UNIVERSITI PUTRA MALAYSIA
BIOLEACHING AND BIOSORPTION OF HEAVY METALS FROM GOLD MINE TAILINGS BY ASPERGILLUS FUMIGATUS IN PENJOM,
MALAYSIA
BAHI JALILI SEH BARDAN
FP 2012 2
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BIOLEACHING AND BIOSORPTION OF HEAVY METALS FROM GOLD MINE TAILINGS BY ASPERGILLUS
FUMIGATUS IN PENJOM, MALAYSIA
By
BAHI JALILI SEH BARDAN
Thesis submitted to the School of Graduate Studies, Universiti Putra Malaysia, in Fulfilment of the Requirements for the Degree
of Doctor of Philosophy
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DEDICATION
This work is dedicated to:
My husband, Fardin, for his endless patience and encouragement.
My father, Amir, who is a true scholar, my favorite storyteller, and always
encourages my interests in science.
My mother, Ameneh, whose love, nurturing, and motivation during my
graduate career were invaluable, and who I always admire for her endless
energy.
My sisters, brothers, sister-in-law and brothers-in-law for their understanding
and love that has supported me throughout the various challenging and
rewarding experiences of my life.
My wonderful nephew, Ahura, who changed my life when he was born.
My best friend, Mahtab, for her sisterly moral support.
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Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfilment of the requirement for the degree of Doctor of Philosophy
BIOLEACHING AND BIOSORPTION OF HEAVY METALS FROM GOLD MINE TAILINGS BY ASPERGILLUS FUMIGATUS IN PENJOM,
MALAYSIA
By
BAHI JALILI SHE BARDAN
January 2012
Chairman: Radziah Othman, PhD
Faculty: Agriculture
Mine tailings is an important source of heavy metal contamination in the
environment. Decontamination of the mine tailings is necessary for the
protection of the environment. In recent years, bioleaching process has
gained increasing attention for extraction of metals from solid substrate, since
it is simple, environmentally friendly and economical. The objectives of this
study were (i) to characterize some physicochemical properties of gold mine
tailings and to isolate and identify indigenous fungi from the tailings, (ii) to
evaluate the ability of the indigenous fungus to bioleach heavy metals in step
bioleaching and column bioleaching processes using distributed and surface
applied techniques, and (iii) to assess the efficiency of A. fumigatus biomass
to biosorb and remove metal from synthetic solutions and from leachate
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derived from a gold mine. Bioleaching experiments were conducted for the
removal of heavy metals from mine tailings using a fungal strain isolated from
the gold mine tailings and it was identified as A. fumigatus based on its 18S
rDNA analysis. The bioleaching processes were carried out in one-step and
spent medium leaching at 1%, 2%, 4% and 8% (w/v) tailing concentrations.
Column bioleaching experiments were carried out to compare the
effectiveness of the fungus to bioleach heavy metals from the tailings using
distribution technique (DT) and surfaced applied technique (SAT). The ability
of A. fumigatus to remove As, Fe, Mn, Pb and Zn from synthetic solutions
and from leachate derived from gold mine during bioleaching was assessed.
Batch sorption experiments were carried out to characterize the capability of
fungal biomass (FB) and iron-coated fungal biomass (ICFB) to remove metal
ions in single and multi-solute systems. Results showed that in the one step
bioleaching process, production of oxalic acid was the highest among the
other organic acids; while in the spent medium leaching citric acid was
dominant. The removal of As, Fe, Mn and Zn was higher in the one step than
the spent medium leaching but the reverse was observed for Pb. Heavy
metals removal efficiency decreased with increasing concentration of tailings
in both bioleaching processes. The column bioleaching study showed that DT
produced higher oxalic acid than the SAT, therefore more As, Fe, Mn, and Zn
were removed by the DT method compared to the SAT. However, Pb
removal was low by DT than the SAT probably due to the precipitation of Pb
as its oxalates. The DT was a more promising method for removal of metal
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ions from the mine tailings because distributing the fungus throughout the
entire soil column improved the bioleaching efficacy of the heavy metals. The
biosorption study showed that sorption data for all systems fitted well the
Langmuir isotherm equation. The maximum sorption capacities of metals by
both FB and ICFB were higher in the single metal system than the multi-
solute system. It indicates that the presence of multiple metal ions in as
solution suppressed adsorption of the individual metal ions. Sorption capacity
of ICFB was higher than FB for all metal ions because the net surface charge
of ICFB was more negative than FB at the pH range of 6 to 9, therefore the
ICFB could adsorb more cationic metals. As for As adsorption, the As formed
complexes with the Fe oxide of the ICFB surface sites. The FT-IR analysis
showed that the functional groups of the fungal biomass were involved in the
metal ions sorption. Overall, the results suggest that bioremediation process
using A. fumigatus was effective for the leaching of heavy metals from mine
tailings especially in the one-step bioleaching process using the DT method.
The ICFB was found to be effective in adsorbing the metals present in the
leachate of the tailing bioleaching process.
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Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai memenuhi keperluan untuk mendapat ijazah Doktor Falsafah
BIOLURUHLARUTAN DAN BIOJERAPAN LOGAM BERAT DARIPADA SISA LOMBONG EMAS OLEH ASPERGILLUS FUMIGATUS
DALAM PENJOM, MALAYSIA
Oleh
BAHI JALILI SEHBARDAN
Januari 2012
Pengerusi: Radziah Othman, PhD
Fakulti: Pertanian
Tahi lombong adalah punca penting pencemaran logam berat dalam
persekitaran. Dekontaminasi tahi lombong adalah perlu bagi perlindungan
alam sekitar. Pada tahun-tahun kebelakangan ini, proses larutlesap telah
mendapat perhatian yang semakin meningkat bagi pengekstrakan logam
daripada substrat pepejal, kerana ia mudah, mesra alam dan ekonomi.
Objektif kajian ini adalah (i) untuk mencirikan beberapa sifat tahi lombong
emas dan untuk mengasingkan dan mengenal pasti kulat asli dari tahi
lombong tersebut, (ii) untuk menilai kemampuan kulat asli A. fumigatus untuk
melarutlesapbio logam berat dalam proses larutlesapbio di dalam kolum
menggunalan teknik edaran dan teknik taburan pada permukaan, dan (iii)
untuk menilai keberkesanan biojisim A. fumigatus untuk menjerapbio dan
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mengeluarkan ion logam dalam air yang terhasil dari proses
larutlesapbio.Eksperimen larutlesapbio telah dijalankan untuk penyingkiran
logam berat dari tahi lombong dengan menggunakan strain kulat yang
diasingkan daripada tahi lombong emas dan ia telah dikenal pasti sebagai A.
fumigatus berdasarkan analisis 18S rDNA. Proses larutlesapbio telah
dijalankan dalam satu langkah, dengan kepekatan media tahi lombong pada
kepekatan 1%, 2%, 4% dan 8% (w / v).Eksperimen larutlesapbio lajur
dijalankan untuk membandingkan keberkesanan kulat untuk
menglarutlesapbiologam berat daripada tahi lombong menggunakan teknik
pengagihan (TP) dan teknik taburan permukaan (TTP). Keupayaan A.
fumigatus untuk menghapuskan As, Fe, Mn, Pb dan Zn daripada larutan
sintetik dan daripada hasil larutlesap yang berasal dari lombong emas
semasa larutlesapbio dinilai. Eksperimen jerapan sekumpulan telah
dijalankan untuk mencirikan keupayaan biojisim kulat (BK) dan biojisim ferum
bersalut kulat (BFBK) untuk mengeluarkan ion logam dalam sistem tunggal
dan pelbagai bahan larut.Hasil kajian menunjukkan bahawa dalam proses
larutlesapbio satu langkah, pengeluaran asid oksalik adalah yang tertinggi di
kalangan asid organik yang lain, sedangkan dalam medium hasil larut lesap,
asid sitrik adalah dominan. Penyingkiran As, Fe, Mn dan Zn adalah lebih
tinggi dalam proses langkah salah satu daripada medium hasil larut lesap
tetapi sebaliknya dilihat untuk Pb. Kecekapan penyingkiran logam berat
menurun dengan peningkatam kepekatan tahi lombong di kedua-dua proses
larutlesapbio. Kajian larutlesapbio kolum menunjukkan TP menghasilkan asid
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oksalik yang lebih tinggi daripada TTP, oleh itu, lebih banyak As, Fe, Mn, dan
Zn telah dikeluarkan dengan kaedah DT berbanding SAT. Walau
bagaimanapun, penyingkiran Pb adalah rendah oleh TP berbanding TTP
mungkin disebabkan oleh pemendakan Pb sebagai oksalat. Kaedah TP
adalah lebih berpotensi untuk penyingkiran ion logam daripada tahi lombong
kerana agihan kulat sepanjang kolum tanah meningkatkan keberkesanan
larutlesapbio logam berat. Kajian biojerapan menunjukkan bahawa data
jerapan untuk semua sistem dilengkapi boleh disesuaikan dengan
persamaan isotherma Langmuir. Kapasiti maksima jerapan logam oleh
kedua-dua BK dan BFBK dalam sistem logam tunggal adalah lebih tinggi
daripada sistem pelbagai bahan larut. Ia menunjukkan bahawa kehadiran
pelbagai ion logam menekan penjerapan penjerapan ion logam secara
individu. Kapasiti jerapan BFBK adalah lebih tinggi daripada BK untuk semua
ion logam kerana caj bersih permukaan BFBK adalah lebih negatif daripada
BK pada julat pH 6-9, oleh itu BFBKB boleh menjerap logam yang lebih
kationik. Bagi penjerapan As, ia membentuk kompleks dengan oksida Fe
pada permukaan BFBK. Analisa FT-IR menunjukkan bahawa kumpulan
berfungsi biojisim kulat terlibat dalam jerapan ion logam.Secara keseluruhan,
keputusan menunjukkan bahawa proses bioremediasi menggunakan A.
fumigatus berkesan untuk larutlesap logam berat dari tahi lombong
terutamanya dalam proses larutlesapbio satu langkah menggunakan kaedah
TP. BFBK didapati berkesan dalam menjerap logam yang hadir dalam hasil
larutlesap proses larutlesapbio tahi lombong.
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ACKNOWLEDGEMENTS
I wish to express my deepest appreciation to Assoc. Prof. Dr. Radziah
Othman for providing valuable academic advices, critical support and
guidance during my graduate study. Her friendship and monitoring helped
guide me throughout my graduate program. I would also like to thank my
committee members, Dr. Samsuri Abd Wahid and Assoc. Prof. Dr. Aminuddin
Husin who provided valuable assistance and advice to ensure the quality of
my research.
I am grateful for the assistance and support of my husband, Dr. Fardin
Sadegh-Zadeh, who provided experimental advice.
I am thankful to the members of the Land Management Department
especially to Mr Ramley Sulaiman and Mr. Alias Tahar for providing me the
equipment to conduct the research.
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I certify that a Thesis Examination Committee has met on 5 January 2012 to conduct the final examination of Bahi Jalili Seh Bardan on his Doctor of Philosophy thesis entitled “Bioleaching and biosorption of heavy Metals from gold mine tailings in Penjom, Malaysia, by Aspergillus fumigatus” in accordance with the Universities and University Colleges Act 1971 and the Constitution of the Universiti Putra Malaysia [P.U. (A) 106] 15th March 1998. The Committee recommends that the student be awarded the Doctor of Philosophy. Members of the Thesis Examination Committee were as follows: Shamsuddin Jusop, PhD Professor Faculty of Agriculture Universiti Putra Malaysia (Chairman) Sariah Meon, PhD Professor Faculty of Agriculture Universiti Putra Malaysia (Internal Examiner) Ahmad Ismail Professor Faculty of Science Universiti Putra Malaysia (Internal Examiner) Yen Peng Ting Associate Professor Department of chemical and bimolecular engineering National University of Singapore (External Examiner)
________________________
Seow Heng Fong, PhD
Professor and Deputy Dean School of Graduate Studies
Universiti Putra Malaysia Date:
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This thesis was submitted to the Senate of Universiti Putra Malaysia and has been accepted as fulfilment of the requirement for the degree of Doctor of Philosophy. The members of the Supervisory Committee were as follows:
Radziah Othman, PhD Associate Professor Faculty of Agriculture Universiti Putra Malaysia (Chairman) Samsuri Abd Wahid, PhD Lecturer Faculty of Agriculture Universiti Putra Malaysia (Member) Aminuddin Husin, PhD Associate Professor Faculty of Agriculture Universiti Putra Malaysia (Member)
______________________________
BUJANG BIN KIM HUAT, PhD Professor and Dean
School of Graduate Studies Universiti Putra Malaysia
Date:
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DECLARATION
I declare that the thesis is my original work except for quotations and citations
which have been duly acknowledged. I also declare that it has not been
previously and is not concurrently, submitted for any other degree at
Universiti Putra Malaysia or other institutions.
Bahi Jalili Seh Bardan
Date: 5 January 2012
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TABLE OF CONTENTS
Page
DEDICATION II ABSTRACT III ABSTRAK VI ACKNOWLEDGEMENTS X APPROVAL XI DECLARATION XIII LIST OF TABLES XVII LIST OF FIGURES XVIII LIST OF ABBREVIATIONS XXI 1 INTRODUCTION 1
2 LITERATURE REVIEW 5 2.1 Heavy metals 5 2.2 Mine tailings material 6 2.1.1Definition and characteristics of mine tailings 6 2.1.2 Metals species in mine tailings 6 2.2 Remediation technology 9 2.2.2 Bioleaching process 9 2.3 Remediation of metal-contaminated leachate using fungal 28 2.3.1 Use of fungal biomass for biosorption 29 2.3.2 Fungal structure 29 2.3.3 Effect of pre-treatment on biosorption by fungal biomass 30 2.3.4 Ecology and biology of Aspergillus fumigatus 32
3 MICROBIAL AND PHYSICOCHEMICAL ANALYSES OF GOLD MINE TAILINGS 34
3.1 Introduction 34 3.2 Materials and Methods 36 3.2.1 Mine tailing site 36 3.2.2 Microbial isolates from mine tailings 38 3.2.3 Molecular identification of fungal isolate 40 3.2.4 Mine tailings characterization 41 3.3 Result and Discussion 46 3.3.1 Selection of microbial isolates for bioleaching 46 3.3.2 Mine tailings characterization 50 3.4 Conclusions 60
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4 BIOLEACHING OF HEAVY METALS FROM MINE TAILINGS
BY ASPERGILLUS FUMIGATUS USING ONE-STEP AND TWO-STEP PROCESS 61
4.1 Introduction 61 4.2 Materials and Methods 62 4.2.1 Microorganisms 62 4.2.2 Bioleaching experiments 63 4.2.3 Chemical leaching experiment 65 4.2.4 Determination ofpH, organic acids and heavy metals and
concentration 65 4.2.5 Bioleaching yield 67 4.2.6 Statistical analyses 67 4.3. Results and Discussion 67 4.3.1 Bioleaching experiments 67 4.3.2 Chemical leaching experiment 80 4.4. Conclusions 84
5 BIOLEACHING OF HEAVY METALS FROM GOLD MINE TAILINGS BY ASPERGILLUS FUMIGATUS USING COLUMN TECHNIQUE 85
5.1 Introduction 85 5.2 Materials and methods 86 5.2.1 Medium for fungus 86 5.2.2 Column bioleaching designs 87 5.2.3 Statistical analyses 89 5.2.4 determination of heavy metals and organic acis
concentration 90 5.3 Results and Discussion 90 5.3.2 Bioleaching experiments 90 5.4 Conclusions 106
6 BIOSORPTION OF HEAVY METALS FROM LEACHATE DERIVED FROM BIOLEACHING OF GOLD MINE TAILINGS USING ASPERGILLUS FUMIGATUS 107
6.1 Introduction 107 6.2 Materials and methods 109 6.2.1 Chemicals preparation 109 6.2.2 Preparation of A. fumigatus biomass 110 6.2.3 Surface charge of fungal biomass 111 6.2.4 Preparation of metal solutions 111 6.2.5 Biosorpation experiments in single-soulte system 112 6.2.6 Biosorption experiments in multi-solute system 115 6.2.7 Preparation of leachate 115
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6.2.8 Biosorption of metal ions from leachate samples using ICFB 115
6.2.9 Fourier transforms infrared (FTIR) spectroscopy of UFB and ICFB before and after biosorption 116
6.2.10 Scanning electron microscopy of fungal biomass 116 6.2.11 Statistical analyses 116
6.3 Results and discussion 117 6.3.1 surface charge of fungal biomass 117 6.3.2 Biosorpation experiments in single system 118 6.3.3 Biosorption experiments in multi-solute system 133 6.3.4 Removal of metal ions from leachate using ICFB 136 6.3.5 FT-IR analysis 138 6.3.6 Scanning electron microscopy 143
6.4 Conclusions 144
7 SUMMARY, GENERAL CONCLUSIONS 145
REFERENCES 150 BIODATA OF STUDENT 164 LIST OF PUBLICATIONS 165
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