Masters thesis of applied sciences the use of the integrated soil microcosms to assess accumulation of caesium (cs) and lead (pb) from contaminated soils by earthworms (eisenia andrei) and the sunflower (helianthus annuus)

THE USE OF THE INTEGRATED SOIL MICROCOSMS TO ASSESS ACCUMULATION OF CAESIUM (Cs) AND LEAD (Pb) FROM CONTAMINATED SOILS BY EARTHWORMS (Eisenia andrei) AND THE SUNFLOWER (Helianthus annuus) HO VU VUONG School of Applied Sciences RMIT University, Melbourne, Australia March 2013 Master by Research Thesis Declaration I certify that except where due acknowledgement has been made, the work is that of mine alone; the work has not been submitted previously, in whole or in part, to qualify for any other academic award; the content of the thesis is the result of work which has been carried out since the official commencement date of the approved research program; and editorial work, paid or unpaid, carried out by a third party is acknowledged HO VU VUONG Date: 28 March 2013 ii Acknowledgements I would like to particular thank to my supervisor Professor Dayanthi Nugegoda and co-supervisor Doctor José M L Rodrigues, for their continued and enthusiastic help and support, encouragement, discussions, reading and correcting drafts and guidance throughout my project I would like to thank RMIT University and the Ministry of Education and Training Vietnam (MOET) in supporting me with the scholarship for my program I would like to forward my appreciation to Professor Andrew Smith, head of School of Applied Sciences in giving me tuition fee for my extended semester I also appreciate the enthusiastic help from all staffs and officers of RMIT Vietnam and Australia during my English courses and my Master program I received a lot of in time help from my first steps till the end I particular thank to Kathryn Thomas who helped me a lot with the paper works of the scholarship for the enrolment, visa application and advices for my first days in Australia I also thank to Prof Ngoc Tuan Nguyen, the coordinator of the program with his enthusiastic help as a bridge connected with MOET I would like to thank to Professor Dinh Don Le who was my supervisor in Vietnam I thank to all staffs from Research Institute of Biotechnology and Environment, Nong Lam University, Vietnam I would also thank to all of my friends and colleges in Vietnam, Australia and overseas (especially Thi Thu Hao Van and Ana Miranda) who always stay by my side to give a hand and give me advices I would like to thank my grandparents, my parents, brothers, sisters and relatives for their love, unconditional support and encouragement Last but not least, thank you to my beloved wife Thi Bich Thao Nguyen for her love, understanding and for being always beside me iii Table of contents Declaration ii Acknowledgements iii Table of contents iv List of figures viii List of tables x Summary .xi Common abbreviations xiii List of communications xv Chapter Introduction 1.1Contamination of soil by trace metals 1.1.1 Soil resources 1.1.2 World-wide soil contamination 1.1.3Sources of heavy metals in soil 1.2 Caesium 1.2.1 Characteristics of Cs 1.2.2 Sources of Cs pollution 1.2.3 Caesium in soils 1.2.4 The Chemical behavior of Cs in soils 11 1.2.5 Caesium and health effect in humans 12 1.2.6 Caesium and environmental effects 12 1.2.7 Remediation of Cs polluted soils 13 1.3 Lead 13 1.3.1Characteristics of Pb 13 1.3.2 Sources of Pb 14 1.3.3 Lead in soils 15 1.3.4 The chemical behavior of Pb in soil 16 1.3.5 Lead and health effects in humans 17 1.3.6 Lead and environmental effects 17 1.3.7 Remediation of Pb polluted soils 18 1.4 Remediation of contaminated soils 19 1.4.1 Remediation of heavy metals in soils 20 1.4.2 Remediation of radionuclides in soils 23 iv 1.4.3 Phytoremediation of heavy metals and radionuclides 24 1.5 Sunflowers for phytoremediation of trace metals in soils 28 1.6 Earthworms for remediation of heavy metals and radionuclides 29 1.7 Use of toxicity tests for environmental risk assessment of contaminated soils 33 1.8 Integrated soil microcosms 35 1.9 Aims of the project 36 Chapter Materials and methods 37 2.1 Soil collection 37 2.2 Soil physical and chemical characterization 37 2.3 Integrated Soil Microcosms 39 2.4 Experimental design 41 2.4.1 Experiment 1: Caesium 43 2.4.2 Experiment 2: Cs and Pbas a mixture of contaminants 43 2.5 Soil sampling, digestion and chemical analysis 44 2.6 Leachate assessment 44 2.7 Earthworm sampling, digestion and chemical analysis 44 2.8 Plant sampling, wet digestion and chemical analysis 45 2.9 Data analysis 46 Chapter Results 48 3.1 Physical and chemical characteristics of soil 48 3.2 Caesium experiment 48 3.2.1 Soil parameters after and weeks in experimental ISMs 48 3.2.2 Caesium in soils after and weeks 51 3.2.3 Effects of Cs on leachates 53 3.2.3.1 Leachate pH 53 3.2.3.2 Caesium concentration in leachates 54 3.2.4 Effects of Cs on earthworms 55 3.2.4.1 Earthworm survival 55 3.2.4.2 Effects of Cs on earthworm distribution 56 3.2.4.3 Effects of Cs on earthworm biomass 58 3.2.4.4 Caesium accumulation in earthworms 60 3.2.4.5 Caesium transfer factor (TF) 63 3.2.4.6 Total uptake of Cs by earthworms 65 3.2.5 Effects of Cs on plants 67 3.2.5.1 Effects of Cs on plant germination rate 67 3.2.5.2 Effects of Cs on plant growth (shoot length) 68 3.2.5.3 Effects of Cs on plant biomass 69 v 3.2.5.4 Caesium accumulation by plants 70 3.2.5.5 Caesium transfer factor (TF) in plants 73 3.2.5.6 Comparison of shoot and roots -translocation factor 74 3.2.5.7 Caesium total uptake in plants 75 3.3 Lead and Caesium+Lead as a mixture experiment 76 3.3.1 Soil parameters after the week experiment 76 3.3.2 Lead concentration in soils 77 3.3.3 Caesium concentration in soils 78 3.3.4 Potassium concentration in soils 79 3.3.5 Effects of Pb only and Cs+Pb on leachates 80 3.3.5.1 pH of leachates 80 3.3.5.2 Lead concentration in leachates 81 3.3.5.3 Caesium and potassium concentration in leachates 82 3.3.6 Effects of Pb only and Cs+Pb as a mixture on earthworms 83 3.3.6.1 Effects of Pb only and Cs+Pb on earthworm survival 83 3.3.6.2 Effects of Pb only and Cs+Pb on earthworm distribution 84 3.3.6.3 Effects of Pb only and Cs+Pb on earthworm fresh biomass 85 3.3.6.4 Lead accumulation in earthworms 86 3.3.6.5 Lead transfer factor in earthworms 87 3.3.6.6 Lead total uptakein earthworms 88 3.3.6.7 Caesium in earthworms 89 3.3.6.8 Caesium transfer factor in earthworms 90 3.3.6.9 Caesium total uptake in earthworms 91 3.3.6.10 Potassium in earthworms 91 3.3.7 Effects of Pb only and Cs+Pb as a mixture on plants 92 3.3.7.1 Plant germination 92 3.3.7.2 Plant growth (shoot length) 93 3.3.7.3 Above ground dried biomass of plants 94 3.3.7.4 Pb accumulation in plants 94 3.3.7.5 Lead transfer factor in sunflowers 96 3.3.7.6 Lead translocation factor in sunflowers 97 3.3.7.7 Total uptake of Pb in sunflowers 97 3.3.7.8 Caesium accumulation by plants 98 3.3.7.9 Caesium transfer factor of sunflowers 99 3.3.7.10 Caesium translocation factor 99 3.3.7.11 Total uptake of Cs in sunflowers 100 3.3.7.12 Potassium in plants 101 3.4 Estimation of the time required for effective phytoremediation 101 vi Chapter Discussion 103 4.1 Integrated soil microcosms 103 4.2 Change of soil properties and metals in soil during the experiments 104 4.2.1 Caesium in soils: 105 4.2.2 Lead in soils: 107 4.3 Leachates 108 4.4 Earthworms 108 4.4.1 Mortality and mobility of earthworms 108 4.4.2 Caesium and Pb accumulation by earthworms 110 4.5 Plants 111 4.5.1 Plant germination, growth and biomass 111 4.5.2 Caesium and Pb accumulation by plants: 113 4.6 Remediation efficiency 116 Conclusions .118 Future work 120 References 121 vii List of figures Figure Page Figure 1.1 Soil classifications Figure 1.2 Dependence of the integrated activity concentration of radionuclides during March and April 2011 on the distance from Fukushima 10 Figure 1.3 Comparison between Third Airbone Monitoring Results and Map of Cs-134 Concentration 11 Figure 1.4 Sunflowers (Helianthus annuus) 29 Figure 1.5 Eisenia andrei 33 Figure 1.6 Main components of an Integrated Soil Microcosm (ISM) 35 Figure 2.1 Integrated soil microcosms 40 Figure 2.2 Earthworm Eisenia andrei 40 Figure 2.3 Sunflowers (Helianthus annuus) 41 Figure 3.1 Cs concentrations in soils after and weeks 52 Figure 3.2 Caesium concentrations in leachates after and weeks 55 Figure 3.3 Earthworm survival after and weeks 56 Figure 3.4 Effects of Cs on earthworm distribution after and weeks 57 Figure 3.5 Effects of Cs on earthworm biomass 59 Figure 3.6 Concentration of Cs in earthworms and soil after and weeks 62 Figure 3.7 Caesium transfer factor in earthworms after and weeks 63 Figure 3.8 Total uptake of Cs by earthworms after and weeks 65 Figure 3.9 Effects of Cs on plant germination rate 67 Figure 3.10 Effects of Cs on shoot length after and weeks 68 Figure 3.11 Effects of Cs on above ground dried biomass after and weeks 69 Figure 3.12 Concentration of Cs in plants after and weeks 72 Figure 3.13.Caesium transfer factor in plants after and weeks 73 Figure 3.14 Caesium translocation factor in plants after and weeks 74 Figure 3.15 Caesium total uptake in plants after and weeks 75 Figure 3.16 Lead concentration in soils after weeks 78 Figure 3.17 Caesium in soils after weeks 79 Figure 3.18 Potassium concentrations in soils after weeks 79 viii Figure 3.19 Lead concentrations in leachates after weeks 81 Figure 3.20 Caesium concentration in leachates after weeks 82 Figure 3.21 Potassium concentrations in leachates after weeks 83 Figure 3.22 Effects of Pb and Cs+Pb on earthworm survival after weeks 83 Figure 3.23 Effects of Pb and Cs+Pb on earthworm distribution after weeks 85 Figure 3.24 Effects of Pb only and Cs+Pb on earthworm biomass after weeks 85 Figure 3.25 Lead concentration in soil and earthworms after weeks 86 Figure 3.26 Lead transfer factor in earthworms after weeks 88 Figure 3.27 Lead total uptake in earthworms after weeks 88 Figure 3.28 Caesium concentration in earthworms after weeks 89 Figure 3.29 Caesium transfer factor of earthworms after weeks 90 Figure 3.30 Caesium total uptake in earthworms after weeks 91 Figure 3.31 Potassium concentration in earthworms after weeks 92 Figure 3.32 Effects of Pb only and Cs+Pb on plant germination rate 93 Figure 3.33 Effects of Pb only and Cs+Pb on plant growth (shoot length) after weeks 93 Figure 3.34 Effects of Pb only and Cs+Pb on plant biomass (above ground dried biomass) after weeks 94 Figure 3.35 Lead concentrations in plants after weeks 95 Figure 3.36 Lead transfer factor in plants after weeks 96 Figure 3.37 Lead translocation factor in plants after weeks 97 Figure 3.38 Lead total uptake in sunflowers after weeks 98 Figure 3.39 Caesium concentration in plants after weeks 98 Figure 3.40 Caesium transfer factor in plants after weeks 99 Figure 3.41 Caesium translocation factor of plants after weeks 100 Figure 3.42 Caesium total uptake in plants after weeks 100 Figure 3.43 Potassium concentration in plants after weeks 101 ix List of tables Table Page Table 1.1 The cleanup market sites in the United States Table 1.2 Organic amendments for heavy metal immobilization 21 Table 1.3 Inorganic amendments for heavy metal immobilization 21 Table 1.4 Substances amenable to the phytoremediation process 24 Table 2.1 Parameters measured at each sampling time 42 Table 2.2 Experimental design used for Cs 43 Table 2.3 Cs and Pb accumulation experimental design 44 Table 3.1 Properties of clean soil collected near the Ecotoxicology laboratory, RMIT Bundoora west campus 48 Table 3.2 Soil pH after and weeks in experimental ISMs 49 Table 3.3 Soil conductivity (μS/cm) after and weeks in experimental ISMs 49 Table 3.4 Soil moisture (%) after and weeks in experimental ISMs 50 Table 3.5 Soil Organic Matter Content (%) after and weeks in experimental ISMs 50 Table 3.6 pH of leachates after and weeks 53 Table 3.7 Soil pH after weeks 76 Table 3.8 Soil conductivity (μS/cm) after weeks 76 Table 3.9 Soil moisture (%) after weeks 77 Table 3.10 Soil organic matter content (%) after weeks 77 Table 3.11 Leachate pH after the four week experiment 80 Table 3.12 Estimation of time required for the phytoremediation of Cs and Pb 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research)-2012 conference, in Melbourne, Australia, in October 2012, Topic: The use of the integrated soil microcosms to assess accumulation of lead (Pb) from contaminated soil by earthworms. .. Cs from soil contaminated with Cs only by sunflowers with and without earthworms The accumulation of Pb from contaminated soil by sunflowers with and without earthworms The accumulation of Cs and