Assessment in Rocky Shore Organisms and Surface Sediment Along Peninsular Malaysia Coastal Waters

RARE EARTH ELEMENTS (LANTHANIDES) ASSESSMENT IN ROCKY SHORE ORGANISMS AND SURFACE SEDIMENT ALONG PENINSULAR MALAYSIA COASTAL WATERS

MOHD FUAD MISKON

A thesis submitted in fulfillment of the requirement for the degree of Doctor of Philosophy in Bioscience

Kulliyyah of Science International Islamic University Malaysia

MARCH 2016 ABSTRACT

This study emphases on the assessment of REEs in rocky shore ecosystem along Peninsular Malaysia coastal waters, describing their partitioning between surface sediment and selected body parts of Saccostrea cucullata, Thais clavigera and Nerita chameleon, and deliberating on interspecies, inter-tissue and interspatial variability. Samples was digested using Teflon Bomb technique and concentrations of 14 naturally occurring REEs were measured using ICP-MS technique, along with selected trace metals for additional data. The REEs fractionation patterns normalized to chondrite or shale were remarkably similar, indicating a common source of the REEs for the whole region, however, mean concentration suggested that east Peninsular Malaysia provides higher REEs compared to west peninsula area. Consistent REEs abundance patterns were found in all samples, with enrichment of LREEs over HREEs, which implies that REEs are transported as a coherent group through aquatic ecosystems. There are dissimilarities in the REEs abundance for each site, but they demonstrate similarities in their REEs distribution patterns, which propose that they are of parallel origins. The contaminant metals As, Mn, Cu, and Cd were significantly correlated with REEs (p<0.05 and p<0.01), consequently suggests that these metals are probably non-anthropogenic in origin as the REEs are geogenic in origin. Quantification of anomalies calculated using values normalized to chondrite, PAAS and NASC data presented ratios higher than unity for Ce and ratios lower than unity for Eu in all sites. Filter feeder S. cucullata is considered to be highly potential as a bioindicator for REEs, undoubtedly due to its feeding behaviour that is much related to particulates as REEs sources. Consistent chondrite-normalized pattern strongly suggests that the REEs accumulated by S. cucullata, T. clavigera and N. chameleon are derived mostly from indigenous rocks. Results showed that the values are considerably below the safety limit, with the exception of Ce and Nd in the soft tissue of S. cucullata. With regards to sediment, comparative analysis resolved that the concentration of surface sediment in this study is very low compared to limit sets.

خالصة البحث

ّسوضث ٘زٖ اٌذساست ػٍى حمٍٍُ اؼٌٕبصش األسضٍت إٌبدسة فً اٌصخٛس اٌشبطئٍت ػٍى طٛي اٌٍّبٖ اٌسبدٍٍت ٌشبٗ جضٌشة ِبٌٍضٌب، حُ ٚصف أخشبس٘ب ِب بٍٓ اٌشٚاسب اٌسطذٍت ٚأجضاء ِخخبسة ٌىً ِٓ Thais clavigera ٚ Saccostrea cucullata ٚ Nerita chameleon ٚدساسخٙب ِٓ بٍٓ األٛٔاع اٌّخخٍفت، ٚبٍٓ األٔسجت ٚاٌخببٌٓ اٌّىبًٔ ٌٙب. حُ اخخٍبس اؼٌٍٕبث بئسخخذاَ حمٍٕت ِضخت اٌخفٚ ٍْٛبئسخخذاَ أسبؼت ػشش حشوٍضا حذذد بشىً طبٚ .ًؼٍحُ لٍبط اؼٌٕبصش األسضٍت إٌبدسة ببسخخذاَ حمٍٕت ICP-MS، جٕبب إٌى جٕب ِغ اؼٌّبدْ إٌبدسة اٌّخخبسة وبٍبٔبث إضبفٍت. حُ حسٌٛت أّٔبط حجضئت اؼٌٕبصش األسضٍت إٌبدسة اٌى وٛٔذسٌج أٚ اٌصخش اٌضٌخً ٚوبٔج ِخشببٙت بشىً ٍِذٛظ، ِّب ٌذي ػٍى اٌّصذس اٌّشخشن ؼٌٍٕبصش األسضٍت إٌبدسة فً إٌّطمت وٍٙب، ِٚغ رٌه، ٌشٍش ٌبذذ إٌى أْ ِٕطمت ششق شبٗ جضٌشة ِبٌٍضٌب ٌٛفش حشاوٍض أػٍى ِٓ اؼٌٕبصش األسضٍت إٌبدسة ِمبسٔت ِغ ِٕطمت شبٗ اٌجضٌشة اٌغشبٍت. حُ اؼٌزٛس ػٍى أّٔبط حٛاجذ اؼٌٕبصش األسضٍت إٌبدسة رببخت فً جٍّغ اؼٌٍٕبث، ِغ خصٛبت ػ LREEsٍى HREEs، ِّب ٌذي ػٍى أٔٗ حُ حٛصٌغ اؼٌٕبصش األسضٍت إٌبدسة وّجػّٛت ِخّبسىت ِٓ خالي إٌظبَ اٌبٍئً اٌّبئً. ٕ٘بن إخخالفبث فً حٛاجذ اؼٌٕبصش األسضٍت إٌبدسة فً وً ِٛلغ ٌىٕٙب ِخشببت فً أّٔبط حٛصٙؼٌب ِّب ٌذي ػٍى أٙٔب ِٓ ِٕشأ ِخٛاصي. وبٔج اؼٌّبدْ اٌٍّٛرت اسسٍٕسٓ، إٌّغٍٕض، إٌذبط، ٚاٌىبدٍَِٛ ِشحبطت بشىً وبٍش ِغ اؼٌٕبصش األسضٍت إٌبدسة ) p<0.05 and p<0.01(، ببٌخبًٌ ٌشٍش إٌى أْ ٘زٖ اؼٌّبدْ ً٘ ػٍى األسجخ غٍش بششٌت إٌّشأ ِزً اؼٌٕبصش األسضٍت إٌبدسة اٌجٍٛجٕه فً األصً. اٌخذًٍٍ اٌىًّ ٌٍذبالث اٌشبرة اٌّذسٛبت ببسخخذاَ لٍُ حسٌٛت اٌىٛٔذسٌج. ٚلذِج بٍبٔبث NASCٚ PAAS ٔسب أػٍى ِٓ اٌسٍشٚ ،ٌَٛٔسب ألً ِٓ Eu فً وبفت اٌّٛالغ. ؼٌخبش ِششخ اٌّغزٌت .S cucullata ٌذٌٗ إِىبٍٔبث ػبٌٍت ببػخببسٖ ِؤششاث بٌٍٛٛجٍت ؼٌٍٕبصش األسضٍت إٌبدسة، ِّب ٌذي ػٍى سٍٛن اٌخغزٌت اٌخً ٌٙب اسحببط وبٍش ٌٍجسٍّبث وّب فً اؼٌٕبصشاألسضٍٗ إٌبدسٖ. ٌٚشٍش ّٔظ حىٌٛٓ اٌىٛٔذسٌج اٌّخٕبسك أْ اؼٌٕبصش األسضٍت إٌبدسة اٌّخشاوّت N. chameleon ٚ S. cucullata, T. clavigera ِسخّذة فً ؼِظّٙب ِٓ اٌصخٛس األصٍٍت. أظٙشث إٌخبئج أْ اٌمٍُ ً٘ ألً بىزٍش ِٓ دذ اٌسالِت، ببسخزٕبء اٌسٍشٚ ٌَٛإٌٍٛدٌٍَّٛ فً األٔسجت إٌٍٍت فً S. cucullata ٚفٍّب ٌخؼٍك فً اٌشٚاسب، فمذ أظٙش اٌخذًٍٍ اٌّمبسْ ٌٙزٖ اٌذساست أْ حشوٍض اٌشٚاسب اٌسطذٍت ِٕخفض جذا ِمبسٔت فً اٌذذٚد اٌّسّٛح فً اٌّجػّٛبث.

iii

APPROVAL PAGE

The thesis of Mohd Fuad Miskon has been approved by the following:

______Kamaruzzaman Yunus Supervisor

______Md. Mokhlesur Rahman Co-Supervisor

______Md. Zahangir Alam Internal Examiner

______Noor Azhar Mohamed Shazili External Examiner

______Ahmad Ismail External Examiner

______Wan Mohd Azizi Wan Sulaiman Chairman

DECLARATION

I hereby declare that this dissertation is the result of my own investigations, except where otherwise stated. I also declare that it has not been previously or concurrently submitted as a whole for any other degrees at IIUM or other institutions.

Mohd Fuad Miskon

Signature ...... Date ......

DECLARATION OF COPYRIGHT AND AFFIRMATION OF FAIR USE OF UNPUBLISHED RESEARCH

RARE EARTH ELEMENTS (LANTHANIDES) ASSESSMENT IN ROCKY SHORE ORGANISMS AND SURFACE SEDIMENT ALONG PENINSULAR MALAYSIA COASTAL WATERS

I declare that the copyright holders of this dissertation are jointly owned by the student and IIUM.

No part of this unpublished research may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording or otherwise without prior written permission of the copyright holder except as provided below

1. Any material contained in or derived from this unpublished research may be used by others in their writing with due acknowledgement.

2. IIUM or its library will have the right to make and transmit copies (print or electronic) for institutional and academic purposes.

3. The IIUM library will have the right to make, store in a retrieved system and supply copies of this unpublished research if requested by other universities and research libraries.

By signing this form, I acknowledged that I have read and understand the IIUM Intellectual Property Right and Commercialization policy.

Affirmed by Mohd Fuad Miskon

……..…………………….. ……………………….. Signature Date

ACKNOWLEDGEMENTS

Alhamdulillah, all praises to Allah for the strengths and His blessing. Only by His grace and mercy this thesis has been completed.

I would like to take this opportunity to express my grateful thanks and deep appreciation to my supervisor, Prof. Dr. Kamaruzzaman Yunus, for his invaluable guidance and encouragement throughout my research. Not forgotten, my appreciation to my co-supervisor, Assoc. Prof. Dr. Mokhlesur Rahman for his guidance, support and knowledge. I am also grateful to the Deputy Dean Research and Postgraduate, Prof. Dr. Ahmed Jalal Khan Chowdhury for his support and help towards my postgraduate affairs.

Special thanks are extended to all the technicians, office staffs and laboratory assistants of Kuliyyah of Science especially to Br. Ahmad Faezal for his kind assistance during sampling session. Sincere thanks to all my friends, especially Br. Zaini, Br. Faiz, Sr. Hafizah, Sr. Atikah and Br. Fadhli for helping me through my research period. Also to those who indirectly contributed in this research, your help and kindness means a lot to me.

My deepest gratitude goes to my beloved parents; Allahyarham Hj. Miskon Kasmat and Hjh Zaiton Talkah, and my siblings, to my beloved parents-in-law; Hj. Faudzi Ahmad and Allahyarhamah Hjh Aziah Othman for their endless love, prayers, support and encouragement in letting me pursue my dreams.

Finally, I am greatly indebted to my beloved wife, Fikriah Faudzi for her endless love, patience, encouragement, sacrifices and prayers throughout the study. Also, I am grateful for the greatest gift from the Almighty Allah, my son, Muhammad Fayyadh Aufaa, who opening up an exciting world and bringing us joy during the stages of my research.

Thank you very much.

vii

TABLE OF CONTENTS

Abstract …………...... ii Abstract in Arabic …………………………...………………………………….....iii Approval Page ……………………………………………………………………..iv Declaration …………………………………………………………………………v Copyright Page …… ...... vi Acknowledgements ...... vii List of Tables …………………………………………...………………………....xiv List of Figures ………………….……………………………….………………....xviii

CHAPTER ONE: INTRODUCTION ...... 1 1.1 Research Background ...... 1 1.2 Problem Statement ...... 4 1.3 Research Questions ...... 4 1.4 Research Scope ...... 5 1.5 General Objectives...... 5 1.6 Specific Objectives ...... 5 1.7 Research Hypothesis ...... 6 1.8 Significance of Research ...... 6 1.9 Limitations of Research ...... 7 1.10 Chapter Summary ...... 7

CHAPTER TWO: LITERATURE REVIEW ...... 9 2.1 Background ...... 9 2.2 Rare Earth Elements ...... 9 2.3 Rare Earth in Nature ...... 13 2.4 REEs Fate in the Environment ...... 16 2.5 REEs as a Pollutant Tracer ...... 18 2.6 Rocky Shore...... 22 2.7 Pollutant Biomonitoring Using Molluscs ...... 23 2.8 Molluscs as Bioindicator in Malaysia ...... 25 2.9 Biology of Studied Organisms...... 27 2.9.1 Saccostrea cucullata (Born, 1778)...... 27 2.9.2 Thais clavigera (Kuster, 1860) ...... 28 2.9.3 Nerita chameleon (Linnaeus, 1758) ...... 29 2.10 Inductively Coupled Plasma Mass Spectrometry (ICP-MS) ...... 30

CHAPTER THREE: MATERIAL AND METHODS ...... 35 3.1 Introduction...... 35 3.2 Sampling Sites ...... 36 3.3 Sampling Sites Description...... 41 3.3.1 Langkawi Island, Kedah ...... 41 3.3.1.1 Pasir Tengkorak Beach ...... 41 3.3.1.2 Tengah Beach ...... 42 3.3.1.3 Beringin Beach ...... 42 3.3.2 Pulau Sayak, Kedah ...... 43

3.3.3 Pulau Betong, Penang ...... 43 3.3.4 Batu Feringghi, Penang ...... 44 3.3.5 Teluk Senangin, Perak ...... 44 3.3.6 Teluk Batik, Perak...... 45 3.3.7 Taman Tanjung, Negeri Sembilan ...... 46 3.3.8 Teluk Kemang, Negeri Sembilan ...... 46 3.3.9 Blue Lagoon, Negeri Sembilan ...... 47 3.3.10 Tanjung Bidara, Malacca ...... 47 3.3.11 Tanjung Kling, Malacca ...... 48 3.3.12 Batu Pahat, Johor ...... 48 3.3.13 Sungai Rengit, Johor ...... 49 3.3.14 Batu Layar, Johor ...... 50 3.3.15 Tanjung Balau, Johor ...... 50 3.3.16 Sedili Kecil, Johor ...... 51 3.3.17 Mersing, Johor ...... 51 3.3.18 Kampung Tanjung Batu, Pahang ...... 52 3.3.19 Teluk Cempedak, Pahang ...... 52 3.3.20 Tanjung Gelang, Pahang ...... 53 3.3.21 Cherating, Pahang ...... 53 3.3.22 Telaga Simpul, Terengganu ...... 54 3.3.23 Teluk Kalong, Terengganu ...... 54 3.3.24 Kijal, Terengganu...... 55 3.3.25 Kemasik, Terengganu ...... 55 3.3.26 Kerteh, Terengganu ...... 56 3.3.27 Tanjung Jara, Terengganu ...... 57 3.3.28 Chendering, Terengganu ...... 57 3.3.29 Bari Kecil, Terengganu ...... 58 3.3.30 Tioman Island ...... 58 3.3.30.1 Dalam Bay ...... 59 3.3.30.2 Tanjung Sekeliling ...... 59 3.3.30.3 Tekek village ...... 60 3.4 Sampling Design ...... 60 3.4.1 Timed Search Method ...... 61 3.4.2 Sample collection ...... 61 3.5 Studied Species ...... 63 3.6 Sample Preparation before Digestion ...... 66 3.6.1 Moisture Content in Biota ...... 67 3.6.2 Conversion Factor (CF) ...... 67 3.7 Digestion Procedure...... 73 3.7.1 Biological sample ...... 74 3.7.2 Sediment sample ...... 75 3.8 Sample Measurement...... 76 3.8.1 Inductively Coupled Plasma Mass Spectrometry (ICP-MS) ...... 76 3.8.2 Detection Limit ...... 78 3.8.3 Handling of Standard Solution ...... 78 3.9 Quality Control and Quality Assurance ...... 79 3.9.1 Laboratory Interferences ...... 79 3.9.2 Procedural Blank ...... 80 3.9.3 Recovery Test ...... 80

3.9.4 Calculation of REEs in Sample ...... 83 3.10 Data Handling ...... 83 3.10.1 Calculation of REE ratios ...... 84 3.10.2 Normalization using Reference Value ...... 85 3.10.3 Quantification of REE anomalies ...... 86 3.10.4 Enrichment factor ...... 88 3.10.5 Statistical Analysis ...... 89 3.10.5.1 ANOVA with post hoc test ...... 89 3.10.5.2 Correlation analysis ...... 90 3.10.5.3 Hierarchical Cluster Analysis ...... 90

CHAPTER FOUR: RESULTS AND DISCUSSION ...... 91 4.1 Introduction...... 91 4.2 Sediment analysis ...... 92 4.2.1 REEs concentration in surface sediment ...... 92 4.2.2 Inter-elemental variation in sediment ...... 97 4.2.3 Enrichment factor in sediment ...... 99 4.2.4 REEs behaviour in sediment ...... 101 4.2.5 REEs normalization in sediment ...... 103 4.2.6 REEs anomalies quantification sediment ...... 108 4.3 Biota analysis ...... 111 4.3.1 REEs concentration in S. cucullata ...... 111 4.3.2 REEs concentration and distribution in different tissues of T. clavigera ...... 116 4.3.3 REEs concentration and distribution in different tissues of N. chameleon ...... 126 4.3.4 REEs Behaviour ...... 136 4.3.5 REEs Anomalies ...... 140 4.4 Inter-Tissue Variation ...... 153 4.5 Interspecies Variation ...... 159 4.6 Interspatial Variation ...... 165 4.7 Assessment with Maximum Permissible Concentrations ...... 169

CHAPTER FIVE: CONCLUSION AND RECOMMENDATION ...... 174 5.1 Conclusion ...... 174 5.2 Recommendation ...... 176

REFERENCES ...... 177

APPENDIX A: CORRELATION MATRIX BETWEEN REES AND TRACE METAL CONCENTRATIONS IN SEDIMENT ...... 196 APPENDIX B: COEFFICIENT OF CORRELATION MATRIX BETWEEN REES CONCENTRATIONS IN SOFT TISSUE, OPERCULUM AND SHELL OF T. CLAVIGERA FROM PARTICULAR SAMPLING SITES ALONG THE PENINSULAR MALAYSIA COASTAL WATERS ...... 197 APPENDIX C: COEFFICIENT OF CORRELATION MATRIX BETWEEN REES CONCENTRATIONS IN SOFT TISSUE, OPERCULUM AND SHELL OF N. CHAMELEON FROM PARTICULAR SAMPLING

xii

SITES ALONG THE PENINSULAR MALAYSIA COASTAL WATERS ...... 199 APPENDIX D: COEFFICIENT OF CORRELATION MATRIX BETWEEN REES CONCENTRATIONS IN SOFT TISSUE VS REES CONCENTRATION IN OPERCULUM AND REES CONCENTRATIONS IN SOFT TISSUE VS REES CONCENTRATION IN SHELL OF T. CLAVIGERA FROM PARTICULAR SAMPLING SITES ALONG THE PENINSULAR MALAYSIA COASTAL WATERS ...... 201 APPENDIX E: COEFFICIENT OF CORRELATION MATRIX BETWEEN REES CONCENTRATIONS IN SOFT TISSUE VS REES CONCENTRATION IN OPERCULUM AND REES CONCENTRATIONS IN SOFT TISSUE VS REES CONCENTRATION IN SHELL OF N. CHAMELEON FROM PARTICULAR SAMPLING SITES ALONG THE PENINSULAR MALAYSIA COASTAL WATERS ...... 202 APPENDIX F: SMARTTUNE PROGRAM RESULTS PRIOR TO EACH MEASUREMENT WORK TO ENSURE ICP-MS IN PERFECT CONDITION ...... 203 APPENDIX G: CALIBRATION CURVES OBTAINED FROM SERIAL MULTI- ELEMENT CALIBRATION STANDARD FOR REES ...... 204 APPENDIX H: TUKEY’S POST HOC TEST COMPARISONS OF ELEMENTS CONCENTRATIONS BETWEEN SAMPLING SITES IN SURFACE SEDIMENT ...... 209 APPENDIX I: TUKEY’S POST HOC TEST COMPARISONS OF ELEMENTS CONCENTRATIONS BETWEEN SAMPLING SITES IN S. CUCULLATA ...... 210 APPENDIX J: TUKEY’S POST HOC TEST COMPARISONS OF ELEMENTS CONCENTRATIONS BETWEEN SAMPLING SITES IN T. CLAVIGERA ...... 212 APPENDIX K: TUKEY’S POST HOC TEST COMPARISONS OF ELEMENTS CONCENTRATIONS BETWEEN SAMPLING SITES IN N. CHAMELEON ...... 214 APPENDIX L: TOTAL CONCENTRATION (µG G-1 DRY WEIGHT) OF ΣREE IN SEDIMENT FROM PARTICULAR SAMPLING SITES ALONG PENINSULAR MALAYSIA COASTAL WATERS .. 216 APPENDIX M: TOTAL CONCENTRATION (µG G-1 DRY WEIGHT) OF ΣREE IN SOFT TISSUE OF S. CUCULLATA FROM PARTICULAR SAMPLING SITES ALONG PENINSULAR MALAYSIA COASTAL WATERS ...... 217 APPENDIX N: TOTAL CONCENTRATION (µG G-1 DRY WEIGHT) OF ΣREE IN SOFT TISSUE, OPERCULUM AND SHELL OF T. CLAVIGERA FROM PARTICULAR SAMPLING SITES ALONG PENINSULAR MALAYSIA COASTAL WATERS ...... 218 APPENDIX O: TOTAL CONCENTRATION (µG G-1 DRY WEIGHT) OF ΣREE IN SOFT TISSUE, OPERCULUM AND SHELL OF N. CHAMELEON FROM PARTICULAR SAMPLING SITES ALONG PENINSULAR MALAYSIA COASTAL WATERS .. 220

xiii

LIST OF TABLES

Table No. Page No.

2.1 REEs chemical properties and abundances in earth's upper crust 14

2.2 Main usages of REEs (from USEPA, 2012, Gonzales et al., 2014) 17

2.3 Maximum permissible concentration and maximum permissible addition in surface water and sediment (Sneller et al., 2000) 21

2.4 REEs and possible interferences in ICP-MS measurement (Dulski, 1994; Dresler et al., 2007) 33

3.1 List of sampling sites and their locations determined by coordinate along the Peninsular Malaysia coastal waters 40

3.2 Availability of study organism for each sampling site 65

3.3 Biometric data (mean and standard deviation) of S. cucullata, T. clavigera and N. chameleon 68

3.4 Means of wet weight, dry weight, water content (%) and conversion factor (CF) of S. cucullata, T. clavigera and N. chameleon 71

3.5 Standard running conditions of ICP-MS 77

3.6 REEs isotope measured using ICP-MS 78

3.7 Analytical results of measured BCR 668 standard reference material, detection limit and percentage of recovery for each element (µg g-1 dry weight) 82

3.8 Analytical results of measured BCR 667 standard reference material, the detection limit and percentage of recovery for each element (µg g-1 dry weight) 83

3.9 Concentration of REEs in NASC, PAAS shales (Taylor and McLennan, 1985) and chondrite (Mcdonough and Sun, 1989) 86

3.10 Contamination categories based on the enrichment factor values 89

4.1 Total concentration (µg g-1 dry weight) of ΣREE in sediment from particular sampling sites along Peninsular Malaysia coastal waters 93

4.2 Correlation between elements in surface sediment from Peninsular Malaysia coastal waters 98

xiv

4.3 Correlation matrix between REEs and trace metal concentrations in sediment 99

4.4 Enrichment factor (EF) values calculated for sediment along Peninsular Malaysia coastal waters 100

4.5 Concentration of light and heavy REEs and ratios of REEs in the sediment 102

4.6 Indicator ratio of Ce/Ce*, Eu/Eu*, and (La/Yb)N in surface sediment of particular sites from Peninsular Malaysia coastal waters using values of PAAS, NASC and chondrite 109

4.7 Total concentration (µg g-1 dry weight) of ΣREE in soft tissue of S. cucullata from particular sampling sites along the Peninsular Malaysia coastal waters 112

4.8 Decreasing pattern of REEs occurrences (µg g-1 dry weight) in soft tissue of S. cucullata from particular sampling sites along Peninsular Malaysia coastal waters 115

4.9 Pearson’s correlation between REEs concentration in soft tissue of S. cucullata from particular sampling sites along the Peninsular Malaysia coastal waters 116

4.10 Total concentration (µg g-1 dry weight) of ΣREE in soft tissue of T. clavigera from particular sampling sites along the Peninsular Malaysia coastal waters 118

4.11 Total concentration (µg g-1 dry weight) of ΣREE in operculum of T. clavigera from particular sampling sites along the Peninsular Malaysia coastal waters 121

4.12 Total concentration (µg g-1 dry weight) of ΣREE in shell of T. clavigera from particular sampling sites along the Peninsular Malaysia coastal waters 124

4.13 Total concentration (µg g-1 dry weight) of ΣREE in soft tissue of N. chameleon from particular sampling sites along the Peninsular Malaysia coastal waters 127

4.14 Total concentration (µg g-1 dry weight) of ΣREE in operculum of N. chameleon from particular sampling sites along the Peninsular Malaysia coastal waters 131

4.15 Total concentration (µg g-1 dry weight) of ΣREE in shell of N. chameleon from particular sampling sites along the Peninsular Malaysia coastal waters 134

4.16 Concentration of ΣREE, ΣLREE, ΣHREE, LREE/HREE, Ce/La, Eu/Sm, Yb/Sm, and La/Yb in soft tissue of S. cucullata 136

4.17 Mean concentration of total REEs (∑REE), ∑LREE, ∑HREE, ratios of LREE/HREE, La/Yb, La/Sm, Ce/La, Eu/Sm and Yb/Sm in soft tissue, operculum and shell of T. clavigera 137

4.18 Concentration of total REEs (∑REE), ∑LREE, ∑HREE, ratios of LREE/HREE, La/Yb, La/Sm, Ce/La, Eu/Sm and Yb/Sm in soft tissue, operculum and shell of N. chameleon 138

4.19 Indicator ratios of Ce/Ce*, Eu/Eu* and (La/Yb)N in soft tissue of S. cucullata 142

4.20 Concentration of Ce/Ce*, Eu/Eu* and (La/Yb)N in soft tissue, operculum and shell of T. clavigera 146

4.21 Concentration of Ce/Ce*, Eu/Eu* and (La/Yb)N in soft tissue, operculum and shell of N. chameleon 151

4.22 Coefficient of correlation matrix between REEs concentrations in soft tissue, operculum and shell of T. clavigera from particular sampling sites along the Peninsular Malaysia coastal waters 153

4.23 Coefficient of correlation matrix between REEs concentrations in soft tissue, operculum and shell of N. chameleon from particular sampling sites along the Peninsular Malaysia coastal waters 154

4.24 Coefficient of correlation matrix between REEs concentration in soft tissue vs REEs concentration in operculum and REEs concentration in soft tissue vs REEs concentration in shell of T. clavigera from particular sampling sites along the Peninsular Malaysia coastal waters 155

4.25 Coefficient of correlation matrix REEs concentration in soft tissue vs REEs concentration in operculum and REEs concentration in soft tissue vs REEs concentration in shell of N. chameleon from particular sampling sites along the Peninsular Malaysia coastal waters 156

4.26 Mean ratios of operculum REEs to soft tissue REEs (OTR) and shell REEs to soft tissue REEs (STR) in T. clavigera and N. chameleon 158

4.27 Bioaccumulation Factors (BAF) calculated in studied organisms 160

4.28 Available comparative reports on the BAF of REEs in aquatic organisms 162

4.29 Maximum permissible limits on selected heavy metals (in µg g-1) for food safety set by different countries 169

4.30 Mean values of REEs concentration (in µg g-1) in wet and dry states of soft tissue of S. cucullata, T. clavigera and N. chameleon, to compare with permissible limits data 170

xvi

4.31 Comparison of sediment data in this study with the maximum permissible concentration (mg kg-1 dry weight) in marine sediment (Sneller, 2000) 172

xvii

LIST OF FIGURES

Figure No. Page No.

2.1 The Periodic Table with REEs, in red colour, in group 3 11

2.2 The distribution pattern of REEs in soil (Markert, 1987) 15

2.3 Schematic representation of an ICP-MS instrument (LPM = L/min). (Beauchemin, 2010) 31

2.4 Schematic representation of the sample introduction system that is standard on most current ICP-MS instruments. (Beauchemin, 2010) 32

3.1 Map showing sampling locations along the Peninsular Malaysia coastal waters. Sites are specified in Table 3.1 39

3.2 Rocky shore at Pasir Tengkorak Beach, Langkawi 41

3.3 Study site at Tengah Beach, Langkawi 42

3.4 Study site at Beringin Beach, Langkawi 42

3.5 Study site at Pulau Sayak, Kedah 43

3.6 Study site at Pulau Betong, Penang 43

3.7 Study site at Batu Feringghi, Penang 44

3.8 Study site at Teluk Senangin, Perak 44

3.9 Study site at Teluk Batik, Perak 45

3.10 Study site at Taman Tanjung, Negeri Sembilan 46

3.11 Study site at Teluk Kemang, Negeri Sembilan 46

3.12 Study site at Blue Lagoon, Negeri Sembilan 47

3.13 Study site at Tanjung Bidara, Malacca 47

3.14 Study site at Tanjung Kling, Malacca 48

3.15 Study site at Batu Pahat, Johor 48

3.16 Study site at Sungai Rengit, Johor 49

3.17 Study site at Batu Layar, Johor 50

xviii

3.18 Study site at Tanjung Balau, Johor 50

3.19 Study site at Sedili Kechil, Johor 51

3.20 Study site at Mersing, Johor 51

3.21 Study site at Kg. Tg. Batu, Pahang 52

3.22 Study site at Tlk. Cempedak, Pahang 52

3.23 Study site at Tg. Gelang, Pahang 53

3.24 Study site at Cherating, Pahang 53

3.25 Study site at Telaga Simpul, Terengganu 54

3.26 Study site at Tlk. Kalong, Terengganu 54

3.27 Study site at Kijal, Terengganu 55

3.28 Study site at Kemasik, Terengganu 55

3.29 Study site at Kerteh, Terengganu 56

3.30 Study site at Tg. Jara, Terengganu 57

3.31 Study site at Chendering, Terengganu 57

3.32 Study site at Bari Kechil, Terengganu 58

3.33 Study site at Dalam Bay, Tioman Island 59

3.34 Study site at Tg. Sekeliling, Tioman Island 59

3.35 Study site at Tekek, Tioman Island 60

3.36 Pictures of (1) S. cucullata (Born, 1778); (2) Soft tissue of S. cucullata on its shell; (3) S. cucullata in the natural environment of rocky shore; (4) T. clavigera (Kuster, 1860); (5) Operculum of T. clavigera at the mouth of its shell; (6) Extracted shell of T. clavigera; (7) T. clavigera in the natural environment of rocky shore; (8) N. chameleon (Linnaeus, 1758); (9) Operculum of N. chameleon at the mouth of its shell; (10) Extracted shell of N. chameleon; (11) N. chameleon in the natural environment of rocky shore 64

3.37 Pictures of (12) Teflon steel jacket and Teflon vessel; (13) Teflon vessel fits inside the Teflon steel jacket 74

3.38 Inductively Coupled Plasma-Mass Spectrometry that used for this research 77

xix

3.39 Pictures of Standard reference material (SRM), (14) BCR 668 Mussel Tissue; (15) BCR 667 Estuarine Sediment 81

4.1 Pattern of REEs in sediment normalized to the concentration of chondrite value 105

4.2 Pattern of REEs in sediment normalized to the concentration of PAAS value 106

4.3 Pattern of REEs normalized to the concentration of NASC value 108

4.4 Chondrite-normalized plots for the REEs in the soft tissue of S. cucullata from particular sites of Peninsular Malaysia coastal waters, divided by region 141

4.5 Chondrite-normalized plots for the REEs in the soft tissue of T. clavigera from particular sites along Peninsular Malaysia coastal waters, divided by region 143

4.6 Chondrite-normalized plots for the REEs in the operculum of T. clavigera from particular sites along Peninsular Malaysia coastal waters, divided by region 144

4.7 Chondrite-normalized plots for the REEs in the shell of T. clavigera from particular sites along Peninsular Malaysia coastal waters, divided by region 145

4.8 Chondrite-normalized plots for the REEs in the soft tissue of N. chameleon from particular sites along Peninsular Malaysia coastal waters, divided by region 147

4.9 Chondrite-normalized plots for the REEs in the operculum of N. chameleon from particular sites along Peninsular Malaysia coastal waters, divided by region 148

4.10 Chondrite-normalized plots for the REEs in the shell of N. chameleon from particular sites along Peninsular Malaysia coastal waters, divided by region 149

4.11 Bioaccumulation Factor (BAF) for REEs in studies species based on surface sediment in dry soft tissue vs. REEs 161

4.12 REEs bioavailabilities comparison across species 163

4.13 Dendrogram of the HCA on soft tissue of S. cucullata collected from particular sites based on their ∑LREE and ∑HREE concentrations 166

4.14 Dendrogram of the HCA on soft tissue of T. clavigera collected from particular sites based on their ∑LREE and ∑HREE concentrations 167

4.15 Dendrogram of the HCA on soft tissue of N. chameleon collected from particular sites based on their ∑LREE and ∑HREE concentrations 167

4.16 Dendrogram of the HCA on sediment collected from particular sites based on their ∑LREE and ∑HREE concentrations 168

xxi

CHAPTER ONE

INTRODUCTION

1.1 RESEARCH BACKGROUND

Rare Earth Elements (REEs) consist of a group of fifteen homologous metals, lanthanides, composed with scandium (Sc) and yttrium (Y). Antagonistic to its name suggests that they are not rare at all and could be found abundantly in the earth crust comparable to Pb or Cd. Naturally occurring REEs show a typical pattern in environmental samples due to its similar physical and chemical properties (Weltje et al., 2002). This pattern will display that the odd-numbered elements will have lower concentrations than the nearby even-numbered ones, acknowledged as Oddo-Harkins rule (Oddo, 1914; Harkins, 1917). The logarithm of the abundance against atomic number presented a saw-tooth pattern, which is strongly conserved as the REEs disperse from indigenous rocks to soils and sediments, water, and eventually biota.

Research on REEs has an increasing trend in Malaysia as REEs have found applications in high and green technology which is important in Malaysia’s aspiration to become a high-income nation. Green technology has become significant criteria nowadays as this technology could mitigate global warming due to climate change.

Malaysia now has been recognized as a strategic place to develop green technology industries. Studies have highlights that mining and processing of REEs do pose risks to health, safety and environment (Chen and Zhu, 2008; Aquino et al., 2009) but recent technologies could manage this risk. Still, there is a need to investigate REEs concentration in their natural environment for monitoring purpose.

REEs are used in enormous quantities in the manufacturing of industrial products comprises lasers, computers, catalysts for the oil industry, pigments for glass

and plastics, and additives (Riondato et al., 2001; Cui et al., 2012). Extraction and processing of REEs ores may produce multiple waste streams that require complicated environmental management systems, which improper waste management may modify the quality of the local environment (USEPA, 2011; 2012). For instance, environmental alterations to surface soils, surface waters, and soil–plant systems have been documented in REE mining areas of China as reviewed by Liang et al. (2014).

Processing of REE compounds for petroleum refining or utilization of REE-containing products such as gadolinium contrast agents or nanomaterials might also end up in the discharge of REEs to the environment (Karn, 2011; Kulaksız and Bau, 2011a, 2011b).

USEPA (2012) has recognized that there is restricted evidence with regard to health and environmental issues connected to REEs.

In a global scale, attention has been drawn due to the wide occurrence of REEs pollution in an aquatic system. It has become a common practice to use aquatic organisms as bioindicators in many monitoring programs throughout the world.

Goldberg et al. (1978) were the first to initiate the ``Mussel Watch Program'' in which the bivalves were used as a surveillance tool of coastal pollution. They are often circulated in response to concern about contamination on seafood or to employ marine organisms as biological monitors (Oehlmann and Schulte-Oehlmann, 2003; Zhou et al., 2008). Marine ecosystems may receive anthropogenic pollutants originating from various sources at the surrounding area or from distant places. Yet, there are occurrences of purely natural concentration in some cases. REEs generally enter the environment in much more soluble form and more reactive ionic forms due to anthropogenic rather than naturally occurring, thus producing them to be more bioavailable (Kumar et al., 2014).

The increasing industrial expenditures of REEs are accompanied by growing emissions, which primarily end up in the aquatic environment. Such occasion leads to a need for an investigation to gain insight whether these emissions could cause effects on aquatic biota. A study by Riondato et al. (2001) indicated that REEs tend to be accumulated by biota and have toxic effects similar to those of Cd, Zn, and Ni. The study of REEs distribution may also provide basic information on the geochemistry of these elements in the coastal marine environment related to their sources. All aquatic invertebrates are found to accumulate REEs in their tissues, regardless whether these elements are essential to metabolism or not. Increasing emissions lead to increasing percentage of REEs to be accumulated in biota and producing toxic effects. No established biological functions of REEs were reported, thus, they may be considered as non-essential elements in biota (Sneller et al., 2000; Weltje et al., 2002).

Various bivalve and gastropod species show a high presence and play an important role as biomonitors for pollution in global monitoring programs throughout the world (Wang et al., 2005). Molluscs exhibit greater spatial sensitivity, thus, are the most reliable tool for identifying sources of bioavailable contamination compare to sediments (Goldberg et al., 1978; Hamed and Emara, 2006). Much attention related to aquatic pollution biomonitoring using molluscs has been carried out in Malaysia by numerous researchers (Shazili et al., 1995; Amin et al., 2006; Yap et al., 2009a,

2009b; Yap et al., 2010), primarily investigating on trace or heavy metal, yet little information exist on REEs distribution and behaviour in biota. However, in terms of sediment study, several studies has been done (Wood et al., 2004; Sultan and Shazili,

2009; Rezaee et al., 2009; Antonina et al., 2013), presenting that REEs studies are gaining attention to understand its behaviour in Malaysian environment.

1.2 PROBLEM STATEMENT

The increasing demand for REEs led to anthropogenic emission, but knowledge of the environmental distribution of these elements in Malaysian environment is limited.

Much attention were focused on sediment while only a narrow consideration has been paid to the REEs accumulation in tissues of living organisms, due to their direct exposure, and related to biomagnification effects along the trophic chain. REEs accumulation in the environment is usually considered as a good tracer of contamination processes. Although some toxicology test has been conducted to gain insight on bioconcentration of REEs in biological organism, the extent of REEs on the environment and the potential health risks for species living in aquatic environment are not well acknowledged, perhaps as a magnitude of low REEs concentrations in living tissues and analytical difficulties associated to their determinations at ng L-1 levels. Thus, ICP-MS is considered to be most reliable technique for REEs analysis.

Determination of REEs using local bioindicator also related to human consumption.

Possible worries regarding effects of exposure to low levels of REEs on human health have been ascending recently. It has been testified that REEs are significantly accumulated in blood, brain and bone after entering human body.

1.3 RESEARCH QUESTIONS

i. How ample possibility that REEs bioavailability in Saccostrea cucullata, Thais

clavigera and Nerita chameleon could attribute to prove to be suitable

bioindicator?

ii. How far the extent of REEs accumulation in our environment with regards to

natural and anthropogenic input?

iii. How considerably REEs variation could be inspected in our various rocky

shore environment features?

iv. How complex the behaviour of REEs across organism tissues and species?

1.4 RESEARCH SCOPE

REEs measurement on selected bioindicators with specific sizes was conducted to evaluate the bioavailability of REEs in their bodies. Sediment analysis was also taken into account as to understand the behaviour of such elements in sediment and their relationship with studied organisms as well. The investigation was done in various sites characteristics such as remote and secluded area, recreational area and heavy industry area. The possible nearby river was also identified to relate the natural or anthropogenic input of REEs to the particular sites.

1.5 GENERAL OBJECTIVES

To investigate the distribution of REEs in selected rocky shore organisms and surface sediment along the coastal waters of Peninsular Malaysia.

1.6 SPECIFIC OBJECTIVES

i. To determine the behaviour of REEs in term of anomalous deviation.

ii. To clarify the partitioning of REEs in different parts of studied organisms body

and relationship among them.

iii. To justify the sediment role as REEs sink to the studied organisms.

iv. To explain spatial variation related to REEs accumulation.