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UNIVERSITI PUTRA MALAYSIA
ACCUMULATION AND DISTRIBUTION OF HEAVY METALS IN GREEN-LIPPED MUSSEL PERNA VIRIDIS (LINNAEUS) FROM
THE WEST COAST OF PENINSULAR MALAYSIA
YAP CHEE KONG
FSAS 1999 42
ACCUMUlATION AND DISTRIBUTION OF HEAVY METALS IN
GREEN-LIPPED MUSSEL PERNA VIRIDIS (LINNAEUS) FROM THE WEST COAST OF PENINSULAR MAlAYSIA
YAP CHEE KONG
MASTER OF SCIENCE UNIVERSITI PUTRA MALAYSIA
1999
ACCUMUlATION AND DISTRIBUTION OF HEAVY METALS IN GREEN-LIPPED MUSSEL PERNA VIRIDIS (LINNAEUS) FROM
THE WEST COAST OF PENINSULAR MALAYSIA
By
YAP CHEE KONG
Thesis Submitted in Fulfilment of the Requirements for the Degree of Master of Science hi the Faculty of
Science and Environmental Studies Universiti Putra Malaysia
August 1999
ACKNOWLEDGEMENTS
I am most indebted and grateful to my supervisor, Associate Professor Dr.
Ahmad Ismail, who has guided me along the way and given me a lot of constructive
comments, advice and ideas no matter how busy he is. He is certamly the person
responsible for the successful completion for my project. I certamly cannot make it this
far without his consistent 'inputs'. Sincerely, thank you.
I would also like to extend my sincere thanks to my co-supervisors Dr. Abdul
Rahim Ismail for commenting and advising on my statistical analysis, and Mr.
Mohamad Pauli Zakaria for his comments on my project and help during the field-trip.
Thank you. Special thanks are also given to Miss Rosniza RamH, Mr. Mansor Rashid,
Mr. Navies Maisin, Mr. Hidir Hashim, Mr. Zahar Jaafar, Mr. Ali Mashinchian and all
the staff in the Biology department. Also thanks to my friends whom I know Kale Leba,
Hena, Latif, Nik, Katty, Sem and my housemates.
Not forgetting my family and my girl mend, Miss Choh Mew Seong, who have
become my constant source of inspiration and motivation. I could not have made it
without you all. Thanks and I love you all.
ii
TABLE OF CONTENTS
Page
ACKNOWLEoo.EMENTS...................................................................... .... ii LIST OF TABLES ......................... ... ............................. .................. ............ vi LIST OF FIGlJR.ES...................................................................................... Xl LIST OF PLATES........................................................................................ xiii LIST OF ABBREVIATIONS....................................................................... xiv ABSmACT.................................................................................................. xv ABSTRAK.......... ........................................................................................... xvii
CHAPTER
I
n
INTRODUCTION ............................................................................ . 1
LITERATlJR.E REVIEW ................................................................. . 6 Marine Ponution ..................................... ........ ............................ ....... .. 6 Studies on Heavy Metals in Malaysia..... ................................................ 8 Biology of Perna viridis (Linnaeus)....................................................... 10
Taxonomy................................................................ ............ .... 10 Morphology................... ... ................... . . . ........... ....................... 12 IIabitats.................................................................................... 13 Food... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Reproduction............. .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 Predators and Diseases.............................................................. 16 Importance............ .................................................................... 17
Mussel Culture in Malaysia...................................... ............................. 18 Rasionale of Using Mussels as Bioindicator............................................ 20 Past Studies of Mussels as Bioindicators for Heavy Metals..................... 22 Past Studies of Mussels as Bioindicators for Other Pollutants.................. 25 Other Studies on Mussel P. viridis .. . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . 27 The Role of Mussel Shells. ................................................................... 29 Mussel Shells as Bioindicator for Heavy Metals..................................... 29 Heavy Metals Studies in Barnacles.......... . ......... .. . .... ............................. 30 Heavy Metals Accumulation in Bivalves................... ............................. 31 Physiological Responses In Musssels..................... ................................ 33 Mussel Watch Programme...... ........................ .............. ........................ 35 Inherent Variability in Metal Concentrations in Bivalves .......................... 36
Page
ill MATERIALS AND METHODS.......................................................... 37 Study Area........................................................................................... 37 Scunpling and Storage.............. .... ... ................. .............. ... ............. ....... 37 Mussel Sex.......................................................................... ........ ... ...... 41 Scunple Preparations............................................................................. 41
Mussel Soft Tissues................................................................. 41 Dissection of Different Tissues in Soft Body Mussel P. viridis... 43 Mussel Shell. ....... ...... ......... ... ........ .................. ... .... ............ ..... 43 Different Layers and Sections in Mussel Shell........................... 46 Byssus..................................................................................... 47 Suspended Particulate Materials and Other Sticky Materials....... 47 Sediment ....................... '" ... .... ..... .......... ...... .... ... ........... .... ..... 48
Sample Digestion ............................. , ............................ , ....... ..... .......... 48 Soft Tissue, Shell and Byssus................................................... 48 SedUnent ......................... .............. ..... ............... .... ................. 49
Water Content and Conversion Factor.. . . . . . . . . . . . . . . . . . . . . . . . . ... . . . . . . . . . . . . . . . . . . . . . . 49 Oeaning Procedures ............ '" ................................................ ,. ... ......... 50 Blank................................................................................................... 50 Recovery Test...................................................................................... 50 Calculations of Indices and Ratios......................................................... 53
Allomenic Ratios....................................................................... 53 Shell lbickness..... ... ..... .... ... ...... ... .... .......... ...... ...... ...... ....... ..... 53 Condition Index........................................................................ 54 Boyden's Equation.................................................................... 55 Coefficients of V ariation............................................................ 56 Metal Bioavai1abi1i.ty Index......................................................... 56
Statistical Analytical Procedures............................................................ 57 Outliers..................................................................................... 57 Pearson's Correlation Coefficient and Stepwise Regression Analysis ................................................................................... 61
IV RESULTS AND DISCUSSION....... ....... ....... ............ ....... ..... ............ 62 Heavy Metals in Total Soft Tissue and Shell ofMusselP. viridis............ 62
Water Content and Conversion Factors... . . ... . . . . . . . . . . . . . . . . . . . . . . .. . . .. . . 80 Comparison with Other Regional Studies.................................... 83 Comparison with Other Biological Scunples................................ 85
iv
V
Distribution of Heavy Metals in Mussel P. viridis ... .... ........................... . Different Tissues ...................................................................... . Different Layers and Sections in Mussel SheD ............................ .
Factors Affecting Accmnulation of Heavy Metals in Mussel P. viridis . . . . . Sediment. ................................................................................ . Suspended Particulate Materials ................................................ . Physico-Chemical Parameters ................................................... . Size - SheD Length and SheD Thickness ................................... . Size - Total Soft Tissue Weigllt. .. .............................................. . Condition Index ....................................................................... . Population Structure ................................................................. . Sex .......................................................................................... . Most Influential Known Factors by Stepwise Regression Analysis .................................................................. .
Mussel SheDs as a Potential Biomonitoring Material ............................. . . Shell - Flesh Ratios ................................................................... . Inter-Metals Relationships ........................................................ . Metal Concentration Variability in Total Soft Tissue and Shell. .. . .
Byssus as a Potential Biomarker for Heavy Metals ............................... . . Heavy Metals in Byssus and Total Soft Tissue .......................... .. Inter-Metals Relationships ......................................................... .
Shell Thickness as a Function of Age ................................................... . . Population Variability .......................................................................... .
Metal Variability ...................................................................... . All .
V--=-L!t!, ometric iUliIUlIlty ............................................................... . Inter- and Intra-station Variability ............................................. . .
CONCLUSION .. ...... . . . . . . . . . . . ..... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
87 87 100 103 103 107 109 1 12 1 17 124 133 137
139 146 146 153 1 59 166 166 171 175 179 179 185 187
194
REFERENCES.. . . . . . . . . . . . . . . . ... . . . . . . . . . . . ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 199
APPENDIX A Programme Statements in SAS.... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232
VITA... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... . . . . . . . . . . . . . . . . . . . . 235
PUBLICATION.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 235
v
LIST OF TABLES
Table
1. Heavy Metals Concentrations in Green-Lipped Mussel P. viridis from
Page
Other Regional Studies..................................................................... 24
2. Other Contaminants and Their Respective Concentrations Using Green-Lipped Mussel P. viridis as Bioindicators from Various Studies............................................................................................ 26
3. Percentage of Recoveries in Cd, Cu, Ph and Zn................................. 53
4. Comparison of Transfonned and Non-Transfonned Data According to the Empirical Rule........................................................................ 60
5. Mean ± Standard Error of Shen Length, Shell Width, Shell Weight, Fresh Dry Weight and Shell Dry Weight Per Individual of Mussel P. viridis from 15 Sampling Stations..................................................... 63
6. Mean ± Standard Error of Shell Thickness, Shell Width : Height Ratios, Shell Width: Length Ratios, Shell Length : Height Ratios and Condition Index Per Individual of Mussel P. viridis from 15 SaIllpling Stations............................................................................ 64
7. Results of ANOV A for Shell Length, Condition Index and Cd, Cu, Ph and Zn Concentrations (Untransfonned) in Total Soft Tissue and Shell in Male and Female of Mussel P. viridis from 15 Populations from Sabah and Peninsular Malaysia.................................. ............... 65
8. Student-Newrnan-Keuls Comparisons of Means for Cd Concentrations in Shell and Total Soft Tissue of Mussel P. viridis.............................................................................................. 68
9. Student-Newman-Keuls Comparisons of Means for Cu Concentrations in Shell and Total Soft Tissue of Mussel P. viridis...... 69
10. Student-Newman-Keuls Comparisons of Means for Ph Concentrations in Shell and Total Soft Tissue ofMusselP. viridis...... 70
11. Student-Newman-Keuls Comparisons of Means for Zn Concentrations in Shell and Total Soft Tissue of Mussel P. viridis...... 71
vi.
Table
12. Percentage of Water Content in Total Soft Tissue and Shell of
Page
Mussel P. viridis from Selected Stations............................................ 81
13. Comparison of Water Contents and Conversion Factors of Different Species of Bivalves.......................................................................... 82
14. Conversion Factor for Each Metal in Total Soft Tissue of Mussel P. viridis.............................................................................................. 83
15. Zn, Cd, Cu and Ph Concentrations in Total Soft Tissue of Mussel P.
viridis Recorded from AlliS Sampling Stations................................ 84
16. Comparisons of Heavy Metals' Levels in Total Soft Tissue of Mussel P. viridis with Other Studies on Freshwater Snails, Shellfish and Fish Reported from Peninsular Malaysia. ...... , .. , ................ .... ..... ... ............ 86
17. Student-Newman-Keuls Comparisons of Cd, Cll, Ph and Zn
Concentrations in Different Tissues of Mussel P.
viridis.............................................................................................. 89
18. Percentage of lipid Content from a Population of Mussel P. viridis at Pasir Panjang................................................................................... 96
19. Student-Newman-Keuls Comparisons of Cd, Cu, Ph and Zn Concentrations in Different Sections of the Shell of Mussel P. viridis.............................................................................................. 102
20. Pearson's Correlation Coefficients Analysis between Cd, Cu, Ph and Zn in the Total Soft Tissue and Byssus of Mussel P. viridis and Sediment......................................................................................... 104
21. Comparisons of Heavy Metals Concentrations in Total Soft Tissue, Shell and Byssus of Mussels P. viridis, Shell and Tissues of Barnacles B. amphitrite, Coastal Sediment and Sediment Found Inside the Mussel's Shells................................................................. 106
vii
Table
22. Estimation of Suspended Particulate Materials that are Attached on
Page
the Byssal Threads of Mussel P. viridis............................................. 108
23. Physico-Chemical Parameters in Waters of Three Cultured Sites........ 11 0
24. Comparisons of Cd, Cu, Ph and Zn Concentrations in Sea Water from Other Studies........................................................................... III
25. Pearson's Correlation Coefficients between Cd, Cn, Pb and Zn Concentrations in Total Soft Tissue with Shell Length, Shell Thickness and Metal-Shell Weight Indices of Mussel P. viridis........... 116
26. Comparisons of Correlation Coefficients for Metal Content Versus Total Soft Tissue Weight from Other Studies.................................... 119
27. Comparison of Condition Indices from Other Studies................... ..... 125
28. Pearson's Correlation Coefficients Analysis between Condition Index and Cd, Cu, Ph and Zn in Total Soft Tissue and between Condition Index and Metal-Shell Weight Indices of the Mussel P. viridis............................................................................................. 130
29. Coefficient of Variation of Various Variables on Mussel P. viridis from Pasir Panjang and Pantai Lido............ ...................................... 135
30. Results of t-Test to Elucidate the Differences in Both Females and Males of Mussel P. viridis................................................................ 138
31. Inter-Statton's Multiple Linear Stepwise Regression Analysis of Dependent Variahles in Flesh and Shell of Mussel P. viridis on All Ratios and Non-Ratios as Independent Variahles................. .............. 141
32. Inter-Station's Multiple Unear Stepwise Regression Analysis of Dependent Variables in Flesh and Shell of Mussel P. viridis on All Ratios as Independent Variables.. .............................. ........ ................ 142
viii
Table
33. Intra-Station's Multiple Linear Stepwise Regression Analysis of Dependent Variables in Flesh and Shell of Mussel P. viridis on All
Page
Ratios and Non-Ratios as Independent Variables.. ............................. 144
34. Concentrations of Cd, en, Pb and Zn within the Flesh and Shell of Male Mussel P. viridis and Ratios of Shell to Flesh from 15 Sampling Stations.................................................................................. ......... 147
35. Concentrations of Cd, cu, Ph and Zn within the Flesh and Shell of Female Mussel P. viridis and Ratios of Shell to Flesh from 15 Sampling Stations............................................................................ 148
36. Comparisons of Shell-Metal to Flesh-Metal Ratios for Cd, Cu, Pb and Zn concentrations from Other Bivalves....................................... 149
37. Pearson's Correlation Coefficients Analysis between Heavy Metals in the Flesh and Shell of Mussel P. viridis............................................. 154
38. Comparison between Coefficient of Variation of Heavy Metals Concentration in Total Soft Tissue and Shell of Female Mussel P.
viridis...... . ...... .. . ...... . ... ....... .... ..... ... . ....... .. ............ ................. .... ...... 162
39. Comparison between Coefficient of Variation of Heavy Metals Concentration in Total Soft Tissue and Shell of Male Mussel P.
viridis. . . . ................... ....................................................... ..... .......... 163
40. Heavy Metals Concentrations in Byssus and Total Soft Tissue, and Ratios of Byssus-Metal to Total Soft Tissue-Metal of Mussel P. viridis.............................................................................................. 167
41. Comparisons of Byssus-Metal to Total Soft Tissue -Metal Ratios from Other Studies........................................................................... 168
ix
Table
42. Pearson's Correlation Coefficients Analysis between Cd, Cu, Pb and Zn Concentrations in the Byssus, Total Soft Tissue and Shell of
Page
Mussel P. viridis. . . . . . . . . . . . . . . . . . . . . . .. . . .. . . . . . . . . . . . . . . . . . .. . . .. . . . . . . . .. . . . . . . . . . . . . . . . . . . 172
43. Pearson's Correlation Coefficients Analysis Among Shell Thickness, Shell Length, Shell Width, Shell Height, Total Soft Tissue and Shell Dry Weight of Mussel P. viridis. .. . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177
44. Coefficient of Variation of the Female Mussel P. viridis from All Sampling Stations . . . . . . . . . . . . . . . . . . . . . . . , . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . .. . . . . . 180
45 Coefficient of Variation of Male Mussel P. viridis from All Sampling Stations. . . . ... ... . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .............. . . . . . . . . . . . . . . . . . ..... ... .. 181
46 Coefficient of Variation of Heavy metal Concentrations in Total Soft Tissue of Other Bivalves from Various Studies. . . . . . . . . . . . . . . . . . . . . ... . . . . . . . . . . 183
47 Pearson's Correlation Coefficients Analysis between Heavy Metals in Total Soft Tissue and Metal-Shell Weight Indices in Intra-station and Inter-station of Mussel P. viridis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . 188
x
LIST OF FIGURES
Figure Page
1 . Modiolus modiolus, Mytilus edulis and Perna viridis . . . . . .. .. .. . . .. . . . .. . .. . 11
2. A 1ripeptide fonned by the linking of three amino acids which is the basic structure of metallothionein ..................................................... .
32
3. Map showing the 15 sampling stations .............................................. . 38
4. Inside view of mussel P. viridis flesh and shell ................................. .. 45
5. The musculature and organs of the visceral mass of mussel P. viridis as seen from the right side .............................................................. . . 45
6. Different sections in the shen of mussel P. viridis used in analysis ..... . 47
7. Recoveries tests used in detennining the actual values of Cd, en, Ph and Zn in samples ........................................................................... .
52
8. Shell morphology of green-lipped mussel P. viridis .......................... .. 54
9. Concentrations of Cd, Cll, Pb and Zn in total soft tissue of mussel P. viridis sampled from 15 sampling sites ............................................ . . 66
10. Concentrations of Cd, Cll, Pb and Zn in shell of P. viridis sampled from 15 sampling sites ..................................................................... . 67
11. Cd, Cll, Pb and Zn concentrations in various tissues of mussel P. viridis ................................................................. ............................ . 88
12. Relative percentages of Cd, en, Pb and Zn concentrations in soft tissue of mussel P. viridis ...... ...................... . . . . ........ . .... ...... ............ ..
90
13. Comparisons of Cd, ell, Pb and Zn concentrations in different layers and sections in the shell of mussel P. viridis .... . . . .. ....... ... .......... .. . . . . .. . 101
xi
Figure
1 4.
1 5.
1 6.
17.
1 8.
1 9
20.
21 .
Relationships between Cd, Cu, Pb and Zn concentrations with shell thickness and shell length of mussel P. viridis from Pasir Panjang, Port Dickson ............... '" ............................. ................. ........ . . ... . .... ..
Relationships between Cd, Cu, Pb and Zn content with total soft tissue dry weight and shell length of mussel P. viridis from Pasir PaJUang, Port })ickson ..................................................................... .
Comparison of condition indices of mussel P. viridis from 1 5 l�""R • • Malay ' sampll.l15 stations m Sla .......................................................... ..
Relationships between condition index with shell and shell length ............................................................................................. .
Relationships between condition index with heavy metals concentrations and metal contents of mussel P. viridis from Pasir Panjang, Port Dickson ..................................................................... .
Relationships between ratios of byssus-metal to soft tissue-metal and heavy metals concentrations in total soft tissue of mussel P.
viridis .......................... , ................................... ................... ........... ..
Comparisons of shell length and shell in mussel P. viridis from 1 5 '!- . samPWlg stations ............................................................................. .
Relationships between total tissue dry weight with shell thickness and shell length of mussel P. viridis in Pasir Panjang, Port Dickson. ......................................................................................... .
Page
1 13
1 1 8
1 24
1 25
132
174
1 76
177
LIST OF PLATES
Plate Page
1. A sampling site at Langkawi Island............ ......... ...... ............... ......... 39
2. A sampling site at Kg. Pasir Putih, Tebrau Straits, Johore. ....... .......... 39
3. A sampling site at Pantai Udo, a jetty at Tebrau Straits, lohore.......... 40
4. A sampling site of cultured mussels at Pasir Panjang, Port Dickson..... 40
5. The different sexes in green-lipped mussel P. viridis.......................... 42
6. Soft tissues of green-lipped mussel P. viridis..................................... 42
7. A green-lipped mussel P. viridis...... ................ .............. .......... ......... 44
8. Substrates of mussels P. viridis........................................................ 44
9. Multilayer population structures of mussel P. viridis found at Pantai lido, Tebrau Straits, lohore............................................................. 134
10. A scattered and monolayer population structure of mussel P. viridis found at Pasir Panjang, Port Dickson...... ............. ............................. 134
xiii
LIST OF ABBREVIATIONS
pg.g.l microgram per gram
ANOVA analysis of variance
CI condition index
CV coefficient of variation
nnw double distilled water
g.cm"2 gram per centimeter square
g.cm"3 gram per centimeter cubic
H2SO4 acid sulphuric
HCl asid hydrochloric
HCLO .. acid perchloric
HN� acid nitric
ill sheD length : sheD height
mg. r1 microgram per litre
ml mililitre
STI soft tissue or flesh
SNK Student-Newman-Keuls
SPM suspended particulate materials
ST sheD thickness
WH sheD width: sheD height
WL sheD width : sheD length
xiv
Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfilment of the requirements for the degree of Master of Science.
ACCUMULATION AND DISTRIBUTION OF HEAVY :METALS IN GREENLIPPED MUSSEL PERNA VIRIDIS (LINNAEUS) FROM TIIE WEST COAST
OF PENINSULAR MALAYSIA
By
YAP CHEE KONG
August 1999
Chairman: Associate Professor Ahmad Ismail, Ph.D.
Faculty: Science and Environmental Studies
The main ann of this study is to establish mussel Perna viridis as a
biomonitoring agent along the west coast of Peninsular Malaysia. In this study, the levels
of heavy metals had been studied in the mussel P. viridis collected from 13 stations
along the coastline of Peninsular Malaysia and 2 stations in Sabah. The results showed
that, in general, the levels of heavy metals in total soft tissue of mussel P. viridis were
within or lower than the levels found elsewhere. The ranges varied widely from 0. 19 to
1.56 JJg.g'l dry weight for Cd, 5.78 to 15.14 JJg.g'l dry weight for Cu, 1. 17 to 8.27
J.!g.g.l dry weight for Pb and 46.78 to 145.95 Jlg.gl dry weight for In.
Mussels P. viridis collected from one population were dissected into eight parts
(byssus, mantle and gills, posterior adductor muscle, retractor byssal muscle, foot,
crystalline style, gonad and remainder visceral mass) and the concentrations of heavy
metals were measured. The highest concentration of Cu was obtained in the crystalline
style while elevated Cu, Pb and Zn concentrations were noted in the byssus. Female's
gonad generally accumulated higher metals levels than male's gonad. In the study of
xv
different parts in the shen of mussel P. viridis, eu and Zn levels were found elevated in
the periostracum layer while Cd and Ph levels were highly elevated in the inner nacreous
shenlayer.
The relationships between the concentrations (jlg.g,l) and contents
(jlg.individual'l) of heavy metals and body size (total STI dry weight, shell thickness and
sheD length) were also investigated. The results indicated that smaller mussels have
higher metals accumulation especially for Cd, Zn and Pb. The metal accumulations in
total STI of P. viridis were also affected by metal concentrations in sediment and
suspended particulate materials. Besides, physico-chemical parameters, condition in4ex,
population structure and sex also played a role in affecting heavy metal accumulation in
mussel P. viridis.
Apart from Sn mussel sheD and byssus also can be considered as good
bioindicators for Cu, Zn, Cd and Cu. Pearson's correlation coefficients in these metals
in the organs showed positive correlations and suggested the organ to be a goosd
bioindicator. The study on coefficient of variation indicated that if analysis of individuals
are used, a very limited size range of samples from all stations should be selected to
increase the ability of biomonitoring programmes.
In general, this study showed that ell, Pb, Zn and Cd accumulated in the STI of
P. viridis and showed some agreements that P. viridis is a good bioindicator for ell, Pb,
Zn and Cd. Tissue distribution of Cll, Ph, Zn and Cd varied in the organs and this
phenomenon probably depends on physiology, ecology and biology of the mussel P.
viridis.
xvi
Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai memenuhi syarat Wltuk ijazah Master Sains.
AKUMULASI DAN TABURAN LOGAM BERAT DI DALAM KUPANG PERNA VIRIDIS (LINNAEUS) DARI PERAIRAN PANTAI BARAT
SEMENANJUNG MALAYSIA
Oleh
YAP CHEE KONG
Ogos 1999
Pengerusi : Profesor Madya Ahmad Ismail, Ph. D.
Fakulti : Sains dan Pengajian A1am Sekitar
Tujuan utama kajian ini adalah mencadangkan dan menggunakan kupang Perna
viridis sebagai agen penWljuk biologi di sepanjang perairan pantai barat Semenanjung
Malaysia. Dalam kajian ini, tahap logam berat telah dikaji dalam kupang P. viridis yang
disampel dari 13 stesen di sepanjang perairan pantai Semenanjung Malaysia dan 2
stesen di Sabah. Keputusan menunjukkan bahawa secara amnya, tahap logam berat
daJam seluruh isi lembut kupang P. viridis adalah dalam lingkungan atau lebih rendah
daripada kajian di tempat-tempat lain. Lingkungannya berubah daripada 0.19 ke 1.56
jJg.g-l herat kering untuk Cd, 5.78 ke 15. 14 )lg.g-l herat kering untuk Co, 1.17 ke 8.27
J,lg.g-l herat kering untuk Pb dan 46.78 ke 145.95 J,lg.g-t herat kering untuk Zo.
Kupang P. viridis yang disampel dari satu populasi telah dibedah kepada 8
bahagian (bisus, mantel dan insang, otot aduktor posterior, otot retraktor bisus, kaki, 8til
berhablur, gonad dan tinggalan jisim viseral) dan kepekatan logam berat telah
ditentukan. Kepekatan tertinggi bagi Cu telah diperolebi di dalam 81il berhablur
manakaIa kepekatan yang tinggi bagi. logam Co, Ph dan Zn telah diperha1ikan di daIam
xvii
bisus. Gonad betina secara amnya menimbun logam lebih 1inggi daripada gonad jan tan.
Dalam kajian mengenai bahagian berlainan dalam cangkerang kupang P. viridis, tahap
Cd dan Zn adalah lehili tinggi dalatn lapisan periostrakwn manakala tahap Cd dan Pb
adaIah sangat Unggi da1am Iapisan da1aman nakreous cangkerang.
Perkaitan di antara kepekatan (�.g'l) dan kandungan �.individu·l) bagi logam
herat dan 8aiz badan (jmnJah isi lembut berat kering, ketebalan cangkerang dan
kepanjangan cangkerang) tcJah dikaji. Keputusan menunjukkan kupang bersaiz kecil
lebih banyak menimbum logam herat terutamanya bagi Cd, Zn dan Pb. Kandungan
logam daJam seluruh isi lembut P. viridis adaJah dipengaruhi oleh kepekatan logam
daIam sedimen dan partikel terampai. Selain itu, parameter fizikokirnia, indeks
'condition', struktur populasi dan jantina juga memainkan peranan dalam
mempengaruhi kandungan logam herat di dalam kupang P. viridis.
Selain Usu lembut, cangkerang and bisus juga boleh dianggap sebagai penunjuk
biologi kepada Cu, Zn, Cd and Cu. Korelasi koefisien Pearson bagi logam-logam ini
dalam organ-organ tersebut menunjukkan keputusan positif yang mencadangkan organ
organ ini menjadi penunjuk biologi yang baik bagi logam-logam tersebut. Keputusan
bagi koefisien variabiliti menunjukkan bahawa sekiranya analisis sampel secara individu
digunakan, sampel dalam julat saiz yang terhad harus dipilih bagi meningkatkan
keupayaan program pengawasan bioawas.
Secara umwnnya, kajian ini menunjuldcan Co, Pb, Zn and Cd menimbun di P.
viridis dan mempersetujui P. viridis sebagai penunjuk biologi yang baik kepada logam
logam tersebut. Taburan Co, Pb, Zn and Cd berubah-ubah di dalam organ dan keadaan
ini mungkin bergantung kepada fisiologi, ekologi dan biologi bagi kupang P. viridis.
xviii
CHAPTER I
INTRODUCTION
For the last fifty to a hundred years in particular, marine organisms have been
faced with an increasing input of a wide variety of potentially toxic chemicals
(Livingstone, 1992). Many of these chemicals are bioaccumulated within the tissues of
biota to concentrations significantly above ambient levels in the environment (phillips et
al., 1982; Rainbow, 1993) and can in some cases be up to 100 or 1000 times higher
than in sea water (Farrington et al., 1983). The rapid increase in inputs of chemicals,
either mobilized or synthesized by man into the estuarine and coastal environment was
due to growth in industrial activities during the twentieth century (Krishnakumar et ai.,
1987). With the steady growth of industrial activities in South East Asia over the
decades, there has been increasing concern over the environmental degradation of
coastal waters and the diverse and unique environments which they support (Brown,
1985). N.lalaysia, as a fast-developing country, should be made aware of the seriousness
that may arise if toxic chemicals were continually discharged into the coastal
environments due to rapid industrialization. In this study, coastal areas were focused
because these areas are hotspots of human activities (Ismail et al., 1993) and have been
the most productive areas for all sort of aquaculture activities (Thomas, 1988).
2
Aquatic contamination by heavy metals is extremely harmful since these
elements are not degradable in the marine environments (Jordao et al., 1997) and keep
accwnulating in the shellfish. Heavy metals, notably, Cll, Zn, Cd and Pb, have been
common by-products of anthropogenic activities since the earliest civilized times.
Furthennore, these heavy metals are potentially hannful to most organisms at a level of
exposure and absorption above a minimwn threshold and thus induce adverse biological
effects. In addition, it is wen-known that the consumption of fish and shellfish
containing excessive amount of heavy metals can be harmful to man. For instance, the
Minamata Disease outbreak in Japan in 19508 was caused by the conswnption of fish
and shellfish contaminated by methylmercury (Forstner and Whittmann, 1981).
Studies on heavy metals' ponution in Malaysian coastal waters have been done
(Law and Singh, 1988; Din, 1992; Ismail, 1993a; Ismail et al., 1993; Ismail and Ramli,
1997). Some of these studies have shown elevated concentrations of heavy metals in
these areas. This information has certainly inferred the need for regular monitoring of
heavy metals along the Malaysian coastlines. The west coast of Peninsular Malaysia has
been targetted because more than 600;6 of the Malaysian population is concentrated here
and most of the areas are being developed heavily (Ismail et al., 1993). Beck (1995)
suggested that the spreading and accwnulation of persistent chemicals, and the
destruction of oceans, are two of the most pressing environmental problems by the year
2020. This is exactly the year by which Malaysia has been expected to reach the status
of a developed countty. Therefore, intensive and regular monitoring of heavy metals'
levels in Malaysia's coastal waters should be done in order to provide comprehensive
baseline information for future reference.
3
A study of mussels is of interests to the public (Cheung and Wong, 1992)
because these animals are important sources of food (Vakily, 1992). Elevated levels of
heavy metals in mussels could mean contamination of this marine food delicacy.
Mussels are among the hardiest and most easily gathered organisms and the mussel
culture is the most productive fonn of saltwater aquaculture (Bardach et al., 1972).
Marine mussels are recognized as a good bioindicator of heavy metals' ponution
(Goldberg, 1975; Viarengo et al., 1995). Mussels have been used for the 'Mussel
Watch' monitoring programmes in which the mussels' species such as M. edulis and M.
galloprovincialis had been studied since the late 19608 (Morita, 1988). Until today,
there are reports on using mussels as bioindicators of heavy metals' levels
(Boonchalennkit et aL, 1998; Senthilnathan et al., 1998). An outstanding feature of
using mussels as bioindicator, when compared to other environmental components, is
that it is capable of assessing the biological availability of the pollutants measured
(Kristensen and Tyle, 1990). Mussels being inhabitants of the coastal waters are more
prone to be affected by pollutants derived from many sources (prabhudeva and Menon,
1987).
Are there any mussels abundantly fOlmd along the coastlines of Malaysia? If
mussels are found, are they as effective as a good bioindicator as described in the
system developed by 'Mussel Watch'? These questions can be answered if a swvey is
conducted and the samples are analysed in order to compare with other environmental
variables. In Malaysia, reports on heavy metals' levels in the tissues of mussel P. viridis
are scant (SivaIingam and Bhaskaran, 1980; Ismail, 1993b).
4
This study also examined the bioavailability of shell and byssus of mussel P.
viridis as better bioindicators, if possible, of heavy metals' levels on the basis of heavy
metals' concentration in the shell and total STI, and byssus and total STI. These two
s!udies were prompted by the questions arising in the usual assessment of the total STI
results in blue mussels M edulis. Do aD the tissues in the mussels accumulate the same
amoWlt of heavy metals? Do the shells or perhaps the byssus has different accumulation
patterns as compared to heavy metals' levels in the total STI of the mussels? The first
question could be answered through the investigation and comparisons of heavy metals'
levels in the byssus and the different soft tissues in the mussel P. viridis.
This study was conducted to :-
1. assess the current status of heavy metals' (Cu, Cd, Pb and Zn) levels and
contamination in the west coast of Peninsular Malaysia,
2. investigate the distribution of heavy metals in different tissues of STI in mussel P.
viridis as well as different layers and sections in the mussel's shell,
3. examine the uses of byssus and sheD as biomarkers and biomonitoring materials for
heavy metals,
4. find out if sediment, suspended particulate materials, physico-chemical parameters,
size, CI, population structure and sex, could be the factors affecting the accumulation
of heavy metals in the total STI of mussel P. viridis, and
PERPUSTAKAAN JNlVEUSlTI PUTRA MALA'iSIA
5. to investigate the concentrations of heavy metals' variability among-individuals,
referred to here as inter-station and intra-station's variability, so as to increase the
tmderstanding of this aspect.