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121 | (1) |
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2.5.7.3 Toxicity or inhibition |
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121 | (12) |
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133 | (44) |
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133 | (3) |
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3.1.1 Basics of respiration |
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134 | (1) |
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3.1.2 Basics of respirometry |
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135 | (1) |
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3.2 General Methodology Of Respirometry |
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136 | (5) |
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3.2.1 Basics of respirometric methodology |
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136 | (1) |
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3.2.2 Generalized principles: beyond oxygen |
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136 | (1) |
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3.2.2.1 Principles based on measuring in the liquid phase |
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136 | (2) |
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3.2.2.2 Principles based on measuring during the gas phase |
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138 | (3) |
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141 | (9) |
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3.3.1 Equipment for anaerobic respirometry |
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141 | (1) |
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3.3.1.1 Biogas composition |
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141 | (1) |
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3.3.1.2 Measuring the gas flow |
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142 | (1) |
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3.3.2 Equipment for aerobic and anoxic respirometry |
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143 | (1) |
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143 | (1) |
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3.3.2.2 Measuring arrangement |
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143 | (1) |
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3.3.2.3 Practical implementation |
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144 | (6) |
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3.4 Wastewater Characterization |
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150 | (19) |
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3.4.1 Biomethane potential (BMP) |
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150 | (1) |
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150 | (1) |
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150 | (1) |
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151 | (1) |
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151 | (1) |
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3.4.2 Biochemical oxygen demand (BOD) |
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152 | (1) |
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152 | (1) |
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152 | (1) |
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153 | (4) |
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3.4.3 Short-term biochemical oxygen demand (BDDst) |
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157 | (1) |
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158 | (2) |
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160 | (1) |
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3.4.4 Toxicity and inhibition |
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160 | (1) |
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160 | (1) |
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160 | (1) |
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161 | (1) |
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3.4.4.4 Biodegradable toxicants |
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162 | (1) |
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3.4.5 Wastewater fractionation |
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163 | (3) |
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3.4.5.1 Readily biodegradable substrate (SB) |
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166 | (1) |
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3.4.5.2 Slowly biodegradable substrate (XCB) |
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167 | (1) |
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3.4.5.3 Heterotrophic biomass (XAHO) |
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168 | (1) |
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3.4.5.4 Autotrophic (nitrifying) biomass (XANO) |
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168 | (1) |
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168 | (1) |
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3.4.5.6 Organic nitrogen fractions (XCB,N and SB,N) |
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168 | (1) |
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3.5 Biomass Characterization |
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169 | (8) |
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3.5.1 Volatile suspended solids |
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169 | (1) |
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3.5.2 Specific methanogenic activity (SMA) |
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169 | (1) |
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169 | (1) |
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169 | (1) |
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169 | (1) |
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170 | (1) |
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3.5.3 Specific aerobic and anoxic biomass activity |
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171 | (1) |
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3.5.3.1 Maximum specific nitrification rate (AUR) |
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171 | (2) |
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3.5.3.2 Maximum specific aerobic heterotrophic respiration rate (OUR) |
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173 | (1) |
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3.5.3.3 Maximum specific denitrification rate (NUR) |
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173 | (4) |
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177 | (24) |
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177 | (1) |
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4.2 Selecting The Sampling Strategy |
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178 | (1) |
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178 | (1) |
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4.2.2 Seasonal variations in emissions |
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178 | (1) |
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179 | (1) |
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4.3 Plant Assessment And Data Collection |
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179 | (6) |
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4.3.1 Preparation of a sampling campaign |
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179 | (1) |
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4.3.2 Sample identification and data sheet |
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180 | (1) |
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4.3.3 Factors that can limit the validity of the results |
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181 | (1) |
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4.3.4 Practical advice for analytical measurements |
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181 | (1) |
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4.3.5 General methodology for sampling |
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182 | (1) |
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4.3.6 Sampling in the framework of the off-gas measurements |
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183 | (2) |
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4.3.7 Testing and measurements protocol |
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185 | (1) |
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4.4 Emission Measurements |
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185 | (1) |
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4.5 N2O Measurement In Open Tanks |
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186 | (5) |
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4.5.1 Protocol for measuring the surface flux of N2O |
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188 | (1) |
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4.5.1.1 Equipment, materials and supplies |
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188 | (1) |
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4.5.1.2 Experimental procedure |
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188 | (1) |
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4.5.1.3 Sampling methods for nitrogen GHG emissions |
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189 | (2) |
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4.5.1.4 Direct measurement of the liquid-phase N2O content |
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191 | (1) |
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4.6 Measurement Of Off-Gas Flow In Open Tanks |
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191 | (1) |
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4.6.1 Protocol for aerated or aerobic zone |
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192 | (1) |
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4.6.2 Protocol for non-aerated zones |
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192 | (1) |
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4.7 Aqueous N2O and CH4 Concentration Determination |
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192 | (7) |
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4.7.1 Measurement protocol for dissolved N2O measurement using polarographic electrodes |
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193 | (1) |
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193 | (1) |
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4.7.1.2 Experimental procedure |
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193 | (1) |
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4.7.2 Measurement protocol for dissolved gasses using gas chromatography |
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194 | (1) |
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4.7.3 Measurement protocol for dissolved gas measurement by the salting-out method |
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194 | (1) |
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195 | (1) |
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4.7.3.2 Sampling procedure |
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195 | (1) |
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4.7.3.3 Measurement procedure |
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195 | (1) |
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196 | (1) |
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4.7.4 Measurement protocol for dissolved gas measurement by the stripping method |
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196 | (1) |
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4.7.4.1 Operational principle |
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196 | (1) |
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197 | (1) |
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4.7.4.3 Calibration batch test |
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198 | (1) |
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4.7.4.4 Measurement accuracy |
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198 | (1) |
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4.7.4.5 Calculation of the N2O formation rate in the stripping device |
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198 | (1) |
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4.8 Data Analysis And Processing |
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199 | (2) |
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4.8.1 Determination of fluxes |
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199 | (1) |
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4.8.2 Determination of aggregated emission fractions |
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199 | (1) |
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4.8.3 Calculation of the emission factors |
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200 | (1) |
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5 Data Handling And Parameter Estimation |
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201 | (34) |
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201 | (1) |
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202 | (9) |
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5.2.1 Data handling and validation |
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202 | (1) |
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5.2.1.1 Systematic data analysis for biological processes |
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202 | (1) |
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5.2.1.2 Degree of reduction analysis |
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203 | (1) |
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5.2.1.3 Consistency check of experimental data |
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204 | (1) |
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5.2.2 Parameter estimation |
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205 | (1) |
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5.2.2.1 Manual trial and error method |
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205 | (1) |
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5.2.2.2 Formal statistics methods |
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205 | (4) |
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5.2.3 Uncertainty analysis |
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209 | (1) |
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5.2.3.1 Linear error propagation |
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209 | (1) |
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5.2.3.2 The Monte Carlo method |
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209 | (1) |
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5.2.4 Local sensitivity analysis and identifiability analysis |
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210 | (1) |
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5.2.4.1 Local sensitivity analysis |
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210 | (1) |
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5.2.4.2 Identifiability analysis using the collinearity index |
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210 | (1) |
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5.3 Methodology And Workflow |
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211 | (3) |
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5.3.1 Data consistency check using elemental balance and a degree of reduction analysis |
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211 | (1) |
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5.3.2 Parameter estimation workflow for non-linear least squares method |
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212 | (1) |
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5.3.3 Parameter estimation workflow for the bootstrap method |
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212 | (1) |
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5.3.4 Local sensitivity and identifiability analysis workflow |
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213 | (1) |
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5.3.5 Uncertainty analysis using the Monte Carlo method and linear error propagation |
|
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213 | (1) |
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214 | (18) |
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5.5 Additional Considerations |
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232 | (3) |
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235 | (28) |
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235 | (1) |
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6.2 Measuring Sludge Settleability In SSTs |
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236 | (10) |
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6.2.1 Sludge settleability parameters |
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237 | (1) |
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6.2.1.1 Goal and application |
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237 | (1) |
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237 | (1) |
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6.2.1.3 The sludge volume index (SVI) |
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237 | (1) |
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6.2.1.4 The diluted sludge volume index (DSVI) |
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237 | (1) |
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6.2.1.5 The stirred specific volume index (SSVI3.5) |
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238 | (1) |
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6.2.2 The batch settling curve and hindered settling velocity |
|
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238 | (1) |
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6.2.2.1 Goal and application |
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238 | (1) |
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239 | (1) |
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6.2.2.3 Experimental procedure |
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239 | (1) |
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6.2.2.4 Interpreting a batch settling curve |
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240 | (1) |
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6.2.2.5 Measuring the hindered settling velocity |
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241 | (1) |
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241 | (1) |
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6.2.3.1 Goal and application |
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241 | (1) |
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242 | (1) |
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6.2.3.3 Experimental procedure |
|
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242 | (1) |
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6.2.3.4 Determination of the zone settling parameters |
|
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243 | (1) |
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6.2.3.5 Calibration by empirical relations based on SSPs |
|
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244 | (1) |
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6.2.4 Recommendations for performing batch settling tests |
|
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245 | (1) |
|
6.2.4.1 Shape and size of the batch reservoir |
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|
245 | (1) |
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6.2.4.2 Sample handling and transport |
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245 | (1) |
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6.2.4.3 Concentration range |
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245 | (1) |
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6.2.4.4 Measurement frequency |
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245 | (1) |
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6.2.5 Recent advances in batch settling tests |
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245 | (1) |
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6.3 Measuring Flocculation State Of Activated Sludge |
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246 | (4) |
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246 | (1) |
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6.3.1.1 Goal and application |
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246 | (1) |
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246 | (1) |
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246 | (1) |
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247 | (1) |
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6.3.1.5 Interpretation of a DSS/FSS test |
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248 | (1) |
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249 | (1) |
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6.3.2.1 Flocculation conditions |
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249 | (1) |
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6.3.2.2 Temperature influence |
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249 | (1) |
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6.3.2.3 Supernatant sampling |
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249 | (1) |
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6.3.3 Advances in the measurement of the flocculation state |
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250 | (1) |
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6.4 Measuring The Settling Behaviour Of Granular Sludge |
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250 | (5) |
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6.4.1 Goal and application |
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250 | (1) |
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251 | (1) |
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6.4.3 Density measurements |
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251 | (1) |
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6.4.4 Granular biomass size determination |
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252 | (1) |
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252 | (1) |
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253 | (1) |
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6.4.5 Calculating the settling velocity of granules |
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253 | (1) |
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254 | (1) |
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6.4.6.1 Validation of results |
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254 | (1) |
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6.4.6.2 Application for flocculent sludge |
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255 | (1) |
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6.5 Measuring settling velocity distribution in PSTs |
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255 | (8) |
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255 | (1) |
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255 | (1) |
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6.5.3 Sampling and sample preservation |
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256 | (1) |
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256 | (1) |
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6.5.5 Analytical protocol |
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257 | (1) |
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6.5.6 Calculations and result presentation |
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258 | (1) |
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6.5.6.1 Mass balance check |
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258 | (1) |
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6.5.6.2 Calculation of the settling velocity distribution |
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258 | (1) |
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259 | (4) |
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263 | (22) |
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263 | (1) |
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263 | (6) |
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7.2.1 Standard applications of light microscopy |
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265 | (1) |
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7.2.2 Low power objective |
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265 | (1) |
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7.2.3 High power objective |
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265 | (1) |
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7.2.4 Immersion objective |
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265 | (1) |
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7.2.5 Important considerations |
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266 | (1) |
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7.2.6 Bright-field and dark-field illumination |
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266 | (1) |
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7.2.7 Fluorescence microscopy |
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267 | (2) |
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7.2.8 Confocal laser scanning microscopy |
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269 | (1) |
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7.3 Morphological Investigations |
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269 | (3) |
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7.3.1 Microscopic `identification' of filamentous microorganisms |
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270 | (1) |
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7.3.2 `Identification' of protozoa and metazoa |
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271 | (1) |
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7.4 Examining Activated Sludge Samples Microscopically |
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272 | (4) |
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7.4.1 Mounting the activated sludge sample |
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272 | (1) |
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273 | (1) |
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7.4.2.1 Reagents and solutions for Gram staining |
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273 | (1) |
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274 | (1) |
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274 | (1) |
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7.4.3.1 Reagents and solutions for Neisser staining |
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274 | (1) |
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275 | (1) |
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275 | (1) |
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7.4.4.1 Reagents and solutions for DAPI staining |
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275 | (1) |
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275 | (1) |
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276 | (1) |
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7.4.5.1 Reagents and solutions for CTC staining |
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276 | (1) |
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276 | (1) |
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7.5 Fluorescence In Situ Hybridization |
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276 | (4) |
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7.5.1 Reagents and solutions for FISH |
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277 | (1) |
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278 | (2) |
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7.6 Combined Staining Techniques |
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280 | (5) |
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281 | (1) |
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7.6.1.1 Reagents and solutions for DAPI staining |
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281 | (1) |
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281 | (1) |
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282 | (1) |
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7.6.2.1 Reagents and solutions for PHA staining |
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282 | (1) |
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282 | (3) |
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285 | |
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285 | (1) |
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286 | (3) |
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8.2.1 General considerations |
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286 | (1) |
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286 | (1) |
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286 | (1) |
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286 | (1) |
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8.2.3.2 Nuclease activity inhibition and protein removal |
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287 | (1) |
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287 | (1) |
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8.2.3.4 Elution and storage |
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287 | (1) |
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8.2.4 Quantification and integrity |
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287 | (1) |
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8.2.5 Optimised DNA extraction from wastewater activated sludge |
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288 | (1) |
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288 | (1) |
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288 | (1) |
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8.3 Real-Time Quantitative PCR (qPCR) |
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289 | (8) |
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8.3.1 General considerations |
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289 | (2) |
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291 | (1) |
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292 | (2) |
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294 | (1) |
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8.3.5 Data output and interpretation |
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294 | (1) |
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295 | (1) |
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295 | (1) |
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295 | (1) |
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8.3.7.2 qPCR reaction setup |
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296 | (1) |
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296 | (1) |
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297 | (22) |
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8.4.1 General considerations |
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297 | (1) |
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8.4.2 The 16S rRNA gene as a phylogenetic marker gene |
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297 | (2) |
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299 | (1) |
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299 | (1) |
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300 | (1) |
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300 | (1) |
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301 | (1) |
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8.4.4.1 Sequencing platform |
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301 | (1) |
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301 | (1) |
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8.4.5 Bioinformatic processing |
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301 | (1) |
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8.4.5.1 Available software |
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301 | (1) |
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302 | (1) |
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8.4.5.3 Quality scores and filtering |
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303 | (1) |
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8.4.5.4 Merging paired end-reads |
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303 | (1) |
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303 | (1) |
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8.4.5.6 Chimera detection and removal |
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304 | (1) |
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8.4.5.7 Taxonomic classification |
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304 | (1) |
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304 | (1) |
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304 | (1) |
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8.4.6.1 Defining the goal of the data analysis |
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304 | (1) |
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8.4.6.2 Data validation and sanity check |
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305 | (1) |
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8.4.6.3 Communities or individual species? |
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305 | (1) |
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8.4.6.4 Identifying core and transient species |
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306 | (1) |
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8.4.6.5 Explorative analysis using multivariate statistics |
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306 | (1) |
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8.4.6.6 Correlation analysis |
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307 | (1) |
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8.4.6.7 Effect of treatments on individual species |
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307 | (1) |
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8.4.7 General observations |
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|
307 | (1) |
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8.4.7.1 A relative analysis |
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307 | (1) |
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307 | (1) |
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307 | (1) |
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308 | (1) |
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8.4.7.5 Impact of the method |
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308 | (1) |
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8.4.8 Protocol: Illumina V1-316S rRNA amplicon libraries |
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308 | (1) |
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308 | (1) |
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308 | (1) |
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309 | (2) |
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8.4.9 Interpretation and troubleshooting |
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311 | (1) |
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8.4.9.1 Sample DNA quality control and dilution |
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311 | (1) |
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312 | (1) |
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312 | (1) |
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8.4.9.4 Library quality control |
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313 | (1) |
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314 | (1) |
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8.4.9.6 Pool quality control and dilution |
|
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314 | (1) |
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314 | (1) |
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8.4.10 Protocol: Illumina V1-3 16S amplicon sequencing |
|
|
314 | (1) |
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314 | (1) |
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314 | (1) |
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314 | (1) |
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8.4.10.4 Interpretation and troubleshooting |
|
|
315 | (2) |
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8.4.11 Design of Illumina 16S amplicon sequencing adaptors |
|
|
317 | (2) |
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319 | |
| List Of Symbols and Abbreviations |
|