Persistent, Bio-Accumulative and Toxic Contaminants in Coastal Marine Environment of Pakistan

JAMSHORO

Ph.D Thesis Persistent, Bio-accumulative and Toxic Contaminants in Coastal Marine Environment of Pakistan

Nuzhat Khan

Submitted to University of Sindh Jamshoro towards fulfillment of the requirement to award of the degree of Doctor of Philosophy in Analytical Chemistry

National Centre of Excellence in Analytical Chemistry University of Sindh, Jamshoro, Pakistan

2012

Persistent, Bio-accumulative and Toxic Contaminants in Coastal Marine Environment of Pakistan

A thesis submitted in fulfillment of the requirements for the degree of

DOCTOR OF PHILOSOPHY

ANALYTICAL CHEMISTRY

Nuzhat Khan

National Centre of Excellence in Analytical Chemistry University of Sindh, Jamshoro, Pakistan

2012

In the name of Allah Subhanutallah

He is Allah, other than whom there is no deity, the Sovereign, the Pure, the Perfection, the Bestower of Faith, the Overseer, the Exalted in Might, the Compeller, the Superior. Exalted is Allah above whatever they associate with Him. sūrat l-ḥashr 59 (59:23).

My humblest thanks to Allah, He who is my acquaintance and my greatest strength.

Thanks for the courage and determination to pursue my dreams.

DEDICATION

This work is dedicated to My mother A pillar of strength, my best friend, my buddy, my inspiration, my protector, my elation, wind beneath my wings without her in it, my life would be zero. My mother remains the love of my life. She is always there for me even before I am there myself, When she is around I am never alone. I thank her for believing in me and always making me see good, even at the most unpleasant times, and helping me make my weakness my greatest strengths. She always encouraged me to peruse my dreams. To my mother I owe the person I am.

Thanks for not just believing, but knowing that I could do this! Love You Maa

ACKNOWLEDGEMENTS

This PhD was researched while I was enrolled as a candidate, as well as employed in a full-time as a Senior Research Officer at National Institute of Oceanography, Karachi. It included many hours of sea-time collecting data and samples over Pakistan coastal area. The intensive field work and samples pre-treatment was carried out with the financial assistance provided by Pesticide Action Network of N. America (PANNA) and WWF-Pakistan Grant WWF-50033101 is greatly appreciated. The opportunity to carry out my research work at National Research Centre for Environmental Toxicology (EnTox), Brisbane Australia would have not been possible without the fellowship, granted by Partnership for Observation of the Global Oceans POPGO, Intergovernmental Oceanographic Commission (IOC) and the Scientific Committee on Oceanic Research (SCOR).

Special appreciation to Prof. Dr M. Iqbal Bhanger, Prof. Dr. Shahid Amjad and Dr. Jochen Müller, it would not have been possible to complete this doctoral thesis without your help and support. Prof Dr.Des Connell, Chairman, School of Public Health, Griffith University to accept me as visiting researcher at National Research Centre for Environmental Toxicology (EnTox), Brisbane Australia, where I have conducted most part of my PhD analytical work. Olaf Päpke facilitate me to obtain very first report on dioxins from Pakistan and to the ERGO laboratories, where these analysis were performed. Mary Hodge provided access to the laboratory at Queensland Health Scientific Services, where I used methodology and techniques, which allowed the analysis of OCPs and PAHs samples that would have been otherwise possible.

My sincere thank to Director General, National Institute of Oceanography, Pakistan for the continued institutional support and encouragement. Thanks to Prof. Dr. Shafi M. Nizamani, University of Sindh for his editorial advice and support.

I would like to acknowledge my team members Miss Saira Ishaq (RO) for providing me assistance in identification and biometric measurements and for all the lab and field work. Mr. Danish (RO) for the geological study, I would also like to thank NIO supporting Field and laboratory staff.

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ACKNOWLEDGEMENTS

A very sincere and extra special thanks to my dear friend Ms Tanya, EnTOX, Australia, I thank her also for the support, company and positive attitude always - a truly sincere person. I am so lucky for her friendship. I always remember my stay at Brisbane just because of you Tanya.

My thanks are extended to the many people who helped along the way from National Research Centre for Environmental Toxicology (EnTox), Brisbane Australia and Centre of Excellence in Analytical Chemistry University of Sindh Jamshoro.

Finally my very especial thank to my lovely family, my Sisters, my Neisse, my Nephews and my Khala for their love and support. They are the one who always there for me.

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ABSTRACT

The present work is the first of its kind to describe in detail the fate and distribution pattern of Persistence Bio-accumulative and Toxic (PBTs) in the coastal area of Pakistan bordering North Arabian Sea. The PBTs included OCPs, PAHs and Dioxins and Dioxin like PCBs.

Despite their widespread use and injurious effect of PBTs, little information is available on contamination levels of these pollutants in the coastal marine environment of Pakistan. Mangrove swamps, intertidal mudflats and Indus Deltaic creek represent main feature of the coastal marine environment. The coastal areas are of significance as there are spawning, breeding and nursery grounds of commercially important fishery resources. These living resources are under continuous threat of untreated industrial effluents and sewages discharges into coastal areas via several sources.

To evaluate the contamination levels of PBTs, 61 sampling sites were selected along coastal Pakistan, 09 sites were identified along the Balochistan coast, while 47 sites were identified along the coastal Sindh, and 5 sites were located in the Indus River.

In the current study, the analytical method routinely used in Environmental Toxicological Lab and Queensland Health Lab was adopted for identification quantification of organochlorine Pesticides, PCBs and PAHs. Analysis was carried out by using Varian 3400 Gas Chromatograph equipped with a Finnigan A 200S auto sampler and Finnigan SSQ710 Single Stage Quadrapole Mass Spectrometer. Sediments were sent to the ERGO Laboratory in Hamburg for the determination of 2,3,7,8-substituted PCDD/PCDF and dioxin-like PCB. Identification of PCDD/Fs was carried out by using retention times of the 13C-labelled standard and isotope ratios.

The results on the contamination levels of PBTs revealed that residual concentration of Organochlorine Pesticides were considerably higher (17.5 ng g-1 dry wt.) in semi-enclosed area (Creeks and Karachi Harbour) in the effluent discharge areas, this was attributed to low tidal flushing. However, in the areas where port and harbour

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AN ABSTRACT OF THE THESIS activities and untreated effluents from industrial and domestic sources are discharged, are reflective of highest PAHs contamination levels (2610.812 ng g-1 dry wt.) in the vicinity of Kemari jetty, Karachi Harbour area. Dioxin and Dioxin-like chemicals were detectable in all samples collected from Pakistan coastal environment. However, OCPs contamination levels in the sediment collected from Balochistan coastal environment were found below detection limit at almost all the sampling sites.

Spatial distribution pattern were significantly different among the localities sampled (p<0.004). Distribution pattern of most of the PBTs were well correlated with total sediment organic carbon contents (p>0.767 & R2=0.66).

A larger variation of ∑OCPs contamination levels (>0.002-17.5 ng g-1 dry wt. with a mean concentration of > 4.5 ng g-1 dry wt.) was observed in the samples collected from various localities of coastal area of Pakistan. The DDT’s metabolites DDE were found in soft tissue of the marine biota (fishes, crab, shrimps and molluscs) collected from Coastal area. The residue of DDT mainly its metabolites DDE and DDD were detected in most of the samples in relatively higher concentrations, compared with the concentration obtained for other OCPs. The high proportion of pp′-DDE at most of the site (41–95%) and ratio of ∑DDT and DDT (0.04 –0.24) suggests old inputs of DDTs in the environment, it is restriction have been made on the use of these chemicals. Pakistan has also switched over to natural pest control or using safer formulas.

The concentrations of ∑16PAHs varied amongst the localities, highest concentrations (2610.81 ng g-1 dry wt.) were detected in sediment samples collected in vicinity of Karachi harbour. Relatively lower levels (>400ng/g) observed in Korangi creek area south-east of Karachi. Higher concentrations of higher molecular weight PAHs such as (Benz(b+k) Fluoranthenen (>442.5 ng g-1 dry wt.), Ind(123cd)pyr (>270 ng g-1 dry wt.) and B(ghi) pryl (242.1 ng g-1 dry wt.) were detected near the discharge points of Lyari and Malir River. The Phen/Anth and Flth/Pyr concentration ratios indicated that mixture of pyrolytic and petrogenic PAHs sources at most of the site along the coast.

First time water borne PAHs contaminations were estimated using Triolein- passive sampler Semi-Permeable Membrane Devices (SPMDs) as an alternative

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AN ABSTRACT OF THE THESIS monitoring tool for coastal waters of Pakistan. The estimated water concentration was 2 found to be highest in the harbour area (CwSPMDs 4.6ng/l) that is well correlated (R =0.5) with the evaluated contamination levels (CwSed. 35.67ng/l) using the levels observed in the sediment.

According to the sediment quality standards of the USEPA and Canadian Council of Ministers of the Environment, observed levels of OCPs, PAHs and Dioxin contamination levels were generally lower than the threshold known to harm wildlife by OCPs. However, PAHs levels demonstrate moderate to low risk. Overall PCDD contributed to about 50 % of the TEQ in the samples with concentrations above 2 pg TEQ g-1 dwt and TCDD together with 1,2,3,7,8-PeCDD and 3,3',4,4',5-Penta-CB were the key contributors to the TEQ.

The results clearly indicate the pollution problem regarding these contaminants was found localized and much lower than the concentrations reported from neighbouring and regional countries.

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TABLE OF F CONTENTS

Summary of Contents Persistent, Bio-accumulative and Toxic Contaminants in Coastal Marine Environment of Pakistan S.No. Page No. Dedication Statement Acknowledgement I Abstract III Table of Contents VI List of Tables X List of Figures XII Abbreviations and Definitions XVII Contents CHAPTER-1

1 INTRODUCTION 1.1. General Introduction 2 1.2. Pakistan Coast 7 1.2.1. Sindh Coast 7 1.2.2 Balochistan Coast 8 1.3. Environmental Condition of the Area 9 1.4. Sources of Persistent, Bio-Accumulative and Toxic Substances (PBTs) 10 along Coastal Marine Environment of Karachi Pakistan 1.4.1. Pesticides 11 1.4.2. Polychlorinated Biphenyls (PCBs) 12 1.4.3. Un-intentional by-products (Dioxin and Dioxin like PCBs) 13 1.4.4. Poly Aromatic Hydrocarbon (PAHs)/Oil Pollution 16

CHAPTER- 2

2 REVIEW OF LITERATURE 2.1 Organochlorine Pesticides (OCPs) 22 2.2 Dioxin and Dioxin like PCBs 23 2.3 Poly Aromatic Hydrocarbons (PAHs) 24

CHAPTER-3 Distribution and Fate of Chlorinated Pesticides in Coastal 3 Marine Environment of Pakistan 3.1 Abstract 28 3.2 Introduction 29 3.3 Materials and Methods 34

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TABLE OF F CONTENTS

3.3.1 Sampling Location in the Study Area 34 3.3.2 Sample Collection 39 3.3.3 Sample Preparation and Transportation 40 3.3.4 Estimations of Sediment Organic Carbon 40 3.3.5 Analysis of Organochlorine Pesticides 40 3.3.6 Recovery of Organochlorine Pesticides 41 3.3.7 Statistical Analyses 41 3.4 Results and Discussion 42

3.4.1 Spatial Distribution of Organochlorine Pesticides (OCPs) in the 42 Sediment 3.4.1.1 Spatial Distribution Pattern of Organochlorine Pesticides in Sediment 44 of Karachi Harbour Area 3.4.1.2 Distribution Pattern of Organochlorine Pesticides in the Sediment of 51 Rocky Beaches (Bulleji) 3.4.1.3 Distribution Pattern of Organochlorine Pesticides in the Sediment of 51 Clifton Beach 3.4.1.4 Distribution Pattern of Organochlorine Pesticides in the Sediment of 51 Indus Deltaic Creek System 3.4.1.5 Distribution of Organochlorine Pesticides in the Sediment of 61 Balochistan Coastal Area 3.4.2. Fate of Organochlorine Pesticides in Coastal Environment of Pakistan 61

3.4.2.1 Distribution, Fate and Sources of DDT and its metabolites (DDE and 62 DDD) in the Coastal Sediment 3.4.2.2 Distribution, Fate and Sources of Hexachlorocyclohexanes (HCHs) and its isomers α-HCH, β-HCH, Lindane (γ-HCH) in Coastal 66 Environment 3.4.3 Correlations of Organochlorine Pesticides and Sediment Organic 68 Carbon 3.4.4 Organochlorine Pesticides Contamination in Marine Biota Collected 72 from Pakistan Coastal Area 3.4.4.1 Organochlorine Pesticides Contamination in Molluscs (Bivalves) 72

3.4.4.2 Residue of Organochlorine Pesticides Contamination in Crabs 77 Collected from Coastal Area. 3.4.4.3 Residue of Organochlorine Pesticides contamination Levels in 78 Shrimps (Penaeid shrimp) 3.4.4.4 Organochlorine Pesticides Contamination in Fishes (Nematalosa nasus (gizzard-shad/Daddi Palli) and Johnius glaucus (croaker/ Mushka)) 79 Collected from Coastal Area 3.4.4.5 Residual Contamination of Organochlorine Pesticides in Cephalopods 80 (Sepia pharaonis (cuttlefish/ Mayya) Collected from the Coastal Area 3.4.4.6 DDT and its metabolites in the Marine Biota Collected from Coastal 81 Area 3.5 Ecotoxicological Concerns /Potential for Biological/Ecological Effects 82

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TABLE OF F CONTENTS

3.6 Comparative (Neighboring, Regional and International) 83 Distribution of Organochlorine Pesticides (OCPs) Contamination 3.7 Conclusions 86 CHAPTER-4 Distribution of PCBs, Dioxin & Dioxin-like Chemicals in 4 River and Coastal Marine Environment of Pakistan 4.1 Abstract 88 4.2 Introduction 89 4.2.1 Un-intentional by-products (Dioxin and Dioxin like PCBs) 89 4.3 Material and Method 91 4.3.1 Sampling Area 91 4.3.1.1 Indus River Sampling Sites 91 4.3.1.2 Sampling sites along the Coast 92 4.3.1.2.1 Karachi Harbour 93 4.3.1.2.2 Gizri Creek 93 4.3.1.2.3 Korangi Creek 93 4.3.1.2.4 Gharo/Phitti Creek System 93 4.3.1.2.5 Khobar Creek 93 4.3.1.3 Sediment Sample Collection 94 4.3.1.4 Sample Preparation and Transportation 94 4.3.1.5 Analysis of Dioxin and Dioxin like PCBs 94 4.3.1.5.1 Pre- treatment /Sample Preparation for the Identification and 94 Quantification 4.3.2 Instrumental Condition 95 4.3.3 Estimation of EQF and TEQ 96 4.4 Results and Discussion 98

4.4.1 Spatial Distribution Pattern of PCBs, Dioxin and Dioxin-like 102 Contamination in Coastal Sediment 4.4.2 Spatial Distribution Pattern of PCBs, Dioxin and Dioxin-like 106 Contamination in Indus River Sediment 4.5 Evolution of Observed Contamination Levels with respect to EQF 107 and TEQ 4.6 Sources of PCBs, Dioxin and Dioxin-like Contamination in the 108 Aquatic Environment of Pakistan 4.6 Compassion of the Observed Contamination Levels with the 110 Published Information from Coastal Countries around the World 4.5 Conclusions 111

CHAPTER-5

Distribution and Fate of Polycyclic Aromatic 5 Hydrocarbons (PAHs) in Coastal Marine Environment of Pakistan 5.1 Abstract 113 5.2 Introduction 114

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TABLE OF F CONTENTS

5.3 Material and Method 116 5.3.1 Study Area 116 5.3.2 Sediment Sampling 116 5.3.3 Semi Permeable Devices (SPMDs) Preparation 116 5.3.4 SPMDs Deployment for the Estimation of Water Borne PAHs 117 Contamination Levels 5.3.5 Sample Preparation and Transportation 118 5.3.6 Analysis of Poly-Aromatic Hydrocarbons (PAHs) 118 5.3.6.1 Sample Pre-treatment/Preparation for the Identification and 118 Quantification of PAHs Contamination Levels 5.3.6.2 Instrumental Condition for the Determination of PAHs (Sediment and 120 SPMDs) 5.3.7 Recovery of PAHs 120 5.3.8 Estimation of Water Concentration 121 5.3.9 Statistical Analyses 122 5.4 Result and Discussion 122 5.4.1 Spatial Distribution of Polycyclic Aromatic Hydrocarbons (PAHs) 122 Contamination along Karachi Coast 5.4.2 Contamination Profiles of Prenatal PAHS in the Coastal Sediment 128 5.4.3 Correlation with total Organic Carbon (TOC) and PAHs levels in 132 Coastal Sediment 5.4.4 Evaluation of PAHs Carcinogenic Risk Potency 134 5.4.5 Estimated Water Concentrations of PAHs in the Coastal Environment 138 5.4.6 Source of PAHs Contamination in Coastal Environment 141 5.4.7 Comparison of Contamination Levels of Sedimentary PAHs with 144 other Coastal Region Around the World 5.5 Conclusion 145

CHAPTER-6 KEY FINDINGS, IMPLICATIONS AND FUTURE 6 RESEARCH Persistent, Bio-accumulative and Toxic Chemicals in the Coastal Environment of Pakistan Key Findings 148 Recommendation/Future Research and Monitoring 156

7 BIBLIOGRAPHY 157

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LIST OF TABLES

Particulars Page Table No. No. Targeted Organochlorine Pesticides (OCPs) for the Present Table-3.1 33 Study Table-3.2 Sampling Locations in Karachi Harbour Area 35 Table-3.3 Sampling Locations along the Indus Deltaic Creek System 38 Organochlorine Pesticide Levels (ng g-1 sed. dry wt.) in Table-3.4 44 Sediment Samples of Karachi Harbour Area Organochlorine Pesticide mean Levels (ng g-1) in Sediment of Table-3.5 53 Indus Deltaic Creek System Summary of WHO 1998 and WHO 2005 TEF Values (Berg Table-4.1 97 et.al 2006 ) Table-4.2 Distribution of PCBs pg/g sed. dry wt. 98 PCDD Congener Profile was obtained in the Coastal and River Table-4.3 99 Sediment (pg g-1 sed. dry wt.) PCDF Congener Profile was obtained in the Coastal and River Table: 4.4 100 Sediment (pg g-1 sed. dry wt.) Concentrations of PBDE Congeners in Sediment Samples from Table 4.5 101 Pakistan, (pg/g dry wt.) Concentrations Expressed on a Toxicity Equivalency basis pg Table-4.6 107 TEQ g-1 dwt. Concentration of Dioxin-like Chemicals in Sediment from Table-4.7 110 different Regions/Countries Sampling sites for Sediment Sampling and SPMDs Deployment Table-5.1 118 along Karachi Coast. Table-5.2 Targeted PAHs for the Present Study 119 Table-5.3 Recovery of Deuterated PAHs 121 Distribution of PAHs Contamination Levels in the Sediment Table-5.4. 123 Collected Coastal Localities. Table-5.5. Distributions of PAHs in the Sediment of Karachi Harbour Area 125 Distributions of PAHs in the Sediment of Indus Deltaic Creek Table-5.6 128 Environment Concentration of parental PAHs found in the Sediment Collected Table-5.7 129 from Study Area. Pearson Correlation matrix for the Coastal sed. Parental Table-5.8 132 Individual PAHs and ∑16PAHs, ∑CARC PAHs ∑Sed.OC PAHs Nonparametric Correlations (Spearman's rho) between Sediment Table-5.9 134 Organic Carbon and Individual PAHs Toxic Equivalent Factor of Individual PAHs Defined by Nisbet Table-5.10 137 and LaGoy. Total PAH Concentration and benzo(a)pyrene, B(a)P, Table-5.11 137 Equivalent Concentration at the Different Study Area.

Table-5.12 Worldwide PAHs 144

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LIST OF TABLES

Persistent, Bio-accumulative and Toxic Chemicals in the Coastal Environment of Pakistan Page xi

LIST OF FIGURES

Page Figure No. Particulars No. Figure-1.1 Map of Pakistan Coast 7 Figure-1.2 Map of Sindh Coast 8 Figure-1.3 Map of Balochistan Coast 9 Figure-1.4 Picture showing Stock Pile of Obsolete Pesticide in Pakistan 12 Figure-1.5 Uncontrolled Burning of all Sort Of Solid Waste Without Adequate Facilities at Unplanned Landfill Site close to Korangi 14 Creek Area. Figure-3.1 Sampling location of Sindh Coastal Area 34 Figure-3.2 Sampling location along Balochistan Coast 34 Figure-3.3 Map showing Sampling Location in the vicinity of Karachi 36 Harbour Figure-3.4 Sampling location in Indus Deltaic Creek System 37 Figure-3.5 Box plot showing Distribution of total Organochlorine Pesticides (ΣOCPs) median, inter-quartile range, range values 42 and outliers in the Sediment collected from selected Coastal localities. Figure-3.6 Box-plots showing Distribution of total Σ DDTs, ΣHCHs, ΣCYCs and HCB median, inter-quartile range, range values 43 and outliers in the Sediment collected from selected Coastal Localities. Figure-3.7 Distribution of ΣOCPs in Sediment Samples of Karachi 45 Harbour Area. Figure-3.8 Distribution of ΣDDTs, ΣHCHs and ΣCyclodienes in the 46 Sediment of Karachi Harbour Area. Figure-3.9 Distribution of HCB in Sediment Samples of Karachi 47 Harbour Area. Figure-3.10 Cyclodienes in Sediment Samples of Karachi Harbour Area. 48 Figure-3.11 Distribution of HCHs and its isomers in the Sediment of 49 Karachi Harbour Area Figure-3.12 Distribution of DDT isomers in Sediment Samples of Karachi 50 Harbour Area. Figure-3.13 Distribution of ΣOCPs in Sediment of Indus Deltaic Creek 51 System Figure-3.14 Total DDTs and Total OCPs Distribution in the Sediment of 54 Indus Deltaic Creek System Figure-3.15 Distribution pattern of Hexachlorobenzene (HCB) in the 55 Sediment of Indus Deltaic Creek System Figure-3.16 Distribution pattern of Cyclodienes in the Sediment of Indus 56 Deltaic Creek System Figure-3.17 Distribution pattern of Hexachlorocyclohexane (HCHs) and 58 its isomers α-HCH, β-HCH, Lindane (γ-HCH) in the

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LIST OF FIGURES

Sediment of Indus Deltaic Creek System. Figure-3.18 Relative concentration of ∑DDTs and ∑OCPs in the 59 Sediment from Creek Environment Figure-3.19 Distribution pattern of DDT and its metabolites (DDE and 60 DDD) in the Creek Sediment Figure-3.20 Correlation between DDD and ∑DDTs (R2 = 0.94), DDE and ∑DDTs (R2 = 0.98) indicated common sources of pollution in 61 the Area. Figure-3.21 Ratios of DDD / DDE in Sediment Samples of Karachi 64 Harbour Area Figure-3.22 Ratios of DDD / DDE in Sediment Samples of Indus Deltaic 64 Creek System Figure-3.23 Ratios of (DDD + DDE) / ΣDDTs in Sediment Samples of 65 Karachi Harbour Area Figure-3.24 Ratios of (DDD + DDE) / ΣDDTs in Sediment Samples of 65 Creek Area. Figure-3.25 Ratios of α-HCH/γ- HCH in Sediment Samples of Karachi 67 Harbour Area Figure-3.26 Ratios of α-HCH/γ- HCH in Sediment Samples of Indus 67 Deltaic Creek System Figure-3.27 Dendogram showing rich Sediment Organic Carbon stations within low flushed Areas in the vicinity of waste discharge points and seaward stations with low Organic Carbon 69 contents well flushed Areas are clearly distinct from all other localities along the Coast Figure-3.28 Correlation between Sediment Organic Carbon and total 69 OCPs along the Coast. Figure-3.29 Comparative levels of ∑OCPs estimated on Sediment dry wt. 70 and Sediment Organic Carbon in Creek environment. Figure-3.30 Comparative levels of ∑OCPs estimated on Sediment dry wt. 70 and Sediment Organic Carbon in Harbour Area. Figure-3.31 Correlation with total Sediment Organic Carbon and ∑DDTs 71 in the Coastal Sediment. Figure-3.32 Residue of Organochlorine Pesticides in Perna viridis 73 Figure-3.33 Residue of Organochlorine Pesticides in Crassostrea sp. 74 (Oyster) Figure-3.34 Residue of Organochlorine Pesticides Concentrations in 75 Turbo sp. Figure-3.35 Residue of Organochlorine Pesticides contamination in the Crabs (Scylla serrata and Portunus pelagicus) collected from 76 Coastal Area.

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LIST OF FIGURES

Figure-3.36 OCPs Contamination in Metapenaeus affinis (shrimp Jinga 77 /Kalri) & Penaeus merguiensis (Banana Shrimp/ Jaira) Figure-3.37 OCPs Contamination in Fishes (Nematalosa nasus (gizzard- shad/Daddi Palli) and Johnius glaucus (croaker/ Mushka)) 78 collected from Coastal Area Figure-3.38 OCs level in Sepia pharaonis (cuttlefish/ Mayya) 79 Figure-3.39 DDE levels in the marine bio collected from the Coastal Area 80 Figure-3.40 Comparivate study on the levels of contamination in marine 81 biota arround the world Figure-4.1 Indus River sampling location 91 Figure-4.2 Location map of Sediment sampling along the Coast of 93 Pakistan Figure-4.3 A GCMS Chromatogram of PCBs analysis 96 Figure-4.4 Dioxin-like chemicals detected (pg g-1 sed. dry wt.) in the 99 Sediment of Indus River and Coastal Area Figure-4.5 Spatial Distribution of Dioxin-like chemicals (pg g-1 sed. dry 101 wt.) Figure-4.6 ∑OCDD and ODDF levels in the Coastal and Indus River 102 Sediment (pg/g dry wt.). Figure-4.7 ∑PCBs levels (pg g-1 sed. dry wt.) in the Coastal and Indus 103 River Sediment (pg/g dry wt.). Figure-4.8 ∑PBDE concentrations in Sediment Samples from Pakistan, 103 (pg/g dry wt.). Figure-4.9 Mono-ortho–substituted PCBs congener profile along the 104 Coast. Figure-4.10 Non-ortho–substituted PCBs congener profiles in the Coastal 104 and Indus River Sediment Figure-4.11 OCDD congener profile in the Coastal environment (pg g-1 105 sed. dry wt.) Figure-4.12 OCDFs congener profile in the Coastal environment (pg g-1 106 sed. dry wt.) Figure-4.13 Relative Distribution pattern of the PCBs, PCDF and PCDD 108 Concentrations expressed on total WHO-TEQ Figure-4.14 Distribution of the PCBs, PCDF and PCDD Concentrations 108 expressed on total WHO-TEQ Figure-4.15 OCDD congener profile obtained from the Sediment of Indus 109 River Figure-4.16 OCDF congener profile for Indus River Sediment (pg g-1 . sed 110 dry wt.) Figure-5.1 Location map of SPMDs deployment sites along the Coast 117 Figure-5.2 SPMDs specification and preparation for development in the 119 Coastal Environment. Figure-5.3 PAHs identification and quantification showing in 120

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LIST OF FIGURES

chromatogram obtained from GCMS Figure-5.4 Box plot showing Spatial Distribution Pattern (mean median and inter-quarterlies) of ∑16PAH levels in the 123 Sediment of selected localities of Coastal Area. Figure-5.5 Hierarchical dendogram for sampling localities represented by three major group obtained by Ward’s Hierarchical 125 clustering method Figure-5.6 Distribution pattern of ∑ PAHs in the Sediment of Karachi 16 126 harbour Area Figure-5.7 Distribution pattern of ∑ PAHs in the Sediment of Indus 16 127 Deltaic Creek System. Figure-5.8 Correlations between ∑16PAHs and ∑CARC PAHs 130 Figure-5.9 Ratio between ∑ PAHs and ∑ PAHs in the Sediment 16 CARC 131 of Karachi Harbour Area. Figure-5.10 Pattern of CARC Distribution in the Sediment of Indus 131 Deltaic Creek System. Figure-5.11 PAHs concentrations normalized to TOC contents 133 Figure-5.12 ∑ PAHs (ng g−1 dry wt.) against total Sediment Organic 16 133 Carbon from the Coastal Area. Figure-5.13 ∑ PAHs (ng g−1 dry wt.) against total Sediment Organic CARC 134 Carbon from the Coastal Area. Figure-5.14 Concentration (ng g-1 dry wt.) of max.. Levels obtained from various localities of the Coastal Area relative to biological effect 135 levels (ERM and ERL). Figure-5.15 Concentration (ng g-1 dry wt.) of max. Levels obtained from various localities of the Coastal Area relative to Threshold Effect 136 Level (TEL) and Probable Effect Level (PEL). Figure-5.16 PAHs concentration (median, interquartile range, range values and outliers) in Sediment and SPMDs along Karachi 138 Coast. Figure-5.17 Estimated water concentration (median, interquartile range, range values and outliers) by SPMDs “Cw " & Sediment “Cw 140 " along Karachi Coast. Figure-5.18 Correlation between log of water concentrations estimated by 141 Sediment and SPMDs. Figure-5.19 The ratios Phe/Ant (phenanthrene/anthracene) verses Flu/Pyr (Fluoranthene/ Pyrene) to evaluate PAHs sources (petrogenic 142 and pyrolytic) along the Coast

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LIST OF FIGURES

Figure-5.20 Dendogram showing input of PAHs from various oThe ratios Phe/Ant (phenanthrene/anthracene) verses Flu/Pyr 143 (fluoranthene/ pyrene) to evaluate PAHs sources (petrogenic and pyrolytic) along the Coast Figure-6.1 Spatial Distribution Pattern of ∑Organochlorine Pesticides in 149 Coastal Sediment of Pakistan

Figure-6.2 Spatial Distribution Pattern of ∑Organochlorine Pesticides normalized on Sediment Organic Carbon from Pakistan 150 Coastal Figure-6.3 Spatial Distribution Pattern of ∑DDTs in Sediment Samples 151 collected from Pakistan Coastal Area Figure-6.4 Spatial Distribution Pattern of ∑HCHs in the Coastal 152 Sediment of Pakistan Figure-6.5 Spatial Distribution Pattern of ∑PAHs in Sediment Samples 153 Collected from Study Area. Figure-6.6 Spatial Distribution Pattern of ∑PAHs s in Sediment CARC 153 Collected from Coastal Area. Figure-6.7 Spatial Distribution Pattern of ∑PAHs normalized on 154 Sediment Organic Carbon

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Definitions, Acronyms, Abbreviations and Units

Definitions

ADI Average daily intake

Congener Compound member of the same chemical family; e.g. there are 75 PCDD congeners

Dioxins General term used here to include PCDDs and PCDFs

Homologue Group of structurally related chemicals that have the same degree of chlorination; e.g. there are 8 PCDD homologues

HpCDD/F Heptachlorodibenzodioxin/furan

HxCDD/F Hexachlorodibenzodioxin/furan

Isomer Substance that belong to the same homologue class; e.g. there are 22 isomers in the homologue group TCDD

Kow Octanol-water partition coefficient

LOAEL Lowest observed adverse effect level

NOAEL No observed adverse effect level

OCDD/F Octachlorodibenzodioxin/furan

PCDDs Polychlorinated dibenzo-p-dioxins

PCDFs Polychlorinated dibenzofurans

PnCDD/F Pentachlorodibenzodioxin/furan

POP Persistent organic pollutant; e.g. PCDD/Fs, PCBs, DDT, Hexachlorobenzene, Mirex, Toxaphene, Aldrin, Dieldrin, Endrin, Chlordane, Heptachlor

TCDD/F Tetrachlorodibenzodioxin/furan

TDI Tolerable daily intake

TEF WHO Toxic equivalency factor, toxicity of a compound relative to that of 2,3,7,8-TCDD (Van den Berg et al. 1998)

TEQ Toxic equivalency defined as the concentration of a compound multiplied by its toxic equivalency factor (TEF) (Van den Berg et al. 1998)

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Definitions, Acronyms, Abbreviations and Units

OCPs Sum of all analyzed Organochlorine Pesticides

Acronyms and abbreviations

ACE Acenaphthene

ACL Acenaphthylene

ADC Adjacent Indus Deltaic Creeks

AET Apparent effects threshold,

ANOVA Analysis of variance

ANT Anthracene

BaA Benzo[a]anthracene

BaP Benzo[a]pyrene

BbF Benzo[b]fluoranthene

BkF Benzo[k]fluoranthene

BPE Benzo[g,h,i]perylene

CB Clifton beach

CCME Canadian Council of Ministers of the Environment

CEE Central and Eastern European

CHR Chrysene

CLRTAP Convention on Long-Range Transboudary Air Pollution

Cwsed Sediment-based aqueous concentrations

Cwspmd SPMD-based aqueous concentrations

D - Di,

DBA Dibenzo[a,h]anthracene

DDD Dichlorodiphenyldichloroethane

DDE Dichlorodiphenyldichloroethylene dw Dry weight

Page xvii

Definitions, Acronyms, Abbreviations and Units

EF Emission Factor

EIA Environmental Impact Assessment

EnTox National Research Centre for Environmental Toxicology

ER-L Effects range-low

ER-M Effects range-median

FLR Fluorene

FLU Fluoranthene

GC Gizri creek

GCMS Gas Chromatograph Mass Spectrometer

GEF Global Environmental Facility

GPC Gel Permeation Chromatography

GPS GCMS retention times (RT)

HCB Hexachlorobenzene

HCH Hexachlorocyclohexane

HITE Hub Trading Estate

Hp- Hepta,

HRMS High Resolution Mass Spectrometry

HW Hazardous waste

Hx- Hexa,

IA Impact Assessment

INP Indeno[1,2,3-cd]pyrene

I-TEQs International Toxic Equivalents

KANUPP Karachi Nuclear Power Plant

KC Korangi creek

KH Karachi Harbour

Page xviii

Definitions, Acronyms, Abbreviations and Units

KIA Korangi Industrial Area

KOC Partition coefficient octanol-water

Kow Octanol/water partition coefficient

KWSB Karachi Water and Sewerage Board

LEC50 50% effect concentration

LITE Landhi Industrial Trading Estate

MoST Ministry of Science and Technology

MU Masaryk University of Brno, Czech Republic

NAP Naphthalene

NEQS National Environmental Quality Standards.

NIP National Implementation Plan Stockholm Convention

NRL National Refineries Ltd

O– Octa

OC Organochlorine

OCCs Organochlorine Compounds

OCPs Organochlorine Pesticides

PAHs Polycyclic Aromatic Hydrocarbons

PBDE Polybrominated diphenyl ether

PBTs Persistent, Bioaccumulative, Toxic substances

PCBs Polychlorinated biphenyls

PCBzs Polychlorinated benzenes

PCDDs Polychlorinated dibenzo-p-dioxins

PCDFs Polychlorinated dibenzofurans

PCPs Polychlorinated phenols

Pe- Penta,

Page xix

Definitions, Acronyms, Abbreviations and Units

PeCP Pentachlorophenol

PEL Pprobable effect level

PHE Phenanthrene

POPs Persistent Organic Pollutants

PPOPs Priority Persistent organic pollutants

PRL Pakistan Refineries Ltd

PYR Pyrene

RECETOX Research Centre for Environmental Chemistry and EcoTOXicology

RT Retention times

SEL Severe Effects Level

SIM Selected Ion Mode

SITE Sindh Industrial Trading Estate

SPMDs Permeable Membrane Devices

SPSS Statistical Product Service Solutions

SQGs Sediment quality guidelines

SW Solubility coefficient in water

T- Tetra,

TEF Toxic Equivalent Factor

TEFs Toxic Equivalent Factors

TEL Threshold effect level

TEQ Toxicity Equivalent

TET Toxic effect threshold

TOC Total Organic Carbon

Tr- Tri,

Page xx

Definitions, Acronyms, Abbreviations and Units

UH Upper harbour

UN ECE United Nations European Commission for Economy

UNEP United Nations Environment Program

U-POP Un-wanted by-products

WB World Bank

WHO World Health Organisation

Units of Measurement g gram

µg g-1 micro-grams per gram ng g-1 nanogram per gram pg g-1 picogram per gram

Page xxi

Certificate

This is to certify that this Ph D thesis “PERSISTENT, BIO-ACCUMULATIVE AND TOXIC CHEMICALS IN COASTAL MARINE ENVIRONMENT OF PAKISTAN” has been completed by Nuzhat Khan under my supervision. This is original research work performed at National Research Centre for Environmental Toxicology (EnTox), Brisbane Australia, the ERGO laboratories, Hamburg Germany and National Centre of Excellence in Analytical Chemistry, University of Sindh, Jamshoro. This work has not been submitted in any form for another degree at any University or other institution of tertiary education. The information derived from the published or unpublished work of others has been acknowledged in the text and a list of references is given. The entire thesis is composed by Nuzhat Khan. Her dissertation is worthy for presentation to the University of Sindh for award the Doctor of Philosophy in Analytical Chemistry.

Prof. Dr. Muhammad Iqbal Bhanger Supervisor National Centre of Excellence in Analytical Chemistry University of Sindh, Jamshoro.

CHAPTER 1 INTRODUCTION

INTRODUCTION CHAPTER-1

1.1 General Introduction

The increase in human population has driven the world to enhance industrial and agricultural practice. Increased demand and continued efforts to meet ever-growing needs of mankind has resulted in use and produce wide range of chemicals. Thus increasing anthropogenic intervention has adversely affected the global environment (Moor, 2002).

The ocean is a natural gift to mankind a huge treasure, covering 140 million square miles and 72 per cent of the earth's surface. Pakistan is one of the countries which are bestowed by the Almighty with the bounties of the ocean. Ocean is a source of present and future food supply. Safe, healthy and productive seas and oceans are vital to human well-being, economic security and sustainable development. Coastal marine areas receive variety pollutants from land base sources such as industrial effluents, agriculture and sewage discharges, surface run-off and deposition from the atmosphere (Sander, 1994).

Most widespread contaminates are synthetic organic chemicals, persistent in nature with the tendency to bio-accumulate and posse toxic effect on human health or environmental are called Persistent Bio-accumulative Toxic substances (PBTs) (Holoubek, et al., 2000). The PBTs could be a single or group of chemicals or a mixture with identical properties and release into the environment (Barrie, et al., 1992).

The subclass of PBTs is Persistent Organic Pollutants (POPs). POPs are group of synthetic chemicals that include agrochemicals (Organochlorine Pesticides), industrial substances (Polychlorinated Biphenyls) and Unintentional by-products (Polychlorinated dibenzo-p-dioxins and Polychlorinated dibenzofurans) (Iuni et al., 2007). The Organochlorine Pesticides (OCPs) include DDTs, HCHs (hexachlorocyclohexanes), CHLs (chlordanes), cyclodienes and HCB (hexachlorobenzene), Aldrin, Dieldrin, Chlordane, Heptachlor, and Mirex Toxaphene.

These groups of synthetic chemicals are among the most dangerous pollutants released into the environment by human industrial and agricultural activities (Harald, et al., 2000). These chemicals persist in the environment and degradation by biological,

Page 2 INTRODUCTION CHAPTER-1 photochemical or chemical means is very poor (Khan, 2004). These contaminates are semi-volatile in nature hence their persistence and movement relate to the environmental condition of an area. They can also travel thousands of miles from their original source through a process of evaporation called “grasshopper effect” (greenpeaceindia.org, 2003). The unique properties of PBTs create local, regional or global concern for human safety (Petr and Holoubek, 2001, Wallack et al., 1998).

Polycyclic Aromatic Hydrocarbons (PAHs) are produced by combustion of organic compounds and are classified as persistent organic compounds. Their occurrences are related to anthropogenic activities and contamination in aquatic environment is serious problem especially in high-density industrial areas (Wong, et al., 2003). Polycyclic Aromatic Hydrocarbons (PAHs) has been recognized as environmental contaminants found in almost all compartment of the global system (Verweij et al., 2004). Preliminary results of PAHs contamination in the coastal environment from human included activities, although small concentrations of PAHs are also produced by the natural processes. PAHs are undesirable group of substances produced as a result of incomplete/inefficient combustion of organic material. They are non-polar, lipophlic aromatic compounds containing two or more fused arenes structures. The persistence increased with increase in aromatic rings. However low molecular weight PAHs are more soluble and have less affinity for surfaces with significant acute toxicity than high molecular weight PAHs. According to the Neff, 1979; Witt, 1995, the higher molecular weight PAHs with a 4, 5 or 6 ring structure are more carcinogenic in nature. These are produced during high temperature combustion of organic matter, whereas low temperature burning results in PAHs with a 2 or 3 ring structure (Readman et al., 2005). It is also reported by Nemr et al., 2006 et al., that PAHs also synthesized naturally by organisms, such as bacteria, algae and fungi.

Page 3 INTRODUCTION CHAPTER-1 concentration in sediment, intermediate in biota and lowest in the water column is expected for the coastal marine environment (CCME, 1992). It is significant to quantify the level of PAHs pollution in the marine environment and their impact on the marine organisms and human health (Wurl et al., 2005).

It seems to be difficult to manage wide spread use and unintentional generation of these chemicals. The PBTs, synthetic chemicals has become an ever-present contaminates in the environment. They are volatile in nature and can be transported worldwide, moving from tropical climate to colder region in many cases (Anu, at el., 2010. They have tendency to bio-accumulate and bio-magnify in the food chain and ultimately pose adverse effect on human health, even in very small quantities (CECS.org, 2003).

United Nations Environment Programme with the cooperation of International Forum for Chemical Safety (UNEP, 1996) identified group of most hazardous persistent organic pollutants. These are synthetic organic chemicals more commonly known as POPs. The Stockholm Convention on Persistent Organic Pollutants (POPs) list twelve priority pollutants that include eight agrochemicals (aldrin, dieldrin,endrin, DDT, chlordane, heptachlor, mirex and toxaphene), two types of industrial chemicals (PCBs and hexachlorobenzene) and two families of unintentionally produced by-products during incomplete combustion of chlorine and chlorine-containing substances (dioxins and furans) called un-wanted POP (U-POPs) (Wong et al., 2003). The PAHs are also unintentionally produced as by-products (U-POPs) of incomplete combustion, mostly waste incinerators and chemical processes that involve organic matter and chlorine (Khan, at al., 2004).

In view of wide spread use and toxicological impact of these contaminants the experts around the globe strongly recommend to monitor their fate and distribution in the environmental compartment for the prediction of their behaviour in the various environmental conditions, deposition/emission processes, transformation processes and bioavailability in the ecosystems (Holoubek, et al., 2000). The pollutants do not recognise national or international boundaries. During the May 22, 2001 Stockholm

Page 4 INTRODUCTION CHAPTER-1 convention on Persistent Organic Pollutants (POPs) was drafted to protect of human health and environment and entered into force on May 17, 2004. The convention is signed by 151 countries and 122 countries have ratified it (CEC.org, 2003). Pakistan signed the Convention on December 6, 2001.

Pakistan is going through a process of growth and industrialization. This, coupled with ineffective policy regulations and weak urban development planning, has resulted in excessive levels of pollutant being discharged in the marine environment. The lack of efficient production and post-production technologies in the industrial sector, and the increasing quantity of untreated industrial effluent has degraded the environment. As a result of this, there is a constant threat to both human health and biodiversity.

Karachi city with population of about 18 million is the biggest trade & economic center/industrial hub of the country. The main source of pollution in the coastal marine environment of Sindh is large industrial activity that is about 6000 large and small industrial units located around the coastal area. It is reported that about 400 million gallons per day (MGD) of untreated industrial effluents (approximately 60 %) and domestic waste (about 40%) is discharged into coastal environment of Karachi (Beg, 1994, Masroor, 2005). These untreated effluents drained into the coastal marine environment via several sources, mainly through Lyari River that is approximately 59% and 25% via Malir River of the total pollution load are being discharges into the coastal area and the rest (15%) of the pollution load is directly introduced into the adjacent open sea coast, Gizri/Korangi and Gharo creeks area (Beg, 1994). The Lyari River carries large quantity of industrial effluents and urban waste into the Karachi harbour. The organic fraction of the waste from domestic sources often contribute more than 50% of the total waste (Ahmed and Zurbrugg 2002), that have great affinity to absorbed/accumulate almost all kind of pollutants particularly hydrophobic organic chemical, like polycyclic aromatic hydrocarbons, organochlorine pesticides, PCBs and dioxin and serve as continuous source of pollution in an environment. Thus, the coastal areas, which are the spawning and breading ground of most of the fishes, may have elevated levels of pollution from land based sources.

Page 5 INTRODUCTION CHAPTER-1

As with the diversity of sources for PBTs/POPs, the factors influencing persistence are also unique in the country. Typical tropic hot humid climate of the country may influence bio-availability, biological uptake, persistence and effect different from other climate country. Evaporation and photo POPs transformation are also more and the native biota involved in primary conversion are also more diverse. The environmental half-lives in the country are much different than in temperate countries. Very little data from field studies are available on this issue.

The restriction have been made on use and manufacture of these chemicals, however case of the poisoning by these chemicals are still being reported mostly from developing countries (Wong, et al., 2003, Aguilar and Borrell 2005, Noël et al. 2009).

There is need for assessment of chemical-contaminants at trace level concentrations in the marine environment and their impact on the marine organisms and human health. In this context the current study was designed to investigate the sources and status of contamination of Persistent, Bioaccumulative, Toxic substances (PBTs) in Pakistan. The focus was placed in particular on the coastal area with potential sources of contaminations since this system is a sink for lipophilic compounds such as OC pesticides, PAHs, PCBs and PCDD/Fs. Due to scarcity of data and information available on PBTs in the coastal environment, this study aimed to provide information on the sources, distribution and fate of these contaminates into and within the coastal environment. Such knowledge is fundamental to assessing the risks associated with the presence of PBTs in the environment.

The final overview on the key findings of this thesis on PBTs and their Eco- toxicological impact were evaluated for marine environment of Pakistan boarding Northern Arabian Sea. Following were the core objectives of the present study:

 Assess the spatial distribution of PBTs contamination along the coastline,

 Provide information on the possible sources  Identify the fate of PBTs within the coastal environment,  Provide information necessary for the assessment of risks to the near shore marine environment and biota.

Page 6 INTRODUCTION CHAPTER-1

1.2. Pakistan Coast

Coastline of Pakistan is about 990 km long along the Arabian Sea (Figure-1.1). It comprises of two distinct zones, the Sindh coastal and the Balochistan Coast (SCAP 1996).

Figure-1.1 Map Showing Pakistan Coastal Area

1.2.1 Sindh Coastline

The Sindh coastal area about 330 km long is located south-eastern part of the country that is about, between Sir Creek on the east to Hub River coast on the west. Sindh Coast can be further divided into two parts, Indus Deltaic Creek system and Karachi Coast (UNESCAP, 1996, Yoshiki and Porfirio, 2008).

1.2.1.1 Indus Deltaic Creek System

The Indus delta is between Korangi Creek and the Runn of Kutch. The Indus River delta was formed by drains of Indus River into the north eastern Arabian Sea. A network of creeks is spread over the entire Sindh coast southeast of Karachi.

Page 7 INTRODUCTION CHAPTER-1

Figure-1.2 Map of Sindh Coastal Area (UN-ESCAP, 1996)

1.2.1.2 Karachi Coast

The coastline of Karachi metropolitan is about 70 km long situated between the Cape Monze, a high cliff projecting into the Arabian Sea and Korangi creek, is relatively well developed as compared to the rest of the Pakistan coast. It is generally oriented NW- SE. Western side is bounded by the Hub River and mangrove swamps and Indus Deltaic Creeks on the east (UN ESCAP, 1996, Yoshiki and Porfirio, 2008). The Lyari and Malir River are the seasonal streams which flow during SW monsoon. Four major inlets Manora Channel (Karachi harbour), Korangi creek, Phitti creek and Khuddi creek invigilate the coastline Khan, at el., 2006).

1.2.2 Balochistan Coast

Balochistan coast is about 660 km, extended from mouth of the Hub River in the east to middle of Gawatar Bay in the west (UN ESCAP, 1996). The cliffs, rocky head-

Page 8 INTRODUCTION CHAPTER-1 lands, sand dunes, lagoons and mangroves marsh are the main features of the area (Figure-1.3).

Figure-1.3 Map showing Balochistan Coastal area

1.3 Environmental Condition of the Area

There are a number of environmental issues in the coastal zone of Pakistan such as disposal of domestic wastes and untreated industrial effluent causing marine pollution problems along the urban centers are the most significant (Khan et al., 2006). The pollution problems have arisen mainly due to the indiscriminate discharges of effluent from coastal industries and agricultural sources together with untreated liquid and solid wastes generated from domestic sources into the coastal environment that could be sources of deterioration of coastal environmental, depletion of marine resources, human health risks and loss of bio-diversity (Masroor, 2005).

There is approximately over 6000 small and large industrial units the in vicinity of Karachi coastal area. In addition to the domestic and industrial waste disposal there are

Page 9 INTRODUCTION CHAPTER-1 some other equally important sources of pollution in the coastal waters. These include maintenance dredging of the navigational channels of the ports and harbours and land reclamation practices along the coast that also carry out dredging of the near-shore areas for providing landfill material. Dredging loosens and re-suspends the settled fraction of silt and sand to produce very high load of suspended solids in the coastal area under the influence of mixing processes. Dredging also disturbs the hydrogen sulfide-laden upper layers of sediments forcibly mixing and dissolving hydrogen sulfide toxic substances in seawater. Karachi harbour is also exposed to wastes from ship and boat, oil spills and other port & oil terminal activities. Therefore, a high incidence of oil and oily waste is expected in the area.

1.4 Sources of Persistent, Bio-accumulative and Toxic substances (PBTs) along coastal marine environment of Karachi Pakistan

In establishing the sources of the PBTs is important to have better understanding of all possible sources including manufacturing/production facilities, use and disposal and stock of unwanted or obsolete material that includes obsolete pesticides stocks and PCBs material in used and emission sources. Most of the PBTs that include PCBs and POPs pesticides have never been manufactured in Pakistan (Khan, 2006). However, DDT, HCB and Dieldrin have been formulated locally at various places along Karachi coast.

The organo-chlorine compounds such as pesticides, insecticides, PCBs and plastic waste materials are amongst prominent persistent pollutants along the coast of Pakistan. The use of chlorinated pesticides and insecticides has been considerably reduced compared with their use about 3-decades ago. However, these are still being used by the agriculture sector particularly for the cotton cash crop.

The plastic pollution has become very prominent on the beaches because of the indiscriminate use and uncontrolled disposal of polyethylene shopping bags which litter the common beaches of Karachi. The other important form of plastic pollution is the presence of plastic pellets (polyethylene and polystyrene balls) of plastic products. These

Page 10 INTRODUCTION CHAPTER-1 balls are common in beach sands along the high water mark. However, no study has yet been made on the plastic pollution in the marine environment of Pakistan.

Hazardous industrial municipal, agricultural and other waste burning is in large scale practice traditionally in the country, without adequate incineration facilities that could be great source of Dioxin in pollution in the area other than industrial sources. Activities like the Gulf War also added to the problem and possible build up of POPs. All this make the issues regarding POPs in this country peculiar and more serious.

In view of the above considerations a critical appraisal of the POPs of priority in the country has been identified.

1.4.1 Pesticides

In establishing the sources of the PBTs is important to have better understanding of all possible sources including manufacturing/production facilities, use and disposal and stock of unwanted or obsolete material that includes obsolete pesticides stocks and PCBs material in used and emission sources. Most of the PBTs that include PCBs and POPs pesticides have never been manufactured in Pakistan (POP-NIP, 2007). However, DDT, HCB and Dieldrin have been formulated locally at various places along Karachi coast. However these pesticides have been deregistered and the registration and/or officially used is bane completely.

However, there are reports of continuous illegal use of DDT for example for the protection of the roots of fruit trees. The origin of this DDT is unclear; illegal import from neighbouring countries could be a probable source.

There are very large stockpiles of obsolete pesticides in Pakistan. Sindh is estimated to have approximate 2000 tons of unsorted and mostly obsolete pesticides in Stock (Khan, 2006) The problem of unwanted and expired pesticides pose the greatest danger to the natural environment and people (POP-NIP, 2007).

Page 11 INTRODUCTION CHAPTER-1

1.4.1.1 Pesticide Stockpiles

Pakistan holds one of the largest obsolete pesticides including POPs pesticide stocks piles of approximately 5000 metric tons or more. These stocks were accommodated in various storage sites up until 1980 in extremely hazardous conditions in more than a thousand sites all over Pakistan (POP-NIP, 2007).. The Stocks Piles is the result of change in Government of Pakistan policy regarding free aerial spray, due to the change of policy the pesticides in Government stores were no longer distributed. The districts of Sindh contain large stocks of obsolete pesticides that included POP Pesticides, more than 2000 tons of unsorted and mostly obsolete formulated pesticides stored in extremely hazardous condition in 210 stores throughout the province Sindh for more than two decades (POP-NIP, 2007). The obsolete pesticides storage sites may be a source of contamination that adversely affects the local communities and the environment.

One of the biggest piles of obsolete Pesticide is location in Karachi at the 15 km distance from Korangi Creek area that is Malir storage Site. This is site contained over 410 tons expired/obsolete pesticides which were accumulated from several decades (Figure-1.4).

Figure-1.4. Picture Showing Stock Pile of Obsolete Pesticide in Sindh, Pakistan

1.4.2 Polychlorinated Biphenyls (PCBs)

The Polychlorinated Biphenyls (PCBs) was introduced in 1929 as a very popular industrial chemical, mainly used as a flame-retardant, cooling agent in transformers and

Page 12 INTRODUCTION CHAPTER-1 capacitors, also use in many industrial process as lubricants, hydraulic fluids, cutting oils, adhesives, liquid seals, coating wood and plastics to prevent them from flame. Also used in paints, varnishes, inks and pesticides and carbonless copy paper (Khwaja and Petrlik, 2005). Soon after it was recognize as dangerous chemical by Soren Jensen a Danish chemist.

The PCBs has been used in Pakistan, however it has never been manufactured/synthesize in the country. This nasty chemical (PCBs) sold in market with various trade names (Clophen, Arocler, Kanechlor, Santotherm, Phenoclor and Pyralene). It has been in used in transformer oil in power sector that could be possible sources of PCBs contamination along the coast. The other sources may be automotive and electric machinery contaminating PCBs oils. The PCBs contamination may be introduced in the environment due to the leakage from the electrical equipment and/or dealing with the wastes containing PCBs. There is hardly any published information available on the contamination of PCBs in various environmental compartments of Pakistan. The use of PCBs-oil has been prohibited in Pakistan since 1974. PCBs contamination is listed as “Phenolic Compounds” in the NEQS-Pakistan (PEPA, 1997).

There are many activities that produce PCBs wastes along Karachi coast. Industrial, automotive and electric machinery are the main sources of PCBs contaminated oils. The release of PCBs into the environment is generally expected to be due to (a) leakage from PCBs containing electrical equipment (transformers, capacitors, circuit breakers, voltage regulators etc. and (b) handling and processing of wastes containing PCBs (Muller et al., 1999). In Pakistan little documented information is available in any form on PCBs use and contaminations.

1.4.3 Un-intentional by-products (Dioxin and dioxin like substances)

Page 13 INTRODUCTION CHAPTER-1 such as forest fires or volcanic activity. Un-intentional by products include the highly toxic dioxins and furans are produced by human activities and are also occurring naturally. The natural sources could be combustion of forest fires and volcanic activity. These contaminations are not industrial substances or involved in industrial processes, except in small amounts produced for research purposes (green-facts.org, 2006). However these are unintentionally produced during industrial processes dealing with organic chemicals and chlorine, they are introduced in an environment via municipal and domestic incineration and combustion processes (green-facts, 2010).

Since 1947 Pakistan industrial activities based on secondary production like refilling, formation and assembling etc. primary production activities are not very well promoted that could be main sources of the pollution. The incineration activities is not present in recent past, however this is going to be promoted day by day in Pakistan. However Practices like open pit burning of waste are very common all over country. There are over 100 truck/day of all kind of waste collected from Karachi city dumped and uncontrolled burned near Korangi Creek area for land unplanned filled (Figure-1.5).

Figure-1.5. Uncontrolled burning of all sort of solid waste without adequate facilities at unplanned landfill site close to Korangi Creek area.

Page 14 INTRODUCTION CHAPTER-1 the environmental problems in the country. Pakistan has a number of industries in the fields such as metallurgy, pulp and petrochemicals, which might be substantial emitters of unintentional POPs. Furthermore, practices like open pit burning of waste are very common.

1.4.3.1 Industrial Activities

Since 1947 Pakistan industrial activities based on secondary production like refilling, formation and assembling etc. primary production activities are not very well promoted that could be main sources of the pollution. However, lack of awareness and poor implementation of the environmental legislation and irresponsible and casual attitude towards environment issues may augment the environmental problems in the country.

1.4.3.2 Incineration

Second main source of the Dioxin pollution that is the domestic incineration activities which were not present in recent past in the current; however this is going to be promoted day by day. At present four hospitals operating incinerator in Sindh province, Civil Hospital, JPMC, Zia uddin and Aga Khan University Hospital, one general purpose Private Incinerator at Korangi Industrial Area (GEL) and City government incinerator at Maywashah is being used to burn only hospital waste collected from 130 local hospitals, with following specifications

Waste introduce to burn in primary chamber: 215kg/15 min Type: dual chamber Primary chamber: 10000C Secondary Chamber: 8000C and retention time: 2sec Capacity: 1000kg/h Routine Practice: Operating temp: Running time: 2-3 hrs/day

Page 15 INTRODUCTION CHAPTER-1

Waste generated: 5-10% (being dumped at KMC land filled site)

1.4.3.3 Open Pit Burning of Solid Waste

Practices like open pit burning of waste are very common all over country. There are over 100 truck/day of all kind of waste collected from city dumped and uncontrolled burned near Koragi Creek area for land unplanned filled (Figure-1.4).

1.4.3.4 Electronic Waste Burning at Shershah

There are reports on the activities of uncontrolled open burning of huge amount of electronic waste at Shershah. Sites has been visited to collect the information on the burning of electronic waste in the area It is hard to locate electronic waste burning site because electronic waste burning is not a routine practices and not a specific site or single site to burn waste. The routine practice is to dump and burn with general waste. The general waste burning is everywhere in the city.

As no legislation covering dioxins or other unintentional POPs exists in Pakistan, little attention has been given to the issue. No inventories on the potential sources have been compiled (Khan 2005). Lack of awareness and poor implementation of the environmental legislation and irresponsible and casual attitude towards environment issues may augment the environmental problems in the country. Pakistan has a number of industries in the fields such as metallurgy, pulp and petrochemicals, which might be substantial emitters of unintentional POPs.

1.4.4 Poly Aromatic Hydrocarbon (PAHs)/Oil Pollution

Major sources of PAHs pollution along Pakistan coast are effluent discharges from oil refineries, mechanized fishing boats and the cleaning of bilges and tank washing by the large number of merchant vessels as well as oil tankers that pass through the EEZ of Pakistan yearly 2500 oil tankers carry 33 million tons of crude oil (UNSCAP, 1996), oil terminals at Karachi Harbour, Port Qasim, and occasional oil spills.

Page 16 INTRODUCTION CHAPTER-1

The two oil refineries; National Refineries Ltd (NRL) and Pakistan Refineries Ltd (PRL); both located near Korangi Creek, Karachi discharge their oily waste waters in to the Korangi creek area. The volume of the effluent discharged from NRL is about 350,000 to 400,000 Million gallons /day (Ahmed, 1979). An equal amount is estimated for the effluent discharged by PRL on per day basis. The available data on the analyses of the final effluent from Sewage Treatment Plant No.1 and 2 show that 39.1 mg/l (or 3,558 kg/day) of Grease in the discharged sewage is contributing to the oily wastes of Karachi Harbour while 80 mg/l (or 7,280 kg/day) is discharged to Gizri Creek from domestic sources.

The quantitative estimate of oil pollution in Karachi coast and adjacent creeks is lacking. IUCN (1997) have reported some values conducted during their short term pollution survey programme. The survey found oil concentrations in seawater for the Karachi coastal waters ranging between 0.9 to 12.5 g/l with highest value from Gizri Creek Mouth indicating influence of Refinery Effluent. Similarly, the oil residue concentrations for the sediments from coastal areas ranged between 0.7 to 208 g/l with highest values reported from Karachi Harbour (167 g/l) and near ship anchorage area offshore 208 g/l, (IUCN, 1997) indicating the influence of oily wastes from oil tanker traffic, shipping and, oil terminals in Karachi Harbour and oily wastes from Karachi City.

Visual observations, however, indicated considerable amount of oil and tar ball resulting from shipping traffic in the Karachi Harbour, parts of Korangi Creek and in the Port Qasim. Most affected areas include Chinna Creek, Backwaters of Mauripur and Sandspit within Karachi Port Trust area and Boat Basin area associated with Karachi Harbour and parts of Gizri Creek due mainly to lack of adequate flushing of the waters in those areas. This has resulted in virtual loss of the habitats within the oil polluted parts of coastal area for most of the marine benthic species.

Page 17 CHAPTER 2 REVIEW OF LITERATURE

REVIEW OF LITERATURE CHAPTER-2

The Persistent, Bio-accumulative and Toxic substances (PBTs), specific group of organic chemicals, are persistent in environment, bio-accumulative in food chain and posses toxic characteristics. These distinctive properties pose a unique threat to human health and environment (Holoubek, et al., 2000).

These groups of synthetic chemicals are among the most dangerous pollutants released into the environment by human industrial and agricultural activities (Harald, et al. 2000). These chemicals persist in the environment and degradation by biological, photochemical or chemical means is very poor (Khan 2004). They are semi-volatile in nature hence their persistence and movement depend up the environmental condition of an area. They can also travel thousands of miles from their original source through a process of evaporation. The process of movement and settlement in remote area scientists are called “grasshopper effect” (eces.org). The unique properties of PBTs create local, regional or global concern (Petr and Holoubek, 2002, Wallack et al., 1998).

Polycyclic Aromatic Hydrocarbons (PAHs) are produced by combustion of organic compounds also classified as persistent organic compounds. Their occurrences are related to anthropogenic activities and contamination in aquatic environment is serious problem especially in high-density industrial areas (Wong, et al., 2003). Polycyclic Aromatic Hydrocarbons (PAHs) has been recognized as environmental contaminants found in almost all compartment of the global system (Verweij et al. 2004).

United Nations Environment Programme with the cooperation of International Forum for Chemical Safety (UNEP, 1996) identified group of most hazardous persistent

Page 19

REVIEW OF LITERATURE CHAPTER-2 organic pollutants. These are synthetic organic chemicals more commonly known as POPs. The Stockholm Convention on Persistent Organic Pollutants (POPs) list twelve priority pollutants that include eight agrochemicals (aldrin, dieldrin,endrin, DDT, chlordane, heptachlor, mirex and toxaphene), two types of industrial chemicals (PCBs and hexachlorobenzene) and two families of unintentionally produced by-products during incomplete combustion of chlorine and chlorine-containing substances (dioxins and furans) called un-wanted POP (U-POPs) (Wong et al., 2003). The PAHs are also unintentionally produced as by-products (U-POPs) of incomplete combustion, mostly waste incinerators and chemical processes that involve organic matter and chlorine (Khan, at al., 2004).

Most of the PBTs were originally made for noble purposes like combating malaria but due to unique qualities persistence and toxic make them potentially hazardous for human health and environment. These compounds having low water solubility and high lipid solubility therefore they are prone to bioaccumulate and causing toxic and harmful impact (UNEP, 2002).

In the view of wide spread use and toxicological impact of these contaminates, the experts around the globe strongly recommend to monitor the fate and distribution of these chemicals in the environmental compartment for the prediction of their behaviour in the various environmental conditions, deposition/emission processes, transformation processes and bioavailability in the ecosystems (Holoubek, et al., 2000). The pollutants do not recognise national or international boundaries. During the May 22, 2001 Stockholm convention on Persistent Organic Pollutants (POPs) was drafted to protect of human health and environment and entered into force on May 17, 2004. The convention is signed by 151 countries and 122 countries have ratified it (CEC.org, 2003). Pakistan signed the Convention on December 6, 2001.

From the early work of Jensenet al., 1969 has elaborated that the numerous chlorinated and other compounds are present in substantial quantities, not only in marine sediments, but also in invertebrates and animals around the world. During the last decades the scientists around the world have revealed global contamination of aquatic

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REVIEW OF LITERATURE CHAPTER-2 and terrestrial environments by these toxic contaminates (Tanabe et al., 1994; Brydon et al., 1995).

The PBTs have been associated with adverse impact on marine ecosystem, human health and environmental. These contaminate may impair reproduction, endocrine disruption, immune suppression and cancer (UNEP, 1996). PBTs linked to various cancers (breast, prostate, endometriosis, etc.), reproductive discrepancy (infertility and sex linked disorders, declining sperm counts, fetal malformations) and neurobehavioral impairment and immune system disfunction.

The harmful effects of PBTs in various compartment of coastal Marine environment have been studies by various scientists around the world. Exposure of PBTs contamination may influence population decline in marine mammals (Tanabe et al., 1994), Elevated levels of DDT, dieldrin, endrin and other pesticides inhibit growth of many phytoplankton and increased mortality rate has also been observed (Bonsdorff et al., 1997), Plankton accumulates 200,000 times more PCBs than the surrounding water, POPs are known to be growth retardant for oysters and other mollusks (Bishop Paul L. 1983), Shrimp mortality can occur at DDT concentration up to 0.2pbb (ESCAP Task Force 1971). Lethal concentration for fish is 0.06 to 0.94 ppb. Decline in fish population in many area have been correlated with PCBs contamination (Bishop Paul L. 1983), According to National Academy of Science (NAS) USA, 1999 POPs (DDT) adverse impact on the immune systems of seals and dolphins, POPs can also affect the chemical messenger system in organisms, compromising reproduction and development (WWF- report), Several pesticides kill marine life, even at very low concentrations (Bishop Paul L. 1983), marine mammals, particularly cetaceans, are among the most vulnerable organisms to long-term toxicity of hazardous man-made chemicals (Tatsukawa et al., 1992). POPs are non-polar molecules that can accumulate in fatty tissues and biomagnify in the higher trophic levels of the food chain (Shaw and Connell, 1986a, b), several other studies worldwide reported high levels of POPs in marine biota and their impacts.

The atmospheric deposition and surface run-off are main transport routes into coastal marine environments. These organochlorines are relatively volatile and liable to

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REVIEW OF LITERATURE CHAPTER-2 transport long-distance through atmospheric pathways (Wallack et al., 1998). These contaminants are detection in areas such as the Arctic, where they have never been used or produced, at levels posing risks to both wildlife and humans (Barrie et al., 1992 and Mulvad et al., 1996).

1.1 Organochlorine Pesticides

Organochlorine pesticides (DDTs, HCHs, CHLs, HCB and cyclodienes) with a circular structure in which hydrogen molecules substituted by chlorine give them a unique potteries as described earlier (Haynes and Johnson, 2000). These compounds having low water solubility and high lipid solubility are liable to bio-accumulate and pose toxic effects. These are considered to act as hazardous chemicals for humans and wildlife and/or environmental health (Colborn and Smolen, 1996).

Coastal sediments have tendency to sobered wide range of contaminates and act as continuous source of anthropogenic pollutants find its way into the ocean and biota. The applied pesticides reach to the coastal marine environment through surface run-off, leaching and vapor phase ultimately accumulate and settle in the bottom sediments. Several studies around the world have reported the presence of organochlorine pesticides sources and contaminations in the coastal marine environment (Fowler 1990, Rubinstein et al. 1990, Vartiainen et al. 1995, Husøy et al. 1996, Rose et al. 1997, Smokler et al. 1979, Shailaja & Sarkar 1997, Brown 1997, 1988c, 1991, Sericano et al. 1990, Mangani et al. 1991, Takeoka et al., 1991; Iwata et al. 1994, Pereira et al. 1994, Sarkar et al., 1994, Van Der Oost et al.1996, 1997, Pearson et al. 1998, Nhan et al. 1999, Kang et al. 2000, DouAbul and Al-Shiwafi 2000, Behnisch et al. 2001, Eljarrat et al. 2001, Frignani et al. 2001, Guruge and Tanabe 2001, Khim et al. 2001, Liu et al. 2001, 2003, Pandit et al., 2001, 2002, 2006; Ma et al. 2001, Zhou et al. 2001, 2002; Billiard et al. 2002, Choi et al. 2002,Mai et al. 2002, Doong et al. 2002; Assem O. Barakat et al, 2002, de Mora et al. 2001, 2004, Fillmann et al. 2002, Fillmann et al. 2002, Torres et al. 2002, Fent 2004, Koh et al. 2004, Galanopoulou et al. 2004, Wurl and Obbard 2005, Rajendran et al 2005, Chen et al. 2006, Hong et al. 2006, Menone et al. 2006, Mostafa et al 2007, Chi et al. 2007, Domingo and Bocio 2007, Götz et al. 2007, Zennegg et al. 2007, Hong et

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REVIEW OF LITERATURE CHAPTER-2 al. 2008, Wang et al. 2008, Pan et al. 2008, Chi et al. 2009, Moon et al. 2009, de Mora et al. 2010, Moon et al. 2010, Piazza et al. 2010, Marin et al. 2011, Pazi et al. 2011, Storelli et al. 2011, Kuranchie-Mensah et al. 2012, Leonel et al. 2012).

1.2. Dioxin and Dioxin-like PCBs

The Polychlorinated Biphenyls (PCBs) was introduced in 1929 as a very popular industrial chemical, mainly used as a flam-retardant, cooling agent in transformers and capacitors, also use in many industrial process as lubricants, hydraulic fluids, cutting oils, adhesives, liquid seals, coating wood and plastics to prevent them from flam. Also used in paints, varnishes, inks and pesticides and carbonless copy paper (Khwaja and Petrlik, 2005). Soon after it was recognize as dangerous chemical by Soren Jensen a Danish chemist

Un-intentional by-products include the highly toxic dioxins and furans are produced by human activities and are also occurring naturally (IPEP, 2005). They are introduces into the environment naturally by incomplete combustion of organic matter such as forest fires or volcanic activity. Un-intentional by products include the highly toxic dioxins and furans are produced by human activities and are also occurring naturally. The natural sources could be combustion of forest fires and volcanic activity. These contaminations are not industrial substances or involved in industrial processes, except in small amounts produced for research purposes (green-facts.org, 2006). However these are unintentionally produced during industrial processes dealing with organic chemicals and chlorine, they are introduced in an environment via municipal and domestic incineration and combustion processes (green-facts, 2010).

As with the diversity of sources for PBTs/POPs, the factors influencing persistence are also unique in the country. Typical tropic hot humid climate of the country may influence bio-availability, biological uptake, persistence and effect different from other climate country. Evaporation and photo POPs transformation are also more and the native biota involved in primary conversion are also more diverse. The scientists around the world are addressing the issues related with contaminants (Dannenberger and Lerz 1996, Brown

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REVIEW OF LITERATURE CHAPTER-2

1997, Holoubek 1999, Müller et al. 1999, Nhan et al. 1999, Haynes et al. 2000, Zhou et al. 2000, Gaus et al. 2001, Khim et al. 2001, Lee et al. 2001, Yuan et al. 2001, Barakat et al. 2002, Chen et al. 2002, Fillmann et al. 2002, Magi et al. 2002, Moore et al. 2002, Readman et al. 2002, Zulin et al. 2002, Carvalho et al. 2003, Fu et al. 2003, Lombaert et al. 2003, Monirith et al. 2003, Sanpera et al. 2003, Zhou and Maskaoui 2003, de Mora et al. 2004, Doong and Lin 2004, Kilemade et al. 2004, Stein et al. 2004, Verweij et al. 2004, Zhang et al. 2004, Boehm et al. 2005, Chen et al. 2005, Guzzella et al. 2005, Ke et al. 2005, Liu et al. 2005, Peris et al. 2005, Rajendran et al. 2005, Wurl and Obbard 2005a, b, Xue et al. 2005, Yang et al. 2005a, Yang et al. 2005b, Barlas et al. 2006, Bhosle et al. 2006, Bícego et al. 2006, Hong et al. 2006, Pocar et al. 2006, Wurl and Obbard 2006, Zhang et al. 2006, Zhou et al. 2006, Feng et al. 2007, Minh et al. 2007, Mostafa et al. 2007, Ohji et al. 2007, Uneyama et al. 2007, Wang et al. 2007, Xu et al. 2007, Zaghden et al. 2007, Zhang et al. 2007, Liu et al. 2008, Malakhova and Voronov 2008, Mansour 2008, Saito and Alino 2008, Sarkar et al. 2008, Tremolada et al. 2008, Arai and Harino 2009, Hu et al. 2009, Jiang et al. 2009, Kim et al. 2009, Lü et al. 2009, Malik et al. 2009, Shen et al. 2009, Tan et al. 2009, de Mora et al. 2010, Hu et al. 2010a, Hu et al. 2010b, Wang et al. 2010, Yang et al. 2010, Zhao et al. 2010, Chen et al. 2011, Cui et al. 2011, Eqani et al. 2011, Hiller et al. 2011, Kohušová et al. 2011, Okay et al. 2011, Sudaryanto et al. 2011, Yang et al. 2011, Mishra et al. 2012, Sarkar et al. 2012, Yang et al. 2012).

1.3 Poly Aromatic Hydrocarbon (PAHs)

The industrial revolution in late 1700s started the increasing use and dependency of mankind on fossil fuels to generate electricity, industrial processes and power transportation. As a result the oil wastes and products can be found in different compartments of the environment (Vilanova et al., 2001).

Polycyclic Aromatic Hydrocarbons (PAHs) has been recognized as environmental contaminants found in almost all compartment of the global system (Verweij et al. 2004). Preliminary results of PAHs contamination in the coastal environment from human included activities, although small concentrations of PAHs are also produced by the natural processes. PAHs are undesirable group of substances produced as a result of

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REVIEW OF LITERATURE CHAPTER-2 incomplete/inefficient combustion of organic material. They are non-polar, lipophlic aromatic compounds containing two or more fused arenes structures. The persistence increased with increase in aromatic rings. However low molecular weight PAHs are more soluble and have less affinity for surfaces with significant acute toxicity than high molecular weight PAHs. According to the Neff, 1979; Witt, 1995, the higher molecular weight PAHs with a 4, 5 or 6 ring structure are more carcinogenic in nature. These are produced during high temperature combustion of organic matter, whereas low temperature burning results in PAHs with a 2 or 3 ring structure (Readman et al 2005). It is also reported by Nemr et al. 2006 et al., that PAHs also synthesized naturally by organisms, such as bacteria, algae and fungi.

Coastal marine areas receive these pollutants from land base sources such as industrial effluents, sewage discharges, surface run-off and cleaning of wharfs and pilings and power boating activities oil spillages and deposition from the atmosphere (Sander, 1994). The PAHs in the marine environment are incorporated into the sediments and suspended matter and it gets assimilated into the marine biota. Therefore, highest concentration in sediment, intermediate in biota and lowest in the water column is expected for the coastal marine environment (CCME 1992). It is significant to quantify the level of PAHs pollution in the marine environment and their impact on the marine organisms and human health (Wurl et al., 2005).

The presence and detection of PAHs in food items like fishes, meat, milk, vegetables and fruits increasing the potential for dietary exposure of human to these compounds. The sources of compounds are predominantly from environmental pollution and some food processing. The presence of PAHs on food, mainly like fruits and vegetables due to contaminated soil, air and water mainly by processes of PAHs adsorption, their deposition and bioaccumulation (Martens et al., 1997; Chung et al., 2008). In 2003, Camargo and Toledo, reported the presence of different PAHs in the fruits and vegetables including; cabbage, lettuce, tomato, grape, pear and apple. Among different PAHs, benzo(a)anthracene was the most abundant being found in 89% of the samples. DouAbul et al., (1997) reported the presence of 40 different un-substituted and alkyle substituted PAHs in the fishes from Red sea. The concentration of the total PAHs

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REVIEW OF LITERATURE CHAPTER-2 was 422.1 ng/g of dry weight edible muscles of collected fish samples. The considerable amounts of different PAHs were detected in the gall bladder and liver of the two different species of fish (Pointet and Milliet, 2000). Reinik et al., (2007) studied presence of different PAHs including benzo(a)pyrene in meat and meat products and reported the high concentration of these compounds in smoky and grilled cook meats.

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CHAPTER 3 Distribution and Fate of Chlorinated Pesticides in Coastal Marine Environment of Pakistan

Chlorinated Pesticides CHAPTER-3

3.1 Abstract

Surficial sediment and selected marine biota samples were collected from coastal area of Pakistan for the assessment of Organochlorine Pesticides (OCPs) contamination levels, sources and fate of the pollutants. Evaluations were also made with respect to the Sediment Quality Guidelines (SQGs) defined by CCME, (2002) for biological effects of these contaminants.

The result reveals that the residual concentration of sum of all analyzed Organochlorine Pesticides (OCPs) varied between >0.002-17.5 ng g-1 dry wt. with a mean concentration of > 3.2175 ng g-1 dry wt. in the sediment samples collected from various localities of coastal area. The prevalent DDTs, HCH, CYC and HCB concentrations were found significantly different in seaward stations of KH (H14 to H16) and sampling sites within the creek environment (C4to C6). However, contamination levels in the vicinity of pollution discharge area (H1 to H3, C1 & C2, C8, C11, C15 and C20) reflecting localized pollution effects along the coast.

The residue of DDT, mainly its metabolites DDE and DDD were detected in most of the sediment samples in significantly higher concentrations. It was observed that total DDTs concentration (>0.01 to 12.3 ng g-1 dry wt.) contributed highest amongst all analyzed OC pesticides contaminants. The higher proportion (>40–95%) of pp′-DDE in most the sediment samples suggests old inputs of DDTs in the environment. Ratio of ∑DDT and DDT were observed >1 in the sediment of all locations which also reflect the discharges of DDT were minimum or were not discharged in the area. That may be attributed to the restriction being put into practice to the use of DDTs, as Pakistan switched over to natural pest control or using safer formulas. Measured concentrations of HCHs were lower than DDTs that might be due to disparity in their physicochemical properties and stability in the marine environment.

It was observed that mean residual concentration of various OC Pesticides are considerably higher in semi-enclosed areas of the Gizri creek and upper Harbour area. The elevated levels of OC pesticides were related to untreated effluents discharges from

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Chlorinated Pesticides CHAPTER-3 coastal industries, domestic sewage and land run off in the coastal environment. However these elevated concentration did not exceed the levels of Severe Effects Level (SEL) and concentration of individual Organochlorine Pesticides were also found much lower than the levels defined by Canadian Sediment Quality Guidelines (CCME, 2002) for the protection of Aquatic Life.

The assessments were also made on the residues contamination levels of Organochlorine Pesticides in the variety of marine biota samples collected from selected localities of the coastal area. It was observed that DDT and its metabolites were detected in most marine biota samples collected from coastal area. Although organochlorines were detectable in Mollusc Perna viridus, oysters, shrimps, crab, Sepia sp. and a variety of fish collected from selected sampling sites along the coast, however, overall concentrations were found much lower than levels reported from other Asian countries.

Present results reveal the declining trend on the environmental burden of OC pesticides in coastal marine environment of Pakistan.

3.2. Introduction

Organochlorine pesticides (DDTs, HCHs, HCB and cyclodienes) with a circular structure in which hydrogen molecules have been substituted by chlorine ((Monirith et al., 2003, Galanopoulou et al., 2005) makes them highly persistent in the environment and degradation by biological, photochemical or chemical means is very poor (Haynes and Johnson, 2000). These compounds having low water solubility and high lipid solubility are liable to bio-accumulate and pose toxic effects on the living organisms. These are considered as hazardous chemicals for humans and wildlife and/or environmental health (Colborn and Smolen, 1996). Most are prone to long-range transport (G Fillman., 2002). Organochlorine pesticides have been of great concern due to their occurrence in high concentration even in remote ecosystems where these chemicals never been used (Iwata et al., 1994; Guruge and Tanabe, 2001, Sarkar et al., 2008).

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Chlorinated Pesticides CHAPTER-3

The organochlorine pesticides have been associated with adverse human health impact and environmental effects, including impaired reproduction, endocrine disruption immuno-suppression and cancer (UNEP, 1996, Sarkar et al., 2008). The population decline in a number of marine mammals has related to the organochlorine contamination (Tanabe et al., 1994). The sources of these contaminants in the coastal marine environment could be surface run-off and atmospheric deposition because most of the organochlorine are volatile in nature, therefore they are liable to transport long-distance through atmospheric pathways and redistributed in an area where these contaminates may have not been in used (G Fillman., 2002).

It has been reported by Tanabe et al., (1994) that due to the low cost and versatility in industry and agriculture applications these chemicals are still in use for sanitation purpose in the developing countries. Over three million people are poisoned and about 200,000 die each year around the globe from pesticide poisoning and a majority of case registered from developing nations (WHO, 1990; FAO, 2000). It is also alleged that the incidence of pesticide poisoning may even be larger than reported, it is due to under-reporting, lack of data and misdiagnosis at most places (Forget, 1991, Sarkar et al., 2008).

Environmental pollution by toxic chemicals is a global problem, particularly Persistent Organic Pollutants (POPs), such as Organochlorine Compounds (OCs) (Monirith et al., 2003). The growing concern about these substances it is due to their bio accumulative nature and toxic biological impacts on wildlife and humans. Elevated levels of OCs have been detected in environmental compartment and biota (Iwata et al., 1993; Kannan et al., 1997; Tanabe, 2000; Tanabe et al., 2000; Monirith et al., 2003). The injurious effects most of these chemicals are linked to the occurrence of immunologic and teratogenic dysfunction, reproductive impairments and endocrine disruption at lower and higher trophic levels (Colborn and Smolen, 1996; Monirith et al., 2003).

The Organochlorine Pesticides were mainly used for agriculture purposes and industrial and domestic use for controlling harmful pests and micro-organisms (Zhou et al., 1996). These pesticides were effectively used to control the damages from insects,

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Chlorinated Pesticides CHAPTER-3 disease and weed and they also enhance the production of agricultural commodities. Despite its effective and diverse applications, the people are now aware of the hazardous health impact and environmental damage from overuse of the pesticides. These substances are persistence in environment and have tendency to accumulate in the living organism (Zhang et al., 2002).

Little attention has been made on monitoring and assessing the levels of contamination by these pollutants in the coastal marine environment of Pakistan. Although range of Organochlorine Pesticides (OCPs) have been used extensively for agriculture purposes in Pakistan. Almost all targeted chlorinated Pesticides have never been produced along the coastal area of Pakistan. However some of them such DDT, HCB and Dieldrin were formulated locally at various places along Karachi coast (Khan 2003). Moreover the large scale use of these pesticides have been banned or restricted in Pakistan for over 15 years (Khan, et al., 2003). The long half lives and potential ecotoxicological impacts of these chemicals warrant essential to monitor these contaminants in the coastal environment (Wong, 2006; Li et al., 1999; Dragan., 2006).

To examine the contamination levels of organochlorine pesticides 63 sediment samples were collected from various localities along the Pakistan coast. For the present study sediments were chosen as main environmental media to assess the contamination levels because it act as temporary or long-term sinks for most of the anthropogenic contaminants (Zhou and Maskaoui, 2003). The applied pesticides can be transported through surface run-off, leaching and vapour phase, which ultimately find its way into the coastal marine environment where these chemicals bind and settle in the bottom sediments and eventually accumulate in biota (Sarkar et al., 2008). Hence bottom sediments is continuous source of particle bound contaminants that have deposited over a longer period of time. Several studies have reported the contamination of OCs in the sediments around the world (Smokler et al. 1979; Brown et al. 1998; Marchand et al. 1988; Sarkar and SenGupta, 1987, 1988c, 1991; Fox et al. 1988; Sericano et al. 1990; Mangani et al. 1991, Takeoka et al., 1991; Iwata et al. 1994; Pereira et al. 1994; Sarkar et al., 1994, 1997; Van Der Oost et al.1996; Connell et al. 1998, Connell et al. 1998; Nhan

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Chlorinated Pesticides CHAPTER-3 et al. 1999; Nhan et al. 1999; Kang et al. 2000; DouAbul and Al-Shiwafi 2000; Guruge and Tanabe 2001; Kang et al. 2000; Khim et al. 2001; Liu et al. 2001, 2003; Pandit et al., 2001, 2002, 2006; Ma et al. 2001, Zhou et al. 2001, 2002;, Mai et al. 2002, Doong et al. 2002; Hong et al. 2006, Assem O. Barakat et al, 2002, de Mora et al. 2001, 2004, Fillmann et al. 2002, Fillmann et al. 2002, Torres et al. 2002, Galanopoulou et al. 2004, Wurl and Obbard 2005, Rajendran et al 2005, Alaa R. Mostafa et al 2007) indicating the presence of their sources and contaminations. FAO/WHO ADI standards: 0.01 mg/kg of DDE oral LD50 is 300 to 500 mg/kg.

Elevated levels of DDT, dieldrin, endrin and other pesticides inhibit growth of many phytoplanktons and increased mortality rate has also been observed. Plankton accumulates 200,000 times more PCBs than the surrounding water. POPs are known to be growth retardant for oysters and other mollusks (Bishop Paul L. 1983). Shrimp mortality can occur at DDT concentration up to 0.2pbb, (ESCAP Task Force 1971). Mortality of many Crustaceans, such as shrimps and crabs and zooplanktons has been observed at PCBs levels in the range of part per billion. Lethal concentration for fish is 0.06 to 0.94ppb. Decline in fish population in many area have been correlated with PCBs contamination (Bishop Paul L. 1983). According to National Academy of Science (NAS) USA, 1999. POPs (DDT) adverse impact on the immune systems of seals and dolphins. POPs can also affect the chemical messenger system in organisms, compromising reproduction and development (WWF- report). Several pesticides kill marine life, even at very low concentrations (Bishop Paul L. 1983). Several other studies worldwide reported high levels of POPs in marine biota and their impacts.

The present work is the first of its kind to describe in detail the fate and distribution pattern of the residual concentration of Organochlorine Pesticides (OCPs) along the Karachi coast. The objective was to provide a comprehensive account of the distribution of organochlorine pesticides in coastal environment.

Surface sediment and marine biota (Mollusc Perna viridus, oysters, fishes, shrimps, crab, Sepia sp) samples were collected from selected sites along the coast (Figure-1.1) representing various environmental conditions for the estimation of residual

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Chlorinated Pesticides CHAPTER-3 levels of Organochlorine Pesticides (OCPs). The investigated area included Balochistan Coast (BC), coastal area (Karachi Harbour (KH), Clifton beach (CB), Gizri creek (GC), Korangi creek (KC) and Gharo/Phitti Creek system). These localities were selected due to diversity of potential inputs from various sources such as untreated industrial effluents, wastewater and sewage discharges as well as agricultural runoff. The coastal sediments were expected to have elevated levels of wide variety of Organochlorine Pesticides (Wurl et al., 2005). Therefore collected sediment samples were analyzed for a broad range of Organochlorine Pesticides described in Table-3.1.

Table-3.1 Organochlorine Pesticides (OCPs) Selected for the Present Study

 DDT (1,1,1-trichloro-2, 2-bis (p-chlorophenyl) ethane) and it metabolites o (DDD) 1,1-dichloro-2, 2-bis (p-chlorophenyl) ethane o (DDE) 1,1-dichloro-2, 2-bis (chlorophenyl) ethylene,

 -HCH (Hexachlorocyclohexane and its isomers) o α—HCH o β-HCH o Lindane (γ-HCH)

 HCB (Hexachlorobenzene)

 Polychlorocyclodienes o Aldrin o Dieldrin o Heptachlor o Heptachlor epoxide The risk assessments were performed by comparing the analyzed concentrations of OC pesticides in the sediment with the corresponding guide-lines promulgated by sediment quality standards of the USEPA and Canadian Council of Ministry of the Environment. The levels of OCPs were compared to available data for other countries in Asia, Europe, USA, Canada, Australia and neighbouring country India and UAE sharing Indian Ocean.

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Chlorinated Pesticides CHAPTER-3

3.3 Materials and Methods

3.3.1 Sampling Location in the Study Area

For the present study to evaluate the contamination status of PBTs 61 sampling sites were selected in the vicinity Coastal area of Pakistan Boarding Northern Arabian Sea (Figure-3.1). The study area included Karachi Harbour and Indus Deltaic Creek system. GPS (GARMIN 12 XL) was used to mark the geographical location of the sampling site.

Figure-3.1 Sampling Location of Sindh Coastal Area

Figure-3.2 Sampling Location along Balochistan Coast

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Chlorinated Pesticides CHAPTER-3

3.3.1.1 Sampling Location along Sindh Coast

3.3.1.1.1 Sampling Location in Karachi Harbour Area

Tidal creeks, mangrove swamps and intertidal mudflats represent main feature of the harbour area that are expected to be under the influence of considerable urban and industrial pollution. Major coastal Industries are located in Karachi harbour area, and all their untreated effluents together with domestic sewage find their way into the west of Karachi harbour and pollutants find their way into the main navigable channel called Manora channel.

Table-3.2. Sampling Locations in Karachi Harbour Area

Sampling LAT LONG DEPTH (meters) locations B1 66.8142 24.8386 Hand pick sample B2 66.8216 24.8415 hand pick sample

B3 66.8236 24.8460 hand pick sample B4 66.7738 24.8405 hand pick sample B5 66.7738 24.8405 hand pick sample

H1 66.9849 24.8711 1.0 H2 67.0245 24.8818 1.5 H3 66.9503 24.8554 1

H4 66.9234 24.8369 hand pick sample

H5 66.9406 24.8348 4 H6 66.9580 24.8352 4.2

H7 66.9743 24.8461 3

H8 66.9842 24.8411 7

H9 66.9658 24.8323 5

H10 66.968 24.8178 4

H11 66.9743 24.8092 10

H12 66.9801 24.7991 4

H13 66.9965 24.8131 5

H14 66.9975 24.7966 7

H15 66.9923 24.7890 7

H16 66.9763 24.7715 12.5

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Chlorinated Pesticides CHAPTER-3

Sixteen (16) sampling station that included upper, lower and navigation channel were selected for present investigation (Figure-3.2 and Table-3.2). Sampling site were located Upper harbour area (stations H1, H2 & H3), these sites area under the influence of effluent discharge, Mid harbour area (stations H4, H5, H6, H7 & H8) few km away from contaminated area within the upper harbour zone: Lower harbour area (stations H9, H10, H11 & H12) sites between jetty, upper harbour and main navigational channel, and the open sea (stations H13, H14, H15 & H16).

Figure-3.3. Map showing Sampling Location in the Vicinity of Karachi Harbour

3.3.1.1.2. Sampling Location in Indus Deltaic Creek System

Organochlorine Pesticides evaluations were made in the sediment samples collected from 25 (station C-1to C-25) were selected sides of Indus Deltaic creek system along Karachi coast (Table-3.3, Figure-3.3).

3.3.1.1.2.1. Sampling Location in Gizri Creek:

Three (3) locations (station C-1 to C-3) in the vicinity of Gizri Creek Area (extreme end, mid/centre, mouth (Gizri Korangi Creek junction).

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Chlorinated Pesticides CHAPTER-3

Figure-3.4 Sampling Location of Indus Deltaic Creek System

3.3.1.1.2.2. Sampling Location in Korangi Creek Area:

Twelve (12) sampling sites (stations C4 to C15) were selected from within a radius of 20 km in length Korangi Creek. The creek is located in the south-east of Karachi at a distance of about 12 km from Karachi Harbour. At the north-eastern end KC is connects with the Phitti Creek and Kadiro Creek, as well as with Rehri fishing village whereas, south-western side drain in open sea. KC received large quantities of liquid wastes from the southern parts of Karachi via The Gizri Creek close to its connection with the open sea. It has well-established coastal fishing village at Goth Ibrahim Hyderi and Goth Rehri with a fishing jetty.

3.3.1.1.2.3. Sampling Location in Gharo/Phitti Creek System

The upper Indus deltaic creek system consists of three creeks Gharo Creek, Kadiro Creek and Phitti Creek. All three are connected in a series starting from Gharo Creek at the north-eastern end to the Phitti Creek at the south-western end; these creeks are located at 22.3 km from Karachi. Nine (8) sampling station (C15 to C22) were

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Chlorinated Pesticides CHAPTER-3 randomly selected within the 28 km long and 250 to 2,500 m wide creek system. Chara Creek was also sampled at two locations (C23 to C24).

Table-3.3 Sampling Locations along the Indus Deltaic Creek System Location Lat Long Depth (meter) Clifton Beach B1 24.7810 67.0431 Hand pick B2 24.7604 67.0606 Hand pick B3 24.8001 67.0193 Hand pick Gizri Creek C1 24.7974 67.0824 4.5 C2 24.7800 67.1000 7 C3 24.7533 67.1000 10 Korangi Creek C4 24.7610 67.1043 12 C5 24.7640 67.1211 10 C6 24.7802 67.1608 7.5 C7 24.7420 67.1657 6.5 C8 24.7838 67.1827 10 C9 24.7995 67.2042 5 C10 24.7913 67.2140 3.5 C11 24.8082 67.2156 4 C12 24.8123 67.2282 3 C13 24.7660 67.2413 5 C14 24.7898 67.2618 13 Gharo/Phitti C15 24.7653 67.3102 3.5 Creek System C16 24.7683 67.3888 7 C17 24.7607 67.4658 2.1 C18 24.8123 67.2282 9 C19 24.7660 67.2413 3 C20 24.7660 67.2413 14 C21 24.7898 67.2618 9 C22 24.7653 67.3102 7 C23 24.7683 67.3888 10 C24 24.7683 67.3888 6

3.3.1.1.3. Sampling Location of Open Sea Area

The open sea area of coast west consist Manora, Sands spit and Hawks bay. These are sandy beaches. From Bulleji to Cape Monze, the coastal area consists of hard substrate and shale cliffs. The unconsolidated sandy clays is found at beyond Hawks Bay

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Chlorinated Pesticides CHAPTER-3 towards west up to the Cape Monze. These sites are exposed to coastal weathering and erosion. Four sampling location were selected for the present study as described in the Table-3.1 and Figure-3.1.

3.3.1.1.4. Sampling Location of Clifton Beach

The Clifton beach is largely composed of dark, grey silty materials with minute flakes of mica. Further east of Clifton there is agglomeration of Gizri hills. Three (3) sampling location were selected from the locality for the present study as described in the Table-3.1and Figure-3.1.

3.3.1.2. Sampling Location of Balochistan Coast

3.3.2. Sample Collection

3.3.2.1. Sediment Sample Collection

The Sediment samples were collected from selected localities along the coast by a using Grab sampler (Peterson type) during the year 2002 and 2003; details about the sampling sites are shown in Table-3.1, Figure-3.1. A handheld GPS (GARMIN 12 XL) was used to mark of the geographical location of the sample site.

To minimize site intra-variability and to ensure that the sediment collected was truly representative of investigated site, each sample was homogenized composite made of equal fractions of five sub-samples collected within a radius of approximately 10 m. (Ni Shuilleabhain et al., 2003). All samples were stored in pre-cleaned and solvent rinsed stainless steel container, kept cool on ice and transported to the laboratory and refrigerated within 2 to 4 h.

3.3.2.2.1 Biota Samples

A variety of marine biota was collected from selected coastal area. The biota samples were wrapped in pre clean aluminium foil and kept in polyethylene bags. They were transported to NIO-lab- on ice to prevent further degradation, and stored in a deep

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Chlorinated Pesticides CHAPTER-3 freezer immediately after reaching the laboratory. In the laboratory, the frozen samples were thawed and biometric measurements were made. Three or more biota samples from each location were pooled, homogenized, transferred into clean glass bottles and freeze dried.

3.3.3. Sample Preparation and Transportation

The sediment samples collected were transported to NIO Lab in ice boxes and stored in freezer till further processing. Later on samples were freeze dried by using LABCONCO, Freeze Dry System. Freeze dried samples were homogenized using mortar and pestle and wrapped up in solvent-washed aluminium foil and posted to National Research Centre for Environmental Toxicology (EnTox), Brisbane Australia, for the estimation of PAHs, OCPs.

As standard procedure and protocol all the glass wares and sampling tools were pre cleaned and rinsed with solvent (acetone and hexane)

3.3.4. Estimations of Sediment Organic carbon

Total organic carbon (TOC) was determined at the QHSS laboratory according to a standardised procedure described by Gaus et al., 2001. The acid catalysed digestion (10 o % HCl, 1 % FeCl2 at 70 C) was used to remove inorganic carbonates. After removal of carbonate, sample was dried and combusted in the LECO induction furnace for the determination of carbon dioxide (CO2) using LECO WR12 CO2 detector.

3.3.5.1 Analysis of Organochlorine Pesticides

3.3.5.2 Pre- treatment /Sample preparation for the Identification and Quantification

For the present study the analytical method routinely used in Environmental Toxicological Lab and Queensland Health Lab was adopted for identification and quantification of Organochlorine Pesticides (OCPs) such as DDT and its metabolites

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(DDTs: p,p'-DDT, p,p'-DDD, and p,p'-DDE), chlordane compounds (CHLs: trans- chlordane, and cis-chlordane), hexachlorohexane isomers (HCHs: α-HCH, β-HCH, and γ- HCH).

Briefly 10g of freeze dried sediment sample was extracted using Soxhlet extraction technique for 18hrs with 200ml of acetone and hexane mixture (1:1) in triplicate and one blank for each five set of sediment samples. The extracted samples were filtered over sodium sulphate and concentrated sample run through Florisil column clean up, eluted with the mixture of 120 ml 6% ethyl acetate and hexane and 100ml of acetone and hexane. The extract concentrated under a gentle stream of nitrogen up to 1 ml.

3.3.5.2. Instrumental Condition

GC MS, QP5050 GCMS equipped with a Shimadzu AOC-20i auto-sampler and a DB-5 fused silica capillary column (30 m × 0.32 mm ID, film thickness 0.25 μm). Purified helium was used as the carrier gas with a flow rate of 1.5 ml/min.

3.3.6. Recovery of Organochlorine Pesticides

Recoveries of OC pesticides through the analytical procedure were examined by spiking 1 mg/l of OCs standard into sediment sample. The results were 100±12% for HCHs, 94±5% for HCB, 103±5% for CHLs, 100±7% for DDTs. A procedural blank was run with every set of five samples to check for secondary contamination.

3.3.7. Statistical Analyses

Statistical tests were carried out with SPSS 18.0 for windows (Statistical Product Service Solutions, Chicago, IL, USA). Data were analysed for statistical significance by the Kruskal-Wallis Test and Mann–Whitney rank sum non-parametric test. Significant differences were determined at a 0.05 level of probability, using a 2-tailed test.

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3.4. Results and Discussion

3.4.1 Spatial Distribution of Organochlorine Pesticides (OCPs) in the Coastal Sediment.

Very little information is available on the contamination levels of OCPs in the coastal environment of Pakistan. Present study represents one of the first efforts to quantify sediment borne concentrations of various chlorinated pesticides (Table-3.1) along the coast (Figure-3.1). The result reveals that the residual concentration of sum of all analyzed Organochlorine Pesticides (OCPs) varied between >0.002-17.5 ng g-1 dry wt. with a mean concentration of > 4.5 ng g-1 dry wt. Figure-3.5 described the distribution of total Organochlorine Pesticides (ΣOCPs) median, inter-quartile range, range values and outliers in the sediment collected from selected coastal localities.

The highest mean levels observed within the close proximity of discharged area of Malir River within the Gizri creek (>7-17.5 ng g-1 dry wt.), Karachi Harbour area (>0.002- 7.2 ng g-1 dry wt.) in the discharge vicinity of Lyari River, Korangi Creek environment in the vicinity of Rehri Goth (>0.01-8.5 ng g-1 dry wt.).

Figure-3.5 Box plot showing distribution of total Organochlorine Pesticides (ΣOCPs) median, inter-quartile range, range values and outliers in the Sediment Samples Collected from Selected Coastal Localities.

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Large variability in the spatial distribution pattern of individual OC-pesticides was observed amongst the localities sampled. The DDE & DDD, α.HCH & Endrin and Lindane & Dieldrin showed similar behaviours along coast. Whereas distribution pattern of Aldrin was significantly different. DDTs were predominant contaminants in most of the sediment samples due to its less volatile and slow degradation nature in the aquatic environment (WHO 1989, Carvalhho et al., 1992). DDTs contributed >60% of total analyzed OCPs. Higher levels of CYCs were found in the sediment of Korangi creek station C11, CH-C and KA-C area. Relatively lower levels of ∑HCH levels (bdl-1.7 ng g- 1 dry wt.) were found along almost entire coast which may be due to the lower lipophilicity and particle affinity (Yang et al 2005, Fu et al., 2003). Whereas HCB levels (0.02-0.12 ng g-1 dry wt.) were relatively uniform (f=15.368 p<0.218) contributed lowest (1-20%) in OCPs concentration. The variation in the concentration of various OCPs levels in sediment of Karachi coast at various localities is delineated in the box-plots, median, inter-quartile range, range values and outliers (Figure-3.6).

Figure-3.6 Box-plots showing Distribution of total Σ DDTs, ΣHCHs, ΣCYCs and HCB median, inter-quartile range, range values and outliers in the Sediment Collected from Selected Coastal Localities.

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3.4.1.1. Spatial Distribution Pattern of Organochlorine Pesticide in Sediment of Karachi Harbour Area

A summary of concentrations of OCPs detected in sediment samples of Karachi Harbour area is shown in Table-3.4. The data on the OCPs obtained for Karachi Harbour sediments showed the presence of wide variety of Organochlorine Pesticides (OCPs) that included DDTs and its metabolites; HCH and its isomers (α, β & γ); Cyclodienes compound (mainly Aldrin & Dieldrin) and HCB (Table-3.4).

Table-3.4 Organochlorine Pesticide Levels (ng g-1 sed. dry wt.) in Sediment Samples of Karachi Harbour Area Pesticides Upper Mid Lower Navigation harbour harbour harbour channel (1 to 3) (4-8) (9-12) (13-16) p,p’-DDD 1.678 0.355 0.291 0.001 p,p’-DDE 2.373 0.537 0.386 0.002 p,p’-DDT 0.521 0.148 0.047 0.000 α-HCH 0.021 0.008 0.001 0.000 β-HCH 0.376 0.098 0.019 0.001 γ-HCH 0.085 0.004 0.000 0.001 HCB 0.140 0.064 0.013 0.000 Aldrin 0.273 0.241 0.030 0.000 Dieldrin 0.700 0.466 0.170 0.002 Endrin 0.008 0.001 0.001 0.000 Heptachlor 0.224 0.107 0.075 0.004 Heptachlor 0.116 0.135 0.039 0.000 Epoxide ΣDDTsa 4.573 1.041 0.724 0.003 ΣHCHsb 0.483 0.110 0.020 0.002 ΣCyCc 1.461 1.014 0.328 0.006 ΣOCPsd 6.517 2.165 1.071 0.010

ΣDDTsa= p, p’-DDE + p, p’-DDD + p, p’-DDT. ΣHCHsb= α-HCH + β-HCH + γ-HCH . ΣCycc= Aldrin + Dieldrin + Endrin + Heptachlor + Heptachlor Epoxide ΣOCPsc = ΣDDTs + ΣHCHs + HCB + ΣCyc.

The residual concentration of sum of all analyzed Organochlorine Pesticides (OCPs) varied between <0.013 to 7.22 ng g-1 dry wt. in the sediment samples (Figure- 3.7). The variation in the spatial distribution in the harbour sediment indicated highest concentration at stations H1, H2 and H3; which were in close proximity of the Lyari

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River discharges. The concentrations of almost all analyzed OCPs showed distinct gradient of contamination between upper and lower sites of Karachi harbour. The highest concentrations were found in the vicinity of the discharge area to Lyari River. The distribution of OCPs was strongly correlated with total organic carbon contents in sediments. (Feng, 2007)

Figure- 3.7 Distribution of ΣOCPs in Sediment Samples of Karachi Harbour Area.

As elaborated in Figure-3.8 the total concentration of DDTs (ΣDDTs) in sediment samples was higher than the total concentration of HCHs (ΣHCHs) and Cyclodienes (ΣCYC) in upper harbour area (samples H1 to H3). Concentration of DDT & its metabolites sharply decreased at other sites. The observed lower levels of HCHs/Cyclodienes in sediment sample collected from the coastal area probable it is due to the difference in the physicochemical and biochemical properties. The HCHs/Cyclodienes have higher water solubility, vapour pressure and biodegradability, and lower lipophicility and particle affinity compared to the DDTs (Rui et al., 2005, Yang et. al 2005). DDTs remains in the sedimentary environment longer than HCHs (Nhan et al., 2001).

The average concentration of ΣDDTs, ΣHCHs and ΣCyclodienes in sediment

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Chlorinated Pesticides CHAPTER-3 samples of Karachi Harbour area were found lower than the average concentration reported by Zhang et al, 2006 from Hong Kong (6.19 ng g-1 sed. dry wt. and 0.52 ng g-1 sed. dry wt.) and they were much lower than the ΣDDTs (37.6 ng g-1) and ΣHCHs (12.2 ng g-1 sed. dry wt.) levels found in soils of Pearl River Delta Region (Fu et al., 2003, Lin et al., 2009).

Figure-3.8 Distribution of ΣDDTs, ΣHCHs and ΣCyclodienes in the Sediment of Karachi Harbour Area.

3.4.1.1.1. Distribution of HCB in the Sediment of Karachi Harbour

Hexachlorobenzene (HCB) residues in sediment samples of Karachi Harbour area are presented in Figure-3.9. Maximum HCB residues 0.140 ng g-1 were found in Upper harbour area followed by mid harbour and lower harbour with 0.064 and 0.013 ng g-1 respectively. HCB residues were not detected in Navigation channel. HCB is a prevalent contaminant that has introduce in marine environment as a by-product during production of a variety of chlorinated compounds and other activities relate to pesticides and (PCNB, PCP, DCPA etc.) persist in sediment (Sanpera at. al., 2003, Wang, 2009 and Tobin, 1986). HCB concentration in the coastal sediment were found to be much low in comparison to level reported from coastal areas.

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Figure-3.9 Distribution of HCB in Sediment Samples of Karachi Harbour Area.

3.4.1.1.2. Distribution of Cyclodienes in the Sediment of Karachi Harbour

Residues were detected of five Cyclodienes insecticides namely Aldrin, Dieldrin, Endrin, Heptachlor and Heptachlor Epoxide. Highest residues of these insecticides were found in the Upper harbour area followed by mid harbour and Lower harbour. Most of the samples collected within the navigational channel were found residue free. Amongst Cyclodienes Dieldrin residues were highest because once released in the environment, Aldrin (0.273 ng g-1 sed. dry wt.) is rapidly metabolized to Dieldrin (0.7 ng g-1 sed. dry wt.). Highest level Heptachlor and its metabolite Heptachlor Epoxide were 0.224 and 0.116 ng g-1 sed. dry wt. respectively. Heptachlor Epoxide residues were lower than its parent heptachlor because Heptachlor Epoxide dissolves more easily in water although it can stay in sediment for many years. The highest average amount extractable Endrin, were 0.008 ng g-1 (Figure-3.10). These concentrations were very low compared to Bed sediments of Hawaii (Brasher and Anthony, 1998).

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Figure-3.10 Cyclodienes in Sediment Samples of Karachi Harbour Area.

3.4.1.1.3. Distribution of HCHs and its Isomers in the Sediment of Karachi Harbour Area

By analyzing the individual HCH isomers (Figure-3.11), it was found that β-HCH had the highest level of concentration among all the samples and this HCH isomer was higher because of its persistence in sediment. The persistence of β-HCH in sediment is mainly due to the higher Kow (log Kow =3.78) and lower vapor pressure value (3.6x10-7 mmHg, 20oC) (Zhang et al., 2006). These will make β-HCH easier to be absorbed to the sediment organic matter and less evaporative loss from sediments (Mackay et al., 1997). Furthermore, the spatial arrangement of Chlorine atoms in the molecular structure of β- HCH make it more persistence and resistant to microbial degradation (Middeldorp et al., 1996).

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Figure-3.11. Distribution of HCHs and its Isomers in Sediment of Karachi Harbour Area

3.4.1.1.4. Distribution of DDT and its Metabolites (DDE and DDD) in the Sediment of Karachi Harbour Area

Variability in distribution pattern of the mean concentration of DDTs levels were not much higher in Karachi Harbour area, however unusually higher levels of ∑DDT <3 to 5 ng/g dry wt had been found at stations H1, H2 and H3 in the vicinity of the Lyari River discharge area, the concentrations were several orders of magnitude higher than those detected in the sediment samples collected from lower Harbour and open sea area (Figure-3.11). The lower concentration of DDTs was observed away from the discharge point within the upper harbour, followed by the lowest levels within in the lower Harbour area. However, concentrations in the open sea area that includes navigation channel, area around oyster rock and west coast of the Harbour were found lowest to Non Detectable levels, this may be attributed to the continuous dredging of the channel and scarification of the sediments/dispersion of the pollutant by the wave action other Harbour activities and poor accumulation of the pollutants in the corrosive nature of the sediment.

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Figure-3.12. Distribution of DDT’s isomers in the Sediment of Karachi Harbour Area.

The total DDT concentration ND to >5 ng dry wt. in the surface sediment samples collected from Karachi Harbour area were found much lower than the levels reported from other harbour which are moderately to highly polluted, such as Xiamen harbour in Chin ranged 4.45–311ng/g (Mai et al., 2002); Macau harbour, China, 5–1629 ng/g (Zhang et al., 2003); Victoria harbour in Hong Kong 1.4-97 (Hong et al., 1995; Zhou et al., 2000); Singapore 2.2–11.9 ng/g (Richardson and Zheng,1999); Sydney Harbour, Australia reported 220 ng/g (Iwata et al., 1993); Keratsini Harbour, Greece 9.1-75.6 ug/g dry wt. (McCready, 2006); Alexandria Harbour, Egypt <0.25-885 (Galanopoulou, et al. 2005). However, the observed mean concentration in Karachi Harbour area was somewhat similar to the concentrations reported from Vancouver Harbour, Canada (2.5ng g−1 dry wt., or less) and Manukkau Harbour, New Zealand 1.2-2.3 ng g-1 (Barakat et al., 2002) and relatively higher than the values reported from Gulf & Gulf of Oman that includes marine sediment of Bahrain 0.088-0.430 ng g-1, Oman 0.0007-.0852 ng g-1, Qatar .00063 to 0.0367, UAE and to 0.0519 (Bolton, et al., 2004).

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3.4.1.2 Distribution Pattern of Organochlorine Pesticides in the Sediment of Rock Shore (Bulleji)

∑OCPs levels in the unconsolidated sandy clays are exposed to strong wave action in the Manora breakwater to Bulleji that included Manora, Sands spit and Hawks bay were found below detection limit. Similar condition was also observed in the area of Bulleji to Capemonze including Hawks Bay.

3.4.1.3 Distribution Pattern of Organochlorine Pesticides in the Sediment of Clifton Beach

Lowest levels were found in the sediment sample collected from west coast of Clifton beach. Detectable relatively lowest levels were observed at the station C-3 in the vicinity of drain/open sewers.

3.4.1.4 Distribution Pattern of Organochlorine Pesticides in the Indus Deltaic Creek System

The result reveals that the residual concentration of sum of all analyzed Organochlorine Pesticides (OCPs) varied between >0.001-17.5 ng g-1 dry wt. with a mean concentration of > 4.2 ng g-1 dry wt. (Figure-3.13).

Figure-3.13. Distribution of ΣOCPs in Sediment of Indus Deltaic Creek System

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The highest levels were found within the close proximity of discharged area of Malir river within the Gizri creek area (>15.21 to 17.5 ng g-1 dry wt.) and Korangi Creek (13.05 ng g-1 dry wt.) in the vicinity of Rehri Goth (C11). The observed elevated levels may be attributed to the influx of untreated effluents coming out from the coastal industries together with raw sewage discharged into the area. Since the flushing of Gizri creek is low, it is expected that the discharges remains stagnant during low tide (Akhtar et al., 1995). Significantly (p<0.045) lower levels (2.4 to 8.22 ng g -1 dry wt.) were observed away from the discharge at Korangi/Gizri creek junction downstream, most part of KC (Station C3, C8 to C11 and C13). The area within Korangi-Kadiro creek Junction upstream, Ch-C, Ka-C (C15 and C20), showed similarly levels of contamination (>5.2 to 7.01 ng g-1 dry wt.). OCPs concentration in the sediment collected from various localities of 20 km Korangi creek area were significantly lower as compared to levels found at Gizri creek, although creek is the outfall of effluent from Korangi Industrial complex and also receives domestic sewage through many open sewer, nalas, streams and storm drains. The Korangi Creek has three pronounced bends, some small inlets as well as its connection with Phitti and Kadiro Creek at the north-eastern end. These connections and bends allow the sea water to mix with waste water which is reflected in the relatively lower levels (0.01-8.5 ng g-1 dry wt.) of OCPs in the Korangi Creek sediment samples (Table-3.5).

No significant difference observed for the distribution pattern of OCPs at strong tidal flushed seaward Fareast end of Korangi creek and stations within navigational channel of Port Qasim (Station CC4 to C6, C17 to C19 andC22 to C24) shown in Figure 3.13.

Relatively higher levels were expected in the sample collected from Gizri/Korangi Junction at the station C3 because Gizri Creek carrying liquid wastes from the southern parts of Karachi into the Creek. In contrast lower levels (< 5.2 ng g-1 dry wt.) were observed in the area that may be attributed to the compact and hard bottom sediment which does not favour the accumulation of contamination. Moreover, it is due to the tidal flushing of the area due its connection close to open sea.

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Table-3.5 Mean levels (ng g-1)of Organochlorine Pesticide in sediment of Indus Deltaic Creek System Other Pesticides GC KC CH-C Ph-C KA-C Gh-C area* p,p’-DDD 0.5438 0.0746 0.0066 0.0007 0.0133 0.0017 Nd p,p’-DDE 5.6201 0.6317 0.3990 0.0321 1.0760 0.0111 Nd p,p’-DDT 3.0693 0.3831 0.1567 0.0085 0.8436 0.0049 Nd α-HCH 0.0258 0.1203 0.0673 0.0328 0.0954 0.0140 Nd β-HCH 0.8588 0.5158 0.0527 0.0779 0.6469 0.3387 Nd γ-HCH 0.2095 0.2662 0.0783 0.0010 0.3704 0.0020 Nd HCB 0.3030 0.2398 0.2060 0.2668 0.3000 0.1312 Nd Aldrin 0.1144 0.3950 0.1967 0.0053 0.4360 0.0022 Nd Dieldrin 0.2271 0.1963 0.0000 0.0013 0.0047 0.2116 Nd Endrin 0.0093 0.0366 0.0000 0.0000 0.0137 0.0010 Nd Heptachlor 0.7901 0.4744 0.3165 0.0020 0.4200 0.0082 Nd Heptachlor Nd 0.8791 0.4080 0.4300 0.0050 0.9600 0.0102 Epoxide ΣDDTsa 9.2332 1.0894 0.5623 0.0413 1.9330 0.0177 Nd ΣHCHsb 1.0940 0.9023 0.1983 0.0744 1.1127 0.3546 Nd ΣCyCc 2.0200 1.5103 0.9431 0.0091 1.8344 0.2324 Nd ΣOCPsd 12.6502 3.7418 1.9098 0.3027 5.1800 0.7359 Nd  Other areas include Clifton Beach, Clifton Beach, Buliji, Pacha ΣDDTsa= p, p’-DDE + p, p’-DDD + p, p’-DDT. ΣHCHsb= α-HCH + β-HCH + γ-HCH. ΣCycc = Aldrin + Dieldrin + Endrin + Heptachlor + Heptachlor Epoxide ΣOCPsd = ΣDDTs + ΣHCHs + HCB + ΣCyc.

The present study show relatively lower levels of OCPs concentrations in the sediment samples collected from various localities of Phitti/Gharo Creek System (0.11 to 1.7 ng g-1 dry wt.) as compared to other polluted side of Karachi coast. The lower level may be due to the enormous water exchange within the creek system, it is estimated that about 0.5 - 1 billion m3 of seawater flows in and out of Gharo/Korangi creek system during a tidal cycle (reference). The inflow of seawater in the creek area dilutes and disperses the contaminations received from various sources in the area.

The lower concentrations observed in the sediment collected from Bakran Creek may be attributed to the continuous input of large quantities of waste water from cooling system characterized by higher temperatures (3-6˚C) and flushing characteristics of the Bakran creek have resulted in low level of OCPs levels. Lower levels were also reflected in the sediment samples collected from Gharo creek within the main navigational channel of Port Bin Qasim which has been dredged to maintain a navigable depth of 11.3 meters.

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Significantly higher levels of OCPs in the sediment samples collected from Kariro creek and Korangi-Khudi creek junction (5.18 to 7.006 ng g-1 dry wt.) may be due to the influx of the waste water from adjacent water course carrying all kind of waste into the Kariro creek environment.

Large variability in the spatial distribution pattern of individual OC-pesticides was observed amongst the localities. The DDE & DDD, α.HCH & Endrin and Lindane & Dieldrin is showed similar behaviours in the sediment of Indus Deltaic Creek along Karachi coast, whereas distribution pattern of aldrine was significantly different. DDTs were predominant contaminants in most of the sediment samples due to its less volatile and slow degradation nature in the aquatic environment (WHO 1989, Carvalhho et al., 1992). DDTs contributed >60% of total analyzed OCPs (Figure-3.14). Higher levels of CYCs were found in the sediment of Korangi creek station C11, CH-C and KA-C area. Relatively lower levels of α-HCH levels (bdl-1.7 ng g-1 dry wt.) were found along almost entire coast which may be due to the lower lipophilicity and particle affinity (Yang et al 2005). Whereas HCB levels (0.02-0.12 ng g-1 dry wt.) were relatively uniform (f=15.368 p<0.218) contributed lowest (1-20%) in OCPs concentration.

Figure-3.14 ∑OCPs and ∑ DDTs Distribution Pattern in the Sediment of Indus Deltaic Creek System

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3.4.1.4.1 Distribution of Hexachlorobenzene (HCB) in the Sediment of Indus Deltaic Creek System

The sources of HCB is it use as fungicide and also introduced in an environment as by-product of various production processes of various agrochemical and industrial chemicals and waste incineration (Sarkar, 2008 and van-Birgelen, 1998). BHC is persists in the environment because of its thermodynamic stability (Breivik et al., 2004).

Measured residues concentration (BDL to <1 ng g-1 dry wt.) of HCB were found lowest amongst all analyzed OCPs in the sediment of Indus deltaic Creek System inspite of storage of large quantity of BHC in the one of the largest Obsolete Pesticide stock pile situated few km distance from Korangi Creek area (Khan, 2003). There were no distribution pattern of HCB reflected in our results (Figure-3.15) may be due to the volatile nature of the pollutant which exhibit a more even concentration pattern in the Creek environment. The statistical variability in distribution of HCB in the sediment of Karachi coast was not observed (p>0.001).

Figure-3.15 Distribution Pattern of Hexachlorobenzene (HCB) in the Sediment of Indus Deltaic Creek System

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3.4.1.4.2 Distribution of Cyclodienes in the Sediment of Indus Deltaic Creek System

The sum of all analyzed cyclodienes (aldrin, dieldrin, endrin, Heptachlor and Heptachlor Epoxide) concentration was found relatively lower (bdl->3.5 ng g-1 dry wt.) in the sediment of creek environment. Lower levels may reflect deregistration of most of the cyclodienes compounds during early 80’s except heptachlor which was in used up until 1997. Aldrin and Mirex never registered in Pakistan. However aldrine was available in open market and stock piles of obsolete pesticides (Khan, 2002).

There was virtually no pattern of cyclodienes concentrations in the surface sediment of creeks (Figure-3.16). Concentrations of cyclodienes varied considerably with location, significantly higher concentrations (>1-3.5 ng g-1 dry wt.) were found in the sediments samples collected from Indus deltaic creek system. Maximum levels (7.5 ng g- 1 dry wt.) were found in the sediment of Korangi creek in the vicinity of Rehri Goth station C11 and Kadiro creek (5.9 ng g-1 dry wt.). Elevated levels in the creek environment may attributed to the leaching of the CYCs compounds from one of the biggest piles contained over 410 tons expired/obsolete pesticides located few km distance from Korangi Creek area. Large quantity of Heptachlore, Dieldrin and BHC were stored together with other obsolete pesticide in the storage since several decades (Khan, 2002).

Figure-3.16 Distribution Pattern of Cyclodienes in the Sediment of Indus Deltaic Creek System

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Present results revealed that the total cyclodienes (CYC) levels were mostly contributed by Heptachlor (bdl->1.8 ng g-1 dry wt.), Heptachlor Epoxide (bdl->4.5 ng g-1 dry wt.), and dieldrin (bdl -> 2.74 ng g-1 dry wt.). Aldrin was only detected in few places within the creek environment. The contamination levels of Heptachlor Epoxide, a degradation product of Heptachlor was found to be highest by factors of 2-8 than Heptachlor, where the concentration ratio of Heptachlor Epoxide to Heptachlor is less than 1 (Wurl et al., 2005) in the creek environment. The data can be interpreted on the basis that there has been no recent introduction of Heptachlor into the creek environment. Lower concentrations of endrin (bdl-<0.2 ng g-1 dry wt.) were mostly observed in the creek sediment. Aldrin was found lowest in all samples because it is more volatile and readily degrades to dieldrin in the environment. Aldrin has never been officially used or manufacture in Pakistan reflects in the low levels along Karachi coast. The detectable amount at few locations may be attributed to the aerial deposition/transportation from other sources or illegal used.

3.4.1.4.3. Distribution Pattern of Hexachlorocyclohexane (HCHs) in the Sediment of Indus Deltaic Creek System

HCH and its isomers α-HCH, β-HCH, Lindane (γ-HCH) concentrations were detected in most of the sediment samples in relatively lower levels (below detection limit to 4.5 ng g-1dry wt.) as compared to DDTs concentration in the sediment samples collected from Indus deltaic creek system. Relatively higher concentrations were found in the sediment of Gizri creek area (station C-2; 1.4 ng g-1dry wt.) and Korangi creek stations C9 and C11 (1.2 to 2.3 ng g-1dry wt.). Highest concentration (4.5 ng g-1dry wt.) was observed in the sediment of Kadiro Creek (Figure-3.17). The residual concentration of HCHs in the Creek environment may be originated from industrial waste discharges in the area (Zhenwe et al., 2007).

Statistical variability in the distribution pattern of ∑HCH concentration in the sediment samples collected from various creek under the study was observed (p<0.001) amongst the localities. Over all distribution of -HCH concentrations in the sediment of

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Indus deltaic creek were close to mean concentration, however distribution pattern in Korangi creek were squeezed more towards lower level (Figure-3.17).

Figure-3.17 Distribution Pattern of Hexachlorocyclohexane (HCHs) and its Isomers α-HCH, β-HCH, Lindane (γ-HCH) in the Sediment of Indus Deltaic Creek System.

Amongst ∑HCHs, β-HCH was the most abundant isomer with a mean concentration >0.3 ng g-1dry wt. and ranged between below detection limit to < 4.00 ng g- 1dry wt.; whereas Lindane (γ-HCH) and α-HCH were below detection limit to >0.96 ng g- 1dry wt. and 0.78 ng g-1dry wt. The mean percentage composition of α-, β- and γ-HCH to ∑HCHs were 5-25%, 30-90% and 3-15% respectively for the creek sediments. Technical HCH has been used as a broad spectrum pesticide for agricultural purpose (Yang et. al 2005), which has been officially banned and deregistered since 1997 in Pakistan (Khan- 2005). The technical grade HCH mostly contains of α-HCH (55–80%), β-HCH (5–14%), γ-HCH (8–15%) and other (2–16%) of ∑HCH respectively (Lee et al., 2001). Relatively higher concentrations of β-HCH in the coastal sediments as compared to lindane (γ-HCH) and α-HCH may be due to the natural transformation of α- and γ-HCH to β-HCH (Hayes 1982, Wu et al., 1997; Walker, 1999). The β-HCH isomer is more persist with comparatively low microbial degradation (Ramesh et al., 1991) and lower vapour pressure (Wu et al., 1997 and Willett et al., 1998), it is therefore elevated levels of β-

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HCH in the creek sediment were found to be higher as compared to the contamination levels observed for α- and γ-HCHs isomers (Figure-3.17).

3.4.1.4.4. Distribution Pattern of DDT and its metabolites (DDE and DDD) in the Sediment of Indus Deltaic Creek System

The residual concentration of total DDT (∑DDT) in the surface sediment samples (n= 5 for each station) collected from creek area varied from ND to >12.3 ng g-1dry wt. at various localities of Indus Deltaic Creek system with a mean concentration of >1.5 ng g- 1dry wt. contributed highest in the total OCPs levels (Figure-3.18).

Figure-3.18 Relative Concentration of ∑OCPs and ∑DDTs in the Sediment Collected from Creek Environment

Distribution pattern of total DDT concentration were significantly different (f=20.641 p<0.001) in the sediment sample collected from creek area. Highest levels of ∑DDT > 11.8 to 12.3 ng g-1dry wt. were found to be at stations C1 and C2 in the vicinity of the Malir River discharged area. The concentrations were several orders of magnitude higher than the levels found in the sediment samples collected from other localities along Karachi coast; reflected in the plot (Figure-3.19). However relatively lower levels were

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Chlorinated Pesticides CHAPTER-3 observed in the sediment of Korangi creek station C11 (3.6 ng g-1dry wt.) in the vicinity of Rehri village. The ∑DDT concentrations in the sediment samples collected away from the discharge point were also found lower >3-5 ng g-1dry wt., followed by the lowest levels (BDL >0.5 ng g-1dry wt.) in the sediment collected from seaward stations farthest end of Korangi Creek (C4 to C6), Korangi Fish Harbour (C14), Bakran creek (C21), Gharo Creek (C24) and Phitti Creek (C19) may be due to the scarification of the sediments/dispersion of the pollutant by the wave action and poor accumulation of the pollutants in the corrosive nature of the sediment. Low levels in the Port Qasim area also due to the continuous dredging of the Port channels.

Figure-3.19 Distribution pattern of DDT and its metabolites (DDE and DDD) in the Creek Sediment

It was observed that DDE and DDD were the major component in the total DDTs concentrations in the sediment samples collected from various localities of creek environment (Figure-3.19). The higher concentrations of DDE and DDD (.02 to 7 ng g- 1dry wt. and .007 to 4.4 ng g-1dry wt.) were found in the sediment as compare to the concentration of their parent compound “DDT” in the Creek Environment. This may be due to the slow degradation of DDTs in the marine environment (Li 1999, Tavares et al., 1999; Khim et. al. 2001 and Yuan et al., 2001). Moreover a significant correlation between DDD and ∑DDTs (R2 = 0.94), DDE and ∑DDTs (R2 = 0.98) indicated common

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Chlorinated Pesticides CHAPTER-3 sources of pollution in the area. The positive correlation of ∑DDT and sediment organic carbon (R2 = 0.79) show that the DDT accumulated in the organic carbon rich sediment of Indus Deltaic Creek system.

Figure-3.20 Correlation between DDD and ∑DDTs (R2 = 0.94), DDE and ∑DDTs (R2 = 0.98) Indicated Common Sources of Pollution in the Coastal Area.

In most samples DDE and DDD occupied the predominant percentage (Figure- 3.21). DDE contributed (<20-90%), DDD (<10-55%) and DDT levels were observed lowest as compared to its metabolites (<1-10 % of DDT) shown in Figure-3.20. Technical grade DDT generally contains 75% p,p-DDT, 15% o,p-DDT, 5% p,p0-DDE, and <5% others.

3.4.1.5. Distribution of Organochlorine Pesticide in the Sediment of Balochistan Coastal Area

Organochlorine Pesticides were not detected (below detection limits) in sediment samples collected from ten localities of Balochistan coastal area. Therefore sediment sampling for further investigations was not carried out.

3.4.2 Fate of Organochlorine Pesticides in Coastal Environment of Pakistan

Large variability in the spatial distribution pattern of individual OC-pesticides

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Chlorinated Pesticides CHAPTER-3 was observed amongst the localities. The DDE & DDD, α.HCH & endrine and lindane & dieadin showed similar distribution pattern along the coast. Whereas, aldine concentrations were found significantly different in the coastal sediment. DDTs were predominant contaminants in most of the sediment samples due to its less volatile and slow degradation nature in the aquatic environment (WHO 1989, Carvalhho, et al., 1992). DDTs contributed >60% of total analyzed OCPs. Higher levels of CYCs were found in the sediment of Korangi creek station C11, CH-C and KA-C area. Relatively lower levels of ∑HCH levels (bdl-1.7 ng g-1 dry wt.) were found along almost entire coast which may be due to the lower lipophilicity and particle affinity (Yang et al., 2005). Whereas HCB levels (0.02-0.12 ng g-1 dry wt) were relatively uniform (f=15.368 p<0.218) contributed lowest (1-20%) in OCPs concentration. The variation of various OCPs levels in sediment of coastal at various localities is delineated in the box-plots, median, inter-quartile range, range values and outliers (Figure-3.5).

Strong correlation between OC-pesticides and sediment organic carbon and marked concentrations difference at seaward stations and area in the proximity of pollution discharge outlets suggests that contamination along Karachi coastal area seems to be localized. Further, good correlation amongst various OC pesticides (Figure-3.29) reflects probably common sources of contamination along Karachi coast.

3.4.2.1.1. Distribution, Fate and Sources of DDT and its metabolites (DDE and DDD) in the Coastal Sediment

Highest levels of DDTs and other analyzed OCPs in the sediment samples collected within the semi enclosed area of Karachi Harbour and Gizri creek, attributed to untreated waste from domestic sources and industrial effluents from various industrial zones being carried by Lyari and Malir rivers and discharged into the upper harbour and Gizri creek area where tidal flushing is very poor and waste water remain stagnated for long period of time (Khan, et al., 2006). Restricted water exchange with the open ocean makes the sediments of the area very anoxic. Aerobic nature of the sediment with high

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Chlorinated Pesticides CHAPTER-3 organic contents having great affinity to accumulate organic pollutants and organic matter also plays an important role towards the degradation of DDT to DDE and DDD. DDT can be effectively transformed to DDE and DDD under aerobic conditions (Kalantzi, et al., 2001, Ana, et al., 2005). DDE and DDD were the major component in the total DDTs concentrations in the sediment samples collected from various localities of coastal area. The mean levels of sediment OCPs contents is significantly correlated with DDTs (R2>0.55; p>0.69) as described earlier and in most samples DDE and DDD occupied the predominant percentage distribution pattern (Figure-3.12 & 3.19). DDE contributed (<20-90%), DDD (<10-55%) and DDT levels were observed lowest as compared to its metabolites (<1-10 % of DDT). The observed compositional difference between DDT and its metabolites is well correlated with composition of technical grade DDT.

Different ratios of DDTs and its metabolites were calculated for the identification of sources of the DDTs in the coastal environment of Pakistan (Figure-3.21 and 3.22). The results reveal that DDT and its metabolite (DDE and DDD) corresponding ratios from the coast are much low (0.4–0.5), according to Hong et al., 1999; Zhang et al., 1999; the observed ratio of (DDE+DDD)/t.DDT found much small than 1, that is indicating degradation of in the past use of DDTs would be the sources of the contamination in the coastal environment of Pakistan. The corresponding ratios of parent DDT and its biological metabolites, DDD and DDE (DDE/∑DDTs and DDD/∑DDTs) were also estimated according to the Tolosa et al., 1995; to evaluate fate of DDT deposition in the coastal area. It was also confirmed with these findings that there is no recent input of DDTs in the coastal areas of Pakistan. According to Khan (2004), DDTs have never been produced in Pakistan; however it has been formulated locally at various places along Karachi coast. Although usage of DDTs is more restricted or forbidden in Pakistan and deregistered in Pakistan since 1992, detection of higher amount of DDE and DDD than DDT, suggest that the DDT deposited after ban was more likely to be “weathered” DDT derived from sediment residues other than continuous applied “fresh” DDT which might contain less metabolites.

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Figure-3.21 Ratios of DDD / DDE in Sediment Samples of Karachi Harbour Area

Figure-3.22 Ratio of DDD/DDE in Sediment Samples Collected from Indus Deltaic Creek System

It is also expected that the detectable amount of DDT and its metabolites are the remnants of old input of the DDT in coastal area of Karachi because the reported environmental half-life of DDTs is estimated at 10–20 years (Woodwell et al., 1971; Sericano et al., 1990). Here the mean ratio of (DDE + DDD)/ PDDT in the sediments from the coastal area were close to unity, which indicated that the degradation occurred significantly. The breakdown products of DDE and DDD occupied predominant percent

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Chlorinated Pesticides CHAPTER-3 of ∑DDT along entire coastal area. This may be attributed to the slow degradation of DDTs in this environment (Tavares et al., 1999; Yuan et al., 2001). Moreover a significant correlation between DDD and ∑DDTs (R2 = 0.97), DDE and ∑DDTs (R2 = 0.89) indicated common sources of pollution in the area. The positive correlation of ∑DDT and sediment organic carbon (R2 = 0.79) show that the DDT accumulated in the organic carbon rich sediment of Pakistan coastal area.

Figure-3.23 Ratio of (DDD + DDE) / ΣDDTs in Sediment Samples Collected from Karachi Harbour Area

Figure-3.24. Ratio of (DDD + DDE)/ΣDDTs in Creek Sediment.

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3.4.1.2. Distribution, Fate and Sources of Hexachlorocyclohexanes (HCHs) and its isomers α-HCH, β-HCH, Lindane (γ-HCH) in Coastal Environment

HCH and its isomers α-HCH, β-HCH, Lindane (γ-HCH) concentrations in the sediment samples collected from coastal area were detected in most of the sediment samples in relatively highest levels (1.7 ng g-1 dry wt.) were found in the sample collected from upper Karachi Harbour area (station H1 to H3) in the close proximity of Lyari river discharged area. Relatively lower concentrations were found in the sediment of upper harbour area few kilometres away from discharges area, Gizri creek area and Korangi creek stations as described earlier. Statistical variability in the distribution pattern of ∑HCH concentration in the sediment samples collected from various localities along Karachi coast was observed (p<0.001) amongst the localities. Over all distribution of -HCH concentrations in the sediment of Karachi harbour area and Indus Deltaic Creek were found to be close to mean concentration, however distribution pattern in Phitti Creek environment was squeezed more towards lower level (Figure-3.11 and 3.17).

Amongst ∑HCHs, β-HCH was the most abundant isomer with a mean concentration >0.3 ng g-1 dry wt. and ranged between below detection limit to >1.4 ng g- 1 dry wt.; whereas lindane (γ-HCH) and α-HCH were below detection limit to >0.055 ng g-1 dry wt and 0.18 ng g-1 dry wt. The mean percentage composition of α-, β- and γ- HCH to ∑HCHs were 5-25%, 30-90% and 3-15% respectively for the sediments along Karachi coastal area. Technical HCH was most widely used pesticide for agricultural purpose. It is officially banned and deregistered since 1997 in Pakistan (Khan, 2005). The typical technical grade HCH generally contains 55–80% of α-HCH, 5–14% of β-HCH, 8– 15% of γ-HCH, 2–16% of ᵟ-HCH and others respectively (Lee et al., 2001). Relatively higher concentrations of β-HCH in the coastal sediments as compared to lindane (γ-HCH) and α-HCH may be due to the natural transformation of α- and γ-HCH to β-HCH (Hayes 1982, Wu et al., 1997; Walker, 1999). The β-HCH isomer is also most stable and relatively resistant to microbial degradation (Ramesh et al., 1991) and lower vapor pressure (Wu et al., 1997 and Willett et al., 1998), which is therefore accumulation of β-

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HCH in the sediment of Karachi coast, is expected to be higher than α- and γ-HCHs isomers as reflected in our results (Figure-3.25 and 3.26).

Figure-3.25 Ratio of α-HCH/γ- HCH in Sediment Samples Collected from Karachi Harbour.

Figure-3.26 Ratio of α-HCH/γ- HCH in Sediment Samples Collected from Indus Deltaic Creek System

Therefore, the predominant of α-isomer in some environmental samples reflects the recent use of technical HCH (Kannan et al., 1997) and domination of β-HCH in the

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Chlorinated Pesticides CHAPTER-3 environment reflect the old or weathered input of HCHs which is natural transformed α- HCH, β-HCH (Wu et al., 1997; Walker, 1999). Many studies have also reported that β- HCH was dominant in sediments from the river or estuary environment after long-term migration and transformation (Wu et al., 1999; Lee et al., 2001; Doong et al., 2002). In general, observed relatively higher concentrations of β-HCH probably reflected the decreasing use of technical HCHs and lindane in the Karachi coast. This may be relative to isomerization of HCHs during the process of transport and transformation in a marine ecosystem (Chen et al., 2000; Iwata et al., 1994; Dou and Zhao, 1998). More over elevated levels of ∑-HCH in the creeks environment also reflect the contamination sources not only agricultural or domestic run off but also industrial activities along Karachi coast.

3.4.3. Correlation of Organochlorine Pesticides and Sediment Organic Carbon

The lipophilic properties of many pesticides determines their accumulation in the organic phase of sediments, from where they can be remobilized and cause adverse effects to aquatic living resources (Carvalho et al., 2002).

The lowest concentrations (0.001-0.199 ng g-1dry wt.) correspond to the lowest TOC value of the seaward area. The cluster analyses of the stations support major groups rich sediment organic carbon stations within low flushed areas in the vicinity of waste discharge points and seaward stations with low organic carbon contents well flushed areas are clearly distinct from all other localities along Karachi coast (Figure-3.27).

The organic contaminants tend to be retained within the organic fraction of the sediment due to higher affinities of these liphophilic chemicals to these fractions (Jeffery and Baker 1999). Karickhoff, 1981 also observed that kinetic behaviour of hydrophobic organic pollutants is much influenced by organic carbon contents in sediments. Distribution pattern of OCPs strongly correlated with total sediment organic carbon contents (p>0.767 & R2=0.66), shown in Figure-3.28

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Figure-3.27 Dendrogram using Average Linkage (Between Groups) Showing Rich Sediment Organic Carbon Stations within Low Flushed Area in the Vicinity of Waste Discharge Points and Seaward Stations with Low Organic Carbon Contents Well Flushed Areas are Clearly Distinct from all other Localities along the Coast.

Figure-3.28 Correlation between Sediment Organic Carbon and total Organochlorine Pesticides found in Coastal Sediment.

Pakistan coastal area is a complex, dynamic system having variety of potential sources of environmental contaminants and it is important to have estimates on both sediment dry weight and TOC of the sediment to have better understanding of the

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Chlorinated Pesticides CHAPTER-3 contamination status and sources in the area. Therefore concentrations were expressed on sediment dry weight basis “ng g-1dry wt.” as well as normalized on organic carbon contents “ng g-1TOC” (Figure-3.29 and 3.30 ) .

Figure-3.29 Comparative Levels of ∑OCPs estimated on sed.dry wt. and Sediment Organic Carbon in Creek Environment.

Figure-3.30 Comparative Levels of ∑OCPs estimated on sed. dry wt. and Sediment Organic Carbon in Harbour Area.

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Analysis of variance (ANOVA) of organic contaminations levels normalized to sediment organic-carbon content was used to evaluate differences in concentrations among the various localities along Karachi coast. The test results indicate whether or not the mean normalized concentrations for the organic constituents are equal among all areas. If the mean normalized concentration were found to differ among the localities then Tukey's Significant Difference test was performed to determine which mean normalized concentration differed from one another (Helsel and Hirsch., 1992).

Figure-3.31 Correlation of total Sediment Organic Carbon and ∑DDTs in Coastal Sediment.

No statistically significant difference (p > 0.05) was observed in the levels found in sediment of almost entire creek system along Karachi coast except seaward stations (Figure-3.29). The cluster analyses showed different distribution pattern of OCPs normalized on TOC (Figure-3.30). The highest levels found at the discharged proximities stations of Korangi creek (C11>562, C4-9>300 ng g-1TOC), Korangi-Kadiro creek Junction upstream (C > 225 ng g-1 TOC) and Gizri-Korangi junction downstream (>500 ng g-1 TOC) in contrast to the highest concentration on sediment dry wt. bases at the Gizri creek station (C1 18 ng g-1 dry wt. and 350 ng g-1 TOC). Surprisingly contaminated

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Chlorinated Pesticides CHAPTER-3 upper Harbour area (>8 ng g-1 dry wt.) was comparable with levels estimated at Phitti creek area (>100 ng g-1 TOC) while concentration normalized on sediment organic carbon. This may be due to the hydrophobic nature of chlorinated pesticides tended to be adsorbed to organic carbon in the bed sediments as explained earlier. Thus, sediment samples with greater amounts of organic carbon can absorb higher concentrations of hydrophobic organic compounds. Normalization accounts for differences in concentrations of hydrophobic organic compounds caused by natural variations in the organic-carbon content of sediment samples; therefore, differences observed in normalized concentrations can be attributed to human activities rather than to natural variations in the organic carbon content of the sediment samples.

Strong correlation (R2 0.9 and 0.8) with total sediment organic carbon contents (Figure-3.31) strongly support the hypotheses of contaminants accumulated in organic rich sediments along Karachi coast such as KH (TOC 7%), GC (5%), few place within the other creek environment (TOC >2-3%) several other studies have also stated that organochlorine in the sediment are mainly associated with the organic matter (Neff, 1997; Knezovich et al., 1987, Doong et al., 2002). The result suggests that contamination along Karachi coastal area seems to be localized.

3.4.4. Organochlorine Pesticides Contamination in Marine Biota collected from Coastal area

Marine organisms have tendency to accumulate most of the contaminants from their environment in which they inhabit that include seawater, suspended & particulate matter, sediments and through the food web. Specific variety of marine biota (fishes, crab, shrimps and molluscs) has been collected for the analyses of organochlorine Pesticides (OCPs).

3.4.4.1. Organochlorine Pesticides Contamination in Molluscs (Bivalves)

For the present study bivalves such as mussels (Perna viridis) and oysters (Crassostrea sp.) were collected from coastal area. These animals are known to be

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Chlorinated Pesticides CHAPTER-3 suitable bio-indicators for monitoring trace toxic contaminant levels in coastal waters due to their wide distribution, sessile lifestyle, easy to collect and have a tolerance to a greater range of salinity, they can resist high stress levels and are good indicator of high accumulation of a wide range of chemicals (Goldberg et al., 1978). However according to Bishop Paul L. 1983 POPs are known to be growth retardant for oysters and other mollusks. Therefore mussels and oysters were collected to have estimates the contamination status in the sessile fauna of the coastal area.

3.4.4.1.1. Organochlorine Pesticides Contamination in Perna viridis (Green mussel)

For the present results on the concentration of OCPs in the Perna viridis (Green mussel) collected from Coastal area (Figure-3.32). It was observed that only breakdown product of most of OCPs (DDD,

DDE, Heptachlor and Dieldrin) were detected in the perna viridis. Perna viridis

The DDD concentration was found highest (>10 ng g-1 dry wt.) whereas parent compound DDT did not observed in the sample. The DDT”s metabolites were found in the order of DDD>DDE. High concentration of dechlorane and trans isomer of chlordane were also recorded these were in the range of 20-70 ng g-1 dry wt. and 5 ng g-1 dry wt. of dieldrin were also present, However, aldrin was not detected in Perna viridis collected from Korangi Creek area.

Figure-3.32 Residue of Organochlorine Pesticides in Perna viridis

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3.4.4.1.2. Organochlorine Pesticides Contamination in Crassostrea sp. (Oyster)

Oysters generally have a high capacity to assimilate organochlorines pesticides (OCs) from the marine environment. This is also a good bioindicator or ‘sentinel’ organism for marine organochlorine monitoring. Crassostrea sp (Oyster)

The results revealed that DDT and its metabolites, dechlorane and trans isomer of Chlordane were present in the analyzed tissue of the Crassostrea sp. (oyster) from Korangi creek area (Figure-3.33). These concentrations were relatively higher than concentration in the oysters collected from Pacha (a relatively clean area).

Figure-3.33 Residue of Organochlorine Pesticides in Crassostrea sp. (Oyster)

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3.4.4.1.3. Organochlorine Pesticides Contamination in Turbo

To evaluate the contamination levels 33 specimen of Turbo sp 2.4-3 cm in size were collected from Korangi creek area Pacha. Relatively lower concentration Heptachlor and aldrin were found in sample collected from both the localities, whereas DDT and its metabolites were Turbo sp. only detected in the turbo samples collected from Korangi Creek (Figure-3.34).

Figure-3.34 Residue of Organochlorine Pesticides Concentrations in Turbo sp.

3.4.4.2. Residue of Organochlorine Pesticides contamination in Crabs Collected from Coastal Area.

Crabs are mostly found in the pelagic area but swim close to the Pakistan at present due to the low demand. Some species, however, have a high nutritional value and seem to be abundant enough to have potential commercial value.

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The Portunus pelagicus (Blue swimming crab/ kekra), a coastal marine species found in brackish waters, estuaries and lagoons. Carnivorous, feeds on various fishes and invertebrates. Ten specimens size (width: 14-12.7 cm, length: 6-6.5 cm and weight: 140-190 g) were Portunus pelagicus collected from the coastal area. The residual concentration of DDE Portunus pelagicus and trans-chlordane were found to be highest in the blue swimming crab.

Scylla serrata (mud crab/ Khakua) A coastal marine species, living in brackish waters, estuaries, mangrove areas even in fresh Scylla serrata water. Feeds on shrimp, small fishes, molluscs and small crabs. It has also been observed feeding on carrion, seaweeds and small fragments of wood. Ten specimens (Length: 6-6.5 cm, Width: 14-12.7 cm, Weight: 140-190g) were collected from coastal. The results obtained during the present study indicating highest concentration of DDE in Scylla serrata and heptachlor epioxide only detected in the Scylla serrata amongst all analyzed marine biota samples. The aldrin was also present in both species (Figure-3.35).

Figure-3.35 Residue of Organochlorine Pesticides Contamination in Crabs (Scylla serrata and Portunus pelagicus) Collected from Coastal Area.

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3.4.4.3. Residue of Organochlorine Pesticides Contamination Levels in Shrimps (Penaeid shrimp)

The shrimps family contains the greatest number of commercially important species of shrimps in Pakistan. OCPs Contamination in Metapenaeus affinis (shrimp Jinga /Kalri) & Penaeus merguiensis (Banana Shrimp/ Jaira)

Shrimps are found from the coastline to about 50 m depths, but most abundant at 20m, on sandy and sandy-mud bottoms. They prefer turbid waters. Penaeus merguiensis and Metapenaeus affinis were collected from the creek environment were analysed for the OCPs described in Table-3.1. Penaeuensis sp.

It was observed that DDTs metabolised DDD and DDE only detected in soft tissue of Metapenaeus affinis in relatively higher concentration. Whereas aldrine was found in both the species in relatively lower levels (Figure-3.36).

Figure-3.36 Organochlorine Pesticides Contamination in Metapenaeus affinis (shrimp Jinga /Kalri) & Penaeus merguiensis (Banana Shrimp/ Jaira)

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3.4.4.4. Organochlorine Pesticides Contamination in Fishes (Nematalosa nasus (gizzard-shad/Daddi Palli) and Johnius glaucus (croaker/ Mushka)) Collected from Coastal Area

The Nematalosa nasus and Johnius glaucus were collected from the creek environment for the OCPs estimations. Preliminary results obtained during the study revealed that highest of concentration of DDTs and its metabolites (DDE and DDD) were detected in the flush of Nematalosa nasus. Whereas DDE only observed OCP in the tissues of Johnius glaucus. However other OCPs were not detected in the sample collected from Korangi creek area (Figure-3.37).

Figure-3.37 Organochlorine Pesticides Contamination in Fishes (Nematalosa nasus (gizzard-shad/Daddi Palli) and Johnius glaucus (croaker/ Mushka)) Collected from Coastal Area

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3.4.4.5. Residual contamination of Organochlorine Pesticides in Cephalopods (Sepia pharaonis (cuttlefish/ Mayya) collected from the coastal area.

Cuttlefish are demersal species, occurring from the coastline at shallow depths. The longevity of the cuttlefish is estimated at about 2 years. It feeds on crustaceans and small fishes. The cuttlefish sample was collected from Karachi Harbour.

The present results showed highest concentration of endrin Penaeuensis in the sepia collected from Karachi Harbour area near Oyster Rock. Lindane and heptachlor were also detected. However most occurring DDTs and metabolites did not found in the soft tissues of sepia sample (Figure-3.38).

Figure-3.38 Residual Contamination of Organochlorine Pesticides Level in Sepia pharaonis (cuttlefish/ Mayya)

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3.4.4.6. DDT and its metabolites in Marine Biota Collected from Coastal Area.

Varity of marine biota samples were collected from coastal area to evaluate the contamination levels in commercially important fisheries resources in Pakistan. The present results revealed that DDE a biological break down of DDT was present (8-15 ng/g dry wt.) in all marine biota samples collected from the study area (Figure-3.39). The highest DDE concentrations were recorded in the mud crab Scylla serrata (45 ng/g dry wt.). however the concentration obtained were found much lower than FAO/WHO ADI standards concentrations of DDE (0.01 mg/kg bw, 2000).

Marine Biota

Figure-3.39 DDE Levels in the Marine Bio Collected from the Coastal Area

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The results obtained on the contamination in the marine biota suggested lower levels in compression with the values reported from other coastal countries (Figure-3.40).

Figure-3.40 Compartivate Study on the Levels of Contamination in Marine Biota repoted from other Coastal Countries arround the World

3.5. Ecotoxicological Concerns /Potential for Biological/Ecological Effects

The sediment quality guidelines (SQG) specified by the USEPA (1997) and by Canadian Council of Ministers of the Environment (CCME, 2002) were used to assess the potential ecotoxicological impacts of organic contaminants measured in the surface sediments of coastal area. Hence Effects range-low (ER-L) and effects range-median (ER-M) values are used to predict potential impacts of contaminants in sediments. ER-L represents the value at which toxicity may begin to be observed in sensitive marine species, whereas ER-M represents the concentration below which adverse effects are expected to occur only rarely. Beside the threshold effect level (TEL), the probable effect level (PEL) is used as the criterion for the prediction of toxicity, and corresponds to a level above which adverse effects are frequently expected.

Canada Sediment Quality Guidelines (CCME, 2002), ISQG exceed for ∑DDT (1.22 ng g-1 dry wt.), 50%; DDT (1.19 ng g-1 dry wt.) 10%; DDE (2.07 ng g-1 dry wt.)

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15%, and heptachlor epoxide (0.6 ng g-1 dry wt.) 25% of sediment samples along the coastal area. It was also observed that 5%, 25%, 5% and 25% investigated samples exceed the threshold effect level (the concentration below which adverse effects are -1 expected to occur rarely) for total DDT (7 ng g dry wt /g), DDE (1.42 ng g-1 dry wt.), DDD(3.45 ng g-1 dry wt.) heptachlor epoxide (0.06 ng g-1 dry wt.) respectively. ∑DDT levels found significantly higher than ER-L value for ∑DDT (1.66 ng g-1 dry wt.) in more 33% samples, DDT (0.8 ng g-1 dry wt. ng g-1) 10%, DDE (2.2 ng g-1 dry wt.) 15 %, DDD (2 ng g-1 dry wtng/g) 8% samples along the coast but are significant lower than the ER-M values leading to an intermediate ranking of sediment toxicity. However concentration for ∑HCH and its isomers, cyclodienes and HCB were found significantly lower than ER-L and ER-M levels (Long et al., 1995).

3.6. Comparative (Neighboring, Regional and International) Distribution of Organochlorine Pesticides (OCPs) Contamination

To evaluate the relative degrees of OC Pesticide contamination in the coastal sediments of Karachi, Pakistan, a comparison has been made against the available data in neighbouring, regional, and other world coastal and marine areas context. Although, the direct comparability is somewhat compromised by the fact that different studies considered different DDT metabolites and sediment weight (wet, dry and sediment organic carbon), still, it is important to evaluate the OC pesticide contamination to get a sense of regional and global similarity. It is evident from present study that Karachi coastal areas present low to moderate OC contamination in sediments.

The concentrations of ∑DDT (BDL to <12.5 ng/g dw) in Karachi coastal sediment were found several order of magnitude lower in comparison with the reported levels of coastal marine region of Indian, Eastern coast (0.10–0.97;Sarkar et al., 1994), west coast, mouth of the estuaries (1.47-25.17; Sarkar et al., 1997), Bassim coast (<27.46-464.55; Pandit et al., 2001), Alibagh coast ( bdl-71.9; Pandit et al., 2001), Orissa coast (1.3-12; Pandit et al., 2001), west and east coast (ND–364f; Pandit et al., 2001), and Mumbai coastal area (0.5-9.6 ng/g; Pandit 2006),

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Chlorinated Pesticides CHAPTER-3

The observed DDT levels were also much lower than the reported concentrations from various region of Indian Ocean along India, eastern Arabian Sea (7.38-179.1 ng g-1; Shailaja and Sarkar, 19), Arabian Sea (1.14-25.17; Sarkar et al., 1997) and Indian Ocean (55-12053 ng/g; Sarkar and Gupta--). Relatively higher levels of OCP in the sediment of Indian coast marine area could be attributed to the large scale production and use of these pesticides in agriculture as well as anti-malaria sanitary activities carried out though out the country (Pandit et al., 2002). Comparatively low levels along Karachi coastal area may be ascribed very limited primary production of these pesticides in past and direct source of agricultural runoff in the area is not evidenced.

The coastal area were found relatively more contaminated as compared to Northeastern coast (0.18-1.93 ng g-1 dw; Guzzella et al., 2005) and Bay of Bengal (0.04- 4.79; Rajendran et al., 2005) along India.

The levels from the Gulf and the Gulf of Oman marine sediment reported by Tolosa et al., 2005 (Bahrain, 0.088-0.430ng/g; Oman, 0.0007-.0852 ng/g; Qatar, 0.00063 to 0.0367; UAE, BDL to 0.0519) were also found much lower. The levels reported by Fox et al., 1988 from Manukkau Harbour, New Zealand (1.2-2.3 ng/g;) was also lower than the levels observed from Karachi coastal areas.

Concentrations were also lower than the ranges testimony from Asian estuaries and coastal areas which are moderately to highly polluted, such as Xiamen harbour in Chin (4.45–311 ng/g; Hong et al., 1995); Pearl River Delta and Macau harbour, China (5–1629 ng/g; Mai et al., 2002); Victoria harbour, Hong Kong (1.4-97; Hong et al., 1995), Richardson and Zheng, 1999); Singapore (2.2–11.9 ng/g Wurl and Obbard, 2004); Kyeonggi Bay, Korea (<0.046–32; Lee et al., 2001); Osaka Bay Japan (2.5–11.9 ng/g; Iwata et al., 1994); Sydney Harbour, Australia (220 ng/g; McCready 2006); Keratsini Harbour, Greece (9.1-75.6 µg/g dry wt; Galanopoulou et al. 2004); Alexandria Harbour, Egypt (<0.25-885; Barakat et al., 2002), North coast of Vietnam (6.2-10.4; Nhan et al., 1999) However, the mean concentrations in the sediment of Karachi coastal area were relatively lower or comparable with the concentrations reported from Vancouver Harbour, Canada (2.5 ng g−1 dry wt.), Bohai and Yellow Sea, China (0.37–1417; Ma et

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Chlorinated Pesticides CHAPTER-3 al., 2001), Daya Bay, China (0.14–20.3; Zhou et al., 2001), Lingding Bay, China (2.6– 115; Kang et al., 2000), Pearl River Delta, China (2.6–1629; Mai et al., 2002), Xiamen Harbour, China (0.01-0.06; Zhou et al., 2002) Macao Harbour, China (1630; Kang et al., 2000), Yangtze Estuary, China (ND–18.95; Liu et al., 2003), Victoria Harbour, Hong Kong (1.4–30 Connell et al., 1998), Wu-Shi estuary, Taiwan (BDL–11 .4; Doong et al., 2002), Kyeonggi Bay, Korea (<0.046–32a; Lee et al., 2001), Namayang Bay, Korea (0.088–0.38a; Lee et al., 2001), Ulsan Bay, Korea (0.02–41.9; Khim et al., 2001), Coast of Korea (0.01–135; Hong et al., 2006), Osaka Bay, Japan (2.5–11.9; Iwata et al., 1994), Singapore (2.2–11.9; Wurl and Obbard 2005), North coast of Vietnam (6.2–10.4f; Nhan et al., 1999), West coast of Sri Lanka (0.09–1.6f; Guruge and Tanabe 2001)

The levels were also compared with the contamination levels reported from USA, Europe, Australia and other coastal marine regions. The evaluation reveals that sediment from Karachi coast were relatively less contaminated (Table-3.6). French coast (<1–86; Marchand et al., 1988), Amsterdam area, The Netherlands (39.1; Van Der Oost et al., 1996)a, Italian coast, Adriatic and Tyrrhenian Seas (1.2–13 1; Mangani et al., 1991), Alexandria Harbour, Egypt (<0.25-885 ; Assem O. Barakat et al., 2002), KeratsiniHarbour, Greece (9.1-75.6, Galanopoulou et al., 2004), Caspian Sea (0.01– 13.4c; de Mora et al., 2004), Coast of the Bohai and the Yellow Sea (0.37–1273f; Ma et al., 2001), Coastline, Black Sea (0.2–72c; Fillmann et al., 2002), Caspian Sea, Russia (0.01–1.9; de Mora et al., 2001), Ukarine, Coastline (0.06–0.6; Fillmann et al., 2002), Brazil, Amazon region (24; Torres et al., 2002), Gulf of Mexico (<0.02–454; Sericano et al., 1990), ManukkauHarbour, New Zealand (1.20–2.3; Fox et al., 1988), Caspian Sea, Kazakhstan (0.01–1.9; de Mora et al., 2001), Caspian Sea, Azerbaijan; 0.56–13.4 de Mora et al., 2001), Caspian Sea, Iran (0.06–3.9; de Mora et al., 2001), Red sea and Gulf of Aden (ND; DouAbul and Al-Shiwafi, 2000), Gulf of Aden, ND–0.74; Alaa R. Mostafa et al., 2007), Pacific coast, USA (Brown et al., 1998), San Fransico Estuary, USA (<0.1– 9; Pereira et al., 1994), Palos Verdes, California (1600–100000, Smokler et al., 1979).

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Chlorinated Pesticides CHAPTER-3

3.7. Conclusions

The chlorinated pesticides have great affinity with the organic phase of the sediment due to lipophilic properties of these contaminates (Carvalho et al., 2002). Therefore their concentrations were found much higher in the sediment with high organic carbon such as semi-enclosed area of Karachi Harbour and Gizri Creek/Korangi Creek where most of these contaminates drained via various sources. Transport/remobilization of these contaminated sediment could be the sources of contaminates in the area and may cause adverse effects to marine resources. Effect based guidelines for sediment quality were use to identify the areas in which the potential for biological effects is greater and to assist environmental management. Present contamination results revealed that most of the exceed values were corresponded to the levels found in the discharged proximity of low flushed areas along Karachi coast such as upper-KH, GC, few localities within KC, Ch-C and Ka-C. Whereas most of the investigated area did not reflect the comparable the concentration above adverse effects (probable effect level PEL), except Gizri creek where DDE levels exceed from PEL (6.75 ng g-1 dry wt) and Apparent Effects Threshold (AET) 9 ng/g for benthic organisms for ∑DDT exposure, whereas KH and KC showed closer levels to the AET. However concentrations found for other Indus deltaic creeks and seaward areas along Karachi coast were found much lower than the reported sediment quality criteria and concentration ranges.

The results clearly indicate that elevated concentrations found in the sediment of collected from coastal area were very much localized and may attributed to the discharges of untreated industrial effluents and waste from domestic sources in the semi- closed area of upper harbour and Gizri creek where tidal flushing is very poor and waste water remain stagnated for longer period of time (Khan, et al., 2006).

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CHAPTER 4 Distribution of PCBs, Dioxin & Dioxin-like Chemicals in Indus River and Coastal Environment of Pakistan

PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4

4.1 Abstract

& like compounds were demonstrated very first time for aquatic environment of Pakistan. The present results revealed that chlorinated PCDD/F congeners, OCDD, Chlorinated dibenzofurans (OCDF), Non-ortho–substituted & Mono- ortho–substituted PCBs were present in the sediment collected from various environmental conditions of Indus River and coastal areas of Pakistan.

The significantly different distribution pattern (OCDD>PCBs> OCDF) were observed. Highest levels (300 pg PCDD/PCDF g-1 dry wt.) and ∑ Non-ortho–substituted PCB and Mono-ortho–substituted PCBs (1642 pg g-1 dwt) were found in the sediment collected from effluent discharge vicinity within semi-enclosed area of upper harbour and Gizri Creek. However, Khobar Creek sediment represented relatively cleaner area while comparing Dioxin contamination. Further analysis of river sediment demonstrated relatively lower contamination levels at most of the site with the exception of the sediment from last barrage on the Indus before the river enters the delta area (Station IR- 4). The observed concentrations were expressed on toxicity equivalency of 2,3,7,8-TCDD (0.63 to 4.8 pg TEQ g-1 dwt.).

The results clearly indicate the same distribution pattern of PCDD/F and PCBs individual congener profile which is dominated by higher chlorinated chemicals such as OCDD and PCBs in the samples collected from both the localities. The identical congener profiles may indicate the same source of contamination in the study areas.

The present results from aquatic environment of Pakistan showed relatively lower levels as compared to neighbouring, regional and European coastal countries. However, results are comparable with the reported levels from New Zealand and Australian estuaries.

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4.2 Introduction

Dioxins and dioxins like compounds (furans) and PCBs are introduced into the environment mainly due to incomplete combustion processes industrial as unintentional by-products and waste burning activities. The aquatic environments like coastal, marine and riverine area are an important sink for these substances. The consumption of contaminated food particularly seafood is an important exposure pathway for humans (Bursian, et al., 2007 and Khan et al., 2004). The Polychlorinated Biphenyls (PCBs) was introduced in 1929 as a very popular industrial chemical, mainly used as a flam-retardant, cooling agent in transformers and capacitors, also use in many industrial process as lubricants, hydraulic fluids, cutting oils, adhesives, liquid seals, coating wood and plastics to prevent them from flam. Also used in paints, varnishes, inks and pesticides and carbonless copy paper (Khwaja and Petrlik, 2005). Soon after it was recognize as dangerous chemical by Soren Jensen a Danish chemist.

The PCBs has been used and sold in market with various trade names (Clophen, Arocler, Kanechlor, Santotherm, Phenoclor and Pyralene) in Pakistan. However, it has never been manufactured/synthesize in the country. It has been used in transformer oil in power sector that could be possible sources of PCBs contamination along the coast. The other sources may be automotive and electric machinery contaminating PCBs oils. The PCBs contamination may be introduced in the environment due to the leakage from the electrical equipment and/or dealing with the wastes containing PCBs. There is hardly any published information available on the contamination of PCBs in various environmental compartments of Pakistan. The use of PCBs-oil has been prohibited in Pakistan since 1974. PCBs contamination is listed as “Phenolic Compounds” in the NEQS-Pakistan (PEPA, 1997).

4.2.1 Un-intentional by Products (Dioxin and Dioxin like substances)

Un-intentional by products include the highly toxic dioxins and furans are produced by human activities and are also occurring naturally. The natural sources could be combustion of forest fires and volcanic activity. These contaminations are not

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4 industrial substances or involved in industrial processes, except in small amounts produced for research purposes (green-facts.org, 2006). However these are unintentionally produced during industrial processes dealing with organic chemicals and chlorine, they are introduced in an environment via municipal and domestic incineration and combustion processes (green-facts, 2010).

As no legislation covering dioxins or other unintentional POPs exists in Pakistan, little attention has been given to the issue. No inventories on the potential sources have been compiled. Lake of awareness and poor implementation of the environmental legislation and irresponsible and casual attitude towards environment issues may augment the environmental problems in the country. Pakistan has a number of industries in the fields such as metallurgy, pulp and petrochemicals, which might be substantial emitters of unintentional POPs. Furthermore, practices like open pit burning of waste are very common.

Environmental monitoring of persistent organic pollutants (POPs) such as PCDD/PCDF and the dioxin-like PCBs is quite expensive and requires specialized laboratory facilities ideally equipped with High Resolution Mass Spectrometry (HRMS) instrumentation. Hence, by far the majority of work on PCDD/PCDF and dioxin-like PCB has been carried out in industrialized countries in Europe, North America and Japan. In contrast there is very little – or no information documented on the levels of these chemicals in the environment of developing nations and to our knowledge no data are available on the levels of these chemicals in the aquatic environment in Pakistan (Khan, et al., 2004). The aim of this study was to evaluate dioxin-like chemicals in sediments

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4 collected along a transect in the Indus River to the Arabian Sea as well as in sediments from the most urbanised and industrialised area south of Karachi to obtain a first in-sight on the levels of dioxin-like chemicals in Pakistan.

4.3. Material and Method

4.3.1. Sampling Area

To evaluate Dioxin and Dioxin like PCBs contamination sediment samples were collected following a transect of the Indus River (Figure-4.1) and in the vicinity of major urbanized area of Pakistan’s coastline i.e. Lyari River discharge area within Karachi Harbour and Gizri/Korangi Creek, a key industrialized area along the coast (Figure-4.2). Samples were also taken from Khobar Creek. GPS (GARMIN 12 XL) was used to mark the geographical location of the sampling site.

4.3.1.1. Indus River Sampling Sites

To evaluate Dioxin and Dioxin like PCBs contamination sediment samples were collected following transect of the Indus River (5 Locations) shown in Figure-4.1.

Figure-4.1 Indus River sampling location

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4

Station (IR-1), A few kilometres above the town of Mithankot, the Indus is joined by its most important tributary, the Panjnad River, which carries the waters of five main tributaries-the Jhelum, the Chenab, the Ravi, the Beas, and the Sutlej.

Station (IR-2), the hub of agricultural and commercial activities in upper Sindh. Sukkur has more forests than any other district of Pakistan. About 5, 587, 821 hectares of the area are covered by naturally or artificially irrigated forests. Blind or Indus dolphin, a rare species of Sukkur, is found only in the Indus River. Before the construction of various barrages, the blind dolphin used to cruise some 3500 km course of the river. However, its habitat is now limited to 170 km area between Sukkur and Guddu Barrages of the Indus. The blind dolphin of Sukkur lives in the heavily silted waters of the river.

Station (IR-3), Right bank of Indus River, Sukkur barrage drainage into the Indus River near Sehwan

Station (IR-4), last barrage on the Indus before the river enters the delta area. Kotri to Khobar creek is about 180km, which is dry most part of the year. During the flood it carries lots of sediment into the sea.

Station (IR-5), Left bank (half way to Kotri & Khobar Creek), almost dry throughout the year except monsoon season.

Station (IR-6), Presently Khobar Creek is the only creek that carries the fresh water of Indus River into the Arabian Sea.

4.3.1.2 Sampling Sites along the Coast.

Four sampling sites were chosen along the coast Karachi Harbour, Khobar Creek, Korangi Creek and Gizri Creek (Figure-4.2).

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Figure-4.2 Location Map for Sediment Sampling along the Coast.

4.3.1.2.1. Karachi Harbour: Four (04) samples collected from upper harbour area (Samples H1, H2, H3 and H4) were pooled.

4.3.1.2.2. Gizri Creek: Two sediment samples were pooled, collected from the locations (station C1-C2) in the vicinity of Gizri Creek Area (extreme end, mid/centre).

4.3.1.2.3. Korangi Creek Area: It is located on the eastern side of Karachi – stretched up to Rehri Creek at the north-eastern side. It is about an area of 64,000 hectares, that is which is one-tenth of the Indus River Delta. For the dioxin estimation in the Korangi Creek area sediment from four most polluted sites were collected and pooled to make a composite representative of the area.

4.3.1.2.4 Khobar Creek: Khobar creek is the main creek through which IR flows into the Arabian Sea. It is about 85km in south-east of Karachi with a length of 20km, an average width of 1200m and an average depth of about 8m.

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4.3.2. Sediment Sample Collection

Sediment were collected from selected sides along the coast and Indus River area by using Grab sampler (Peterson type) during the year 2002 and 2003, details about the sampling sites are elaborated in Figure-4.1 and 4.2. GPS (GARMIN 12 XL) was used to mark of the geographical location of the sample site.

All samples were stored in pre-cleaned and solvent rinsed stainless steel containers, kept cool on ice and transported to the laboratory and refrigerated within 2 to 4 h.

4.3.3. Sample Preparation and Transportation

Samples from collection sites were transported to NIO Lab in ice boxes and kept them in freezer till further processing. Later on samples were freeze dried by using LABCONCO, Freeze Dry System. Freeze dried samples were homogenized by using mortar and pestle and wrapped up in solvent-washed aluminium foil and transported to ERGO Laboratory in Hamburg.

All the glass wares and sampling tools were pre cleaned and rinsed with solvent (acetone and hexane)

4.3.4. Analysis of Dioxin and Dioxin like PCBs

4.3.4.1. Pre- treatment /Sample preparation for the identification and quantification

Briefly 10g of freeze dried sediment sample was extracted using Soxhlet extraction technique for 18hrs with 200ml of acetone and hexane mixture (1:1) in triplicate and one blank for each five set of sediment samples. The extracted samples were filtered over anhydrous sodium sulphate and concentrated samples were run through Florisil column clean up, eluted with the mixture of 120 ml 6% ethyl acetate and hexane and 100ml of acetone and hexane. The extract was concentrated under a gentle stream of nitrogen up to 1 ml.

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4.3.5. Instrumental Condition

For analysis freeze-dried, homogenized samples were sent to the ERGO Laboratory in Hamburg, Germany. All analyses were performed following the isotope dilution method. Samples were analysed for 2,3,7,8-substituted PCDD/PCDF and dioxin-like PCB. In brief, the freeze-dried, homogenised samples were soxhlet extracted for 20 hours using toluene. A blank was included. Prior to extraction samples were spiked with a 13C-labelled PCDD/PCDF and PCB standard of known quantity. The extracts were concentrated and subjected to clean up using acid-base (H2SO4/CsSiO) and alumina (Alox B-super, ICN) columns in series. Samples were further purified on activated carbon and an acid-base (H2SO4/CsSiO) clean-up, if required. Samples were concentrated to near dryness and transferred into vials with a known quantity of 1,2,3,4- TCDD, used as recovery standard. Analysis of tetra to octa-CDD/Fs was performed on a GC (DB-5 fused silica column, 60 m, 0.25 mm i.d., 0.1 µm film thickness) interfaced to a VG Autospec Mass Spectrometer operating on a resolution of approximately 10 000. Identification of 2,3,7,8-substituted PCDD/Fs was performed using retention times of the 13C-labelled standard and isotope ratios M+ and M2+.Several criteria had to be fulfilled for quality control: a) the retention times (RT) of the analytes in a sample had to be within 2 s of the RT of the internal standards b) isotope ratios for each congener of the M+ and M+2+ ions had to be within 20% of the respective individual value c) PCDD/F limit of quantification was defined by a signal to noise ratio greater than 3 times the average baseline variation and a substance quantity in the sample greater than 3 times the quantity in the respective blank. Typical PCBs chromatograms obtain from GCM is shown in Figure-4.3.

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Figure-4.3. GCMS Chromatogram of PCBs analysis

4.4 Estimation of EQF and TEQ

2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD) is considered the most toxic of the chemical family of polychlorinateddibenzodioxins (PCDDs) and polychorinateddibenzofurans (PCDFs). It is assigned the toxicity equivalent factor (TEF) of "1", and all other PCDDs and PCDFs are assigned a TEF value less than "1" (NCEA-I- 0836, 2003.) described in Table-4.1.

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Total Toxic Equivalency (TEQ) = ΣkCn ∗TEFn ...... 4.1 n=1

Toxic Equivalent Quantity (TEQ) is the Quantity of each PCDD/PCDF emitted multiplied by its TEF (eq-4.1). In this manner, sources of different relative quantities of the different PCDD/PCDF compounds can be compared to a standard limit (Berg et.al 2006). Table-4.1 Summary of WHO 1998 and WHO 2005 TEF Values (Berg et.al 2006 )

Compound (PCDD/F congeners) WHO 2005 TEF Chlorinated dibenzo-p-dioxins

2,3,7,8-TCDD 1 1,2,3,7,8-PeCDD 1 1,2,3,4,7,8-HxCDD 0.1 1,2,3,6,7,8-HxCDD 0.1 1,2,3,7,8,9-HxCDD 0.1 1,2,3,4,6,7,8-HpCDD 0.01 OCDD 0.0003 Chlorinated dibenzofurans WHO 2005 TEF 2,3,7,8-TCDF 0.1 1,2,3,7,8-PeCDF 0.03 2,3,4,7,8-PeCDF 0.3 1,2,3,4,7,8-HxCDF 0.1 1,2,3,6,7,8-HxCDF 0.1 1,2,3,7,8,9-HxCDF 0.1 2,3,4,6,7,8-HxCDF 0.1 1,2,3,4,6,7,8-HpCDF 0.01 1,2,3,4,7,8,9-HpCDF 0.01 OCDF 0.0003 Non-ortho–substituted PCBs WHO 2005 TEF 3,3#,4,4#-tetraCB (PCB 77) 0.0001 3,4,4#,5-tetraCB (PCB 81) 0.0003 3,3#,4,4#,5-pentaCB (PCB 126) 0.1 3,3#,4,4#,5,5#-hexaCB (PCB 169) 0.03 Mono-ortho–substituted PCBs

2,3,3#,4,4#-pentaCB (PCB 105) 0.00003 2,3,4,4#,5-pentaCB (PCB 114) 0.00003 2,3#,4,4#,5-pentaCB (PCB 118) 0.00003 2#,3,4,4#,5-pentaCB (PCB 123) 0.00003 2,3,3#,4,4#,5-hexaCB (PCB 156) 0.00003 2,3,3#,4,4#,5#-hexaCB (PCB 157) 0.00003 2,3#,4,4#,5,5#-hexaCB (PCB 167) 0.00003 2,3,3#,4,4#,5,5#-heptaCB (PCB 189) 0.0003

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4

4.5 Results and Discussion The monitoring of toxic contaminations such as PCBs, PBDF, Dioxin and Dioxin like chemicals produced unintentionally or by-product of various combustion processes never been addressed in Pakistan. It may due to the lack of expertise and facilities in the country. The present study was the very first attempt to demonstrate the contamination of PCBs (non-ortho-substitute and mono-ortho-substitute) (Table-4.2), PCDD/F congeners (2,3,7,8-chlorine substituted PCDD/PCDF) (Table-4.3 and 4.4) and PBDF congeners (Table-4.5) in the sediment sample taken from various localities of Indus River and coastal area of Pakistan represented various environmental conditions .

Table-4.2 Distribution of PCBs (pg g-1 sed. dry wt.) in River and Coastal Sediment.

Khobar Korangi Gizri Karachi Non-ortho–substituted PCBs IR-1 IR-2 IR-3 IR-4 IR-5 Creek Creek Creek Harbour. 3,3',4,4'-Tetra-CB 77 4 4 4 42 4 4 8.1 41 82 3,4,4',5-Tetra-CB 81 0.4 0.4 0.4 1.4 0.4 0.4 0.4 1.8 3.1 3,3',4,4',5-Penta-CB 126 0.7 0.7 0.7 7 0.7 0.7 1.3 5.6 9.7 3,3',4,4',5,5'-Hexa-CB 169 0.3 0.3 0.3 1 0.3 0.3 0.3 1.1 1.6 ∑ Non-ortho–substituted 5.4 5.4 5.4 51.4 5.4 5.4 10.1 49.5 96.4 PCBs Mono-ortho–substituted PCBs 2,3,3',4,4'-Penta-CB 105 34 27 32 143 17 47 54 129 335 2,3,4,4',5-Penta-CB 114 15 16 14 19 15 14 17 29 34 2,3',4,4',5-Penta-CB 118 148 85 86 218 66 134 162 320 878 2',3,4,4',5-Penta-CB 123 22 21 20 20 21 19 20 22 24 2,3,3',4,4',5,-Hexa-CB 156 16 18 16 60 16 15 22 53 164 2,3,3',4,4',5'-Hexa-CB 157 15 16 14 13 15 14 15 17 31 2,3',4,4',5,5'-Hexa-CB 167 16 16 16 27 15 15 15 34 59 2,3,3',4,4',5,5'-Hepta-CB 189 18 19 17 16 18 17 18 22 21 ∑Mono-ortho–substituted 284 218 215 516 183 275 323 626 1546 PCBs

Over distribution pattern of these contaminates levels are described in the box plot mean, median and interquartile (Figure-4.4). The presence of these chemical could relate to hazardous industrial, municipal, agricultural and other waste burning is in large scale practice in the country without proper disposal facilities other than industrial sources could be sources of Dioxin pollution in the area (www.ipen.org, 2006). The contamination levels in the sediment collected from coastal environment is relatively different from the levels obtained from most of places within the Indus River (Figure- 4.5). Page 98

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-1 Figure-4.4 Dioxin-like chemicals detected (pg g sed. dry wt.) in the Sediment Collected from Indus River and Coastal Area

Table-4.3 PCDF congener profile obtained from Coastal and River Sediment (pg g-1 sed. dry wt.)

Chlorinated Khobar Korangi Gizri Karachi IR-1 IR-2 IR-3 IR-4 IR-5 dibenzofurans Creek Creek Creek Harbour. 2,3,7,8-TCDF 0.43 0.5 0.72 3.3 0.33 0.3 0.52 0.69 0.76 1,2,3,7,8-PeCDF 0.1 0.4 0.36 0.92 0.2 0.2 0.32 0.6 0.70 2,3,4,7,8-PeCDF 0.2 0.5 0.39 0.55 0.2 0.2 0.14 0.7 0.65 1,2,3,4,7,8-HxCDF 0.3 0.9 1.3 1.3 0.56 0.47 0.49 1.1 2.1 1,2,3,6,7,8-HxCDF 0.2 0.4 0.3 0.49 0.2 0.16 0.14 0.9 1.2 1,2,3,7,8,9-HxCDF 0.4 0.7 0.9 0.3 0.2 0.4 0.3 0.39 0.5 2,3,4,6,7,8-HxCDF 0.2 0.7 0.52 0.98 0.3 0.3 0.3 1.5 1.8 1,2,3,4,6,7,8-HpCDF 0.6 0.6 0.86 4.7 0.23 0.79 1.9 4.3 9.7 1,2,3,4,7,8,9-HpCDF 1 1 2 2 0.6 0.8 0.7 0.8 0.7 OCDF 0.6 1 2 13 0.6 3 8 14 36 ∑PCDF 4.03 6.7 9.35 27.54 3.42 6.62 12.81 24.98 54.11

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Table-4.4 PCDD Congener Profile Obtained from Coastal and River Sediment (pg g-1 sed. dry wt.)

Chlorinated dibenzo-p- Khobar Korangi Gizri Karachi IR-1 IR-2 IR-3 IR-4 IR-5 dioxins Creek Creek Creek Harbour

2,3,7,8-TCDD 0.1 0.1 0.1 0.74 0.1 0.24 0.25 0.48 1.2

1,2,3,7,8-PeCDD 0.1 0.2 0.1 0.61 0.1 0.2 0.1 0.46 0.71

1,2,3,4,7,8-HxCDD 0.5 0.3 0.3 0.39 0.1 0.3 0.2 1.1 0.72

1,2,3,6,7,8-HxCDD 0.2 0.2 0.2 1.1 0.1 0.3 0.25 1.5 1.8

1,2,3,7,8,9-HxCDD 0.2 0.2 0.2 0.75 0.1 0.2 0.2 0.99 0.87

1,2,3,4,6,7,8-HpCDD 0.9 0.7 0.7 2.24 0.3 0.8 0.65 3.59 3.39

OCDD 2.9 7.1 6.2 116 5.4 8 18 144 222 121.8 ∑PCDD 4.9 8.8 7.8 6.2 10.04 19.65 152.12 230.69 3

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4

Table-4.5. Concentrations (pg g-1 sed. dry wt.) of PBDE Congeners in Sediment Samples Collected from River and Coastal Area of Pakistan.

Area Total 17 28 47 66 99 100 153 154 183 209 BDE PK 1 n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. - - - 0.08 -0.002 -0.007 -0.024 -0.002 -0.012 -0.002 -0.002 0.002 0.003 0.02 PK 2 n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. - - - 0.07 -0.002 -0.007 -0.02 -0.002 -0.014 -0.002 -0.002 0.002 0.003 0.02 PK 3 n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. - - 0.16 -0.002 -0.007 -0.024 -0.002 -0.014 -0.002 -0.002 -0.1 0.002 0.002 PK 4 n.d. n.d. 0.003 0.025 0.02 0.005 0.003 0.003 0.015 1.74 2.45 -0.007 -0.002 PK 5 n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. - - - 0.06 -0.002 -0.006 -0.024 -0.002 -0.012 -0.002 -0.002 0.002 0.003 0.04 Khobar n.d. n.d. n.d. n.d. n.d. n.d. n.d. Creek 0.013 - 0.003 n.a. 0.06 -0.002 -0.007 -0.024 -0.002 -0.004 -0.003 0.003 Korangi n.d. n.d. n.d. 0.24 0.03 0.003 0.003 0.004 0.005 0.88 1.17 Creek -0.002 -0.005 -0.002 Gizri 0.028 0.08 0.15 0.011 0.22 0.046 0.029 0.031 0.043 5.85 6.49 Creek Karachi 0.018 0.07 0.1 0.007 0.13 0.024 0.02 0.021 0.046 6.68 7.12 Harbour.

Figure-4.5 Spatial Distribution of Dioxin-like chemicals -1 (pg g sed. dry wt.)

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4.4.1 Spatial Distribution Pattern of PCBs, Dioxin and Dioxin-like Contamination in Coastal Sediment

The results showed that Karachi harbour was the most contaminated sites along the coast. It was observed that highest levels of PCDD (230 pg g-1 dwt.), PCDF (24.98 pg g-1 dwt,) shown in Figure-4.6. Whereas the total non-ortho-substitute PCBs (96.40 pg g-1 dwt.), mono-ortho-substitute PCBs (15.46 pg g-1 dwt.) and PBB (7.12 pg g-1 dwt.) elaborated in Figure-4.7 and 4.8 were found in sediment collected from Karachi Harbour area in the vicinity of Lyari River discharged. The elevated levels may be due to the inducement discharges of untreated waste from coastal industries and domestic sources. The levels obtained for GC area is comparable with levels observed in the Indus River sediment collected from last barrage on the Indus before the river enters the delta area (station IR-4). The OCDD concentration was found highest amongst the PCDDs/PCDFs. Whereas levels of OCDF were found relatively lower in the coast sediment. However, Khobar Creek environment represented relatively cleaner area while comparing Dioxin contamination.

250 OCDF OCDD

200

150

100 Concentration

0 IR-1 IR-2 IR-3 IR-4 IR-5 Khobar Cr Korangi Cr Gizri Ck Kara. H. study Area

Figure-4.6 ∑OCDD and ODDF levels in the coastal and Indus River sediment (pg/g dry wt.).

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4

1800 t.PCBs 1600

1400

1200

1000

800

Concentration 600

400

200

0 IR-1 IR-2 IR-3 IR-4 IR-5 Khobar Cr Korangi Cr Gizri Ck Kara. H. Study area

Figure-4.7 ∑PCBs levels (pg g-1 dry wt.) in the Coastal and Indus River Sediment (pg g-1 dry wt.).

8 Total BDE 7

3 Concentration

0 IR1 IR-2 IR-3 IR-4 IR-5 Khobar Cr. Korangi Cr. Gizri Cr. Karachi Hr. Study Area

Figure-4.8 Total PBDE Concentrations (pg/g dry wt.) in Sediment Samples from Pakistan.

The Mono-ortho–substituted PCBs congener profile in the coastal sediment is dominated by 2,3,4,4,5-Penta-CB 123 (Figure-4.9). Whereas, main constitute of Non-

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4 ortho–substituted PCBs congener profile was 3,3,4,4-Tetra-CB 77 in the coastal area sediment (Figure-4.10).

1000 900 2,3,4,4',5-Penta-CB 114 2,3,3',4,4'-Penta-CB 105 800 2,3',4,4',5-Penta-CB 118

700 2,3,3',4,4',5,-Hexa-CB 156 600 2,3,3',4,4',5'-Hexa-CB 157 2,3',4,4',5,5'-Hexa-CB 167 500 2,3,3',4,4',5,5'-Hepta-CB 189

400 2',3,4,4',5-Penta-CB 123 concentration 300 200 100 0 IR-1 IR-2 IR-3 IR-4 IR-5 Khobar Korangi Gizri Ck Kara. H. Cr Cr Study Area

Figure-4.9 Mono-ortho–substituted PCBs congener profile (pg/g dry wt.) along the Coast.

90 3,3',4,4'-Tetra-CB 77 3,3',4,4',5-Penta-CB 126 80 3,4,4',5-Tetra-CB 81 3,3',4,4',5,5'-Hexa-CB 169

Concentration 30

0 IR-1 IR-2 IR-3 IR-4 IR-5 Khobar Korangi Gizri Ck Kara. H. Cr Cr Sampling Location

Figure-4.10 Non-ortho–substituted PCBs congener profiles (pg/g dry wt.) in the Coastal and Indus River Sediment

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4

The OCDD found highest (222-8 pg g-1 sed. dry wt.) amongst PCDD family, contributed more than 89% in sediment of KH and GC area (Table-4.3). Whereas, the concentration of 2,3,7,8-tetrachlorodibenzo-p-dioxin, the most toxic amongst Polychlorinated Dibenzo-dioxins (PCDDs) and polychlorinated Dibenzo-furans (PCDFs) family were observed highest (1.2 pg g-1 sed. dry wt.) in KH sediment, whereas levels recorded within the Gizri Creek environment (0.48 pg g-1 sed. dry wt.), it is due to the more or less same environmental condition together with similar physics of the area with minimum tidal flushing. However, relatively lower levels were found in Khohar creek (0.25 pg g-1 sed. dry wt.), it is due to its connection with Arabian Sea (Figure-4.11).

The OCDF was found highest (0.6 to 36 pg g-1 sed. dry wt.), contributed about 80% of total OCDF (Table-4.4). However, concentration profile of OCDFs is also dominated by 1,2,3,4,6,7,8 Hepta-CDF, concentrations were found in the sediment of Karachi harbour are (0.7 to 0.8 pg g-1 sed. dry wt.). A similar congener profile was obtained from coastal sediment except for the sediment sample collected from Khobar Creek area (Figure-4.12).

2 2,3,7,8-Tetra-CDD 1,2,3,7,8-Penta-CDD 1,2,3,4,7,8-Hexa-CDD 1.8 1,2,3,6,7,8-Hexa-CDD 1,2,3,7,8,9-Hexa-CDD 1.6

1.4

1.2 1

0.8 concentration 0.6 0.4 0.2 0 Khobar Cr Korangi Cr Gizri Ck Kara. H. Study area

-1 Figure-4.11 OCDD Congener Profile in the Coastal Environment (pg g sed. dry wt.)

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4

12 2,3,7,8-Tetra-CDF 1,2,3,7,8-Penta-CDF 10 2,3,4,7,8-Penta-CDF 1,2,3,4,7,8-Hexa-CDF 1,2,3,6,7,8-Hexa-CDF 1,2,3,7,8,9-Hexa-CDF 2,3,4,6,7,8-Hexa-CDF 1,2,3,4,6,7,8-Hepta-CDF 8 1,2,3,4,7,8,9-Hepta-CDF

Concentration 4

0 Khobar Cr Korangi Cr Gizri Ck Kara. H. Study Area -1 Figure-4.12 OCDFs Congener Profile in the Coastal Environment (pg g sed. dry wt.)

4.4.2 Spatial Distribution Pattern of PCBs, Dioxin and Dioxin-like Contamination in Indus River Sediment

Contamination levels in the sediment samples collected from Indus River at various localities were found relatively low (Figure-4.5) as compared to the coastal samples sites. The lower levels may be attributed to the sediment composition which was mostly compact fine grained and mostly ranges between fine sand and clay size fraction. The coarseness and compactness with comparatively less organic carbon of bottom sediments has low affinity to words the accumulation of most substances.

The comparative levels observed within the Khobar creek, it is mainly due to the sediment that Indus River carried to the delta remains within the Khobar Creek. It was also reported by Inam (2007) that sediments in the Khobar Creek is Indus River derived.

The highest levels (121.83 pg g-1 dwt.) of PCDDs were observed at the station IR- 4 (Figure-4.6). The OCDD was found to be the most dominated conger amongst the PCDD family contributing 70 to 88 % in total PCDD concentration levels. The sum of the detectable 2,3,7,8-PCDD/PCDF 3.3 pg ΣPCDD/PCDF g-1 dwt in the sample collected

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4 from the most upstream site in the Indus River (Table-4.3 and 4.4). The 2,3,7,8-TCDD was detectable in 5 of the 9 samples. Overall PCDD contributed to about 50 % of the TEQ in the samples with concentrations above 2 pg TEQ g-1 dwt and TCDD together with 1,2,3,7,8-PeCDD and 3,3',4,4',5-Penta-CB were the key contributors to the TEQ (Figure-4.13).

The levels of PCDF (27.54 pg g-1 dwt,) were observed relatively lower (Figure- 4.7). Whereas, total non-ortho-substitute PCBs (51.4 pg g-1 dwt.) and mono-ortho- substitute PCBs (516 pg g-1 dwt.) was found relatively higher at all location (Figure-4.8). The contamination levels of Polybrominated biphenyls levels were found highest (2.45 pg g-1 dwt.) in the sediment of IR-4 (Table-4.5).

4.4.3 Evolution of the Observed Contamination Levels With Respect to the EQF and TEQ

Toxic Equivalent Quantity (TEQ) is the Quantity of each PCDD/PCDF emitted multiplied by its TEF (Table-4.6). In this manner, sources of different relative quantities of the different PCDD/PCDF compounds can be compared to a standard limit (WHO 2005). If the concentrations are expressed on a toxicity equivalency basis, the concentrations of PCDD (0.3 to 2.35 pg TEQ g-1 dwt.), PCDF (0.23 to 0.967 pg TEQ g-1 dwt.), PCBs (.09 to 1.08 pg TEQ g-1 dwt.) were found in the study area (Figure-4.13). Overall PCDD contributed to about 50 % of the TEQ in the samples with concentrations above 2 pg TEQ g-1 dwt and TCDD together with 1,2,3,7,8-PeCDD and 3,3',4,4',5- Penta-CB were the key contributors to the TEQ (Figure-4.14)

Table-4.6 Concentrations Expressed on a Toxicity Equivalency Basis pg TEQ g-1 dwt.

Khoba Koran Gizri Kara. Compound IR-1 IR-2 IR-3 IR-4 IR-5 r Cr gi Cr Cr H. ∑PCDD 0.3 0.38 0.28 1.63 0.23 0.53 0.43 1.38 2.35 ∑PCDF 0.232 0.498 0.531 0.901 0.234 0.246 0.255 0.741 0.967 ∑PCBs 0.0929 0.0912 0.0906 0.7544 0.09 0.096 0.155 0.623 1.079

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4

Figure- 4.13 Distribution of the PCBs, PCDF and PCDD Concentrations expressed on total WHO-TEQ

100%

75%

50%

Total WHO-TEQ PCB 25% Total WHO-TEQ PCDF Total WHO-TEQ PCDD

Contributionn to TEQ to Contributionn 0%

Hrb

1 2 3 4 5

Karachi

Gizri Ck Gizri

Indus R. Indus

Oceanic

Korangi Ck Korangi

Indus River Indus River Indus River Indus River Indus River Indus Site

Figure-4.14 Relative Distribution Pattern of the PCBs, PCDF and PCDD Concentrations expressed on total WHO-TEQ

4.4.4 Sources of PCBs, Dioxin and Dioxin-like Contamination in the Aquatic Environment of Pakistan

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4 poor implementation of the environmental legislation and irresponsible and casual attitude towards environment issues may augment the environmental problems in the country.

1.2

2,3,7,8-Tetra-CDD 1,2,3,7,8-Penta-CDD 1 1,2,3,4,7,8-Hexa-CDD 1,2,3,6,7,8-Hexa-CDD

0.8 1,2,3,7,8,9-Hexa-CDD

0.6

concentration 0.4

0.2

0 IR-1 IR-2 IR-3 IR-4 IR-5 Sampling Locations

Figure-4.15 OCDD Congener Profile obtained from the Sediemnt of Indus River

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5 4.5 2,3,7,8-Tetra-CDF 1,2,3,7,8-Penta-CDF 2,3,4,7,8-Penta-CDF 1,2,3,4,7,8-Hexa-CDF 4 1,2,3,6,7,8-Hexa-CDF 1,2,3,7,8,9-Hexa-CDF 3.5 2,3,4,6,7,8-Hexa-CDF 1,2,3,4,6,7,8-Hepta-CDF 3 1,2,3,4,7,8,9-Hepta-CDF 2.5

2 Concentration 1.5 1 0.5 0 IR-1 IR-2 IR-3 IR-4 IR-5 Khobar Cr Sampling Area

-1 Figure-4.16 OCDF Congener profile for Indus River Sediment (pg g sed. dry wt.)

4.6 Compassion of the Observed Contamination Levels with the Published Information from Coastal Countries World

A comparison of the results obtained in Pakistan shows that the concentrations found in Pakistan are relatively low compared to many other countries in Asia as well as results from Europe and North America and are similar to results from estuaries in New Zealand and Australia (Table-4.7).

Table-4.7 Concentration (pg g-1 sed. dry wt.) of dioxin-like chemicals in sediment from different regions/countries around the world.

Concentration in pg Area Environment N TEQ g-1 dwt Mean (Min Author – Max) Pakistan Fresh – Marine 9 1.9 (0.63 – 4.8) This study Marine & Hong Kong Harbour 8 12 (4 – 33) 1 Estuarine Japan Marine 205 6.8 (nd – 260) 2 Korea Marine 19 (0.01 – 5.5) 3 Russia, Caspian Sea Marine 17 (0.7 – 28) 4 Australia Estuarine Marine 8 3.0 (0.05 – 9.9) 5 New Zealand Estuarine 26 0.53 (0.08 – 2.7) 6 Marine (Florida) 32 0.5 – 77.8 North America 7, 8 Fresh. (Ran. Riv) 45 0.3 – 34 EU, estuaries Background 22 <1 – 19 but up to > 200 9

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PCBs, Dioxin & Dioxin-like Chemicals CHAPTER-4

4.7 Conclusion

As no legislation covering dioxins or other unintentional PBTs such as dioxin and dioxin like substances exists in Pakistan, little attention has been given to the issue. No inventories on the potential sources have been compiled. Lake of awareness and poor implementation of the environmental legislation and irresponsible and casual attitude towards environment issues may augment the environmental problems in the country.

The results on the dioxin and dioxin like substance revealed that elevated levels in the coastal sediment may be attributed to the untreated effluent continuously discharges from various coastal industries such as metallurgy, pulp and petrochemicals, which might be substantial sources of these unintentional contaminates. Furthermore, practices like open pit burning of all sort of waste are very common practise in the country.

In the light of present results it may be concluded that the contamination along the coast found to be much localized. Furthermore, the contamination sources are same all along the coast. However date collected from Indus River may be not be enough to have realistic view of the contamination situation of the area. It is further recommended that detailed investigation should be carried in an around the River Indus.

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CHAPTER 5 Distribution and Fate of Polycyclic Aromatic Hydrocarbons (PAHs) in Coastal Marine Environment of Pakistan

Polycyclic Aromatic Hydrocarbons CHAPTER-5

5.1 Abstract

Assessments were made on the contamination levels of Polycyclic Aromatic Hydrocarbons (PAHs) in the sediment samples collected from selected coast localities representing diverse environmental conditions of the area. First time for Pakistan coastal waters born PAHs concentrations were estimated by using PAHs sequenced in Semi Permeable Devices (SPMDs). The sediment based aqueous levels were also evaluated for the present study.

The present results reveal that the residual concentration of sum of all analyzed

Polycyclic Aromatic Hydrocarbons (∑16PAH) varied between >5.3 to >2433.3 ng g-1 dry wt. and 8.5 to 1793.52 ng g-1 sed. OC. with a mean concentration of >457 ng g-1 dry wt. and 341.6 ng g-1 sediment OC, whereas PAHs human carcinogen contributed more than

50% (1.92 to 1301.38 ng g-1, mean levels 262.52 ng g-1 dry wt. for PAHsCARC). Significant difference (p<0.001) among the localities was observed along the coast.

Highest concentration of ∑16PAHs (>2433 ng g-1 dry wt.) were found in the sediment sample collected from close proximity of Kamari Jetty. The relatively higher levels (>1081 to 1066 ng g-1 dry wt.) were observed within the effluent discharge vicinity of Malir and Lyari River within Karachi Harbour and Gizri Creek area. Higher levels 1612 ng g-1 dry wt.) were also evidence were found in the sediment sample collected from close proximity of Kamari Jetty and Karachi Fish Harbour (Karachi Harbour area), Ibrahim Hydri Fish Harbour (Korangi creek area). However, contamination levels in the sediment of seaward stations that included navigation channel, area around oyster rock and Far east end of Korangi creek reflect significantly lowest levels (>7 to 14.36 ng g-1 dry wt.).

Distribution parental of individual PAHs were also found significantly (p<0.001) different amongst the localities. The parental PAHs profiles from effluent discharge vicinity were almost uniform, indicating predominance of the higher molecular weight PAHs (4 and 5 rings).

Although the elevated levels of ∑16PAH were related to untreated effluents from coastal Industries and domestic sewage and land run off in the coastal marine

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Polycyclic Aromatic Hydrocarbons CHAPTER-5 environment of the coast but concentrations did not exceed the safe levels range of 4000ng g-1 dry weight described by NOAA sediment Quality Guideline and concentration of individual PAHs. The ∑PAHs levels also did not exceed the concentration levels illustrated by Canadian Sediment Quality Guidelines for the protection of Aquatic Life.

5.2 Introduction

Polycyclic Aromatic Hydrocarbons (PAHs) have been recognized as environmental contaminants found in almost all compartment of the global system (Verweij et al. 2004). Preliminary results of PAHs contamination in the coastal environment from human included activities, although small concentrations are also produced by the natural processes. PAHs are undesirable group of substances produced as a result of incomplete/inefficient combustion of organic material. They are non-polar, lipophlic aromatic compounds containing two or more fused arenes structures. The persistence increased with increase in aromatic rings. However low molecular weight PAHs are more soluble and have less affinity for surfaces with significant acute toxicity than high molecular weight PAHs. According to the Neff, 1979; Witt, 1995, the higher molecular weight PAHs with a 4, 5 or 6 ring structure are more carcinogenic in nature. These are produced during high temperature combustion of organic matter, whereas low temperature burning results in PAHs with a 2 or 3 ring structure (Readman et al., 2005). It is also reported by Nemr et al., 2006 that PAHs also synthesized naturally by organisms, such as bacteria, algae and fungi.

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Polycyclic Aromatic Hydrocarbons CHAPTER-5 the level of PAHs pollution in the marine environment and their impact on the marine organisms and human health (Wurl et al., 2005).

The industrial activities in Pakistan are mostly concentrated along the coastal areas of Karachi. The industrial activities coupled with increasing coastal population are the main source of pollution in the coastal marine environment. There is dirth of “published information available on the PAHs contamination levels for the coastal marine environment of Pakistan. The coastal areas particularly in the vicinity of harbour, port and shipping yard activities are continuously under the influence of oily wastes from ship and boat, oil spills and other port & oil terminal activities (Malik et al., 2007). Therefore, a high incidence of oil and oily waste is expected in the area. Consequently coastal regions which are biologically productive areas and spawning and breading grounds of most of the fisheries may be threatened with the elevated levels of hydrophobic organic chemical, like Polycyclic Aromatic from land based sources. Only few studies have addressed the issues of PAHs in the coastal marine environment of Pakistan (Khan et al., 2002 and 2005).

The distribution pattern and occurrence of PAHs contamination levels is depended on the sources of production and their physio-chemical properties in the coastal environment (Xiang et.al. 2007, Nemr et al., 2006).

The present study is focused on the evaluation of PAHs contamination. Monitoring of these chemicals traditionally rely on the sediment sampling and a prediction of water concentration from sediment analysis due to hydrophobic in nature. For the present study Semi Permeable Devices (SPMDs), an alternative monitoring tool have been used to estimate the water borne contaminations of various PAHs in selected sites along the coast for the first time in Pakistan coastal areas. The water borne concentrations were also estimated through observed PAHs in sediment.

To evaluate the spatial distribution pattern of PAHs along the coast, 40 sampling stations were chosen in the vicinity of creek environment and Karachi

Harbour area (Figure-1.1). The total PAHs (∑16 PAHs) is the sum of priority parental PAHs such as Acenapthylene (Ace), Acenapthene (Act), Fluorene (Fl), Phenanthrene

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Polycyclic Aromatic Hydrocarbons CHAPTER-5

(Phe), Anthracene (Ant), Fluoranthene (Flu), Pyrene (Pyr), Benz(a)Anthracene (BaA), Chrysene (Chr), Benz (b+k) Fluoranthene (BbkF), Benz (e) Pyrene (BeP), Benz (a) Pyrene (BaP) ,Perylene (Pryl), Indeno (123cd) Pyrene (InP) and Benz (ghi) Perylene (BghiP).

The PAHs levels were also estimated on sediment dry weight basis “ng g-1 dry wt.” as well as normalized on organic carbon contents “ng g-1 TOC” of the sediment. The ratios of Phenanthrene/Anthracene (Phen/Anth), Fluoranthene/Pyrene (Fl/Fl + Py) were used to evaluate the sources of PAHs contamination in the area (Nemr et al., 2006).

Assessments were made on the probable eco-toxicological impacts of observed PAHs concentration to marine organisms along the coastal area by following the sediment quality guideline specified by the USEPA (1997) and the CCME (2002).

The results obtained on the level of PAHs contamination in the coastal environment of Pakistan were compared with the reported PAHs pollution levels from other coastal areas around the world.

5.3 Material and Method

5.3.1 Study Area

For the assessment of PAHs in the sediment samples collected from coastal is previously described in detail Chapter-3

5.3.2 Sediment Sampling

Methodology used for the PAHs sediment sampling was same as sample take for OCPs investigation that is elaborated in Chapter-3.

5.3.3. Semi Permeable Devices (SPMDs) Preparation

The clean-up and analyses of SPMDs were performed by using the method described by Huckins et al., 2000; Shaw et al., 2004; Shaw & Muller, 2005. SPMDs were prepared in EnTox Australia by using the method from Huckins et al., (2000).

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Polycyclic Aromatic Hydrocarbons CHAPTER-5

For the present study low density polyethylene (LDPE) 90-95 µm thickness; width 2.5 cm were prepared in the laboratory using pre-extracted LDPE tubing which was obtained from Brentwood Plastics, USA (Figure-5.2). Triolein (1, 2, 3tri [cis-9- octadecenoyl] glycerol) of 95% purity was obtained from Sigma Aldrich (Shaw et al. 2005). SPMDs were posted to Pakistan. Three SPMDs were deployed at each location with one blank as control during May-June 2003. After exposure for 3-4 weeks SPMDs were retrieved. Surface cleaned SPMDs wrapped up in foil and shipped back to EnTox Australia (during the transportation SPMDs were kept refrigerated).

66.7 66.8 66.9 67 67.1 67.2 24.9 24.9

r ive ri R

Lya

)

e r

u Korangi Fish Harbour

r a

a r C

a l

e i K f t o n

d 24.8 24.8

(

e k

a e

c e

h r

e r

t i t al

a d n d

L u n B la Is

24.7 24.7

66.7 66.8 66.9 67 67.1 67.2 Longitude (degree E)

Figure-5.1. Location map of SPMDs deployment sites along the Coast

5.3.4 SPMDs Deployment for the Estimation of Water Borne PAHs Concentration

In order to estimate PAHs contamination levels in the coastal waters of Karachi three selected sites were selected for SPMDs deployment viz Karachi harbour at the mouth of Lyari River, extreme end of the Gizri creek head of Malir river and Korangi Creek (Figure-5.1 and Table-5.1).

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Polycyclic Aromatic Hydrocarbons CHAPTER-5

Table-5.1. Sampling sites for Sediment Sampling and SPMDs Deployment along Karachi Coast.

Stations Description Longitude Latitude Depth S %0

Back side of the Karachi Fish Karachi Harbour (KFH) in the close 66.95031 24.8554 3M 33 Harbour proximate of Lyari river discharged area

Gizri Extreme end of Creek 67.0828 24.8137 1M 31 Creek

Korangi Korangi Fish Harbour (well 67.2156 24.8082 7M 35 Creek flushed area)

5.3.5 Sample Preparation and Transportation

Samples from collection sites were transported to NIO Lab in ice boxes and stand in freezer till further processing. Later on samples were freeze dried by using LABCONCO, Freeze Dry System. Freeze dried samples were homogenized by using morter and pestle and wrapped up in solvent-washed aluminium foil and posted to National Research Centre for Environmental Toxicology (EnTox), Brisbane Australia Australia for the estimation of PAHs, OCPs. All the glass wares and sampling tools were pre cleaned and rinsed with solvent (acetone and hexane).

5.3.6 Analysis of Polycyclic Aromatic Hydrocarbons (PAHs)

5.3.6.1. Sample Pre-treatment/Preparation for the Identification and Quantification of PAHs Contamination Levels

Analysis of priority PAHs (Table-5.2) were performed using techniques routinely used at Environmental Toxicological Lab and Queensland Health Lab. Pre-treatment was also elaborated by Khan et al. (2004). Briefly SPMDs were extracted using dialysis with hexane and sediments were extracted with toluene. Purification of the extracts included Gel Permeation Chromatography (GPC) and silica clean-up. SPMDs (three standard SPMDs combined) were analysed for a range of PAH using methods previously described by Shaw et al., 2004 and Shaw and Muller, 2005 .

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Polycyclic Aromatic Hydrocarbons CHAPTER-5

Figure-5.2. SPMDs specification and preparation for development in the coastal Environment.

Table-5.2. Targeted PAHs for the present Study

PAHs Abb. Acenapthylene Ace Acenapthene Act Fluorene Fl Phenanthrene Phe Anthracene Ant Fluoranthene Flu Pyrene Pyr Benz(a)Anthracene BaA Chrysene Chr), Benz (b+k) Fluoranthene BbkF Benz (e) Pyrene BeP Benz (a) Pyrene BaP Perylene Pryl Indeno (123cd) Pyrene InP Benz (ghi) Perylene BghiP

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Polycyclic Aromatic Hydrocarbons CHAPTER-5

Figure-5.3. Showing PAHs Identification and Quantification showing in Chromatogram obtained from GCMS

5.3.6.2 Instrumental Condition for the Determination of PAHs (Sediment and SPMDs):

The Analysis was conducted in SIM mode on a Varian 3400 Gas Chromatograph equipped with a Finnigan A 200S auto sampler and coupled to a Finnigan SSQ710 Single Stage Quadrapole Mass Spectrometer. A 1µL injection volume was used with split less injection onto a J&W DB-1 column approximately 20 m long with an ID of 0.2 mm and a film thickness of 0.33 lm. The chromatogram obtained from GCMS is shown in Figure-5.3.

5.3.7 Recovery of PAHs

PAHs in sediment and SPMDs were analyzed using isotope dilution techniques, prior to the extraction an internal standard of 10 deuterated PAHs (Table-5.3) was spiked into the sediment and SPMD. Into the final extracts prior to analyze 50μL of deuterated benzo[e]Pyrene was added as recovery standard.

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Polycyclic Aromatic Hydrocarbons CHAPTER-5

Table-5.3. Recovery of Deuterated PAHs

INTERNAL STANDARD Recovery % D10 ACENAPTHENE 73.18 D10 FLUORENE 81.172 D10 PHENANTHRENE 93.543 D10 FLUORANTHENE 88.949 D12 BENZ (a) ANTHRACENE 91.477 D10 CHRYSENE 98.218 D12 BENZ (b) FLUORAN 97.114 D12 BENZ (a) PYRENE 99.87 D12 INDENO (cd) PERYLENE 90.901 D12 BENZ (ghi) PERYLENE 98.66

5.3.8 Estimation of Water Concentration

The water concentration was predicted by using observed PAHs levels in the coastal sediment and SPMDs.

Sediment-based aqueous concentrations: Sediment-based aqueous concentrations (Cwsed) for present study were estimated using the equation 5.1, proposed by Verweij (2004):

-1 Cwsed=Csed(Koc) ...... 5.1

SPMDs-based aqueous concentrations SPMDs designed to measure bioavailable fractions of contaminate in the water phase. The SPMD-based aqueous concentrations (Cwspmd) of PAHs were calculated using the formula (eq-5.2) described by Petty (2004):

Cw(SPMDs) = CSPMDs VSPMD / Rst ...... 5.2

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5.3.9 Statistical Analyses

Statistical tests were carried out with SPSS 18.0 for windows (Statistical Product Service Solutions, Chicago, IL, USA). Data were analysed for statistical significance NAOVA. Hierarchical cluster analysis was also applied.

5.4 Result and Discussion

Very little published information is available on the levels of PAHs in the coastal environment of Pakistan. For the present study the estimations were made on the contamination levels in the sediment samples collected from Pakistan coastal area. This study represents one of the first efforts to quantify water borne concentrations of various PAHs estimated by evaluated PAHs levels in sediment and PAHs compounds sequestered in SPMDs deployed along selected sites of coastal area.

5.4.1 Spatial Distribution of Polycyclic Aromatic Hydrocarbons (PAHs) Contamination along Karachi Coast

Spatial distribution pattern of ∑16PAHs concentration in the sediment collected from various localities of the Pakistan coast were significantly different among the localities (p<0.004). Result obtained from present study revealed that mean -1 concentrations of ∑16PAHs (457.80 ng g sed. dry wt., and 263.34 ng/mg-TOC) were found to be in the surfacial sediment collected from coastal area. Spatial distribution pattern was observed significantly different (p<0.001) among the localities shown in Box plot mean median and inter-quarterlies (Figure-5.4). The concentrations varied between the lowest concentration (5.28 ng g-1 sed. dry wt., 8.50 ng/mg-TOC) was observed in the sediment of farthest end of Korangi Creek (Station C-4) to the highest levels (2433.27 ng g-1 sed. dry wt. 1793.52 ng/mg-TOC) found in the close proximity of Kemari Jetty, Karachi Harbour (Station H-8).

Relatively higher concentrations of various parental PAHs were observed in the sediment collected from Karachi Harbour area (>6.9 to 2433.27 ng g-1 dry wt.) than Gizri/Korangi Creek (>1072.27 ng g-1 dry wt. and 1976 ng g-1 dry wt.). Whereas,

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Polycyclic Aromatic Hydrocarbons CHAPTER-5 lowest levels were found in the sediment of Phitti/Gharo Creek system (>20.06 ng g-1 dry wt. to 216.31 ng g-1 dry wt.) shown in Table-5.4.

Table-5.4. Distribution of PAHs Contamination Levels in the Sediment Collected Coastal Localities.

∑16 PAHs ng g-1 sed. dry wt. ∑16 PAHs ng/mg-TOC Station Mean Minimum Maximum Mean Minimum Maximum

G-Cr 752.577 124.360 1072.210 177.303 123.540 218.040

K-Cr 414.653 5.280 1976.670 279.929 8.500 1080.150

Kd-Cr 183.210 150.110 216.310 194.345 170.200 218.490

Ph-Cr 13.440 9.370 20.060 17.976 9.340 32.350

Gh-Cr 108.763 64.260 165.410 388.848 184.350 827.050

Over all 457.799 5.280 2433.270 263.340 8.500 1793.520

Figure-5.4. Box plot showing Spatial Distribution Pattern (mean median and inter-quarterlies) of ∑16PAH levels in the Sediment of Selected Localities of the Coastal Area.

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The relatively higher concentrations observed in the sediment of Karachi Harbour area may be attributed to the industrial effluent discharges into Karachi harbour as well as higher levels of oil pollution due to various harbour activities. Since the flushing of the harbour and backwaters is low, it is expected that the discharges from the Lyari River remains stagnant during low tide (Akhtar et al 1995).

To have better understanding on the spatial distribution pattern of the PAHs contamination in the sediment collected from various environmental conditions of the coastal area, PAHs levels obtained were demonstrated by the Hierarchical cluster analysis that showed prominent gradient between the area under the influence of harbour and jetty, semi-enclosed area of KH and GC in the close proximity of discharge point of untreated effluents from coastal industrial zones and domestic waste from various sources and the area within the navigational channel of port and harbour and well tidal flushed where total PAHs concentrations showed variation along the coast (Figure-5.5).

The localities (H1 to H3) of semi-enclosed upper harbour and the backwaters are least flushed areas by seawater during normal daily tidal flushing (Akhtar et al 1995). Moreover during low water the oil and oily wastes floating on the surface get attached to the hard substrata in Chinna Creek and boat basin and therefore, at each high-water more, oils recruitments are brought in while less goes back out of the Karachi Harbour into the open sea area during low waters. Furthermore the untreated effluents from the Industries located around Karachi Harbour area together with the domestic sewage discharges within the upper harbour area remain stagnated in the area for long period of time (Khan et al 2006). This results in relatively higher PAHs levels (1081.00 ng g-1sed. dry wt.) in the sediment of upper harbour area (station H1 to H3).

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Figure-5.5. Hierarchical Dendogram for Sampling Localities represented by three major group obtained by Ward’s Hierarchical Clustering Method

Table-5.5. Distributions of PAHs in the Sediment of Karachi harbour Area (ng g-1 sed. dry. wt.)

Upper Mid Lower Navigation STATION KFH JK harboura harbourb harbour channel OC 7.40 2.53 1.98 1.36 1.06 0.50 AcT 0.38 8.23 49.21 53.22 15.20 0.25 Ace 0.66 1.77 12.35 18.70 2.12 0.37 Fl 0.89 2.77 9.04 21.00 0.89 0.13 Phe 94.03 50.04 171.17 328.90 17.62 0.76 Ant 26.73 3.91 7.72 22.20 1.32 0.06 Flu 133.00 52.55 165.36 178.78 5.30 0.35 Pyr 88.01 48.61 235.36 379.43 14.25 1.14 Chry 97.55 42.27 133.80 141.50 2.18 3.36 Pryl 27.94 24.44 39.95 90.78 2.58 0.02 B(a)Ant 53.49 25.32 91.65 110.42 1.69 0.97 B(b+k) flu 138.03 63.27 329.76 442.50 5.09 0.53 B(e) pyr 13.63 13.38 35.24 38.89 13.27 0.60 B(a) pyr 38.49 38.57 91.97 151.97 20.40 0.80 DiB (ah) ant 20.59 2.70 1.15 10.09 0.02 1.23 Ind(123cd)pyr 167.42 28.24 130.59 270.10 12.39 3.04 B(ghi) pryl 180.17 102.57 107.64 174.80 15.55 0.73 ∑PAHs (16) 1081.00 508.65 1611.94 2433.27 129.90 14.36 t-PAHs-Sed. 146.40 200.94 812.76 1793.52 122.84 39.49 OC PAHs CARC 695.73 302.94 886.55 1301.38 57.33 10.67 Ratio(T.PAHs/ 0.64 0.60 0.55 0.53 0.44 0.99 t.PAHs carc) Upper harboura: (Satiation H1 to H3), Mid harbourb: (Satiation H4 to H6), KFHc: Karachi Fish harbour (H8) JKd : Kamari Jetty (H7), Lower harboure: (Satiation H9 to H12), Navigation channelf: Satiation H13 to H16)

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The area around the Kemari Jetty within the Karachi Harbour (station H8) was found to be most contaminated site along the Coast (2433.27 ng g-1 dry wt.). This is could be attributed to the contamination levels were further exaggerated by the fact that Karachi Harbour is continuously under the influence of oily wastes from and oil spills from the port’s oil terminal and other harbour activities. Therefore, the highest concentration PAHs levels were found in the sediment of Karachi Harbour (Figure-5.6).

3000 ∑PAHs (16) 2500

2000

sed.dry. wt.)

1 - 1500

1000

500 Concentration (ngg Concentration 0 H1 H2 H3 H4 H5 H6 H7 H8 H9 H10 H11 H12 H13 H14 H15 H16 Sampling Location

-1 Figure-5.6. Distribution Pattern of ∑16 PAHs (ng g sed. dry. wt.) in the Sediment of Karachi harbour Area

It was reported that over 750,000 litres/Million gallons /day oily waste water from oil refineries (Ahmed, 1979) together domestic waste drained into the Indus Deltaic Creek environment via Gizri Creek (Khan et al 2006). Therefore relatively higher concentrations (1072.217 ng g-1 sed. dry wt.) were observed in the sediment of Gizri creek (station C1 and C2). However, lower levels (124.36 ng g-1 sed. dry wt.) were observed at Korangi/Gizri Creek junction (station C3), this is due its connection with the open sea area. Sediment from Korangi Creek environment showed a wide variation (p<0.001), in the spatial distribution in the PAHs. Highest level (1976.67 ng g-1 sed. dry wt. and 875.19 ng g-1 sed. dry wt.) was recorded in the vicinity of Hyderi fishing jetty (station C-8) and Rehri fishing village (station C-11), the elevated levels attributed to the mechanised boat activities, trafficking and well established repairing facilities in the area (Figure-

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5.7 and Table-5.5). A new fishing Harbour has been built in Korangi Creek, however the harbour activities were not been fully operational at the time of the sampling as shown in relatively lower levels (<172.11 ng g-1 dry wt.) at station C-14. Lowest concentrations (<5.18 ng g-1 dry wt.) were observed in the strong tidal flushed at the farthest end of Korangi Creek close to open sea area (station C4 to C6).

1200

∑PAHs (16)

1000

800

sed. dry. wt.) dry. sed.

1 -

600

400 Concentration (ng g (ng Concentration 200

0 C1 C2 C3 C4 C5 C6 C7 C10 C11 C12 C13 C14 C15 C16 C17 C18 C19 C20 C21 C22 C23 C24

-1 Figure-5.7. Distribution Pattern of ∑16 PAHs (ng g sed. dry. wt.) in the Sediment of Indus Deltaic Creek System.

The observed levels in the sediment of Kadiro Creek and Gharo creek indicate the influence of major industrial complexes including Port Qasim Industrial Zone and Steel Mill Complex that are situated in the Southern site of the coast in proximity to the Indus Deltaic Creeks. The area also received bulk of the domestic sewage (Table-5.6).

Even though navigational channel of Karachi Harbour and Gharo/Phitti Creek at Port Qasim and steel mill area are under influence of shipping traffic, the sediment from these area reflected lower levels, it may be due to the continuous dredging and strong tidal flushing of the area. The lowest concentrations were found in sandy sediments collected from Clifton beach whereas PAHs levels were found to be below detection limits in the sandy sediment of Buleji, Paradise point, Hawkes bay and Cape Monze.

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Table-5.6. Distributions of PAHs in the Sediment of Indus Deltaic Creek Environment (ng g-1 sed. dry. wt.) STATION GC KC-I KC-II IbFJ KC-OS Ka-Cr Ch-Cr Ph-Cr Gh-Cr OC 5.25 1.19 1.84 1.83 0.40 0.88 0.94 0.63 0.35 AcT 2.45 9.46 27.63 23.91 0.00 1.45 1.50 0.02 0.51 Ace 0.23 2.50 2.06 0.00 0.03 1.00 5.95 0.10 0.67 Fl 13.26 18.26 58.07 66.51 0.09 10.30 5.77 0.59 11.50 Phe 38.73 22.59 59.50 126.08 0.11 35.61 33.56 1.31 6.30 Ant 17.86 4.09 14.52 6.35 0.02 7.32 11.72 0.11 0.43 Flu 156.92 30.02 56.08 122.91 0.01 17.65 14.38 0.85 5.83 Pyr 114.78 19.72 43.59 179.58 0.02 11.20 7.71 0.80 3.89 Chry 143.82 19.54 84.36 137.71 0.20 30.54 31.71 0.26 10.28 Pryl 33.98 8.53 46.34 49.00 0.34 1.87 1.48 0.07 5.17 B(a)Ant 32.56 24.35 29.65 183.68 0.34 1.11 8.44 0.01 27.92 B(b+k) flu 134.71 28.43 159.40 260.28 1.40 14.20 13.19 0.29 3.08 B(e) pyr 53.04 22.61 55.76 144.99 0.62 0.00 3.61 0.30 9.17 B(a) pyr 22.61 14.00 50.60 231.69 0.40 7.11 5.66 0.04 16.38 DiB (ah) ant 1.00 2.19 15.31 11.21 0.01 0.02 0.57 0.00 0.69 Ind(123cd)pyr 176.05 17.53 55.91 190.67 1.52 7.60 30.83 0.91 3.22 B(ghi) pryl 124.68 24.26 39.64 242.10 2.06 3.13 7.17 1.10 3.75 ∑PAHs (16) 1066.68 268.07 798.40 1976.67 7.18 150.11 183.21 6.75 108.76 t-PAHs-Sed. OC 204.19 250.57 432.55 1080.15 23.48 170.20 194.34 7.19 388.85 PAHs CARC 635.44 130.29 434.87 1257.34 5.93 63.71 97.55 2.61 65.31 Ratio 0.60 0.49 0.54 0.64 0.82 0.42 0.52 0.25 0.63 (t.PAHs/t.PAHs carc)

5.4.2 Contamination Profiles of Parental PAHs in the Coastal Sediment.

Analytical results on the concentrations of parental PAH in the sediment samples collected from coastal area were found to be significant (Table-5.7). The Benz(b+k) Fluoranthenen was found highest (442.50 ng g-1 sed. dry wt.) in the sediment collected from KH area, Whereas mean concentration (68.671 ng g-1 sed. dry wt.) was observed in the sediment of the study area. The relatively higher levels of all most all individual PAHs were present in the sediment of Kamari Jetty, Karachi Fish harbour and Ibrahim Hydri fishing jetty. The composition of the parental PAH in the majority of the sediments within the Gizri/Korangi creek environment and upper harbour area (H1 to H6) is more or less uniform and is characterized by the predominance of higher molecular weight PAHs. The Highest concentration were found in the waste discharge vicinity of KH, GC and KC area, Benz(b+k)Fluoranthenen (>442.5 ng g-1 dry wt.), Ind(123cd)pyr (>270 ng g-1 dry wt.), B(ghi) pryl (242.1 ng g-1 dry wt.), Dibenzo(a,h)anthracene

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(>231.7 ng g-1 dry wt.), Benz(e)Pyrene; (144.99 ng g-1 dry wt.), Benzo (a) Anthracene (>183 ng g-1 dry wt.) and Chrysene (156.32 ng g-1 dry wt.). whereas lower concentration (>25.22 ng g-1 dry wt.) of Benz(a)Pyrene was found (Table-5.7). The presence of these higher molecular weight PAHs in the sediment of least flushed area it is due to the fact that these PAHs have great affinity to bind with sediment and persist for longer period in the sediment (Feng et al., 2007). It was also observed that these PAHs are well correlated with sediment organic carbon (Table-5.8). However, lowest levels anthracene (Ace) and benzo[a]-pyrene were present at most of sites because photo degradation is much more efficient along the coast (Lui et al. 2008). The ratio between ∑PAHcarc (sums of the carcinogenic PAHs) and ∑16PAH were also evaluated. The highest (0.79-0.85) ratio was observed for within the harbour area (Figure-5.9), while the lowest (0.39) was observed in Phitti Creek area (Figure-5.10).

Table-5.7. Concentration (ng g-1 sed. dry. wt.) of parental PAHs found in the Sediment Collected from Study Area.

PAHs Mean Minimum Maximum

BakF 68.671 .010 442.500 IND 49.525 .030 270.100 BghiP 50.323 .110 242.100 DBA 28.242 .010 231.690 BaA 27.769 .000 183.680 BeP 20.682 .000 144.990 BaP 4.415 .000 25.220 Chr 39.169 .000 156.320 Pyr 44.670 .010 379.430 Phe 43.339 .020 328.900 Flu 44.304 .010 198.770 Ace 6.689 .010 31.050 Act 3.069 .000 18.700 Pyrl 14.891 .000 90.780 Ant 10.628 .000 81.220 Fl 11.042 .000 66.510 T.PAHs 457.80 5.28 2433.27 t.CARC 262.52 1.92 1301.38

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Relatively higher concentrations of Chrysene was investigated in the sediments of Gizri Creek, Korangi Creek and upper harbour localities reflected the land run off in the area. Elevated levels in the terrestrial input were also explained by Wakeham et al., 1980.

A correlation analysis was carried out to have better understanding on the data structure and relative distribution parental of 16 PAHs. A significant correlation was observed between ∑16PAHs and higher molecular weight parental PAHs (Table-5.8). It was also observed that of the ∑CARCPAHs is well correlated with ∑16PAHs (R² = 0.987) (Figure-5.8) and total sediment organic carbon (R² = 0.69) (Figure- 5.13).

1000

900

800

700 PAHs CARC

600 y = 0.5866x sed. dry. wt.) dry. sed.

Linear (PAHs CARC) 1 - R² = 0.9803 500

400

300

t.PAHs (CARC) (ng g (ng (CARC) t.PAHs 200

100

0 0 200 400 600 800 1000 1200 1400 1600 1800 t.PAHs (ng g-1 sed. dry. wt.)

Figure-5.8. Correlations between ∑16PAHs and ∑CARC PAHs

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0.9

0.8

0.7

sed. dry. wt.) wt.) dry. sed.

- 0.6

0.5

0.4

0.3

Concentration (ng g (ng Concentration

0.2

0.1

0 H1 H2 H3 H4 H5 H6 H7 H9 H10 H11 H12 H13 H14 H15 H16

Figure-5.9 Ratio between ∑16PAHs and ∑CARCPAHs in the Sediment Collected from Karachi Harbour Area.

90 % CARC (T.PAHs/t.PAHs carc) 80

concentration ng/g concentration 30

C1 C2 C3 C4 C5 C6 C7 C8 C9

C10 C11 C12 C13 C14 C15 C16 C17 C18 C19 C20 C21 C22 C23 C24

Figure-5.10 Pattern of CARC Distribution in the Sediment of Indus Deltaic Creek System.

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Table- 5.8. Pearson Correlation matrix for the Coastal Sed. Parental Individual PAHs and ∑16PAHs, ∑CARC PAHs ∑Sed.OC PAHs

Correlations Ace Act Fl Phe Ant Flu Pyr Chr Pyrl BaA BakF BeP BaP DBA IND BghiP T.PAHs t.CARC

OC 0.47 0.04 0.29 0.78 0.78 0.80 0.82 0.75 0.78 0.64 0.77 0.72 0.62 0.47 0.77 0.79 0.83 0.79

Ace 0.54 0.59 0.71 0.59 0.67 0.66 0.66 0.67 0.62 0.70 0.66 0.69 0.33 0.64 0.57 0.72 0.71

Act 0.11 0.30 0.24 0.25 0.24 0.18 0.12 0.16 0.27 0.01 0.21 0.15 0.21 0.03 0.19 0.18

Fl 0.52 0.47 0.56 0.52 0.55 0.63 0.60 0.67 0.59 0.54 0.39 0.36 0.49 0.57 0.55

Phe 0.93 0.91 0.91 0.87 0.85 0.79 0.91 0.70 0.85 0.59 0.81 0.89 0.95 0.92

Ant 0.90 0.85 0.88 0.80 0.71 0.86 0.66 0.73 0.63 0.81 0.84 0.90 0.87

Flu 0.96 0.90 0.89 0.84 0.92 0.71 0.73 0.55 0.83 0.86 0.95 0.92

Pyr 0.85 0.87 0.83 0.91 0.73 0.78 0.58 0.80 0.83 0.94 0.91

Chr 0.87 0.78 0.90 0.76 0.74 0.62 0.80 0.80 0.92 0.92

Pyrl 0.84 0.92 0.80 0.80 0.60 0.78 0.84 0.94 0.93

BaA 0.88 0.66 0.70 0.63 0.67 0.76 0.87 0.87

BakF 0.75 0.81 0.59 0.83 0.87 0.95 0.95

BeP 0.79 0.32 0.66 0.66 0.79 0.79

BaP 0.42 0.64 0.73 0.85 0.86

DBA 0.51 0.50 0.61 0.59

IND 0.83 0.86 0.88

BghiP 0.91 0.90

∑PAHs 0.99

5.4.3 Correlation with total Organic Carbon (TOC) and total PAHs in Sediment

Sedimentrolgical studies such as TOC content and particle size distribution showed varied environmental condition for the accumulation of most of the contaminates along the coast (Wang et al, 2001). Therefore, to reduce the effect of sediment properties the observed PAHs levels were normalized to total sediment organic carbon (TOC) contents (Figure-5.11). It was pragmatic that the sediment with high organic reflect the higher levels of PAHs except at the stations in close proximity discharges, Ibrahim

Hyderi (C-7) and Kamari jetty (H8). Distribution pattern of 16PAHs and PAHsCARC strongly correlated (R2=0.836, R² = 0.69) with total sediment organic carbon contents

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(Figure-5.12 & 5.13). It was also reported by Nemr et al. 2006, that sediment organic carbon plays an important role in controlling the PAHs levels in the marine sediment. It was pragmatic that higher molecular weight PAHs are well correlated with the sediment organic carbon than lower molecular PAHs (Table-5.9).

2500

t.PAHs-OC

) 1 - 2000

1500

1000 Concentration (ng sed. OC sed. (ng Concentration 500

0 H1 H3 H5 H7 H9 H11 H13 H15 C1 C3 C5 C7 C9 C11 C13 C15 C17 C19 C21 C23

Stations along the coast Figure-5.11. Distribution Pattern of ∑PAHs Concentrations Normalized to total Sediment Organic Carbon

1600 R² = 0.836 1400

1200

1000 dry wt.) dry

1 1 800 -

600 (ng g (ng 400

200 ∑ PAHs ∑

0 0 1 2 3 4 5 6 7 8 Sed OC

−1 Figure-5.12. ∑16PAHs (ng g dry wt.) against total Sediment Organic Carbon from the Coastal Area.

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1000

900 y = 104.86x + 19.964 800 PAHs CARC R² = 0.6889

700

drywt.) 1 1

- 600 Linear (PAHs CARC) (ngg

500

400

300

200

PAHs (CARC) PAHs ∑ 100

0 0 1 2 3 4 5 6 7 8

Sed. OC

−1 Figure-5.13. ∑CARCPAHs (ng g dry wt.) against total Sediment Organic Carbon.

Table-5.9. The Nonparametric Correlations (Spearman's rho) between Sed. Organic Carbon and Individual PAHs

PAHs Ace Act Fl Phe Ant Flu Pyr Chr Pyrl BaA BakF BeP BaP DBA IND BghiP T.PAHs t.CARC

Correlation .467 .040 .285 .784 .784 .802 .820 .746 .776 .641 .775 .724 .616 .467 .768 .792 .826 .788 Coefficient

Sig.(2- 0.004 .083 0.082 .000 .000 .000 .000 .000 .000 .000 .000 .000 .000 .006 .000 .000 .000 .000 tailed)

5.4.4 Evaluation of PAHs Carcinogenic Risk Potency

Amongst PAHs family the USEPA (IRIS 1994) has defined PAHs (benzo(a)anthracene, benzo(a)pyrene, benzo(b)fluoranthene, benzo(k)fluoranthene, chrysene, dibenzo(a,h)anthracene and indeno(1,2,3-cd)pyrene) are probable human carcinogens. It was observed that PAHs contamination in the most of coastal sediment dominated by higher molecular weight PAHs (BaA, B(bk)P, InP, BaP, B(ghi)P (Table- 5.9). These contaminates are probable and possible human carcinogens (Hassain et al 1998). PAHs CARC contributed about 67 to 71 % in the sediment of KH area, whereas, Gizri Creek dominated (>60%).

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The concentrations observed in the sediment were evaluated with the biological effect levels defined by Long et al. 1995. The results from present study revealed that the concentrations of PAHs CARC were found to be much lower than adverse biological effect levels (ERL and ERM) recommended by Long et al. (1995) for the coastal environment of Pakistan (Figure- 5.14).

2000

1800 ERL KH GC KC ERM 1600

drywt.) 1400

1 1 - 1200 (ng g (ng 1000 800 600 400 Concentrations 200 0 B(a)A B(b+k)F B(e)P B(a)P DBA PAHs

Figure-5.14. Concentration (ng g-1 dry wt.) of max. Levels obtained from various Localities of the Coastal Area Relative to Biological Effect Levels (ERM and ERL).

The similar observations were also noticed for the Threshold Effect Level (TEL) defined by CCME, 2001. Whereas B(a)A and B(a+k)F levels in the sediment of KC, B(a+k)F, B(e)P and B(a)P and Karachi Harbour were higher than and Probable Effect Level (PEL) (Figure-5.15). However, the concentrations Ace, Fl and Ant were found much higher than the defined TEL, ERL and PL for the sediment of KH in the proximity of KFH and KJ and KC at Ibrahim Fishing Jetty. Whereas, Flu concentrations were closed to the corresponding TEL value.

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800

700 TEL1

KH 600

dry wt.) dry 1 1

- 500 KC PEL

(ngg 400

300

200

Concentration 100

0 B(a)A B(b+k)F B(e)P B(a)P DBA

PAHs Figure-5.15. Concentration (ng g-1 dry wt.) of max. Levels obtained from various Localities of the Coastal Area relative to Threshold Effect Level (TEL) and Probable Effect Level (PEL).

For the present study the TEF defined by Nisbet and LaGoy (Table-5.10) have been used to evaluate the toxic potency of each individual PAH compound. The total PAHs concentration observed in the coastal sediment and their relative levels with their corresponding benzo(a)pyrene equivalent concentrations (Table-5.11).

This may be summaries that the CARC-PAHs in the coastal sediments may not create adverse effects on the marine organisms, especially benthic fauna or directly in contact with the sediments. It is also concluded that toxicity of lower molecular weight PAHs need to further investigate;

Amongst PAHs family the USEPA (IRIS 1994) has defined seven PAHs (benzo(a)anthracene, benzo(a)pyrene, benzo(b)fluoranthene, benzo(k)fluoranthene, chrysene, dibenzo(a,h)anthracene and indeno(1,2,3-cd)pyrene) are probable human carcinogens.

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Table-5.10 Toxic Equivalent Factor of Individual PAHs defined by Nisbet and LaGoy. PAH Components TEF Naphthalene 0.001 Acenaphthalene 0.001 Acenaphthene 0.001 Fluorene 0.001 Phenanthrene 0.001 Anthracene 0.01 Fluoranthene 0.001 Pyrene 0.001 Benzo(a)anthracene 0.1 Chrysene 0.01 Benzo(b)fluoranthene 0.1 Benzo(k)fluoranthene 0.1 Benzo(a)pyrene 1.0 Indeno(1,2,3-cd)pyrene 0.1 Dibenzo(a,h)Anthracene 1.0 Benzo(g,h,i)perylene 0.01

Table-5.11. Total PAHs and Carcinogenic PAHs in Coastal Sediment

∑ PAHs ng g-1 sed. dry wt. ∑CARC PAHs ng g-1 sed. dry wt. Station Mean Minimum Maximum Mean Minimum Maximum

KH 639.79 6.94 2433.27 369.00 4.07 1301.38

GC 752.58 124.36 1072.21 435.71 36.25 711.65

KC 414.65 5.28 1976.67 235.01 4.17 1257.34

KdC 183.21 150.11 216.31 97.55 63.71 131.39

PhC 13.44 9.37 20.06 4.39 1.92 5.92

GhC 108.76 64.26 165.41 65.31 44.89 92.83

∑PAHs 457.80 5.28 2433.27 262.52 1.92 1301.38

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5.4.5 Estimated Water Concentrations of PAHs in the Coastal Environment

The highest total PAHs concentrations in water from Karachi Harbour were estimated by sediment as well as SPMDs. Sediment and SPMD estimated water concentration was well correlated (R2 =0.5) Figure-5.16. Higher levels of PAHs observed at Karachi Harbour could be attributed to the industrial effluent discharges into Karachi harbour as well as to the higher levels of oil pollution due to various harbour activities. Since the flushing of the harbour and backwaters is low, it is expected that the discharges from the Lyari River remain stagnant during low tide (Akhter et al 1995). This could create conditions close to the equilibrium between water and sediment and therefore, stable concentrations of PAHs in both, sediment and SPMDs was pragmatic. Similar situation was also observed for Gizri Creek site (R2=0.7). Despite the good correlation for the regression of log total PAH(SPMD) against log total PAH(sed), there were substantial compositional differences between PAH levels in sediment and SPMDs (Figure-5.16) (Boehm et. at. 2005).

Figure-5.16. PAHs Concentration (median, interquartile range, range values and outliers) in Sediment and SPMDs along Karachi Coast.

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It was observed that SPMDs were augmented with low molecular weight PAHs, whereas relatively higher molecular weight PAHs were found in the sediment of Karachi coastal area. The difference in sequencing in the SPMDs and sediment levels were also reported by various authors (Lebo, et al., 1995; Shigenaka and Henry, 1995; Petty, et al., 2004; Huckins, et al., 1999; Bergqvist, et al., 1998; Rantalainen, et al.,1998; Miege, et al.,2000; Zimmerman, et al., 2000; Bruce, et al., 2003; Lu & Wang, 2003; Wang, et al., 2001; Verweij, et al 2003, Wang, et al., 2003; Boehm, et al., 2005; Richardson, et al., 2005; Gourlay, et al., 2005; Vrana, et al., 2005). SPMDs accumulate nonpolar organic compounds in dissolved and colloidal forms by direct adsorption/absorption in the polyethylene membrane (Boehm et al. 2005). The rate of equilibration of the triolein phase of SPMDs with dissolved PAH in the ambient water decreases with increasing

PAH molecular weight and octanol/water partition coefficient (Kow) (Boehm et al. 2005).. Lower molecular weight PAH partition rapidly from sediment into the SPMD and reach equilibration within relatively shorter period of deployment time (Boehm et al., 2005). The SPMDs deployed for 20 to 25 day in the present study may not be efficient for the sequestration of higher molecular weight PAHs because high molecular weight 4- and 5- ring PAH partition into the SPMD more slowly and may require more time to reach equilibrium (Boehm et al. 2005,). That could be reason for the lower levels of estimated water concentrations by SPMDs and low variability in different PAHs levels at all stations (Figure-5.17).

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Figure- 5.17. Estimated Water Concentration (median, interquartile range, range values and outliers) by SPMDs “Cw " & Sediment “Cw " along Karachi Coast.

Higher levels of low molecular weight PAHs were also observed in SPMDs estimated water born concentration. A larger variation in Cwsed was also observed by (Verweij, et al

2003). The differences between Cwsed and Cwspmd, may be attributed to the variability in distribution pattern of various pollutants in the sediments and Koc value used for computing Cwsed were not evaluated from (Boehm et al., 2005). The specific sediment organic carbon composition and the assumptions of sorption equilibrium and mixing with the water column complicates the prediction of aqueous contaminant concentrations from sediment concentrations (Verwej et al. 2004).

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Figure-5.18. Correlation between log of Water born Concentrations Estimated by Sediment and SPMDs.

5.4.6 Source of PAHs Contamination in the Coastal Environment

Present results revealed that Petroleum derived PAHs (three or less aromatic rings) and pyrogenic PAHs (four or more aromatic rings) are both present. This could be due to the variety of PAHs sources in the coastal area (Readmen et al., 2002).

Pakistan coastal area is complex dynamic system with several sources of pollution. The effluent discharges from oil refineries, mechanized fishing boats and the cleaning of bilges and tank washing by the large number of merchant vessels as well as oil tankers that pass through the EEZ of Pakistan, yearly about 2500 oil tankers carry 33 million tons of crude oil (UN SCAPICZM, 1996), oil terminals at Karachi Harbour, Port Qasim, and occasional oil spills together with untreated effluent from coastal industries and domestic sewage could be the sources of PAH pollution in the coastal marine environment of Pakistan. Therefore It is important to categorize the sources of PAH

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Polycyclic Aromatic Hydrocarbons CHAPTER-5 pollution in the coastal marine area. The PAHs introduced in the environment as a result of incomplete combustion of organic matters that include combustion of fossil fuel, smelting, waste incinerators, vehicular engine combustion, forest fire and coal combustion called pyrolytic sources and petrogenic input is due to the unburned petroleum and its product – gasoline, kerosene, diesel, lubricating oil and asphalt.

For the present study Phenanthrene/Anthracene (Phen/Anth), Fluoranthene/Pyrene (Fl/Py) molecular ratio has been used for the identification of PAHs contamination sources in the area. the Phen/Anth ratio in this study are in the range of 1.2 – 1.9. The Lower Phen/Anth concentration (>10) ratios at most of the creek environment along the coast indicated that the PAHs were derived from pyrolytic rather than petrogenic sources at most of the site (Figure-5.19).

The dendrogram represents three major groups (Figure-5.20). The first major group mainly consists of anthropogenic carcinogenic PAHs and can be assigned as combustion and carcinogenic group. The second consists Ant, Pyr, Flu and Chr while the third group consists petroleum origin (Ace, Fl, Naph and Acth).

KH GC KC ADC 30

MIXTURE

PETROGENIC

Phe/Ant Phe/Ant 20

10 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 Flu/Pyr 5 PYROLYTIC

Figure-5.19. The ratios Phe/Ant (phenanthrene/anthracene) verses Flu/Pyr (Fluoranthene/ Pyrene) to evaluate PAHs sources (petrogenic and pyrolytic) in the Coastal Sediment.

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A mixture of pyrolytic and petrogenic PAHs sources were observed in most of the sediments of Karachi harbour area, usually with a slightly pyrolytic predominance within the upper Harbour area. Higher level of perylene at certain location (Karachi fish harbour-H7 and Kemari Jetty H-8) indicated biogenic input of PAHs (Readman et.al. 2005). Whereas, Gizri Creek is enriched with pyrolytic driven PAHs (Figure-5.19).

The present results on the molecular ratio indicate that sediments along coast contain a mixture of petrogenic and pyrolytic sources of PAHs. Stations with greatest pyrolytic inputs include upper harbour area, Gizri Creek and most of part of Korangi creek.

To have better understanding of the source potential of area the values of PHE/ANT were plotted against values of FLTH/PYR (Fig. 5.19) that effluents from Malir and Lyari rivers carries pyrolytic inputs of PAHs, this is confirmed that harbour stations in the close vicinity of discharge points contributes oil to sediments at the station in the close vicinity of Malir River out fall (Readman et.al. 2005). The plot confirms the dominance of the combustion derived material at most Karachi harbour stations (Figure- 20). However, the diagram fails to classify e.g. station in vicinity of open sea area into its clear petrogenic class endorsing the need to use a variety of measurements in order to provide a robust assessment (Readmen et al., 2002).

Figure-5.20. Dendogram showing input of PAHs from various sources

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5.4.7 Comparison of Contamination Levels of Sedimentary PAHs with Other Coast Regions Around the World

The concentration range of total PAHs found in this study was compared with those reported from other countries (Table-5.12). Three Asian locations e Thailand (Boonyatumanond et al., 2006), Malaysia, and Tokyo (Zakaria et al., 2002) are included. Globally, rivers, lakes, estuaries, harbours and coastal areas have been polluted by PAHs, with concentrations ranging from 1 to 760,000 ng g-1 , and with modal concentrations of 103e10,000 ng g-1 (Zakaria et al., 2002). Sedimentary PAH concentrations in Coastal sediment were comparable to those in Thailand. They were one order of magnitude greater than those in Malaysia and i.e. orders of magnitude lower than those in Tokyo. In general, contamination levels of sedimentary PAHs in Karachi coast can be categorized as low to moderate on a global scale. Contamination levels in the Creek environment can be categorized as low, due to the concentrations ranging from 107 to 633 ng g-1. Those in the Harbour area with concentrations of 313-1707 ng g-1 can be categorized as low to moderate.

Table- 5.12. Worldwide PAHs Contamination Levels

Area Total PAHs References Pakistan Coast 8.5-2433 (16 PAHs) Present study Xiamen Harbor, China 2900-61000 Hong et al. (1995) Saudi Arabia, Gulf 11000–6900000 Readman et al. (1996) Kitimat Harbor, Canada ND- 10000000 Simpson et al. (1996) Lazaret Bay, France 1600–48090 Benlahcen et al. (1997) Gironde estuary, France 622–4888 Budzinski et al. (1997) Crete,Eastern 500–5700 Gogou et al. (2000) Mediterranean Boston Harbor, USA 7300–358000 Wang etal. (2001) Santander Bay, Spain 20–25800 Viguri et al. (2002) Sochi, Black Sea, Russia Redman et al. (2002) Hsin-ta Harbor, Taiwan 1155–3382 Fang et al. (2003)

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5.5 Conclusions

Very little information is available on the levels of PAHs in the coastal environment of Pakistan. For the present study the estimations were made on the contamination levels in the sediment samples collected from Pakistan coastal area.

Significant difference (p<0.001) among the localities was observed along the coast. Karachi Harbour in the close proximity of Kamari Jetty was found most contaminant site along the coast whereas, Creek environment around the Ibrahim Hydri Fish Harbour (Korangi creek area) were observed under influence of PAHs pollutants. However, contamination levels in the sediment of seaward stations that included navigation channel, area around oyster rock and Far East end of Korangi creek reflect a cleaner environment while comparing the PAHs pollution levels. Distribution parental of individual PAHs were also found significantly different. The parental PAHs profiles from effluent discharge vicinity were almost uniform, indicating predominance of the higher molecular weight PAHs (4 and 5 rings).

The present results reveal that the residual concentration of sum of all analyzed

Polycyclic Aromatic Hydrocarbons (∑16PAH) were related to untreated effluents from coastal Industries and domestic sewage and land run off in the coastal marine environment of the coast but concentrations did not exceed the safe levels range of 4000ng g-1 dry weight described by NOAA sediment Quality Guideline and concentration of individual PAHs. The ∑PAHs levels also did not exceed the concentration levels illustrated by Canadian Sediment Quality Guidelines for the protection of Aquatic Life.

This study represents one of the first efforts to quantify water borne concentrations of various PAHs estimated by evaluated PAHs levels in sediment and PAHs compounds sequestered in SPMDs deployed along selected sites of coastal area.

The differences between Cwsed and Cwspmd, observed in the present study could be due to the variability in the distribution of various pollutants in the sediments and Koc value used for computing Cwsed values, that do not account for the specific sediment organic carbon

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CHAPTER 6 KEY FINDINGS, IMPLICATIONS AND FUTURE RESEARCH

CONCLUSION CHAPTER-6

The present work is the first of its kind to describe in detail the fate and distribution pattern of Persistence Bio-accumulative and Toxic (PBTs) that included Organochlorine Pesticides (OCPs), PCBs, Dioxins and Dioxin like substances and Poly Aromatic Hydrocarbon (PAHs) in the coastal environment of Pakistan bordering Northern Arabian Sea.

Despite their widespread use and injurious effect of PBTs, little information is documented on contamination of these substances in the coastal marine environment of Pakistan. Mangrove swamps, intertidal mudflats and Indus Deltaic Creek system are the main feature of the coastal area of Pakistan. The coastal areas are of significance as there are spawning, breeding and nursery grounds of commercially important fishery resources. These living resources are under continuous threat of untreated industrial effluents and sewages discharges into coastal areas via several sources.

To evaluate the spatial distribution pattern and potential sources of contamination of PBTs in the Sindh and Balochistan coastal environment were investigated. The influence of Indus River discharges in the coast environment were examined by evaluating unintentional produced contaminates such as dioxin and dioxin like substances. The results from these studies provided fundamental information for a semi quantitative assessment of risks associated with the presence of elevated PBTs.

To obtain a true picture on the spatial distribution pattern of OCPs and PAHs contamination levels in coastal environment of Pakistan the results obtained during the present study was plotted using the Software "ArcGIS Version 10", the extension "Geostatistical Analyst" and "Radius Basis Function" interpolation method.

The Followings are the Review of Key Findings of this Study

The results obtained from the present study on the Organochlorine Pesticides contamination levels revealed that a larger variation of ∑OCPs contamination levels (>0.002-17.5 ng g-1 dry wt. with a mean concentration of > 4.5 ng g-1 dry wt.) was observed in the samples collected from various localities of coastal area of Pakistan. Highest residual concentrations (17.5 ng g-1 dry wt.) were found in semi-enclosed area in the effluent discharge vicinity attribute to low tidal flushing of the area. Whereas, strong tidal flushed vicinities were showed lowest level of contamination. Spatial distribution

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CONCLUSION CHAPTER-6 pattern were significantly different among the localities (p<0.004). Distribution pattern of most of the PBTs were strongly correlated with total sediment organic carbon contents (p>0.767 & R2=0.66). However, OCPs contamination levels in the sediment collected from Balochistan coastal environment were found below detection limit at most of the sampling sites.

The OCPs contamination levels were found localized in the coastal area, higher in the discharged vicinity and found lowest in the strong tidal flushed area. To have better understanding to the situation of the spatial distribution pattern the interpolation models were developed from the data obtained from the present study that is also reflected in the localized pollution effects of OCPs along the coast (Figure-6.1).

Figure-6.1 Spatial distribution pattern of ∑ Organochlorine Pesticides in Coastal sediment of Pakistan

Strong correlation between OC-pesticides and sediment organic carbon and marked concentrations difference at seaward stations and area in the proximity of pollution discharge outlets were also suggests that contamination along Karachi coastal area seems to be localized (Figure-6.2). Further, good correlation amongst various OC

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CONCLUSION CHAPTER-6 pesticides (Figure-3.29) reflects probably common sources of contamination along Karachi coast.

Figure-6.2 Spatial Distribution Pattern of ∑ Organochlorine Pesticides Normalized on Sediment Organic Carbon from Pakistan Coastal

The residue of DDT mainly its metabolites DDE and DDD were widely distributed and have been detected in most of the samples in relatively higher concentrations as compared to other OCPs (Figure-6.3). DDTs were predominant contaminants in most of the sediment samples due to its less volatile and slow degradation nature in the aquatic environment (WHO 1989, Carvalhho et al 1992). DDTs contributed >60% of total analyzed OCPs. The high proportion of pp′-DDE at most of the site (41–95%) and ratio of ∑DDT and DDT (0.04 –0.24) suggests old inputs of DDTs in the environment. This may be attributed to the restrictions being implemented on the use of DDTs and Pakistan has also switched over to natural pest control or using safer formulas. The DDT’s metabolites DDE were also found in soft tissue of the marine biota (fishes, crab, shrimps and molluscs) collected from Coastal area.

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By analyzing the individual HCH isomers, it was found that β-HCH had the highest level of concentration among all the samples and this HCH isomer was higher because of its persistence in sediment. The persistence of β-HCH in sediment is mainly due to the higher Kow (log Kow =3.78) and lower vapor pressure value (3.6x10-7 mmHg, 20oC) (Zhang et al., 2006). These will make β-HCH easier to be absorbed to the sediment organic matter and less evaporative loss from sediments (Mackay et al., 1997). Furthermore, the spatial arrangement of Chlorine atoms in the molecular structure of β- HCH was supposed to be more resistant to microbial degradation in soils (Middeldorp et al., 1996). The spatial distribution pattern of ∑ HCH is elaborated in Figure-6.4.

Figure-6.3 Spatial Distribution Pattern of ∑ DDTs in Sediment Samples Collected from Pakistan Coastal Area

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Figure-6.4. Spatial Distribution Pattern of ∑ HCHs in the Coastal Sediment of Pakistan

The port and harbour activities and untreated effluents from industries and domestic sources reflect highest PAHs contamination levels (2610.812 ng g-1 dry wt.) in the vicinity of Kemari jetty, Karachi Harbour area. The dispersion model of PAHs contamination levels indicating the most affected area were found to be the areas under the influence of port and harbour area and fishing jetties along the coastal area (Figure-

6.5). However, PAHsCACR were also concentrated in the discharged vicinities of Harbour and Creek environment (Figure-6.6).

The concentrations of ∑16 PAHs varied amongst the localities, highest concentrations (2610.81 ng g-1 dry wt.) were detected in sediment samples collected in vicinity of Karachi harbour. Relatively lower levels (>400ng/g) observed in Korangi creek area south-east of Karachi. Higher concentrations (>330ng/g dry wt.) of higher molecular weight PAHs such as, Benz (a) Pyrene (B(a)P) were detected near the discharge points of Lyari river. The Phen/Anth and Flth/Pyr concentration ratios indicated that mixture of pyrolytic and petrogenic PAHs sources at most of the site along the coast.

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Figure-6.5 Spatial Distribution Pattern of ∑PAHs in Sediment Samples Collected from Study Area.

Figure-6.6 Spatial Distribution Pattern of ∑PAHsCARCs in Sediment Collected from Coastal Area.

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To reduce the effect of sediment properties the observed PAHs levels were normalized to total sediment organic carbon (TOC) contents. The spatial distribution pattern were found comparable results found on the total sediment weight indicating same sources of pollution and contamination along the coast related to the environmental condition of the area that elaborated in the spatial distribution plot (Figure-6.7). It was pragmatic that the sediment with high organic reflect the higher levels of PAHs except at the stations in close proximity discharges, Ibrahim Hyderi (C-7) and Kamari jetty (H8). 2 Distribution pattern of 16PAHs and PAHsCARC strongly correlated (R =0.836, R² = 0.69) with ∑ sediment organic carbon contents.

Figure-6.7 Spatial Distribution Pattern of ∑PAHs Normalized on Sediment Organic Carbon

The PBTs were found to be strongly correlated with sediment organic carbon and tendency to accumulate in the organic phase of the sediment. Therefore sediment could be continued sources of contamination and cause adverse impact on the marine resources (Carvalho et al 2002). Effect based guidelines for sediment quality were

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CONCLUSION CHAPTER-6 use to identify the areas in which the potential for biological effects is greater and to assist environmental management. Present contamination results revealed that most of the exceed values were corresponded to the levels found in the discharged proximity of low flushed areas along Karachi coast such as upper-KH, GC, few localities within KC, Ch-C and Ka-C. Whereas most of the investigated area did not reflect the comparable the concentration above adverse effects are predicted to occur frequently (probable effect level PEL), except Gizri creek where DDE levels exceed from PEL (6.75 ng g-1 dry wt) and Apparent Effects Threshold (AET) 9 ng/g for benthic organisms for ∑DDT exposure, whereas KH and KC showed closer levels to the AET. However concentrations found for other Indus deltaic creeks and seaward areas along Karachi coast were found much lower than the reported sediment quality criteria and concentration ranges.

The results clearly indicate that elevated concentrations found in the sediment of Pakistan coastal area were very much localized and may attributed to the untreated waste from domestic sources and industrial effluents discharged nd in the semi-closed area of upper harbour and Gizri creek where tidal flushing is very poor and waste water remain stagnated for longer period of time (Khan et al 2006).

Dioxin and Dioxin-like chemicals were detectable in all samples collected from Pakistan. As no legislation covering dioxins or other unintentional POPs exists in Pakistan, little attention has been given to the issue. No inventories on the potential sources have been compiled. Lack of awareness and poor implementation of the environmental legislation and irresponsible and casual attitude of authorities towards environment issues may augment the environmental problems in the country. Pakistan has a number of industries in the fields such as. metallurgy, pulp and petrochemicals, which might be substantial emitters of unintentional POPs. Furthermore, practices like open pit burning of waste are very common.

First time water borne PAHs contaminations were estimated using Triolein- passive sampler Semi-Permeable Membrane Devices (SPMDs) as an alternative monitoring tool for coastal waters of Pakistan. The estimated water concentration found 2 highest in the harbour area (CwSPMDs 4.6ng/l) that is well correlated (R =0.5) with the

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CONCLUSION CHAPTER-6 evaluated contamination levels (CwSed. 35.67ng/l) using the levels observed in the sediment.

Anthropogenic PAHs were quantitatively analyzed in this study. ∑ PAH concentrations of surface sediments from Karachi coast ranged from 13 to 116 ng/mg- TOC, with an average of 68 ng/mg-TOC. Contamination levels of sedimentary PAHs may be categorized as low to moderate on the global scale.

Source analysis revealed that, the sediment along coast, petrogenic sources played a major role in most part of Karachi harbour whereas Creek environment, both petrogenic and pyrogenic sources both were significant.

According to the sediment quality standards of the USEPA and Canadian Council of Ministers of the Environment, observed levels of OCPs, PAHs and Dioxin contamination levels were generally lower than the threshold known to harm wildlife by OCPs. However, PAHs levels demonstrate moderate to low risk.Overall PCDD contributed to about 50 % of the TEQ in the samples with concentrations above 2 pg TEQ g-1 dwt and TCDD together with 1,2,3,7,8-PeCDD and 3,3',4,4',5-Penta-CB were the key contributors to the TEQ.

The results clearly indicate that elevated concentration of PBTs contamination in the marine sediment of coastal areas was localized and much lower than the concentrations reported from neighbouring and regional countries.

Recommendation for Future Research and Monitoring

The present study was focused on the selected high priority pollutants along the coast. It is recommended that a detailed hydrodynamic study with reference to the residence time of the various pollutants in the area need to be evaluated for better understanding of the fate of pollution and their rate of dispersion.

It is also recommended that monitoring of other PBTs may also be monitored to have comprehensive profile of the pollutants in the marine environment of Pakistan.

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