ABSTRACT. using substrates that are rich in organic matter, such as compost. Methanotrophs are

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ABSTRACT Methane (CH 4 ) is an important greenhouse gas with a global warming potential 25 times higher than carbon dioxide (CO 2 ). Landfill is a one of the major contributor to global CH 4 emission and is estimated to be 500-800 Mt CO 2 eq/year. Previous studies have shown that microbial oxidation of CH 4 in landfill cover soil can be enhanced using substrates that are rich in organic matter, such as compost. Methanotrophs are group of bacteria that utilize CH 4 as its sole carbon and energy source. Therefore this study is aimed to characterize the physiochemical properties of biocover material, while identifying the potential methanotrophic bacteria from landfill cover soil. It is also intended to evaluate a comparative assessment of the CH 4 efficiency of the biocover material under controlled conditions for bottle and column experiments with the addition of potential methanotrophic bacteria. Compost has been identified as best biocover material based on the associated physiochemical properties and their ability to oxidize 4% of CH 4 within 4 days. Three types of methanotrophic bacteria were isolated from landfill soil and identified as Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2. Methylococcus sp 1 showed the highest CH 4 oxidation capacity when compared to Methylomonas sp and Methylococcus sp 2 which took only 24 hours for complete CH 4 oxidation. Batch experiment with addition of individual cultures and mixed cultures to the compost carried out at different parameters indicated higher CH 4 oxidation capacity at 35 C and 40 C, ph 6 and at 60% v/v moisture level. Addition of Methylococcus sp 1 showed the highest CH 4 oxidation activity at the rate of 8.33 X 10 3 µg g -1 h -1 while the CH 4 oxidation rate with addition of Methylomonas sp was 4.16 X 10 3 µg g -1 h -1. Addition of Methylococcus sp 2 ii

showed 75% lower activity compared to Methylococcus sp 1 and 50% lower activity compared to Methylomonas sp. Bacterial count at end of the experiment showed highest count for Methylomonas sp and Methylococcus sp 1. Statistical analysis (P < 0.05) showed significant increase in CH 4 oxidation with the addition of Methylomonas sp and Methylococcus sp 1 to the compost at optimum temperature, moisture and ph when compared to the control. Column experiment carried out with addition of potential methanotrophic bacteria to the compost at different column height showed highest CH 4 oxidation activity at 60cm with the addition of Methylomonas sp and Methylococcus sp 1 and the comparison with control also showed 50% increase in the CH 4 oxidation activity. The experiment carried out at different incubation temperature and moisture content showed highest CH 4 oxidation at the temperature of 35 C to 40 C and 60% moisture level which are similar to batch experiment. Kinetic studies using Michaelis Menten equation for batch experiment at optimum parameters showed highest potential CH 4 oxidation rate with the addition of Methyloccocus sp 1. Addition of methanotrophic bacteria to compost showed an enhancement and significant increase in the CH 4 oxidation under optimum parameters which are also similar to tropical conditions. A biocover with 60cm column height is potentially the best height for optimal CH 4 oxidation. iii

ABSTRAK Gas Metana (CH 4 ) merupakan gas rumah hijau yang penting dengan potensi pemanasan global 25 kali lebih tinggi daripada karbon dioksida (CO 2 ). Tapak pelupusan merupakan salah satu penyumbang utama kepada pembebasan CH 4 global dan dianggarkan 500-800 Mt CO 2 persamaan / tahun. Kajian terdahulu telah menunjukkan bahawa pengoksidaan mikrob CH 4 dalam tanah penutup tapak pelupusan boleh dipertingkatkan dengan menggunakan substrat yang kaya dengan bahan organik, seperti kompos. Bakteria Metanotrofik adalah kumpulan bakteria yang menggunakan CH 4 sebagai karbon tunggal dan sumber tenaga. Oleh itu kajian ini bertujuan untuk mencirikan sifat-sifat physiochemical bahan biocover, di samping mengenal pasti bakteria methanotropik yang berpotensi dari tanah penutup tapak pelupusan. Ia juga bertujuan untuk menilai perbandingan kecekapan CH 4 bahan biocover bawah keadaan terkawal untuk botol dan column eksperimen dengan penambahan bakteria Metanotrofik yang berpotensi. Kompos telah dikenal pasti sebagai bahan biocover yang terbaik berdasarkan sifat-sifat yang berkaitan fisiokimia dan keupayaan mereka untuk mengoksidakan 4% CH 4 dalam tempoh 4 hari. Tiga jenis bakteria Metanotrofik telah diasingkan daripada tanah tapak pelupusan dan dikenalpasti sebagai spesies Methylomonas, spesies Methylococcus 1 dan spesies Methylococcus 2. Spesies Methylococcus 1 menunjukkan kapasiti pengoksidaan tertinggi CH 4 apabila dibandingkan dengan spesies Methylomonas dan spesies Methylococcus 2 yang mengambil masa hanya 24 jam untuk pengoksidaan CH 4 lengkap. Eksperimen kelompok dengan tambahan individu dan campuran kompos dijalankan pada parameter yang berbeza menunjukkan kapasiti pengoksidaan CH 4 iv

yang lebih tinggi pada suhu 35 C - 40 C, ph 6 dan pada tahap kelembapan 60% v / v. Penambahan species Methylococcus 1 menunjukkan aktiviti pengoksidaan CH 4 pada kadar tertinggi, 8.33 X 10 3 μg g -1 h -1 manakala kadar CH 4 pengoksidaan dengan penambahan spesies Methylomonas adalah 4.16 X 10 3 μg g -1 h -1. Penambahan spesies Methylococcus 2 menunjukkan aktiviti 75% lebih rendah berbanding dengan spesies Methylococcus 1 dan aktiviti 50% lebih rendah berbanding spesies Methylomonas. Kiraan bakteria pada akhir eksperimen menunjukkan bilangan tertinggi dengan penambahan spesies Methylomonas dan spesies Methylococcus 1. Analisis statistik (P <0.05) menunjukkan peningkatan yang ketara dalam pengoksidaan CH 4 dengan penambahan spesies Methylomonas dan spesies Methylococcus 1 untuk kompos pada suhu, lembapan, dan ph optimum. Column eksperimen dijalankan dengan penambahan bakteria metanotrofik yang berpotensi untuk kompos pada ketinggian yang berbeza menunjukkan pengoksidaan CH 4 tertinggi pada ketinggian 60cm dengan penambahan spesies Methylomonas dan spesies Methylococcus 1 dan perbandingan dengan kawalan juga menunjukkan peningkatan sebanyak 50% dalam aktiviti pengoksidaan CH 4. Eksperimen yang dijalankan pada suhu berbeza dan kandungan lembapan menunjukkan pengoksidaan CH 4 yang tertinggi pada suhu 35 C hingga 40 C dan kelembapan 60% tahap yang sama kepada kumpulan eksperimen. Kajian kinetik menggunakan persamaan Michaelis Menten bagi eksperimen kelompok pada parameter optimum menunjukkan potensi kadar pengoksidaan CH 4 yang tertinggi dengan penambahan spesies Methylococcus 1. Penambahan bakteria metanotrofik pada kompos menunjukkan peningkatan yang ketara dalam pengoksidaan CH 4 di bawah parameter optimum yang juga sama dengan keadaan tropika. Biocover dengan ketinggian 60cm adalah berpotensi untuk pengoksidaan optimum CH 4. v

ACKNOWLEDGEMENT First of all, I would like to thank God and Guruji for being with me throughout the entire journey in completing my research successfully. My sincere gratitude goes to my supervisor Professor Dr Agamuthu Pariatamby who dynamically guided me throughout my research studies. My special thanks for funding Research project by IPPP (PS297/ 2010A), and UMRG grant (RG143/11SUS) and Ministry of Higher Education (MyBrain15) for providing me scholarship. I am also thankful to Dr. Fauziah Shahul Hamid, Dr Abioye Peter, Lim Boon Tien, Siva Shangari, Emenike Chijioke, Theepa, Arezoo, Siti Zubaidah, Leong Hong Yeng, Nizam, Kasapo and also my other fellow lab mates and friends for their valuable co-operation and advice. Last but not least, I would like to thanks my parents (Mr Barasarathi and Mrs. Vanaja) and my sisters, Puvaneswari and Mangeikarasi for their endless supports in completion of my studies. vi

CONTENTS Abstract Abstrak Acknowledgement Contents List of Figures List of Tables List of Plates List of symbol and abbreviations ii iv vi vii xiv xx xxi xxii Chapter 1.0: Introduction 1.1 Waste generation 1 1.2 Landfill 2 1.3 Landfill Gas 3 1.4 Global Warming 5 1.5 Methane Oxidation 6 1.6 Biocover as Landfill Cover 7 1.7 Methanotrophic bacteria in CH 4 Oxidation 8 1.8 Problem Statement 9 1.9 Research Objective 10 Chapter 2: Literature Review 2.1 Waste 11 2.2 MSW Generation 11 2.3 Source of MSW and Characterization 12 2.4 MSW Composition 13 vii

2.5 Solid Waste Management 15 2.6 Different options for Solid waste treatment 16 2.7 Landfilling as Disposal method 17 2.7.1 Landfilling in Malaysia 17 2.8 Landfill output 20 2.9 Generation of LFG 20 2.9.1 Factors that affect landfill gas production 24 2.10 Landfill gas composition 25 2.11 Landfill CH 4 mass balance 27 2.12 Landfill Gas and the Impacts 28 2.12.1 Greenhouse gas effect 30 2.12.2 Ozone depletion 31 2.12.3 Fire and explosion hazards 31 2.12.4 Health risks due to Toxic VOC s in air 31 2.12.5 Damages to vegetation 32 2.12.6 Odour nuisances 33 2.13 Global landfill gas CH 4 emission 33 2.14 Potential application of LFG 35 2.14.1 Gas collection system 38 2.14.2 Power generation from LFG 39 2.14.3 Direct use of LFG 39 2.15 Issues in LFG use 39 2.16 Methanotrophic activity in CH 4 oxidation 40 2.16.1 Methanotrophic bacteria 41 viii

2.16.2 Factors that affect methanotrophic activity 44 2.16.2.1 Temperature 44 2.16.2.2 Moisture content 46 2.16.2.3 ph 47 2.16.2.4 CH 4 Concentration 48 2.16.2.5 Oxygen supply 49 2.16.2.6 Organic contents and nutrients 49 2.17 Landfill components 50 2.18 Landfill cover 51 2.18.1 Different Cover Material 54 2.19 Compost as Landfill cover/biocover 56 2.20 CH 4 oxidation 57 2.21 Kinetics of CH 4 oxidation 60 2.22 Summary of Literature Review 63 Chapter 3: Materials and Method 3.1 Composting 64 3.2 Isolation of methanotrophs 65 3.3 The bacteria culture preparation 67 3.3.1 Preparation of individual culture 67 3.3.2 Preparation of mixed culture 67 3.4 Batch experiment on CH 4 oxidation activity with Matured compost 68 3.5 CH 4 depletion efficiency by pure culture of methanotrophs 69 3.6 Batch experiment on CH 4 oxidation activity with addition of methanotrophic bacteria individual culture to the compost at ix

different concentration 70 3.6.1 Methylomonas sp 70 3.6.2 Methylococcus sp 1 70 3.6.3 Methylococcus sp 2 70 3.7 Batch experiment carried out at different varying parameters for individual cultures 71 3.7.1 Influence of temperature on the CH4 oxidation rate with the addition of Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2 individually 71 3.7.2 Influence of moisture content on the CH4 oxidation rate with the addition of Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2 individually. 71 3.7.3 Influence of ph on the CH 4 oxidation rate with the addition of Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2 individually 72 3.8 Batch experiment: Methane oxidation rate with the addition of mixed culture of Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2 72 3.9 Batch experiment carried out at different parameters with mixed cultures 72 3.9.1 Influence of temperature on the CH 4 oxidation rate with the addition of mixed culture 72 3.9.2 Influence of moisture content on the CH 4 oxidation rate with the addition of mixed culture 73 x

3.9.3 Influence of moisture content on the CH 4 oxidation rate with the addition of mixed culture 73 3.10 Batch experiment with different concentration of CH 4 73 3.11 Batch experiment: Study with compost and sterilized compost with addition of Individual cultures or mixed cultures at fixed amounts and fixed parameters 74 3.12 Batch experiment with different bacteria type combination 74 3.13 Bacterial counting 75 3.14 Column Experiment 75 3.14.1 Column experiment tested at different incubation temperature 77 3.14.2 Column experiment tested at different moisture content 77 3.15 Column experiment tested with and without daily O 2 input 78 3.16 The calculation on the CH 4 oxidation rate 78 3.17 Kinetic modeling studies 79 3.18 Bacteria characterization and Identification 79 Chapter 4: Results and Discussion 4.1 Physico-chemical properties of Biocover Material 80 4.2 Batch experiment on CH 4 oxidation activity with Matured compost 82 4.3 Isolation and identification of methanotrophic bacteria 86 4.4 CH 4 depletion efficiency by pure culture of methanotrophs (4% CH 4 introduced) 88 4.5 Batch experiment on CH 4 oxidation activity with addition of methanotrophic bacteria individual culture to the compost at different concentration 89 xi

4.5.1 Batch experiment: Influence of temperature on the CH 4 oxidation rate with the addition of individual cultures Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2 to compost. 93 4.5.2 Batch Experiment: The influence of moisture content on the CH 4 oxidation rate with the addition of individual cultures of Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2 to compost. 100 4.5.3 Batch experiment: The influence of ph on the CH 4 oxidation rate with the addition of individual cultures of Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2 to compost 105 4.6 Batch experiment on CH 4 oxidation activity with addition of methanotrophic bacteria mixed culture Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2) to the compost at different concentration 110 4.6.1 The influence of temperature on the CH 4 oxidation rate with the addition of mixed microbial culture (Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2). 111 4.6.2 Batch Experiment: The influence of moisture content on the CH 4 oxidation rate with the addition of mixed microbial culture (Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2) 114 4.6.3 Batch Experiment: The influence of ph on the CH 4 oxidation rate with the addition of mixed microbial culture (Methylomonas sp, Methylococcus sp 1 and Methylococcus sp 2). 116 xii

4.7 Batch experiment with different concentration of CH 4 tested with individual methanotrophic cultures and mixed culture 119 4.8 Batch experiment: The comparative study on compost and sterilized compost with the addition of cultures at fixed amounts and fixed parameters. 121 4.9 Batch experiment with different bacteria type combination 124 4.10 Bioreactor Column Experiments with addition of methanotrophic bacteria 127 4.10.1 Column experiment tested at different incubation temperature 132 4.10.2 Column experiment tested at different moisture content 140 4.10.3 Column experiment with and without daily O 2 daily 147 4.11 Kinetic modeling to evaluate the efficiency of CH 4 oxidation 150 4.11.1 Kinetic modeling to evaluate the efficiency of batch experiments on CH 4 oxidation activity with the addition of methanotrophic bacteria at optimum parameters 151 4.11.2 Kinetic modeling for column experiment calculated for biocover heights of 60cm with addition of methanotrophic bacteria 156 Chapter 5: Conclusion 161 References 163 xiii

LIST OF FIGURES Figure 1.1: LFG production phases and times variation of landfill 4 Figure 2.1: Waste disposal options 16 Figure 2.2: The sectional view of municipal solid waste sanitary landfill 18 Figure 2.3: Major stages of waste degradation in landfills 21 Figure 2.4: Time-dependent methane production and recovery over a landfill lifetime 24 Figure 2.5: Landfill gas compositions over time 27 Figure 2.6: Landfill Methane Balance 28 Figure 2.7: Different scales of landfill effects 29 Figure 2.8: Natural greenhouse gas effect 30 Figure 2.9: Passive gas collection system 36 Figure 2.10: Active gas collection system 38 Figure 2.11: Complete pathway of the microbial oxidation of CH 4 to CO 2 by methanotrophs 42 Figure 2.12: Schematic diagrams of MSW landfill containment systems 51 Figure 2.13: Figure 2.14: Schematic diagram of a methane oxidation layer in a biocover system 54 Conceptual CH 4 oxidation zone in a landfill cover soil 60 Figure 4.1: Headspace gas analysis for CH 4, O 2 and CO 2 83 Figure 4.2: Percentage of CH 4 reduction 86 Figure 4.3: Figure 4.4: CH 4 depletion with addition of pure methanotrophic cultures. (Bar indicates standard error, n=3). 88 Influence of Methylomonas sp bacterial culture concentrations on the CH 4 oxidation rate. (Bar indicates standard error, n=3). 90 xiv

Figure 4.5: Figure 4.6: Figure 4.7: Figure 4.8: Figure 4.9: Figure 4.10: Figure 4.11: Figure 4.12: Figure 4.13: Figure 4.14: Figure 4.15: Figure 4.16: Figure 4.17: Influence of Methylococcus sp 1 bacterial culture concentrations on the CH 4 oxidation rate. (Bar indicates standard error, n=3). 91 Influence of Methylococcus sp 2 bacterial culture concentrations on the CH 4 oxidation rate. (Bar indicates standard error, n=3). 92 Influence of temperature on CH 4 oxidation rate with the addition of Methylomonas sp. (Bar indicates standard error, n=3) 95 Influence of temperature on CH 4 oxidation rate with the addition of Methylococcus sp 1. (Bar indicates standard error, n=3) 97 Influence of temperature on CH4 oxidation rate with the addition of Methylococcus sp 2. (Bar indicates standard error, n=3) 98 Bacterial counts at end of experiment for temperature with the addition of bacteria cultures 99 Influence of moisture on CH 4 oxidation rate with the addition of Methylomonas sp. (Bar indicates standard error, n=3) 101 Influence of moisture on CH 4 oxidation rate with the addition of Methylococcus sp 1. (Bar indicates standard error, n=3). 102 Influence of moisture on CH4 oxidation rate with the addition of Methylococcus sp 2. (Bar indicates standard error, n=3). 103 Bacterial count at end of experiment for moisture content experiments with the addition of bacteria cultures 105 Influence of ph on CH 4 oxidation rate with the addition of Methylomonas sp. (Bar indicates standard error, n=3) 106 Influence of ph on CH 4 oxidation rate with the addition of Methylococcus sp 1. (Bar indicates standard error, n=3) 108 Influence of ph on CH 4 oxidation rate with the addition of Methylococcus sp 2. (Bar indicates standard error, n=3) 108 xv

Figure 4.18: Figure 4.19: Figure 4.20: Figure 4.21: Figure 4.22: Figure 4.23: Figure 4.24: Figure 4.25: Figure 4.26: Figure 4.27: Figure 4.28: Figure 4.29: Figure 4.30: Bacterial count at end of experiment for ph experiments with the addition of bacteria culture 110 CH 4 oxidation rate with addition of different concentration of mixed culture. (Bar indicates standard error, n=3) 111 Influence of temperature on CH 4 oxidation rate with addition of mixed culture to the compost. (Bar indicates standard error, n=3). 112 Bacterial count for influence of temperature on CH 4 oxidation with addition of bacterial culture. (Bar indicates standard error, n=3). 113 Influence of moisture content on CH 4 oxidation rate with addition of mixed culture to the compost. (Bar indicates standard error, n=3). 115 Bacterial count of the influence of moisture content on CH 4 oxidation with addition of bacterial culture. (Bar indicates standard error, n=3). 116 Influence of ph on CH 4 oxidation rate with and without addition of bacteria culture to the compost. (Bar indicates standard error, n=3) 118 Bacterial count for the influence of ph on CH 4 oxidation with addition of bacterial culture. (Bar indicates standard error, n=3) 118 Influence of CH 4 concentration on the methanotrophic activity. (Bar indicates standard error, n=3) 119 Comparison study with compost and sterilized compost with addition of cultures. (Bar indicates standard error, n=3). 122 Bacterial count at end of experiment set up for comparison study with compost and sterilized compost with addition of cultures. (Bar indicates standard error, n=3) 123 CH 4 oxidation activity at different concentration with bacterial combination of Methylomonas sp and Methylococcus sp 1 (ratio of 1:1). (Bar indicates standard error, n=3). 124 CH 4 oxidation activity at different concentration with bacterial combination of Methylomonas sp and Methylococcus xvi

sp 2 (ratio of 1:1). (Bar indicates standard error, n=3). 125 Figure 4.31: Figure 4.32: Figure 4.33: CH 4 oxidation activity at different concentration with bacterial combination of Methylococcus sp 1 and Methylococcus sp 2 (ratio of 1:1). (Bar indicates standard error, n=3). 127 Column Experiment of Control without addition of bacteria (Bar indicates standard error, n=3) 128 Column Experiment with addition of Methylomonas sp to compost. (Bar indicates standard error, n=3). 129 Figure 4.34: Column Experiment with addition of Methylococcus sp 1 to compost. (Bar indicates standard error, n=3). 130 Figure 4.35: Column Experiment with addition of Methylococcus sp 2 to compost. (Bar indicates standard error, n=3). 131 Figure 4.36: Figure 4.37: Figure 4.38: Figure 4.39: Figure 4.40: Figure 4.41: Figure 4.42: Figure 4.43: Bacterial count at the end of experiment for column experiment to compost. (Bar indicates standard error, n=3). 132 Column experiment with addition of mixed culture at 30 C (Bar indicates standard error, n=3). 133 Bacterial population at end of the experiment at 30 C with the addition of methanotrophic mixed culture and control. (Bar indicates standard error, n=3). 134 Column experiment with addition of mixed culture at 35 C (Bar indicates standard error, n=3). 136 Bacterial counting at end of the experiment at 35 C with the addition of methanotrophic mixed culture and control. (Bar indicates standard error, n=3). 137 Column experiment with addition of mixed culture at 40 C (Bar indicates standard error, n=3). 138 Bacterial counting at end of the experiment at 40 C with the addition of mixed culture and control. (Bar indicate standard error, n=3). 140 Column experiment with addition of mixed culture at 50% moisture content. (Bar indicates standard error, n=3). 141 xvii

Figure 4.44: Figure 4.45: Figure 4.46: Figure 4.47: Figure 4.48: Figure 4.49: Figure 4.50: Figure 4.51: Figure 4.52: Figure 4.53: Figure 4.54: Bacterial counting at end of the experiment at 50% moisture content with the addition of methanotrophic mixed culture and control (Bar indicates standard error, n=3). 142 Column experiment with addition of mixed culture at 60% moisture content. (Bar indicates standard error, n=3). 143 Bacterial counting at end of the experiment at 60% moisture content with the addition of methanotrophic mixed culture and control. (Bar indicates standard error, n=3). 144 Column experiment with addition of mixed culture at 70% moisture content. (Bar indicates standard error, n=3). 145 Bacterial counting at end of the experiment at 70% moisture content with the addition of methanotrophic mixed culture and control (Bar indicates standard error, n=3). 146 The column experiment with 8% O 2 supply daily. (Bar indicates standard error, n=3). 147 The column experiment with one time O 2 supply. (Bar indicates standard error, n=3). 148 Bacterial counting at the end of experiment set up for column experiment for daily O 2 supply. (Bar indicates standard error, n=3). 149 Bacterial counting at the end of experiment set up for column experiment for with one time O 2 supply. (Bar indicates standard error, n=3). 150 CH 4 oxidation value of control (Bar indicates standard error, n=3). 152 CH 4 oxidation value with addition of Methylomonas sp. (Bar indicates standard error, n=3). 152 Figure 4.55: CH 4 oxidation value with addition of Methylococcus sp 1. (Bar indicates standard error, n=3). 153 Figure 4.56: CH 4 oxidation value with addition of Methylococcus sp 2 (Bar indicates standard error, n=3). 154 Figure 4.57: CH 4 oxidation value with addition of methanotrophic mixed culture. (Bar indicates standard error, n=3). 154 xviii

Figure 4.58: Figure 4.59: Figure 4.60: Figure 4.61: Figure 4.62: CH 4 oxidation value of column experiment at 60cm height for control (Bar indicates standard error, n=3). 156 CH 4 oxidation value of column experiment at 60cm height with addition of Methylomonas sp. (Bar indicates standard error, n=3). 157 CH 4 oxidation value of column experiment at 60cm height with addition of Methylococcus sp 1. (Bar indicates standard error, n=3). 157 CH 4 oxidation value of column experiment at 60cm height with addition of Methylococcus sp 2. (Bar indicates standard error, n=3). 158 CH 4 oxidation value of column experiment at 60cm height with addition of mixed culture. (Bar indicates standard error, n=3). 159 xix

LIST OF TABLES Table 2.1: Source and Types of Solid Waste 13 Table 2.2: Waste Composition in Malaysia (1995-2010) 14 Table 2.3: MSW Composition of selected countries 15 Table 2.4: Landfills in Malaysia as of 2012 19 Table 2.5: Characteristics of Landfill gas 26 Table 2.6: Global sources of CH 4 to the atmosphere 34 Table 2.7: Table 2.8: The phylogenetic relationship of 11 genera of methanotrophic bacteria identified and their morphological and physiological properties 43 Design parameters and structures of landfill cover for optimization of CH 4 oxidation 53 Table 2.9: Kinetic studies of the CH 4 oxidation 62 Table 3.1: Bacterial Combination 74 Table 4.1: Physiochemical properties of compost used for methane oxidation 81 Table 4.2: Characteristics of methanotrophic bacteria isolated 87 Table 4.3: Table 4.4: Potential CH 4 oxidation rate, Rp is calculated for batch experiment at optimum parameters 155 Potential CH 4 oxidation rate, Rp is calculated for column experiment at optimum heights of 60cm and optimum parameters. 160 xx

LIST OF PLATES Plate 1.1: Passive release of landfill gas to the atmosphere 5 Plate 3.1: Matured Compost from grass clippings 65 Plate 3.2: Methylomonas sp 66 Plate 3.3: Methylococcus sp 1 66 Plate 3.4: Methylococcus sp 2 67 Plate 3.5: Wheaton bottles experiment with compost 68 Plate 3.6: Gas chromatography 69 Plate 3.7: Injection of gas sample to GC 69 Plate 3.8: Schematic picture of a column 76 Plate 3.9: Column 77 xxi

LIST OF SYMBOL AND ABBREVIATIONS ASTM - American Society for Testing and Materials GHG GWP LFG - Greenhouse gas - Global Warming Potential - Landfill Gas NMVOC s - Non methane volatile organic compounds VOC CO 2 CH 4 NH 4 H 2 S O 2 H 2 0 C N NH 3 - Volatile Organic compound - Carbon dioxide - Methane - Ammonium - Hydrogen sulphide - Oxygen - Water - Carbon - Nitrogen - Ammonia xxii