Wai Chin Li Ming-Hung Wong
Effects of bacteria on metal bioavailability, speciation and mobility in different metal mine soils: acolumn study
Wai Chin Li • Ming-Hung Wong
W. C. Li • M. H. Wong ()
Croucher Institute for Environmental Sciences, and Department of Biology, Hong KongBaptistUniversity, Hong Kong SAR, P.R. China
e-mail:
Page 1
Table S1 F-values and significance levels of three-way ANOVA for pH, EC, DOC, water-soluble N, P, olsen-P and acid phosphataseof soil added with different proportions of polluted soil A, QZ or E. Layer (upper and lower layer), treatment (with or without bacterial inoculation) and concentration of polluted soil (0%, 10%, 20% and 40%) are independent variables
Site / Analyses of variance / pH / EC / DOC / Water-soluble N / Water-soluble P / Olsen-P / Acid phosphataseA / Layer (layer) / 0.16 / 28.9*** / 11.7** / 44.2*** / 125*** / 25.6*** / 23.4***
Treatment (treat) / 5.79* / 7.75** / 0.49 / 18.3*** / 14.7*** / 1.17 / 12.4***
Concentration (conc) / 70.6*** / 10.8*** / 18.1*** / 11.1*** / 164*** / 11.0*** / 19.0***
Layer x treat / 1.97 / 0.18 / 4.00 / 1.36 / 19.0*** / 6.80* / 14.5***
Layer x conc / 0.93 / 4.00* / 1.35 / 0.87 / 18.8*** / 6.00** / 20.7***
Conc x treat / 0.29 / 0.53 / 0.11 / 1.31 / 0.34 / 0.01 / 9.26***
Layer x conc x treat / 2.54 / 0.01 / 0.27 / 0.38 / 0.28 / 0.62 / 25.2***
QZ / Layer (layer) / 93.9*** / 19.0*** / 25.6*** / 8.67** / 41.0*** / 16.7*** / 25.7***
Treatment (treat) / 0.01 / 1.35 / 3.79 / 25.0*** / 0.21 / 0.41 / 11.2**
Concentration (conc) / 8.20** / 28.2*** / 0.94 / 2.20 / 45.6*** / 39.1*** / 15.1***
Layer x treat / 1.03 / 0.17 / 0.32 / 3.76 / 1.49 / 6.00* / 35.2***
Layer x conc / 0.95 / 0.73 / 0.03 / 5.56** / 2.17 / 1.15 / 5.69**
Conc x treat / 0.51 / 6.32** / 0.60 / 6.15** / 0.44 / 1.36 / 1.26
Layer x conc x treat / 1.56 / 0.12 / 2.43 / 0.77 / 0.62 / 0.51 / 1.28
E / Layer (layer) / 3.99 / 0.03 / 0.13 / 2.42 / 24.2*** / 4.56* / 165***
Treatment (treat) / 0.73 / 0.75 / 0.71 / 8.23** / 7.65* / 6.84* / 33.1***
Concentration (conc) / 11.0*** / 75.5*** / 2.98 / 6.37** / 66.7*** / 2.09 / 6.77**
Layer x treat / 0.65 / 0.45 / 0.43 / 0.08 / 0.67 / 2.10 / 185***
Layer x conc / 0.68 / 1.18 / 12.8*** / 0.16 / 14.4*** / 1.73 / 83.1***
Conc x treat / 0.05 / 0.68 / 1.07 / 0.01 / 8.10*** / 5.25* / 19.6***
Layer x conc x treat / 0.73 / 0.53 / 0.25 / 0.04 / 3.76* / 0.004 / 3.48*
*** P<0.001, **P<0.01, *P<0.05
Page 1
Table S2 F-values and significance levels of three-way ANOVA for single chemical extracted metals of soil added with different proportions of polluted soil A. Layer (upper and lower layer), treatment (with or without bacterial inoculation) and concentration of polluted soil (A0%, A10%, A20% and A40%) are independent variables. *** P<0.001, **P<0.01, *P<0.05
Analyses of variance / As / Cd / Cu / Pb / ZnWater-soluble
Layer (layer) / 30.12*** / 0.11 / 27.8*** / 8.94** / 15.0***
Treatment (treat) / 0.91 / 9.82** / 50.9*** / 3.41 / 8.27**
Concentration (conc) / 54.4*** / 1.55 / 16.6*** / 17.2*** / 1.63
Layer x treat / 7.47* / 0.61 / 20.2*** / 1.35 / 8.45**
Layer x conc / 0.44 / 0.45 / 3.01 / 0.23 / 0.21
Conc x treat / 3.78* / 1.63 / 0.95 / 0.58 / 0.47
Layer x conc x treat / 0.26 / 2.54 / 2.37 / 0.63 / 0.29
DTPA-extractable
Layer (layer) / 2.06 / 69.2*** / 111*** / 24.6*** / 22.7***Treatment (treat) / 2.57 / 27.6*** / 57.8*** / 30.5*** / 34.0***
Concentration (conc) / 10.6*** / 226*** / 82.3*** / 102*** / 306***
Layer x treat / 0.001 / 28.5*** / 23.7*** / 6.07* / 18.5***
Layer x conc / 2.13 / 1.05 / 4.81* / 0.92 / 0.54
Conc x treat / 0.99 / 3.64* / 2.00 / 1.03 / 1.13
Layer x conc x treat / 0.02 / 3.21 / 0.82 / 0.90 / 0.28
NH4NO3-extractable
Layer (layer) / 40.1*** / 9.54** / 80.9*** / n.a. / 132***
Treatment (treat) / 5.68* / 28.4*** / 15.9** / n.a. / 5.26*
Concentration (conc) / 63.3*** / 35.2*** / 57.8*** / n.a. / 355***
Layer x treat / 29.9*** / 9.76** / 12.5** / n.a. / 40.6***
Layer x conc / 1.51 / 4.31* / 1.13 / n.a. / 28.3***
Conc x treat / 1.67 / 11.0*** / 0.67 / n.a. / 4.21*
Layer x conc x treat / 2.62 / 5.50* / 2.28 / n.a. / 17.6***
NH4OAc-extractable
Layer (layer) / 1.29 / 281*** / 143*** / 58.9*** / 296***
Treatment (treat) / 30.7*** / 111*** / 71.4*** / 3.81 / 83.1***
Concentration (conc) / 265*** / 207*** / 164** / 17.4*** / 208***
Layer x treat / 13.5*** / 75.8*** / 63.4*** / 3.07 / 88.4***
Layer x conc / 46.8*** / 18.3*** / 0.002 / 1.84 / 18.1***
Conc x treat / 3.96* / 16.2*** / 3.57* / 1.65 / 11.0***
Layer x conc x treat / 6.94** / 14.0*** / 3.61* / 1.45 / 17.4***
Table S3 F-values and significance levels of three-way ANOVA for single chemical extracted metals of soil added with different proportions of polluted soil QZ. Layer (upper and lower layer), treatment (with or without bacterial inoculation) and concentration of pollutedsoil QZ0%, QZ10%, QZ20% and QZ40%) are independent variables. *** P<0.001, **P<0.01, *P<0.05
Analyses of variance / As / Cd / Cu / Pb / ZnWater-soluble
Layer (layer) / 8.94** / 0.79 / 10.9** / 4.81* / 3.98Treatment (treat) / 6.07* / 10.1** / 23.9*** / 0.82 / 7.42*
Concentration (conc) / 0.64 / 2.85 / 6.52** / 5.92** / 5.68**
Layer x treat / 0.64 / 0.88 / 5.16* / 0.26 / 0.49
Layer x conc / 1.50 / 2.55 / 2.97 / 5.27* / 14.1***
Conc x treat / 4.18* / 2.91 / 4.57* / 2.25 / 1.40
Layer x conc x treat / 0.22 / 0.01 / 0.30 / 0.33 / 0.06
DTPA-extractable
Layer (layer) / n.a. / 1.62 / 27.3*** / 4.15 / 81.11***Treatment (treat) / n.a. / 0.48 / 0.12 / 2.43 / 60.1***
Concentration (conc) / n.a. / 715*** / 297*** / 336*** / 1089***
Layer x treat / n.a. / 2.37 / 2.59 / 3.79 / 0.77
Layer x conc / n.a. / 0.28 / 10.4*** / 25.1*** / 16.3***
Conc x treat / n.a. / 0.67 / 1.84 / 1.19 / 14.0***
Layer x conc x treat / n.a. / 1.60 / 0.57 / 0.52 / 0.09
NH4NO3-extractable
Layer (layer) / n.a. / 30.3*** / 22.7*** / 10.1** / 2.21
Treatment (treat) / n.a. / 10.5** / 0.002 / 1.14 / 1.76
Concentration (conc) / n.a. / 105*** / 4.12* / 21.6*** / 29.8***
Layer x treat / n.a. / 0.72 / 0.33 / 1.82 / 0.08
Layer x conc / n.a. / 10.4*** / 0.65 / 1.13 / 0.67
Conc x treat / n.a. / 1.99 / 1.35 / 0.17 / 2.09
Layer x conc x treat / n.a. / 2.72 / 2.37 / 0.67 / 2.38
NH4OAc-extractable
Layer (layer) / n.a. / 568*** / 22.0*** / 68.3*** / 563***
Treatment (treat) / n.a. / 48.9*** / 1.76 / 0.02 / 4.17
Concentration (conc) / n.a. / 595*** / 38.6*** / 38.6*** / 524***
Layer x treat / n.a. / 64.2*** / 0.06 / 0.87 / 2.67
Layer x conc / n.a. / 78.8*** / 3.13 / 11.27*** / 107***
Conc x treat / n.a. / 15.2*** / 1.98 / 0.89 / 0.15
Layer x conc x treat / n.a. / 16.5*** / 0.24 / 0.30 / 0.13
Table S4 F-values and significance levels of three-way ANOVA for single chemical extracted metals of soil added with different proportions of polluted soil E. Layer (upper and lower layer), treatment (with or without bacterial inoculation) and concentration of polluted soil (E0%, E10%, E20% and E40%) are independent variables. *** P<0.001, **P<0.01, *P<0.05
Analyses of variance / As / Cd / Cu / Pb / ZnWater-soluble
Layer (layer) / 35.9*** / 7.76* / 46.5*** / 1.21 / 11.2**Treatment (treat) / 37.7*** / 12.4** / 6.38* / 5.44* / 4.81*
Concentration (conc) / 35.4*** / 6.13** / 64.2*** / 36.5*** / 4.09*
Layer x treat / 0.62 / 0.49 / 5.13* / 0.13 / 3.64
Layer x conc / 18.6*** / 0.51 / 0.25 / 0.65 / 2.84
Conc x treat / 2.66 / 1.36 / 1.45 / 3.25 / 1.16
Layer x conc x treat / 3.85* / 0.26 / 2.19 / 0.61 / 1.51
DTPA-extractable
Layer (layer) / 27.0*** / 0.21 / 4.77* / 0.80 / 1.18Treatment (treat) / 10.9** / 0.16 / 0.32 / 1.19 / 0.69
Concentration (conc) / 2.92 / 83.6*** / 35.5*** / 101*** / 84.4***
Layer x treat / 0.65 / 0.49 / 0.01 / 2.16 / 0.98
Layer x conc / 6.31** / 1.20 / 6.24** / 0.67 / 0.89
Conc x treat / 1.66 / 0.64 / 0.71 / 0.32 / 0.62
Layer x conc x treat / 0.31 / 0.19 / 1.39 / 1.01 / 0.27
NH4NO3-extractable
Layer (layer) / 6.00* / 1.56 / 12.9*** / 51.3*** / 0.53
Treatment (treat) / 12.2** / 1.64 / 0.16 / 0.31 / 1.31
Concentration (conc) / 10.5*** / 12.1*** / 1.94 / 3.29 / 10.3***
Layer x treat / 0.52 / 6.03* / 1.84 / 5.86* / 1.01
Layer x conc / 10.5*** / 0.94 / 9.39*** / 0.54 / 3.93*
Conc x treat / 0.87 / 7.99** / 2.70 / 0.98 / 15.3***
Layer x conc x treat / 1.05 / 0.28 / 0.70 / 0.81 / 0.17
NH4OAc-extractable
Layer (layer) / 40.8*** / 33.3*** / 0.71 / 9.67** / 38.7***
Treatment (treat) / 4.06 / 0.08 / 2.56 / 0.18 / 0.98
Concentration (conc) / 4.83* / 40.2*** / 20.6*** / 53.1*** / 49.2***
Layer x treat / 4.06 / 0.03 / 0.01 / 2.37 / 0.27
Layer x conc / 4.83* / 0.36 / 0.70 / 1.88 / 0.36
Conc x treat / 0.20 / 0.23 / 1.34 / 0.11 / 0.54
Layer x conc x treat / 0.120 / 0.69 / 0.04 / 0.18 / 0.19
Page 1
Table S5 F-values and significance levels of three-way ANOVA for sequential extracted metals (HOAc-soluble, Reducible and Oxidisable) of soil added with different proportions of polluted soil A, QZ and E. Layer (upper and lower layer), treatment (with or without bacterial inoculation) and concentration of polluted soil (E0%, E10%, E20% and E40%) are independent variables. *** P<0.001, **P<0.01, *P<0.05
A / QZ / EAnalyses of variance / As / Cd / Cu / Pb / Zn / As / Cd / Cu / Pb / Zn / As / Cd / Cu / Pb / Zn
HOAc-soluble
Layer (layer) / 1.17 / 97.3*** / 70.0*** / 7.50* / 20.7*** / 0.17 / 66.5*** / 0.04 / 137*** / 51.7*** / 15.8*** / 0.04 / 0.01 / 5.82* / 0.67Treatment (treat) / 3.38 / 19.2*** / 0.04 / 0.29 / 21.7*** / 4.58* / 8.76** / 1.22 / 0.58 / 23.2*** / 6.12* / 0.18 / 2.95 / 3.01 / 0.17
Concentration (conc) / 7.14** / 153*** / 1.54 / 7.50** / 107*** / 14.3*** / 607*** / 64.3*** / 218*** / 1140*** / 102*** / 87.9*** / 43.8*** / 155*** / 215***
Layer x treat / 1.14 / 16.8*** / 2.35 / 0.29 / 12.2** / 0.00 / 1.86 / 2.37 / 2.51 / 1.21 / 0.01 / 0.42 / 3.42 / 0.18 / 0.17
Layer x conc / 5.82** / 0.23 / 3.69 / 7.50** / 36.4*** / 1.03 / 7.64** / 1.74 / 43.2*** / 85.2*** / 2.69 / 0.19 / 1.06 / 1.45 / 1.40
Conc x treat / 0.34 / 0.96 / 1.16 / 0.29 / 3.09 / 0.65 / 2.96 / 0.33 / 0.74 / 6.27** / 2.92 / 1.02 / 0.99 / 5.51* / 0.80
Layer x conc x treat / 0.70 / 5.42* / 0.20 / 0.29 / 0.69 / 1.21 / 1.05 / 1.95 / 0.36 / 0.13 / 0.16 / 0.05 / 0.43 / 0.04 / 0.13
Reducible
Layer (layer) / 0.09 / 1.80 / 5.27* / 0.38 / 0.002 / 0.11 / 0.42 / 663*** / 0.19 / 6.68* / 14.0*** / 27.9*** / 4.34* / 32.7*** / 167***Treatment (treat) / 5.49* / 1.06 / 0.23 / 0.62 / 2.05 / 5.10* / 17.4*** / 0.12 / 2.36 / 2.18 / 0.08 / 2.24 / 3.32 / 2.73 / 11.9**
Concentration (conc) / 8.96*** / 71.8*** / 1.76 / 17.0*** / 56.0*** / 83.5*** / 107*** / 92.3*** / 40.1*** / 284*** / 28.4*** / 43.8*** / 4.03* / 65.6*** / 215***
Layer x treat / 2.28 / 1.88 / 0.06 / 0.02 / 0.70 / 1.65 / 1.72 / 0.12 / 1.83 / 0.04 / 0.88 / 0.27 / 4.06 / 2.99 / 0.32
Layer x conc / 2.40 / 1.42 / 4.06* / 3.49 / 1.67 / 1.86 / 0.57 / 91.1*** / 0.12 / 2.53 / 118*** / 8.86*** / 2.71 / 9.06*** / 25.0***
Conc x treat / 0.12 / 0.65 / 1.04 / 0.55 / 0.02 / 1.37 / 3.45* / 2.15 / 0.95 / 0.17 / 2.67 / 0.75 / 3.47* / 0.65 / 4.52*
Layer x conc x treat / 0.09 / 1.37 / 0.68 / 0.47 / 0.73 / 0.85 / 0.02 / 2.03 / 0.48 / 0.17 / 2.27 / 0.23 / 4.36* / 1.51 / 0.23
Oxidisable
Layer (layer) / 0.27 / 57.0*** / 9.79** / 3.79 / 2.48 / 60.7*** / 2.07 / 3.53 / 1.09 / 29.9*** / 13.8*** / 0.20 / 29.5*** / 3.82 / 12.5**Treatment (treat) / 0.09 / 0.89 / 2.78 / 5.03* / 3.49 / 2.13 / 1.85 / 0.74 / 0.93 / 6.70* / 10.4** / 1.64 / 0.24 / 13.7*** / 9.85**
Concentration (conc) / 51.3*** / 184*** / 10.2*** / 209*** / 173*** / 6.07* / 47.7*** / 210*** / 495*** / 253*** / 3.44* / 16.2*** / 76.8*** / 334*** / 312***
Layer x treat / 0.16 / 2.76 / 0.04 / 1.69 / 0.10 / 1.27 / 0.01 / 0.08 / 0.64 / 2.00 / 0.05 / 1.72 / 9.76** / 0.17 / 0.91
Layer x conc / 0.27 / 2.92 / 3.92* / 0.17 / 7.76** / 12.17*** / 1.66 / 0.87 / 1.29 / 8.50** / 0.78 / 0.20 / 2.94 / 1.45 / 0.16
Conc x treat / 0.09 / 1.47 / 0.42 / 2.85 / 0.63 / 2.91 / 0.63 / 0.01 / 0.23 / 5.69** / 0.42 / 0.56 / 7.30** / 3.01 / 2.05
Layer x conc x treat / 0.16 / 0.29 / 3.09 / 1.04 / 0.01 / 0.69 / 0.11 / 0.41 / 0.04 / 0.550 / 0.76 / 2.95 / 4.34* / 4.92 / 2.77
Page 1
Table S6 F-values and significance levels of three-way ANCOVA for water-soluble metals of soil added with different proportions of polluted soil A, QZ or E during 12 weeks’ incubation. Sampling time (week 1 to12), layer (upper and lower layer), treatment (with or without bacterial inoculation) and concentration of polluted soil (0%, 10%, 20% and 40%) are independent variables
Soil A
Sampling Time / 9.29** / 0.11 / 47.1*** / 2.91 / 199***Layer (layer) / 3.95* / 0.16 / 2.75 / 2.94 / 22.2***
Concentration (conc) / 1.34 / 0.05 / 2.11 / 0.13 / 2.52
Treatment (treat) / 8.64** / 1.33 / 1.87 / 0.06 / 65.4***
Layer x conc / 8.58*** / 0.53 / 1.09 / 1.37 / 1.09
Layer x treat / 3.23 / 0.20 / 5.03* / 0.31 / 0.53
Conc x treat / 6.15** / 0.18 / 2.19 / 0.94 / 1.22
Layer x conc x treat / 1.18 / 0.06 / 0.42 / 0.47 / 0.02
Soil QZ
Sampling Time / 10.8*** / 11.1*** / 36.8*** / 10.7*** / 7.07**Layer (layer) / 3.60 / 9.28** / 2.26 / 0.47 / 26.1***
Concentration (conc) / 0.52 / 25.0*** / 0.87 / 4.23* / 25.5***
Treatment (treat) / 0.50 / 11.8*** / 47.7*** / 1.56 / 127***
Layer x conc / 0.27 / 4.04** / 2.52 / 0.14 / 5.76**
Layer x treat / 0.62 / 0.25 / 0.14 / 0.001 / 1.48
Conc x treat / 0.12 / 2.00 / 2.87 / 0.68 / 3.99*
Layer x conc x treat / 0.37 / 0.25 / 0.91 / 0.21 / 0.34
Soil E
Sampling Time / 0.07 / 0.07 / 42.2*** / 13.4*** / 119***Layer (layer) / 4.78* / 1.80 / 15.4*** / 0.004 / 3.939*
Concentration (conc) / 9.97*** / 0.59 / 10.6*** / 0.10 / 0.58
Treatment (treat) / 4.51* / 1.79 / 5.74* / 2.96 / 37.4***
Layer x conc / 4.88** / 0.57 / 2.20 / 2.12 / 2.16
Layer x treat / 0.23 / 0.34 / 0.16 / 1.21 / 0.91
Conc x treat / 1.08 / 0.14 / 0.54 / 0.72 / 0.94
Layer x conc x treat / 0.60 / 0.120 / 0.91 / 0.75 / 0.72
*** P<0.001, **P<0.01, *P<0.05
Table S7 F-values and significance levels of three-way ANCOVA for pH of soil added with different proportions of polluted soil A, QZ and E. Sampling time (week 1 to12), layer (upper and lower layer), treatment (with or without bacterial inoculation) and concentration of polluted soil (0%, 10%, 20% and 40%) are independent variables
A / QZ / E
Sampling time / 3.48 / 0.25 / 5.66*
Layer (layer) / 130*** / 155*** / 86.1***
Treatment (treat) / 3.24 / 3.13 / 10.4***
Concentration (conc) / 6.29** / 4.36* / 0.24
Layer x treat / 0.23 / 0.12 / 0.79
Layer x conc / 0.23 / 1.26 / 3.68*
Conc x treat / 4.17* / 1.30 / 0.91
Layer x conc x treat / 0.002 / 0.41 / 0.32
*** P<0.001, **P<0.01, *P<0.05
Page 1