Oyster Biomonitoring Study€¦ · oysters each were deployed at eight locations with increasing distance from the Burwood Beach outfall. Oysters were deployed in approximate north-eastern

HUNTER WATER

Oyster Biomonitoring Study

Burwood Beach WWTW

301020-03413 – 105

August 2013

Infrastructure & Environment

3 Warabrook Boulevard Newcastle, NSW 2304 Australia PO Box 814 NEWCASTLE NSW 2300 Telephone: +61 2 4985 0000 Facsimile: +61 2 4985 0099 www.worleyparsons.com ABN 61 001 279 812

© Copyright 2013 WorleyParsons

HUNTER WATER

OYSTER BIOMONITORING STUDY

BURWOOD BEACH WWTW

Page ii 301020-03413 : 105 FINAL DRAFT: August 2013

SYNOPSIS

The Burwood Beach Oyster Biomonitoring Study was undertaken to assess the potential for effluent

and biosolids discharges to lead to bioaccumulation of chemicals in marine life, over a range of

spatial scales, using oysters as a biomonitor. A specific requirement of the study was to establish the

zone in which there is a detectable increase in the concentration of chemicals in oysters that is

related to the outfall. Concentrations of a suite of organic and metals/metalloid chemicals were

measured in Sydney rock oyster, Saccostrea glomerata, tissue following three eight week long

deployments in the receiving waters of the Burwood Beach WWTW. Three replicate cages of thirty

oysters each were deployed at eight locations with increasing distance from the Burwood Beach

outfall. Oysters were deployed in approximate north-eastern and south-western directions at

distances < 50 m (outfall), 100 m, 500 m and 2000 m from the outfall biosolids diffuser.

Concentrations of organics and metals/metalloids were compared to the Australian and New Zealand

Food Authority (ANZFA) Standards Maximum Residue Limits (MRLs) (ANZFA 2011). These were

used as a guide (in the absence of other available guidelines) rather than to assess health risks for

oyster consumption, as there are no commercially grown oysters within the boundaries of this study.

Analysis of organic chemicals included a suite of organochlorine (OC) and organophosphate (OP)

pesticides, polychlorinated biphenyls (PCBs) congeners and total PCBs (summation of PCB

congeners). There were detections of organic chemicals in oysters suggesting that Burwood Beach

WWTW was a source during some of the deployment periods. Heptachlor, trans-chlordane, cis-

chlordane and dieldrin were detected in oysters following January - April 2012 at concentrations lower

than the ANFZA MRLs (2011). Concentrations of PCB congeners and total PCB concentrations were

lower than the LOR of 0.01 mg/kg in all oysters following the three sampling events.

Assessment of metals/metalloid concentrations in oyster tissue following deployments demonstrated

that most metals/metalloids were at low concentrations. No metals/metalloids were found to exceed

the available ANFZA MRLs (2011). Concentrations of metals/metalloids were also found to be similar

to those reported as background concentrations for S. glomerata in NSW estuarine locations. There

were no significant differences in the spatial patterns of individual metals/metalloids or the multivariate

suite of metals/metalloids to suggest that oysters deployed closer to the Burwood Beach WWTW

outfalls had accumulated higher concentrations of metals/metalloids than those at sites which were

located further away.

The key objective of the Burwood Beach Oyster Biomonitoring Study was to assess the potential for

effluent and biosolids discharges to lead to bioaccumulation of chemicals in marine biota, over a

range of spatial scales, using oysters as a biomonitor. This study found that there was evidence of

bioaccumulation of OC pesticides in oysters at the outfall during one sampling event. There were no

consistent significant differences in the spatial patterns of metals/metalloids to suggest that oysters

deployed closer to the Burwood Beach WWTW had accumulated higher concentrations of these. It

would be expected that with increases in future discharges of effluent and biosolids at Burwood

Beach WWTW that higher concentrations of organic chemicals and metal/metalloids would be found

via oyster biomonitoring studies.

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Disclaimer

This report has been prepared on behalf of and for the exclusive use of Hunter Water, and is

subject to and issued in accordance with the agreement between Hunter Water and

WorleyParsons. WorleyParsons accepts no liability or responsibility whatsoever for it in respect of

any use of or reliance upon this report by any third party.

Copying this report without the permission of Hunter Water or WorleyParsons is not permitted.

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BURWOOD BEACH WWTW

Page iv 301020-03413 : 105 FINAL DRAFT: August 2013

Internal and Client Review Record

PROJECT 301020-03413 – BURWOOD BEACH OYSTER BIOMONITORING STUDY

REV DESCRIPTION ORIG REVIEW WORLEY- PARSONS APPROVAL

DATE CLIENT APPROVAL

DATE

A Draft issued for internal review

Dr K Newton / Dr M Priestley

S Codi King

11 Sept 2012 N/A

B Draft issued for internal review

Dr M Priestley

Dr K Newton

11 Sept 2012

C Draft issued for internal review

Dr M Priestley

Dr K Newton / S Codi King

16 July 2013

D Draft issued for client review

Dr M Priestley / Dr K Newton

Hunter Water / CEE

18 July 2013

E FINAL DRAFT

Dr M Priestley / Dr K Newton

EPA

August 2013

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CONTENTS

1 INTRODUCTION ................................................................................................................ 1

1.1 Burwood Beach WWTW ..................................................................................................... 1

1.1.1 Treatment Process ................................................................................................. 1

1.1.2 Environmental Protection Licence Conditions ....................................................... 1

1.1.3 Characteristics of Current Effluent and Biosolids Discharges ............................... 4

1.1.4 Effluent and Biosolids Flow Data ......................................................................... 11

1.1.5 Dilution Modeling / Dispersion Characteristics .................................................... 12

1.1.6 Biosolids Deposition ............................................................................................. 13

1.2 Burwood Beach Marine Environmental Assessment Program ......................................... 13

1.2.1 Initial Consultation ................................................................................................ 14

1.3 Study Area ........................................................................................................................ 14

1.4 Scope of Work / Study Objectives .................................................................................... 14

1.4.1 Null Hypothesis .................................................................................................... 14

1.5 Review of Previous Australian Studies ............................................................................. 15

1.5.1 Biomonitors of Chemicals in the Aquatic Environment ........................................ 15

1.5.2 Oysters as Biomonitors of Metals / Metalloids ..................................................... 15

1.5.3 Abiotic and Biotic Factors that can Influence the use of Oysters as Biomonitors of

Metals / Metalloids ............................................................................................................ 17

1.5.4 Oysters as Biomonitors of Organics .................................................................... 19

1.5.5 Abiotic and Biotic Factors that can Influence the use of Oysters as Biomonitors of

Organics ............................................................................................................................ 19

1.5.6 Biomonitoring of the Receiving Environment of Burwood Beach WWTW ........... 21

2 METHODS ........................................................................................................................ 23

2.1 Consultation / Requirements of Stakeholders .................................................................. 23

2.2 Oyster Source ................................................................................................................... 23

2.3 Oyster Deployment ........................................................................................................... 24

2.4 Spatial and Temporal Assessment ................................................................................... 26

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2.4.1 Temporal Assessment ......................................................................................... 26

2.4.2 Sampling Sites ..................................................................................................... 26

2.4.3 Replication Achieved ........................................................................................... 28

2.5 Laboratory Analysis .......................................................................................................... 29

2.5.1 Laboratory Analysis of Organics .......................................................................... 29

2.5.2 Laboratory Analysis Metals / Metalloids .............................................................. 33

2.5.3 Laboratory Quality Assurance / Quality Control ................................................... 33

2.6 Guideline Values and Comparison Criteria for Chemicals in Oyster Tissue .................... 34

2.7 Baseline Concentrations in Source Oysters ..................................................................... 37

2.8 Statistical Analysis ............................................................................................................ 37

2.8.1 Univariate Analysis .............................................................................................. 37

2.8.2 Multivariate Analysis ............................................................................................ 38

3 RESULTS ......................................................................................................................... 39

3.1 Organic Chemicals............................................................................................................ 39

3.2 Metals / Metalloids ............................................................................................................ 39

3.3 Univariate Analysis of Metals / Metalloids ........................................................................ 40

3.4 Multivariate Analysis of Metal Profiles .............................................................................. 57

3.4.1 January - April 2012 ............................................................................................. 57

3.4.2 May - July 2012 .................................................................................................... 58

3.4.3 March - May 2013 ................................................................................................ 59

3.4.4 Overall .................................................................................................................. 60

3.5 Power Analysis ................................................................................................................. 62

4 DISCUSSION .................................................................................................................... 64

4.1 Organics ............................................................................................................................ 64

4.2 Metals ............................................................................................................................... 65

5 CONCLUSIONS ................................................................................................................ 68

6 ACKNOWLEDGEMENTS ................................................................................................. 69

7 REFERENCES ................................................................................................................. 70

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Figures

Figure 1.1 Location of Burwood Beach WWTW.

Figure 1.2 Burwood Beach WWTW and outfall alignment.

Figure 1.3 Effluent and biosolids flow data for the study period (July 2011 - May 2013).

Figure 2.1 Mooring design.

Figure 2.2 Experimental design of moorings at Newcastle.

Figure 2.3 Pictures of oyster deployment moorings, oyster bags and Burwood Beach WWTW.

Figure 2.4 Locations of oyster deployments.

Figure 3.1 Concentrations of arsenic in Saccostrea glomerata tissue.

Figure 3.2 Concentrations of cadmium in Saccostrea glomerata tissue.

Figure 3.3 Concentrations of chromium in Saccostrea glomerata tissue.

Figure 3.4 Concentrations of cobalt in Saccostrea glomerata tissue.

Figure 3.5 Concentrations of copper in Saccostrea glomerata tissue.

Figure 3.6 Concentrations of iron in Saccostrea glomerata tissue.

Figure 3.7 Concentrations of lead in Saccostrea glomerata tissue.

Figure 3.8 Concentrations of manganese in Saccostrea glomerata tissue.

Figure 3.9 Concentrations of mercury in Saccostrea glomerata tissue.

Figure 3.10 Concentrations of nickel in Saccostrea glomerata tissue.

Figure 3.11 Concentrations of selenium in Saccostrea glomerata tissue.

Figure 3.12 Concentrations of silver in Saccostrea glomerata tissue.

Figure 3.13 Concentrations of zinc in Saccostrea glomerata tissue.

Figure 3.14 MDS plot of multivariate suite of metals/metalloids for each sample from the January -

April 2012 deployment.

Figure 3.15 MDS plot of multivariate suite of metals/metalloids for each sample from the May - July

2012 deployment.

Figure 3.16 MDS plot of multivariate suite of metals/metalloids for each sample from the March - May

2013 deployment.

Figure 3.17 MDS plot of multivariate suite of metals/metalloids by distance from the outfall, pooled

over deployment period.

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Figure 3.18 MDS plot of multivariate suite of metals/metalloids by deployment period, pooled over

distance from the outfall.

Figure 3.19 MDS plot of multivariate suite of metals/metalloids by direction from the outfall, pooled

over deployment period.

Tables

Table 1.1 Load limits for effluent and biosolids discharges.

Table 1.2 Summary of physicochemical, metal/metalloid and organics data in effluent collected during

2006 - 2013.

Table 1.3 Summary of physicochemical, metal/metalloid and organics data in biosolids during 2006 -

2013.

Table 1.4 Effluent and biosolids flow data for the study period (July 2011 - May 2013).

Table 1.5 Classification of zones based on prior effluent dilution modelling.

Table 2.1 GPS coordinates of oyster moorings at Burwood Beach.

Table 2.2 Deployment periods and replication achieved at each site.

Table 2.3 Organochlorine pesticides tested in oysters

Table 2.4 Organophosphate pesticides tested in oysters

Table 2.5 Polychlorinated biphenyls tested in oysters

Table 2.6 Metals and metalloids tested in oysters

Table 2.7 Comparison criteria for organic chemicals in oyster tissue.

Table 2.8 Comparison criteria for metals/metalloids in oyster tissue.

Table 3.1 Factorial GLM ANOVAs on metals/metalloids (mg/kg, wet weight) concentrations in oyster

tissue deployed during January - April 2012, May - July 2012 and March - May 2013.

Table 3.2 Regressions of oyster tissue metal/metalloids concentration with distance from the outfall

during each sampling event.

Table 3.3 PERMANOVA analysis of a suite of metals/metalloids in the Sydney rock oyster following

three deployment periods.

Table 3.3 Estimates of sample sizes required to detect a significant difference between

metals/metalloids in oysters based on power analysis of January - April 2012 sampling data

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Appendices

APPENDIX 1: Organic Chemical Concentrations in Oysters

APPENDIX 2: Metal/metalloid concentrations in Oysters

APPENDIX 3: Power Analysis

APPENDIX 4: NMI QA/QC Reports

Abbreviations

ANZFA Australian and New Zealand Food Authority

CEE Consulting Environmental Engineers

EPA Environmental Protection Authority

EPL Environment Protection License

LOR Limit of Reporting

MRL Maximum Residue Limit

MEAP Marine Environmental Assessment Program

NHMRC National Health and Medical Research Council

OC Organochlorine Pesticides

OEH Office of Environment and Heritage

OP Organophosphate Pesticides

PCBs Polychlorinated Biphenyls

WWTW Wastewater Treatment Works

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1 INTRODUCTION

1.1 Burwood Beach WWTW

The Burwood Beach Wastewater Treatment Works (WWTW) is located on the Hunter Central Coast of

New South Wales (NSW) approximately 2.5 km south of the city of Newcastle (Figure 1.1). The plant

treats wastewater from Newcastle and the surrounding suburbs, servicing approximately 185,000 people

and local industry and has an average daily dry weather flow of 44 million litres of wastewater (44 ML/d).

Over the next 30 years these flows are expected to increase to 55 - 60 ML/d, even with water

conservation measures in place.

1.1.1 Treatment Process

The secondary treatment process at Burwood Beach consists of physical screening to remove large and

fine particulates, biological filtration and waste activated sludge (biosolids) processing including aeration

and settling stages. Secondary treated effluent from Burwood Beach WWTW is discharged to the ocean

through a multi-port diffuser which extends 1,500 m offshore, with diffusers at a depth of approximately

22 m (Figure 1.2). Approximately 2 ML/d of biosolids, which is surplus to treatment requirements, is also

discharged to the ocean via a separate multi-port diffuser that extends slightly further offshore than the

effluent outfall. Both outfalls have been operating in their current configuration since January 1994.

1.1.2 Environmental Protection Licence Condi tions

The Environment Protection Licence (EPL) for Burwood Beach WWTW specifies limit conditions for the

operation of the plant. These conditions provide an indication of the characteristics of the effluent and

biosolids discharged into the ocean. Condition L1 specifies that the operation of the outfall must not

cause or permit waters to be polluted (i.e. the licensee must comply with section 120 of the Protection of

the Environment Operations Act 1997). Condition L2 specifies limits relating to total loads discharged to

the ocean (including the effluent and biosolids). These limits are provided in Table 1.1. Condition 3

specifies limits to concentrations of suspended solids and oil / grease in the effluent discharged to the

outfall. The three day geometric mean concentration limit for suspended solids is 60 mg/L and for oil /

grease is 15 mg/L. Condition 4 sets volume and mass limits of effluent and biosolids discharged via the

outfalls. The limit for effluent flow rate is 510 ML/d (to allow for higher flows in wet weather) and for

biosolids the flow limit is 5 ML/d. Daily monitoring of flow is required.

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Figure 1.1 Location of Burwood Beach WWTW.

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Figure 1.2 Burwood Beach WWTW and outfall alignment.

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Table 1.1 Load limits for effluent and biosolids discharges.

Parameter Load Limits

kg/year kg/day

Total suspended solids 4,717,189 12,924

Biochemical oxygen demand - -

Total nitrogen 778,257 2,132

Oil and grease 341,290 935

Total phosphorous - -

Zinc 3,943 11

Copper 2,080 5.7

Lead 1,472 4.0

Chromium 224 0.61

Cadmium 124 0.34

Selenium 14 0.038

Mercury 9 0.025

Pesticides and PCBs 7 0.019

1.1.3 Characteristics of Current Effluent and Biosolids Discharges

The final treated effluent and biosolids from Burwood Beach WWTW has been monitored by Hunter

Water for microbiological indicators of faecal contaminations and for a suite of metals/metalloids and

organic chemicals. A summary of this data during the period 2006 - 2013 is provided in Tables 1.2

(effluent) and 1.3 (biosolids) (data provided by Hunter Water 2013).

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Table 1.2 Summary of physicochemical, metal/metalloid and organics data in effluent collected during 2006 - 2013.

Group Parameter (units) Period N Median Mean Min Max Std

Error 75%ile 90%ile

Physicochemical Suspended solids (mg/L) 2006-13

449 27 33.6 <1 390 1.6 40 60

UV254nm Transmittance (%T) 2006-13

6 59.2 58.4 43.6 68.31 3.4 62.475 65.705

pH 2006-13

224 7.6 7.6 7 8 0.01 7.7 7.8

Total dissolved solids (mg/L) 2006-13

56 440 448.5 276 734 12.9 487.5 545

Biological Oxygen Demand - total (mg/L) 2006-13

239 23 27.4 <2 144 1.3 36 50

Chemical Oxygen Demand - Flocculated (mg/L)

2006-13

19 42 41.8 32 55 1.6 46 51.4

Grease - total high range (mg/l) 2006-13

3 <5 4.7 <5 10 2.7 6 8.4

Grease - total low range (mg/l) 2006-13

444 <2 2.7 <2 60 0.2 3 5

Ammonium nitrogen (mg/L N) 2006-13

70 23.0 21.7 1 33.1 0.8 26.8 29.4

Nitrate + nitrate oxygen (mg/L N) 2006-13

236 1.0 1.6 <0.05 14 0.1 2.1 3.7

Total Kjeldahl Nitrogen (mg/L N) 2006-13

236 26.9 26.1 2.2 48.7 0.6 33.0 36.9

Total nitrogen (mg/L N) 2006-13

236 28.7 27.6 2.45 48.7 0.6 33.6 37.7

Total phosphorus (mg/L P) 2006-13

236 2.3 2.64 0.09 8.2 0.11 3.625 4.8

Metals / Metalloids

Silver-Ag-AAS furnace (µg/L) 2006-13

31 1 3.1 <1 18 0.9 2.5 13

Silver Ag-ICP (µg/L) 2006-13

59 0.5 0.7 <1 7 0.1 0.5 1

Arsenic As-vga (µg/L) 2006-13

90 1.7 1.8 0.05 3.9 0.1 2.1 2.51

Cadmium Cd-furnace (µg/L) 2006-13

5 <1 <1 <1 <1 - <1 <1

Cadmium Cd-ICP (µg/L) 2006-13

59 <1 0.5 <1 1 <1 <1 <1

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Error 75%ile 90%ile

Chromium Cr-furnace (µg/L) 2006-13

31 1 1.9 <1 28 0.9 1.2 2

Chromium Cr- ICP (µg/L) 2006-13

59 <1 0.7 <1 2 0.1 0.75 1

Chromium Cr VI-furnace (µg/L) 2006-13

90 <1 0.7 <1 1 - 1 1

Copper Cu-furnace (µg/L) 2006-13

31 17 21.2 4 115 3.5 21 34

Copper Cu-ICP (µg/L) 2006-13

93 0.25 0.4 0.04 1.7 - 0.47 0.728

Mercury Hg-VGA ug/L) 2006-13

90 <0.1 0.1 <0.1 1.6 - <0.1 0.2

Manganese Mn-furnace (µg/L) 2006-13

31 70 76.0 31 173 6.6 82 105

Manganese-ICP (µg/L) 2006-13

59 61 63.8 27 119 2.0 67.5 80.2

Nickel Ni-furnace (µg/L) 2006-13

90 <1 <1 <1 <1 - <1 <1

Nickel Ni-ICP (µg/L) 2006-13

59 4 5.3 <1 20 0.6 5.5 13.2

Lead Pb-furnace (µg/L) 2006-13

90 3 3.1 <1 17 0.3 4 5

Selenium Se-VGA (µg/L) 2006-13

90 0.1 0.3 <0.1 2 - 0.4 0.6

Zinc Zn (µg/L) 2006-13

31 50 49.4 10 120 4.3 55 70

Zinc Zn-ICP (µg/L) 2006-13

59 24 31.2 4 164 3.2 35 55.8

Organics

Aldrin (µg/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 - <0.01 <0.01

α-BHC Bhc-a (µg/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 - <0.01 <0.01

β-BHC-b (µg/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 - <0.01 <0.01

α Chlordane (ug/L) 2006-13

90 <0.01 0.000 <0.02 0.003 - <0.01 <0.01

Chlordane (ug/L) 2006-13

90 <0.01 0.001 <0.02 0.020 - <0.01 <0.01

λ Chlordane (µg/L) 2006-13

11 <0.01 0.000 <0.02 0.001 - <0.01 <0.01

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Error 75%ile 90%ile

Chlorpyrifos 2006-13

90 <0.01 0.007 <0.05 0.629 0.007 <0.01 <0.01

Lindane (µg/L) 2006-13

90 <0.01 0.000 <0.01 0.005 - <0.01 <0.01

DDT (ug/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 - <0.01 <0.01

DDD (µg/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 - <0.01 <0.01

DDE (µg/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 - <0.01 <0.01

Diazinon (ug/L) 2006-13

90 <0.01 0.000 <0.1 0.030 - <0.01 <0.01

Dieldrin (µg/L) 2006-13

90 <0.01 0.000 <0.01 0.012 - <0.01 <0.01

Endosulfan (µg/L) 2006-13

0 <0.01

Endosulfan-s (µg/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Endosulfan-1 (µg/L) 2006-13

0 <0.01

Endosulfan-2 (µg/L) 2006-13

0 <0.01

Endrin (µg/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Heptachlor (µg/L) 2006-13

90 <0.005 <0.005 <0.005 <0.005 <0.005 <0.005 <0.005

HCB (µg/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Heptachlor-epoxide (µg/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Methoxychlor (µg/L) 2006-13

90 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Parathion (ug/L) 2006-13

90 <0.1 0.000 <0.1 0.010 0.000 <0.1 <0.1

Total PCBs (µg/L) 2006-13

90 <0.1 <0.1 <0.1 <0.1 <0.1 <0.1 <0.1

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Table 1.3 Summary of physicochemical, metal/metalloid and organics data in biosolids during 2006 - 2013.


Error 75%ile 90%ile

Physicochemical

Total solids (%w/w) 2006-13 458 0.41 0.45 0.00 2.42 0.01 0.50 0.67

Volatile solids (%w/w) 2006-13 440 69.12 66.35 20.61 96.72 0.51 72.68 74.60

Ammonium N_Total (mg/L N) 2006-13 440 24.00 25.03 0.01 85.40 0.55 30.13 39.00

Grease – total low range (mg/L) 2006-13 440 153.5 172.0 1.0 841.0 5.5 230.0 328.2

Fluoride (mg/L) 2006-13 3 0.77 0.67 0.42 0.82 0.13 0.80 0.81

Metals / Metalloids

Silver-Ag-AASurnace (µg/L) 2006-13 152 22 23 4 63 1 29 40

Silver Ag-ICP (µg/L) 2006-13 279 11 12 0.5 38 0 15 18

Arsenic As-vga (µg/L) 2006-13 431 14.7 18.33 2.6 130 0.70 19.75 30.5

Cadmium Cd-furnace (µg/L) 2006-13 152 4 5.93 0.5 128 1.04 6 8

Cadmium Cd-ICP (mg/L) 2006-13 279 0.005 0.01 0.005 0.06 0.00 0.01 0.01

Chromium Cr VI-furnace (µg/L 2006-13 152 1 1.00 1 1 0.00 1 1

Chromium Cr_VIi-furnace (µg/L ) 2006-13 279 5 10 5 25 0.00 5 25

Chromium Cr-furnace (µg/L) 2006-13 152 46.5 68.16 1 750 7.41 68.5 105

Chromium cr- ICP (µgLl) 2006-13 279 30 50 5 3200 10 40 70

Copper Cu-furnace (µg/L) 2006-13 152 839 954 125 3930 42.8 1134 1426

Copper Cu-ICP (µg/L) 2006-13 279 830 880 5 3300 20 1000 1300

Mercury Hg- VGA ug/L) 2006-13 431 3.7 3.93 0.005 10.2 0.08 4.8 6.3

Manganese Mn-furnace (µg/L) 2006-13 152 339 360 33 1270 13.73 446.25 512.5

Manganese -ICP (mg/L) 2006-13 279 0.39 0.41 0.06 1 0.01 0.47 0.57

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Nickel Ni-furnace (µg/L) 2006-13 152 40 47.21 13 180 2.49 55 77.7

Nickel Ni-ICP (mg/L) 2006-13 279 0.03 0.04 0.005 0.33 0.00 0.05 0.07

Lead Pb-furnace (µg/L) 2006-13 152 187 224 13 900 11.37 269.25 375

Lead Pb ICP µg/L) 2006-13 279 120 130 10 450 0.01 150 212

Selenium Se-VGA (µg/L)) 2006-13 431 0.1 0.91 0.05 5.9 0.06 1.7 2.7

Zinc Zn (mg/L) 2006-13 152 2.4 3.03 0.78 15.6 0.16 3.515 5.39

Zinc Zn-ICP (mg/L) 2006-13 279 2.2 2.46 0.13 6.9 0.06 2.8 3.7

Organics

Aldrin (µg/L) 2006-13 96 0 0 0 0 0 0 0

α-BHC Bhc-a (µg/L) 2006-13 96 0 0 0 0 0 0 0

β-BHC-b (µg/L) 2006-13 96 0 0 0 0 0 0 0

α Chlordane (ug/L) 2006-13 96 0 0 0 0 0 0 0

Chlordane (ug/L) 2006-13 96 0 0 0 0 0 0 0

λ Chlordane- (µg/L) 2006-13 13 0 0 0 0 0 0 0

Chlorpyrifos (µg/L) 2006-13 96 0 0.003 0 0.239 0.003 0 0

DDT (uµ/L) 2006-13 96 0 0 0 0 0 0 0

DDD (µg/L) 2006-13 96 0 0 0 0 0 0 0

DDE (µg/L) 2006-13 96 0 0 0 0 0 0 0

Diazinon (ug/L) 2006-13 96 0 0 0 0 0 0 0

Dieldrin (µg/L) 2006-13 96 0 0.006 0 0.315 0.004 0 0

Endosulfan-s (µg/L) 2006-13 96 0 0 0 0 0 0 0

Endrin (µg/L) 2006-13 96 0 0 0 0 0 0 0

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HCB (µg/L) 2006-13 96 0 0 0 0 0 0 0

Heptachlor-epoxide (µg/L) 2006-13 96 0 0.0001 0 0.013 0.0001 0 2.8

Heptachlor (µg/L) 2006-13 96 0 0 0 0 0 0 0

Lindane (µg/L) 2006-13 96 0 0 0 0 0 0 0

Malathion (µg/L) 2006-13 96 0 0 0 0 0 0 0

Methoxychlor (µg/L) 2006-13 96 0 0 0 0 0 0 0

Parathion (ug/L) 2006-13 96 0 0 0 0 0 0 0

Total PCBs (µg/L) 2006-13 96 0 0 0 0 0 0 0

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1.1.4 Effluent and Biosolids Flow Data

Effluent and biosolids flow data for the study period was obtained from the Burwood Beach WWTW.

A summary of flow data for the period July 2011 to May 2013 is provided in Table 1.4 and Figure 1.3.

Table 1.4 Effluent and biosolids flow data for the study period (July 2011 - May 2013).

Date

Rainfall (mm)

Secondary Flow (ML)

1

By-Pass Flow (ML)

2

Total Flow (ML)

WAS (ML)

3

July 2011 238.2 2068.14 777.24 2845.38 71.66

Aug 2011 47.8 1775.64 0 1775.64 87.73

Sep 2011 136.0 1731.62 205.9 1937.52 82.86

Oct 2011 161.4 1966.85 301.27 2268.12 94.93

Nov 2011 184.5 2004.51 465.58 2470.09 86.71

Dec 2011 110.8 1825.98 6.37 1832.35 92.83

Jan 2012 53.6 1481.64 22.32 1503.96 93.38

Feb 2012 336.7 2296.60 485.42 2782.02 89.47

Mar 2012 188.0 2083.66 403.74 2487.40 96.36

Apr 2012 174.0 1889.04 306.14 2195.18 88.98

May 2012 26.2 1470.51 0 1470.51 94.01

June 2012 188.0 2255.16 373.09 2628.25 95.01

July 2012 83.5 1839.45 24.17 1863.62 86.77

Aug 2012 71.0 1704.78 62.22 1767.00 93.44

Sep 2012 16.7 1305.15 0 1305.15 87.82

Oct 2012 13.5 1257.72 0 1257.72 76.17

Nov 2012 44.6 1201.80 0 1201.80 86.92

Dec 2012 114.2 1375.59 52.98 1428.57 98.06

Jan 2013 229.0 1488.58 322.25 1810.83 99.86

Feb 2013 175.0 1855.55 397.11 2252.66 87.39

Mar 2013 241.0 1954.00 629.58 2583.58 112.08

Apr 2013 94.5 1702.77 116.92 1819.69 102.98

May 2013 60.0 1538.14 55.7 1593.84 95.64

Note 1. Secondary Flow is total secondary treated flow through the plant (i.e. total volume of screened and degritted sewage

into secondary plant over a 24 hour period from 12 midnight and discharged to ocean).

Note 2. By-Pass Flow is total volume of screened and degritted sewage which bypasses the secondary plant over a 24 hour

period from 12 midnight and is discharged to ocean.

Note 3. WAS is the Volume of Waste Activated Sludge (biosolids) pumped from the clarifier underflow over a 24 hour period

from 12 midnight and is discharged to ocean.

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Figure 1.3 Effluent and biosolids (WAS) flow data for the study period (July 2011 - May 2013).

1.1.5 Dilution Modeling / Dispersion Characteristics

Consulting Environmental Engineers (CEE 2007) calculated a predicted initial dilution for the Burwood

effluent outfall, assuming a discharge rate of 43 ML/d and all duckbill valves in operation. The model

predicted a typical dilution of 219:1 for the effluent field. Allowing for the reduction in dilution due to

the orientation of the diffuser ports parallel to the currents, initial dilution is expected to be in the range

of 180:1 to 220:1. The Water Research Lab (WRL 2007) also carried out field tests of effluent dilution

using rhodamine dye. The dilution of the surface field showed a typical dilution of 185:1. WRL (2007)

reported that the average near-field dilution was 207:1 and the 95th percentile minimum dilution was

78:1. CEE (2010) therefore considers it reasonable to base the environmental risk assessment of the

effects of effluent discharge on an effluent plume near the ocean surface with an initial dilution in the

range of 100:1 to 200:1.

The dilution of a combined biosolids and effluent discharge through the biosolids diffuser was also

calculated (CEE 2007). The CEE model predicted a typical dilution of 475:1 for discharged biosolids if

they rose to the ocean surface, or about 250:1 if trapped by stratification at mid-depth (CEE 2007).

The WRL hydrodynamic computer model showed a median dilution of 300:1, with a minimum dilution

of 100:1 when strong stratification decreases the rise and dilution of the small biosolids plumes, and a

maximum dilution at times of strong currents exceeding 1,000:1 (WRL 2007). The WRL model also

showed the biosolids plume is often trapped well below the surface by the natural stratification of the

ocean water column. WRL field tests of the biosolids plume, with dilution measured using rhodamine

dye, showed a typical dilution of 841:1. WRL reported that the average near-field dilution of the

biosolids plume was 268:1 and the 95th percentile minimum dilution was 205:1, for a submerged

plume (WRL 2007). Based on these results, it is considered reasonable to base the assessment of

the effects of biosolids discharge on two conditions; surface plume with an initial dilution of 300:1 and

submerged plume with an initial dilution of 200:1 (CEE 2010). WRL (1999) modelled the biosolids

plume at 10 m depth and showed that the centre of the plume, at about 10 m depth, the dilution

achieved is between 200:1 and 1,000:1. At a distance of 200 m from the diffuser, the dilution

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exceeds 1,000:1 and increases further with distance travelled. The diluted biosolids extends to the

south of the diffuser, but would be indistinguishable except by the sensitive techniques used in the

field studies. Based on the field tests and dilution modelling undertaken by WRL (1999, 2007) and

CEE (2007), the following mixing zones (Table 1.5) were determined for reporting purposes only.

Table 1.5 Classification of zones based on prior effluent dilution modelling.

Distance from Diffuser Zones

< 50 m outfall impact zone outfall impact

> 50 - 100 m

mixing zone

nearfield mixing zone

> 100 - 200 m midfield mixing zone

> 200 - 2,000 m farfield mixing zone

> 2,000 m reference zone reference

1.1.6 Biosolids Deposition

Previous diver inspections undertaken at the Burwood Beach outfall (i.e. by commercial divers

inspecting the outfall infrastructure) reported that biosolids deposits at the seabed can vary

significantly. In-situ diver observations have reported a biosolids thickness of 0 to 125 mm, with

variation likely a result of weather conditions. Divers have noted biosolids being washed away after

storms with no long-term accumulation on the seabed evident. More protected areas such as small

caves have a greater depth of biosolids and a peak of 750 mm was recorded in 1994/96 (note that at

this time effluent was not mixed with biosolids before discharge). ANSTO (1998) undertook a study of

the movement of seabed sediments 1,100 m south east of the outfall using iridium-radiated glass

beads. The beads were found to disperse over 100 m to the east and west and over 150 m to the

north, providing an indication of the likely expected movement of sandy sediments on the seabed. It

is expected that smaller biosolids particles would disperse at a greater rate and further than sand

particles.

1.2 Burwood Beach Marine Environmental Assessment Program

A number of monitoring programs and studies have previously been undertaken to assess the impact

of treated effluent and biosolids discharge on the marine environment at Burwood Beach (e.g. NSW

Environment Protection Authority (EPA) 1994, 1996; The Ecology Lab 1996, 1998; Australian Water

Technologies (AWT) 1996, 1998, 200, 2003; Sinclair Knight Merz (SKM) 1999, 2000; Ecotox Services

Australasia (ESA) 2001, 2005; BioAnalysis 2006; Andrew-Priestley 2011; Andrew-Priestley et al.

2012). While providing a wealth of data on the marine environment here, it is considered that these

previous studies have not effectively assessed the spatial extent and ecological significance of the

outfalls impact (CEE 2010).

The aim of the Burwood Beach Marine Environmental Assessment Program (MEAP) was to establish

the impact footprint of the existing outfall, establish the gradient of impact with distance to the edge of

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the outfall and predict the potential footprint of future impacts. A key concern of the community and

other stakeholders was the impact that the WWTW effluent and biosolids discharge has on the

bioaccumulation of toxic or harmful chemicals in marine organisms (CEE 2010). The current study

aims to address this issue by providing an assessment of the bioaccumulation of a range of

chemicals in oysters deployed in a gradient (increasing distance) from the outfall.

1.2.1 Initial Consultation

Prior to commencement of the Burwood Beach MEAP, details of the proposed sampling program and

survey methodology were discussed with Hunter Water (Client), CEE and the NSW Environment

Protection Authority (EPA) (then the Office of Environment and Heritage (OEH) (Regulator) on 10

October 2011. This meeting was undertaken to ensure that the proposed MEAP was adequate in

addressing the requirements of both the Client and the Regulator. During this meeting, any concerns

with the proposed program were raised and the methodology of the assessment program was

subsequently altered accordingly.

Prior to deployment of the mooring systems at Burwood Beach consultation with Newcastle Fishing

Co-operative, Newcastle Ports Corporation (NPC) and NSW Fisheries (Port Stephens) was also

undertaken to identify any concerns raised by commercial fisheries that operate at Burwood Beach

and in the vicinity of the study area (further detail of outcomes in Section 2.1).

1.3 Study Area

Burwood Beach is located in Newcastle, on the Hunter Central Coast of NSW (Figure 1.1). The

seabed in the vicinity of the outfall consists of small areas of low profile patchy reef, which is subject

to strong wave action and periodic sand movement, interspersed between large areas of soft

sediment habitat. These low profile reefs extend to approximately 1 m above the sand. Water depth

is approximately 22 m at the outfall diffuser. Mobile sandy sediments occur in the gutters and low-

lying seabed between reef patches. Extensive sandy beaches with intertidal rocky reef habitats occur

along the shoreline adjacent to the outfall. Merewether Beach lies to the north and Dudley Beach to

the south of Burwood Beach.

1.4 Scope of Work / Study Objectives

The key objective of the Burwood Beach Oyster Biomonitoring Study was to assess the potential for

effluent and biosolids discharges to lead to bioaccumulation of chemicals in marine biota, over a

range of spatial scales, using oysters as a biomonitor. A specific requirement of the study was to

establish the zone in which there is a detectible increase in the concentration of chemicals in oysters

that is related to the Burwood Beach outfall.

1.4.1 Null Hypothesis

The hull hypothesis was:

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There is no significant difference in the level of chemicals in the tissue of oysters deployed

at sites located at a range of distances from the Burwood Beach WWTW outfall.

1.5 Review of Previous Australian Studies

1.5.1 Biomonitors of Chemicals in the Aquatic Environment

Environmental chemicals, such as metals/metalloids, polychlorinated biphenyls (PCBs),

organochlorines (OC) pesticides and organophosphorus (OP) pesticides may enter the sewage

treatment process via domestic and industrial sources. Incomplete removal during sewage treatment

can result in entry into the aquatic environment via sewage effluent release. Once released into the

aquatic environment, chemicals may dissolve in seawater or bind to particulates or sediments.

Aquatic biota may then take up chemicals through direct ingestion of water, particulates or food

(Naimo 1995). Uptake by aquatic biota is of concern due to potential toxicity effects on biota but also

associated human health concerns. Thus, routine biomonitoring of metals/metalloids and organic

chemicals in the aquatic environment is required.

Biomonitoring of metals/metalloids, PCBs and pesticides in seawater or sediments can sometimes

pose difficulties. Chemicals in seawater are often at low concentrations, below limits of analytical

detection. Another consideration is that release of chemicals is not constant but instead likely to be

dependent on pulse releases into the aquatic environment. To address these issues, biological

monitors (biomonitors) have been used to monitor chemicals in the aquatic environment. In

particular, molluscs have been shown to be useful and able to bioaccumulate chemicals that reflect

concentrations from the surrounding environment. They have also been used because it is the

chemicals that are bioavailable that are of most concern and can impact aquatic organisms.

Assessing concentrations in oysters provides a direct indication of what chemicals are bioavailable

and have the potential to cause harm.

The use of molluscs to measure chemicals includes the successful development of the Mussel Watch

programme (Goldberg et al. 1983) and, in Australia, the Sydney rock oyster, Saccostrea glomerata, is

the predominant molluscan species used for biomonitoring of heavy metals/metalloids (Avery et al.

1996; Scanes 1996; Lincoln-Smith and Cooper 2004; Robinson et al. 2005). Oysters have also been

used for biomonitoring of organics in the marine environment (Ajani et al. 1999; NSW EPA 1996;

Scanes 1996). This species has also been for biomonitoring of organics in the marine environment

(Ajani et al. 1999; NSW EPA 1996; Scanes 1996) but not as frequently.

1.5.2 Oysters as Biomonitors of Metals / Metalloids

In the aquatic environment, molluscs may be exposed to metals/metalloids through direct ingestion of

water or food sources (Naimo 1995). Following uptake of chemicals, the organism may excrete, bind

to a biomolecule and / or store within tissues. Also, in the case of essential metals (i.e. copper, zinc

and iron)they can be used for essential metabolic processes (Rainbow 2002). One of the concerns of

uptake of metal/metalloids by biota is the potential toxicity effects on the organism. In molluscs,

potential effects due to metal/metalloid exposure includes reproductive and growth impairments,

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behavioural abnormalities and in some cases, mortality (Keller and Zam 1991; Naimo 1995; Norris

and Carr 2006). A second concern is the potential environmental persistence of metals and

metalloids. Metals/metalloids have the potential to bioaccumulate in biota and bio-magnify through

the food chain. Consumption of biota that has elevated concentrations of metals/metalloids presents

a concern for human health. Thus, routine biomonitoring of metals/metalloids in the aquatic

environment is required.

Molluscs, in particular oysters, have been established to be used as biomonitors of metals/metalloids.

There have been a range of laboratory studies undertaken that demonstrate how oysters are capable

of bioaccumulating metals/metalloids. Eisler et al. (1972) found that the Atlantic oyster (Crassostrea

virginica) bioaccumulated cadmium to greater concentrations than was recommended for human

consumption (13 mg/kg), despite residing in waters with a concentration lower than that

recommended for safe drinking (0.1 µg/L). Further, Watling (1983) demonstrated that the Pacific

oyster (Crassostrea gigas), exposed to 100 µg/g (wet weight) of chromium, cadmium and lead for

three weeks bioaccumulated up to 3.5, 11 and 27 fold, respectively, metal concentrations in controls.

In the same study, Watling (1983) found that essential metals, copper and zinc, bioaccumulated in

C. gigas, although the rate of bioaccumulation was lower in comparison to non-essential metals, such

as chromium, cadmium and lead. Following three weeks exposure to 100 µg/g (wet weight) copper

and zinc, concentrations in C. gigas tissue were 2 and 1.1 fold that of controls.

The capacity of oysters to bioaccumulate metals/metalloids, which reflect environmental

concentrations, has resulted in their widespread use as biomonitors of bioavailable metals/metalloids

in aquatic environments. In Australia, S. glomerata, is commonly used for biomonitoring of heavy

metals/metalloids in the marine environment (Avery et al. 1996; Scanes 1996; Spooner et al. 2003;

Lincoln-Smith and Cooper 2004; Robinson et al. 2005; Andrew-Priestley 2011; Andrew- Priestley

2012). For example, Brown and McPherson (1992) used Saccostrea glomerata (then known as S.

commercialis) to assess spatial and temporal changes in copper and zinc throughout the Georges

River (NSW, Australia) and observed that metal concentrations increased with distance from the

mouth of the estuary, with an increase of 40% for copper and 300% for zinc concentrations from 1975

compared to 1987. Further, Spooner et al. (2003) analysed wild Saccostrea glomerata (then known

as S. commercialis) and found that concentrations of zinc and copper were significantly elevated in

oysters which were grown in an Australian contaminated location, Botany Bay, (2600 ± 690 μg/g for

zinc and 170 ± 45 μg/g for copper) compared to oysters grown at reference locations, Jervis Bay and

Batemans Bay (which ranged from 980 ± 400 μg/g to 1793 ± 392 μg/g for zinc and 22 ± 14 μg/g to 65

± 18 μg/g for copper).

Scanes (1996) also demonstrated that S. glomerata, was a useful biomonitor of metal contamination

in waters adjacent to wastewater treatment outfalls in Sydney, NSW. The concentrations of

metals/metalloids were reduced following the commissioning of deepwater offshore discharge.

Where oysters are used to monitor chemicals in the receiving environment of sewage effluent

discharge it is likely that exposure to effluent, and any potential chemicals contained in the effluent, is

not constant but fluctuates with environmental conditions and plume dynamics. Importantly, studies

have shown that oysters are capable of bioaccumulating metals and maintaining their tissue

concentrations (Nielsen and Hrudey 1983; Luoma et al. 1985; Boisson et al. 2003).

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1.5.3 Abiotic and Biotic Factors that can Influence the use of Oysters

as Biomonitors of Metals / Metalloids

Many factors can influence the rate of metal/metalloid uptake by biota and potentially contribute to

variability among or within studies, sites and individuals, even those which employ the same species

as a biomonitors. This could include factors that are biological such as age, sex, size, feeding,

gonadal development and / or pre-exposure history to metals (Ayling 1974; Boening 1999) or

environmental such as organic carbon, sediment composition, temperature, pH, dissolved oxygen

and / or hydrologic features such as oceanic currents and sewage plume dynamics (Elder and Collins

1991).

DEPLOYMENT PERIODS

Deployment periods should be sufficient to allow for metal/metalloid equilibrium in oyster tissue.

Equilibrium is defined as the time that it takes for biomonitors to reach environmental concentrations.

Biomonitors should be deployed for a period which is sufficient for all metals to reach equilibrium in

the tissue. One of the ways to adequately address equilibrium in the tissues is to understand the

rates of uptake and depuration (whereby oysters in clean water remove metals/metalloids from their

tissue). Metals/metalloids have been estimated to take three to twelve weeks to equilibrate in oyster

tissue (Watling 1983; Scanes and Roach 1999).

Essential metals are those which have roles in metabolic functioning, certain quantities of these

metals are required to meet metabolic needs and these metals cannot be immediately detoxified or

excreted. Although oysters are considered to be effective biomonitors of all heavy metals (Phillips

and Rainbow 1989; Scanes 1996), the equilibrium time may be longer for essential metals. Thus,

non-essential metals/metalloids may have higher potential for bioaccumulation and provide relative

concentrations in tissues which more closely reflect environment loads, in comparison to essential

metals including copper, zinc and iron.

Deployment and exposure periods should be selected that are appropriate in terms of timeframe for

metal uptake of both essential and non-essential metals. However, this can sometimes be a trade off

with being able to maintain the experiment. It is difficult to maintain oyster deployments in the marine

environment, due to potential issues relating to sea conditions, tampering or removal of oysters by

humans and interference by marine wildlife (i.e. shark consumption).

BACKGROUND METAL AND METALLOID CONCENTRATIONS

Metals/metalloids are natural elements of the environment. Different oyster populations can have

different metal and metalloid concentrations due to natural variation in environmental background.

Spatial variation in metal/metalloid concentrations poses difficulties in the measurement of metals and

metalloids and the capacity to differentiate between background concentrations and elevated

concentrations due to anthropogenic input (Phillips and Rainbow 1993; Cantillo 1997; Scanes and

Roach 1999). Scanes and Roach (1999) calculated background heavy metal concentrations for

S. glomerata for twelve locations in NSW which were identified as having a low risk of metal

contamination. Care must be taken though, as these locations were estuarine (as opposed to the

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marine location used in the current study), but this provides a baseline indication for metals/metalloids

expected in NSW.

MORPHOLOGY AND GENETIC MAKE-UP

Many factors can influence bioaccumulation of metals in oysters including age, size, reproductive

condition, tissue type and genetic make-up (i.e. such as triploid versus diploid).

Different populations of oysters may have different rates of uptake or responses for metals/metalloids.

For example, Robinson et al. (2005) demonstrated that two different populations of oysters sourced

from two close sites (both within the Clyde River) had significantly different (p < 0.05) zinc

concentrations. This demonstrates that care should be taken comparing different populations and

studies. Oysters should be sourced from a single population or sufficient replication should be used

to assess natural variability between populations.

Robinson et al. (2005) investigated how factors can influence metal uptake. They measured a suite of

metals (including copper, cadmium, zinc, lead and selenium) in S. glomerata from two

uncontaminated estuaries and five estuaries known to have elevated metals. In particular, age and

tissue type were found to be highly influential. Oysters aged three years had significantly higher

tissue metal concentrations compared to those one year old. They also found a significant difference

between tissue types, with the mantle tissue bioaccumulating the highest concentrations of the five

metals.

Reproductive condition can be an issue if oysters spawn during a biomonitoring program. This is

because during peak reproductive condition, a large proportion of an oyster‟s body weight is gonadal

tissue (up to 60%) which is lost during spawning (Cox et al. 1996), however individuals can spawn

different amounts and not always at the same time. Thus, results may have higher variability if

comparing individuals where spawning has occurred for some individuals during the biomonitoring.

Oysters should be selected from a similar size, age and population to help minimise sources of

variations. Oysters that are selected from a single population, and then deployed to the locations, will

eliminate the variability that is associated with different ages and populations. Oysters should be

selected from a similar size class, and additional measurements of weight and condition index can

help to account for variation among sizes. Using a composite sample (pooled sample of individuals)

rather than individuals will also assist with reducing variability. Robinson et al. (2005) recommends

using at least six oysters.

In addition, the timing of deployments should avoid the period when spawning is likely to occur.

Although the timing of spawning varies from year to year, it is usually during mid-February to late

March for S. glomerata along the mid north NSW coast of Australia.

ANALYSIS METHODS

The moisture content of an oyster affects the wet weight of the organism and thus influences the final

calculation of chemicals per unit mass of the tissue. The measurement of chemicals in tissue is

generally undertaken in dried tissue eliminating this issue; otherwise the percentage of moisture can

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be reported or accounted for as an additional factor in analyses. Using composite samples should

also assist with minimising variation.

1.5.4 Oysters as Biomonitors of Organics

Molluscs are also considered to be effective biomonitors of organic chemicals in the aquatic

environment (Scanes 1996, 1997; Ajani et al. 1999; Andrew-Priestley 2011, Andrew-Priestley et al.

2012), although in Australia this has been studied to a much lesser extent compared to oyster

biomonitoring of metals/metalloids due to the high lipid content in oysters which is known to interfere

with organic chemical analyses.

The majority of Australian literature on field of studies biomonitoring for organics has focused on fish

or other invertebrates, such as abalone. There have been several studies using oysters. Scanes

(1997) undertook field studies to demonstrate that the Sydney rock oyster, S. glomerata, is capable of

bioaccumulating OC pesticides (including chlordane, dieldrin, heptachlor epoxide, PCBs, DDT, DDD

and DDE) from contaminated locations. Detectable concentrations of all organics were present in S.

glomerata tissue after three days deployment. Concentrations of organics in tissue were highest

following 209 days, although the large majority of bioaccumulation occurred quickly. After 209 days

the oysters were transported to a clean location to monitor rates of depuration of organics. Biological

half-lives (a measure of depuration) were relatively fast and ranged from four days (heptachlor

epoxide) to 46 days (DDT). Deployment of S. glomerata was also useful in demonstrating a

significant reduction in OC contamination of waters adjacent to nearshore sewage treatment outfalls

in Sydney, NSW, Australia, following commissioning of deep-water offshore discharge (Scanes 1996).

1.5.5 Abiotic and Biotic Factors that can Influence the use of Oysters

as Biomonitors of Organics

DEPLOYMENT PERIODS

Biomonitors should be deployed for a period which is sufficient for organics to reach equilibrium in the

tissue, which varies considerably between different compounds. It is necessary to understand the

rates of uptake and depuration of each organic contaminant. Scanes (1997) deployed S. glomerata

from clean waters into a known contaminated location (Long Bay, Sydney) and measured tissue

organic concentrations (mg/L, wet weight) at numerous intervals throughout the deployment which

lasted 209 days. Based on this information, estimates were provided of the time required for

equilibrium (time required for oyster tissue to reach environmental concentrations) of organics based

on their graphs of tissue organic concentrations. It was reported that equilibrium in S. glomerata took

three to twelve days for dieldrin, twelve to twenty days for chlordane, seventy two days for DDT and

twelve to twenty days for PCBs (Scanes 1997). If oysters are not deployed for a sufficient time to

allow organics to equilibrate then oyster tissue concentrations may not be a true representation of

environmental concentrations or organics may be „missed‟ and below the limit of reporting (LOR).

Another consideration is that if exposure to organics fluctuates then oysters may depurate (i.e.

remove) for some organics. Where oysters are used to monitor organics in the receiving environment

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of sewage effluent discharge it is likely that exposure to effluent, and any potential chemicals

contained in the effluent, is not constant but fluctuates with environmental conditions and plume

dynamics. In the same study outlined above, Scanes (1997) demonstrated that the biological half-life

varied considerably between different organics in S. glomerata. Half-lives were estimated as four

days for heptachlor epoxide, twelve days for dieldrin, twenty-four days for chlordane, forty-six days for

DDT and eighty-one days for PCBs (Scanes 1997). The relatively long half-lives of organics suggest

that they should remain elevated in oyster tissue even if there are fluctuations between contaminated

and clean waters.

Oysters should be deployed for a sufficient time to allow for organic equilibrium in tissues. However,

this can sometimes be a trade off with being able to maintain the experiment. In particular, it is

difficult to maintain oyster deployments in the marine environment, due to potential issues relating to

sea conditions, tampering or removal of oysters by humans and interference by marine wildlife. The

above studies suggest that eight weeks should be an appropriate deployment period for the

equilibrium of the majority of OC and OP pesticides and PCBs.

MORPHOLOGY AND GENETIC MAKE-UP

Many factors can influence bioaccumulation of organics in oysters including age, size, reproductive

condition, tissue type and genetic make-up (i.e. such as triploid versus diploid). There is not an

extensive amount of literature characterising how these factors may influence organic uptake by

oysters. However, it is likely that some or all of these factors would also be influential as sources of

variability on organic bioaccumulation by S. glomerata, i.e. it is likely that oysters of different ages and

sizes have different rates of organic uptake.

Reproductive condition is an important consideration for biomonitoring of organics in oysters. In

comparison to other tissue types, the gonad of an oyster is higher in lipids and likely to bioaccumulate

higher concentrations of organics. During peak reproductive condition, a large proportion of the body

weight of an oyster is gonadal tissue (up to 60%) which is lost during spawning (Cox et al. 1996).

Thus, if oysters spawn then the fraction of the body weight (which is most likely to have higher

organic concentrations compared to other tissues) is lost. Further, within a single population,

individuals can spawn different amounts and not always at the same time. In addition, the results

may have higher variability if comparing individuals where spawning has occurred for only some

individuals during the biomonitoring.

ANALYSIS METHODS

Organics are lipid soluble chemicals and it has been suggested that the proportion of lipids should be

accounted for in biomonitoring studies. This could include measurement of lipids to ensure there is

low variability between individuals/samples or, as recommended by Connell (1988), reporting organic

concentrations per unit weight of lipids (rather than tissue). Scanes (1998) demonstrated that organic

tissue concentrations strongly correlate with the percentage of lipids in S. glomerata. In contrast,

Scanes (1996) found no correlation between lipids and organics and suggested this was due to lower

organic concentrations. The National Oceanic and Atmospheric Administration (NOAA 1989) also

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found that there were no correlations between lipids and PCBs, DDT, lindane and PAHs and weak

correlations for chlordane and dieldrin.

It is suggested that the measurement of lipids could be accounted for through measurement and

inclusion as a co-factor or by reporting results as per unit of lipids, particularly in studies of highly

contaminated areas. Alternatively, using composite samples should also assist with minimising this

variation between samples.

1.5.6 Biomonitoring of the Receiving Environment of Burwood Beach

WWTW

Several studies have been conducted at Burwood Beach WWTW to assess chemical

bioaccumulation in oysters (following short term deployment periods in the receiving waters), resident

fish, final treated sewage effluent and sediments. The Hunter Environmental Monitoring Program

(Hunter EMP) was conducted between 1992 - 1996 (Ajani et al. 1999; NSW EPA 1996) to assess OC

pesticides, PCBs and metals/metalloids in deployed oysters and sediments. Oysters, S. glomerata,

were deployed in the receiving waters of Boulder Bay WWTW, Burwood Beach WWTW and at four

reference locations (Point Stephens, Boat Harbour, Redhead and Terrigal) for three months at a time,

with subsequent measurements of metals/metalloids (arsenic, cadmium, chromium, cobalt, copper,

lead, manganese, mercury, nickel, selenium, silver and zinc) and OCs (aldrin, BHC, lindane, technical

chlordane, Dieldrin, DDD, DDE, DDT, endosulfan, endrin, heptachlor, hexachlorbenzene,

methoxychlor, oxychlordane and PCBs). Deployments were repeated eight times during 1992 - 1994,

with three deployments prior to and five following the commissioning of the extended Burwood Beach

and Boulder Bay WWTWs.

Within oysters, only technical chlordane (a mixture of 23 chlordane isomers and related compounds),

DDE and DDD were detected (out of the seventeen organochlorines monitored) and none of these

were in oysters deployed in waters near Burwood WWTW. Technical chlordane was detected once in

August 1991, in oysters deployed at Boulder Bay, at a concentration of 0.02 mg/kg. Concentrations

ranged from 0 - 0.0142 mg/kg for technical chlordane, 0 - 0.0011 mg/kg for DDE and

0 - 0.0008 mg/kg for DDD. All measurements of DDD were below the LOR, apart from August 1992

where it was detected in samples from all locations. Redhead was the only location where mean

concentrations of DDD exceeded the LOR of 0.02 mg/kg. For sediments, there were no OC

pesticides detected at Burwood Beach, although trace concentrations were detected at some of the

reference locations.

Within sediments, all metals/metalloids were comparable to background levels at the Burwood Beach

location. For oysters, selenium at Burwood Beach was the only metal which was higher than the

ANZFA MRLs (ANZFA 2011). A possible explanation provided was that natural levels of selenium

are higher within this region, however it was concluded that further investigation was required. For

metals/metalloids it was found that there was high variability and there were no clear patterns with

metal/metalloids concentrations and locations. One of the major outcomes of this study was the

recommendation that an impact versus control comparison was not suitable for Burwood Beach.

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Several changes in treatment technology to Burwood WWTW have occurred during and since the

study of NSW EPA (1996).

More recently, BioAnalysis (2007) performed an assessment of OC pesticides and metals/metalloids

in sediments from the Burwood Beach outfall. No OC pesticides were detected in sediments from

Burwood Beach or at any of the reference locations. Metal/metalloid concentrations in sediments

were also low and apart from manganese (which ranged from ~ 45 to 75 mg/kg at Burwood Beach) all

reference locations had higher concentrations in comparison to the Burwood Beach WWTW sampling

location. All metals/metalloids were below the ANZECC trigger guidelines (ISQ-Low) (BioAnalysis

2007).

In 2008, metals/metalloids from the Burwood Beach WWTW were analysed in Sydney rock oysters,

S. glomerata, which were deployed for 6 weeks in effluent receiving waters (Burwood near: < 50 m

and Burwood far: > 150 m) and at reference locations (Redhead and Fingal Island) at depths of 4, 8

and 12 m (Andrew-Priestley 2011). Results of this study showed concentrations of most heavy

metals/metalloids were not significantly different (p > 0.05) in the tissue of S. glomerata deployed at

Burwood Beach compared to those at the reference locations (Fingal Island and Redhead). All

metals fell below the ANFZA MRLs (ANZFA 2011) except for arsenic (1.24 mg/kg compared to

ANZFA MRL of 1 mg/kg). With the exception of nickel, selenium and lead, mean concentrations of

metals in the study were considerably lower than NSW median background concentrations

determined by Scanes and Roach (1999) both at impact and reference locations. Nickel, selenium

and lead concentrations in oyster tissue were higher than those reported by Scanes and Roach

(1999) at both impact and reference locations. Comparisons to historic data (HWC 1990 and NSW

EPA 1996) suggested that, via measurement in oyster tissue, metal concentrations released into the

marine environment via sewage effluent from Burwood Beach WWTW have not changed from earlier

studies; however, further investigation is underway to determine present levels in the marine

environment. Findings suggested that S. glomerata was a suitable biomonitor for heavy

metals/metalloids in Australian waters.

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2 METHODS

2.1 Consultation / Requirements of Stakeholders

Prior to deployment of the mooring systems at Burwood Beach consultation with the EPA (then OEH),

Newcastle Fishing Co-operative, Newcastle Port Corporation (NPC) and NSW Fisheries (Port

Stephens) was undertaken to identify any concerns raised by commercial fisheries that operate at

Burwood Beach and in the vicinity of the study area. A number of requirements were identified and

suggestions made as listed below:

To assess stakeholders‟ concerns, large surface buoys with flashing lights (SLB800 with

SL60 lights) were used for the study and moorings were placed outside shipping channels.

As requested, GPS co-ordinates of all moorings were provided to NSW Maritime, NSW

Fisheries, NPC and Newcastle Fisherman‟s Co-op following the deployment.

Oysters were placed at a depth of ~ 3 m to allow survival of the oysters and reduce the

potential for accidental damage / loss by recreational vessels.

NSW Fisheries and Marine Parks permits were required for this study and have been

obtained for this activity (Fisheries Permit P11/0051; Marine Parks Permit 2011/046).

The EPA suggested that the three replicate cages on each mooring should be separated

by a distance of 30 m (i.e. a long line arrangement would be needed). It was not

considered that this arrangement would be suitable or feasible for this study, firstly due to

the distance of each mooring from each other (i.e. two at the outfall and two within 100 m of

the outfall), the risk to recreational boating and fishing activities and the logistical

constraints in doing this (i.e. issues with keeping all replicate oyster cages at the same

required level / depth, additional surface or subsurface buoys would be required, additional

lines and moorings).

Insurance against losses was addressed by having an extra mooring deployed at the

outfall. For the first sampling event, the extra samples from the outfall were required be

tested for metals only to determine whether there was a difference in exposure to effluent /

sludge around the outfall. If no difference were found these samples would simply act as

insurance for all the following surveys. If significant differences were found, these samples

would be sampled for all parameters in the following rounds.

2.2 Oyster Source

Oysters were obtained from a commercial oyster farmer, XL Oysters (located at Tea Gardens, within

Port Stephens estuary). Oysters were depurated in clean water for two weeks prior to deployment.

Oysters were sourced from the same population and all individuals were of a similar size and

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approximately 2 years of age. Sex was not determined as studies have indicated there is not likely to

be a difference between sexes in S. glomerata bioaccumulation of chemicals (Robinson et al. 2005).

2.3 Oyster Deployment

The deployment of the mooring systems and oysters at Burwood Beach was carried out by

McLennan‟s Dive Services. Oysters were deployed at a depth of 3 m (selected based on the plume

dynamics outlined in WRL 2007) using buoyed moorings of oysters in UV resistant mesh cages.

Oysters were deployed for ~ 8 weeks with biannual sampling events over a 2 year period (resulting in

four sampling periods in total). For each sampling period, three (replicate) mesh cages of oysters

(containing thirty oysters per bag) were deployed at each of the locations for a period of 8 weeks. A

schematic of a mooring system is provided in Figure 2.1, the experimental design is presented in

Figure 2.2 and pictures are shown in Figure 2.3.

Figure 2.1 Mooring design.

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Figure 2.2 Experimental design of moorings at Newcastle.

Figure 2.3 Pictures of oyster deployment moorings, oyster bags and Burwood Beach WWTW.

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2.4 Spatial and Temporal Assessment

2.4.1 Temporal Assessment

Oysters were deployed for a period of eight weeks. Four deployments were undertaken over a two

year period. Deployments were undertaken from (1) 31 January to 2 April 2012, (2) 22 May to 9 July

2012, (3) 16 October to 18 December 2012 and (4) 26 March to 22 May 2013. The exposure period

of eight weeks was selected based on the time required for equilibration but also a suitable

deployment period to minimise the risk of oyster mortalities, loss of moorings and / or oyster bags.

Prior studies have shown that eight weeks has been sufficient deployment / exposure time for oysters

to equilibrate metals (Scanes 1998; Boisson et al. 1998, 2003) and organics (Scanes 1997) to that of

the environmental / exposure concentrations. Due to the low recovery of oyster bags during October

- December 2012, this deployment period was not included in the results (see Section 2.4.3).

2.4.2 Sampling Sites

Oysters were deployed at seven sites at range of distances from the outfall in an approximate NE /

SW direction; 0 m (outfall A and B), 100 m NE and SW, 500 m NE (A and B) and SW and 2,000 m NE

and SW (Figure 2.4). The seven sampling sites were distributed along the known dispersion

pathway (WRL 2007) of the plume in order to establish a gradient of exposure. As insurance against

potential losses, one extra mooring was deployed at the Burwood Beach outfall site and at 500 N (i.e.

a total of nine moorings). GPS co-ordinates of all sampling sites are provided in Table 2.1.

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Figure 2.4 Locations of oyster deployments.

Table 2.1 GPS coordinates of oyster moorings at Burwood Beach.

Location Site Distance from

outfall (m)

Direction Latitude (S) Longitude (E)

Outfall Impact

Outfall A 0 Outfall 32°58.176' 151°45.102'

Outfall B* 0 Outfall 32°58.163' 151°45.104'

Midfield Mixing Zone

100 N 100 NE 32°58.128' 151°45.168'

100 S 100 SW 32°58.209' 151°45.076'

Farfield Mixing Zone

500 N A 500 NE 32°57.978' 151°45.356'

500 N B* 500 NE 32°58.020' 151°45.364'

500 S 500 SW 32°58.369' 151°44.896'

Reference 2000 N 2000 NE 32°57.402' 151°46.040'

2000 S 2000 SW 32°58.981' 151°44.472'

* Added as insurance moorings.

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2.4.3 Replication Achieved

During this study there were a number of oyster bags that went missing following deployment. The

actual replication that was achieved at each site during each sampling event is summarised in Table

2.2. During the first two sampling events, nearly all replicates were recovered. However, this was not

the case during October - December 2012 or March - May 2013.

During the October - December 2012 sampling event, a large number of oyster bags were missing

including all replicates from the outfall and all northern sites. A 1 m length of rope was used to attach

oyster bags to the mooring chain. This rope was woven through the three bags and secured with two

knots and multiple cable ties. At all these oyster moorings, the rope that attached the bags was not

cut and hanging loose (suggesting that the rope was untied from the mooring). Based on this, it was

suspected that these losses were due to tampering with oyster moorings (as it would have been

impossible for the ropes to come loose otherwise). Due to the low level of replication achieved, this

sampling event was not tested or included in the results.

During the March - May 2013 sampling event, the oyster bags were attached in a similar way as

described above, except that a stainless steel wire was intertwined with the rope and knots making it

difficult to remove the bags without stainless steel cutters. At all oyster moorings, except for 100 m N

and 500 m N A, there were only two bags available. For the missing replicates, the bags were

attached but there were holes in the bottom corners. On all these bags, the holes were not a clean

straight cut and this may suggest that these losses were due to interference by a marine mammal or

fish (i.e. chewing open bags to access the oysters inside).

Table 2.2 Deployment periods and replication achieved at each site.

Site Deployment Period & Number of Cages Retrieved

Jan - April 2012 May - July 2012 Oct - Dec 2012 March - May 2013

Outfall A 3 3 0 2

Outfall B 3 3 0 2

100 N 3 3 0 3

100 S 3 3 2 2

500 N A 3 3 0 3

500 N B* 0 3 0 0

500 S 3 3 3 2

2000 N 3 2 0 2

2000 S 3 2 3 0

Note: variations from planned replication are highlighted in bold.

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2.5 Laboratory Analysis

Each bag of oysters was treated as a replicate and a composite sample of ten individual oysters per

bag was used for analysis (n = two or three bags per site). Oysters were removed from sample bags,

opened, composited, homogenized then analysed by the National Measurement Institute (NMI),

which is a NATA accredited laboratory in the processing of biological tissue samples for chemical

analyses. Samples were collected using the following quality control procedures:

Initially oysters (consisting of three composite samples of ten oysters / site) were collected

from the proposed source (XL oysters) and analysed for the suite of chemicals to confirm

the source was suitable – these were named “time zero”;

“Time zero” oysters (three replicate samples of ten oysters each, collected at the start of

each deployment from the oyster population) were included in each sampling round;

During deployments, oysters were collected by a qualified marine scientist, transferred

(using gloves) into pre-labelled snap lock bags, frozen overnight (-20ºC) and placed into a

clean esky on ice for delivery. No shucking or sub-sampling of oysters was undertaken in

the field to prevent contamination of tissue samples;

Samples were sent to the laboratory under standard chain of custody (COC) conditions and

removed from sample bags, sub sampled and analysed by NMI following their sampling

protocols for marine tissue samples.

2.5.1 Laboratory Analysis of Organics

Organochlorine (OC), organophosphate (OP), total polychlorinated biphenyls (PCBs) and PCB

arochlors were analysed by NMI using their method for Determination of Organochlorine Pesticides,

Organophosphorus Pesticides (OPPs) and Polychlorinated Biphenyls (PCBs) in Biota (method NR19,

NMI 2008b) (Tables 2.3 - 2.5). Whole oyster tissue (wet weight) was homogenised in a blender and

mixed with anhydrous sodium sulphate then extracted using dichloromethane. The extract was

cleaned up by Gel Permeation Chromatography (GPC). The final extract was analysed by Gas

Chromatography - Electron Capture Detector (GC-ECD) (dual column) for OC and PCBs and Gas

Chromatography - Nitrogen/Phosphorus Detector (GC-NPD) for OP compounds. For every batch of

twenty samples or less, at least one blank, one duplicate, one blank spike, one sample spike and one

laboratory control sample (CRM or in-house reference) was tested.

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Table 2.3 Organochlorine pesticides tested in oysters.

Sample

and Matrix

Individual Test Test Limit of Reporting

(LOR)

Method Reference

Whole

Oyster

Tissue

(wet

weight,

mg/kg)

Organochlorine

Pesticides (OCs)

Bromophos-ethyl 0.01 mg/kg

NR19

Carbophenothion 0.01 mg/kg

NR19

Chlorfenvinphos (E)

and (Z)

0.01 mg/kg

NR19

Chlorpyrifos 0.01 mg/kg

NR19

Chlorpyrifos-methyl 0.01 mg/kg

NR19

Demeton-methyl 0.01 mg/kg

NR19

Diazinon 0.01 mg/kg

NR19

Dichlorvos 0.01 mg/kg

NR19

Dimethoate 0.01 mg/kg

NR19

Ethion 0.01 mg/kg

NR19

Fenamiphos 0.01 mg/kg

NR19

Fenthion 0.01 mg/kg

NR19

Malathion 0.01 mg/kg

NR19

Azinphos Methyl 0.01 mg/kg

NR19

Monocrotophos 0.01 mg/kg

NR19

Parathion 0.01 mg/kg

NR19

Parathion-methyl 0.01 mg/kg

NR19

Pirimphos-ethyl 0.01 mg/kg

NR19

Prothiofos 0.01 mg/kg

NR19

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Table 2.4 Organophosphate pesticides tested in oysters.

Sample

and Matrix


(LOR)

Method Reference

Whole

Oyster

Tissue

(wet

weight,

mg/kg)

Organophosphat

e Pesticides

(OPs)

Aldrin 0.01 mg/kg

NR19

Alpha- BHC 0.01 mg/kg

NR19

Beta- BHC 0.01 mg/kg

NR19

2, 2- DDD 0.01 mg/kg

NR19

4,4-DDE 0.01 mg/kg

NR19

4,4-DDT 0.01 mg/kg

NR19

DDT (total) 0.01 mg/kg

NR19

Dieldrin 0.01 mg/kg

NR19

Alpha endosulfan 0.01 mg/kg

NR19

Beta endosulfan 0.01 mg/kg

NR19

Endosulfan sulphate 0.01 mg/kg

NR19

Endosulphan (total) 0.01 mg/kg

NR19

Endrin 0.01 mg/kg

NR19

Endrin aldehyde 0.01 mg/kg

NR19

Endrin ketone 0.01 mg/kg

NR19

Heptachlor 0.01 mg/kg

NR19

Heptachlor epoxide 0.01 mg/kg

NR19

Hexachlorobenzene

(HCB)

0.01 mg/kg

NR19

Gamma-BHC 0.01 mg/kg

NR19

Methoxychlor 0.01 mg/kg

NR19

Cis-chlordane 0.01 mg/kg

NR19

Trans-chlordane 0.01 mg/kg

NR19

Chlordane (total) 0.01 mg/kg

NR19

Oxychlordane 0.01 mg/kg

NR19

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Table 2.5 Polychlorinated biphenyls tested in oysters.

Sample

and Matrix


(LOR)

Method Reference

Whole

Oyster

Tissue

(wet

weight,

mg/kg)

Polychlorinated

Biphenyls (PCB)

Congeners

# 8 0.01 mg/kg

0.002 mg/kg for the

April - May 2013

deployment.

NR19

NR19

# 18 0.01 mg/kg

NR19

# 28 0.01 mg/kg

NR19

# 44 0.01 mg/kg

NR19

NR19

# 52 0.01 mg/kg

0.002 mg/kg for the

April - May 2013

deployment.

NR19

# 66 0.01 mg/kg

NR19

# 77 0.01 mg/kg

NR19

NR19

# 101 0.01 mg/kg

NR19

# 105 0.01 mg/kg

0.002 mg/kg for the

April - May 2013

deployment.

NR19

# 118 0.01 mg/kg

NR19

NR19

# 126 0.01 mg/kg

NR19

# 128 0.01 mg/kg

NR19

# 138 0.01 mg/kg

0.002 mg/kg for the

April - May 2013

deployment.

NR19

NR19

# 153 0.01 mg/kg

NR19

# 169 0.01 mg/kg

NR19

# 170 0.01 mg/kg

NR19

NR19

# 180 0.01 mg/kg

0.002 mg/kg for the

April - May 2013

deployment.

NR19

# 187 0.01 mg/kg

0.002 mg/kg for the

April - May 2013

deployment.

NR19

# 195 0.01 mg/kg

NR19

NR19

# 206 0.01 mg/kg

NR19

# 209 0.01 mg/kg

NR19

Total

Polychlorinated

Biphenyls (PCBs)

Total sum of

congeners

0.01 mg/kg

NR19

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2.5.2 Laboratory Analysis Metals / Metalloids

Metals/metalloids were analysed by NMI using their method for Determination of Elements in Food

and Biota (method NT2.46, NMI 2012) (Table 2.6). Whole oyster tissue (wet weight) was

homogenised in a blender. The sample was digested with concentrated nitric acid (or a mixture of

nitric and hydrochloric acids) by heating on top of a boiling water bath. Elements were determined

using Inductively Coupled Plasma - Mass Spectrometry (ICP - MS) and / or Inductively Coupled

Plasma - Atomic Emission Spectrometry (ICP - AES). The choice of analytical technique depended

on the required detection limit and the need to avoid interferences. For every batch of twenty

samples or less, at least one blank, one duplicate, one blank spike, one sample spike and one

laboratory control sample (CRM or in-house reference) was tested.

Table 2.6 Metals/metalloids tested in oyster tissue samples.

Sample

and Matrix


(LOR)

Method Reference

Whole

Oyster

Tissue

(wet

weight,

mg/kg)

Metals/metalloids Arsenic 0.05 mg/kg NT2_46

Inorganic arsenic 0.05 mg/kg NT2_56

Cadmium 0.01 mg/kg NT2_46

Chromium 0.05 mg/kg NT2_46

Cobalt 0.01 mg/kg NT2_46

Copper 0.01 mg/kg NT2_46

Iron 0.01 mg/kg NT2_46

Lead 0.01 mg/kg NT2_46

Manganese 0.01 mg/kg NT2_46

Mercury 0.01 mg/kg NT2_46

Nickel 0.01 mg/kg NT2_46

Selenium 0.01 mg/kg NT2_46

Silver 0.02 mg/kg NT2_46

Zinc 0.01 mg/kg NT2_46

2.5.3 Laboratory Quality Assurance / Quality Control

For every batch of twenty samples or less, at least one blank, one duplicate, one blank spike, one

sample spike and one laboratory control sample (CRM or in-house reference) was tested. Quality

assurance and quality control (QA/QC) procedures employed by the analytical laboratory (National

Measurement Institute, NMI) are listed below:

NMI has National Association of Testing Authorities (NATA) accreditation and Quality

System certification in accordance with ISO 9001 and ISO 17025;

Every twenty samples, an analysis blank (containing solvent used to extract chemicals)

was tested;

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Every twenty samples, a laboratory control sample (spiking of known concentration of

measured analyte) was undertaken. The acceptable range for spike recoveries was 50 -

150%;

Every ten samples, a sample was duplicated to test the replication between sample runs.

Acceptable relative percentage differences on duplicates was 40%;

Every twenty samples a matrix spike (containing analyte concentrations which were

unknown to the analyst and different to that being tested) was undertaken. The acceptable

range for spike recoveries was 50 - 150%. Acceptable relative percentage differences on

spikes were 40%.

The percentage of moisture in oysters was analysed following the December 2012 -

January 2013 and April - May 2012 deployments. Ideally, concentrations of metals should

be adjusted by the percentage of moisture and reported in dry weight concentrations but

this was not possible as this information was not available during all sampling events.

However, the percentage of moisture was similar between sites and sampling events. The

percentage of moisture for December 2012 - January 2013 ranged from 85 - 88.8%, with

an average of 86.86% and the percentage of moisture for April - May 2012 ranged from 84

- 87.7%, with an average of 85.51%.

For each sampling round, a quality control report was provided by NMI. The report was reviewed to

ensure quality control blanks, duplicates and spikes were within acceptable ranges as outlined above.

Copies of the QA/QC reports for each sampling event are provided in Appendix 4.

2.6 Guideline Values and Comparison Criteria for Chemicals in Oyster Tissue

A summary of available guidelines for maximum residues of organic and metal/metalloid chemicals in

a) saltwater used for aquaculture production, and b) oysters sourced from NSW background

concentrations is provided in Tables 2.7 and 2.8.

Maximum residue levels (ANZFA 2011) were used for comparison to assess whether chemicals were

present at concentrations of concern. These guidelines have been used as a point of comparison in

other studies that have used S. glomerata as a biomonitor of oysters in the receiving waters of

Boulder Bay and Burwood Beach (for example, Andrew-Priestley 2011; Ajani et al 1999; NSW EPA

1996).

Note: Maximum residue levels (ANZFA 2011) are not applicable to this study in terms of health risks

for human consumption of oysters as the main aim of this study was to use oysters as a biomonitor

for environmental contamination, not to assess whether chemicals exceed concentrations in oysters

intended as a food source. There are no oysters commercially grown within the boundaries of this

study. However, in the absence of other available guidelines the ANZFA MRLs are used as a point of

comparison to assess whether metals are elevated.

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Metal/metalloid concentrations in oysters were compared to those in time zero oysters to determine

whether concentrations had changed following the eight week deployments.

Background metal/metalloid concentrations reported by Scanes and Roach (1999) were also used for

comparison. Background metal concentrations are expected in environmental samples, such as

oysters, as metals/metalloids are natural constituents of the environment. Background concentrations

of individual metals/metalloids can vary between locations, according to the natural geology of the

region. Scanes and Roach (1999) provide an indication of the background metal concentrations in

S. glomerata found in 20 estuaries throughout NSW.

Note: The study undertaken by Scanes and Roach (1999) was based on measurement of

metals/metalloids in S. glomerata sampled from uncontaminated estuaries through NSW. There may

be differences between estuaries and marine environments in background levels of metals/metalloids.

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Table 2.7 Comparison criteria for organic chemicals in oyster tissue.

a ANZFA (2011),

b Scanes and Roach (1999).

Contaminant ANZFA (2011) Food Standards: Maximum Residue Limits

for molluscs (mg/kg, wet weight) a

Organochlorine Pesticides (OCs)

Aldrin 0.1

Alpha-BHC 0.01

Alpha-Endosulfan NA

Beta-BHC 0.01

Beta-Endosulfan NA

Cis-Chlordane 0.05

Delta-BHC 0.01

Dieldrin 0.1

Endosulfan Sulfate NA

Endrin NA

Endrin Aldehyde NA

Endrin Ketone NA

gamma-BHC (Lindane) 1

Heptachlor 0.05

Heptachlor epoxide 0.05

Hexachlorobenzene (HCB) 0.1

Methoxychlor NA

Oxychlordane 0.05

pp-DDD 1

pp-DDE 1

pp-DDT 1

Trans-Chlordane 0.05

PCBs

Total PCBs < 0.5

Organophosphate Pesticides (OPs)

Azinphos (Ethyl) NA

Azinphos (Methyl) NA

Chlorfenvinphos (E) NA

Chlorfenvinphos (Z) NA

Chlorpyrifos NA

Chlorpyrifos Methyl NA

Demeton-S-Methyl NA

Diazinon NA

Dichlorvos NA

Dimethoate NA

Ethion NA

Fenitrothion NA

Fenthion NA

Malathion NA

Parathion (Ethyl) NA

Parathion (Methyl) NA

Pirimiphos (Ethyl) NA

Pirimiphos (Methyl) NA

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Table 2.8 Comparison criteria for metals/metalloids in oyster tissue.

Metals / metalloids

ANZFA Food Standards: Maximum Residue Limits for molluscs

(mg/kg, wet weight) a

Concentrations in NSW oysters from background / reference locations

(mg/kg, wet weight) b

Arsenic < 1.0 1.88

Cadmium < 2.0 0.54

Chromium NA 0.26

Cobalt NA 0.064

Copper NA 21.6

Iron NA -

Lead < 2.0 0.085

Manganese NA 2.53

Mercury < 0.5 -

Nickel NA 0.13

Selenium 1.0 0.4

Silver NA 0.24

Zinc 1000 277

a ANZFA (2011),

b Scanes and Roach (1999).

2.7 Baseline Concentrations in Source Oysters

Prior to the first oyster deployment, two oyster composite samples (aggregates of ten oysters) from

the proposed oyster source (XL oysters, Port Stephens) were tested for the same suite of analytes to

ensure that the source was appropriate for the study. All OC pesticides, OP pesticides, PCB

congeners and PCBs were below the limit of reporting (LOR) and a summary of the results is

provided in Appendix 1.

Background trace metal concentrations are expected in environmental samples, such as oysters, as

metals are natural elements of the environment. Background concentrations of individual metals can

vary between locations, according to the natural geology of the region. Scanes and Roach (1999)

provide an indication of the background metal concentrations in S. glomerata found in 20 estuaries

throughout NSW. Concentrations of trace metals/metalloids were well below national food authority

maximum residue level for oysters (ANZ MRLs) (ANZFA 2011) and oyster background concentrations

for NSW (Scanes and Roach 1999). The analysis demonstrated that XL oysters were a suitable

oyster source for the Burwood Beach Oyster Biomonitoring Study.

2.8 Statistical Analysis

2.8.1 Univariate Analysis

Statistical analyses were performed using Statistica Version 7. Chemical concentrations in oyster

tissue were examined for normality, using a normal probability plot and homogeneity of variance,

HUNTER WATER


BURWOOD BEACH WWTW


using a means versus standard deviation test and the data transformed (ln x+1) where applicable.

Where chemicals were below the LOR, half the LOR was used. Differences in concentrations of

metals/metalloids in oyster tissue over all deployments were assessed using a general linear model

factorial ANOVA with time and site as the main factors. An interaction between time and site was

also assessed. In the case of some metals (including cobalt, iron, lead, manganese, nickel and

silver), a normal distribution could not be achieved using a log transformation. These metals were

instead analysed via univariate PERMANOVAs in Primer 6, which computes a randomisation p-value

and does not rely on data being normally distributed.

Linear regression was also used for each deployment period to determine if there were significant

gradient relationships (p < 0.05) between chemicals and distance from Burwood Beach WWTW

outfall. While the relationship between oyster metal bioaccumulation is not necessarily a linear

relationship, regression should still show if tissue concentrations of each metal/metalloid was related

to site distance from the outfall. The results for cobalt and nickel in August - September 2012 and

arsenic and cobalt in April - May 2013 did not meet normality assumptions and were unable to be

transformed. These metals were instead analysed using permutational linear regression (Butler et al.

2003) which randomises cases and provides a distribution free test. It was also used when standard

regression p-values were very close to 0.05 and more evidence was required to confirm a statistical

relationship. In all cases 999 randomisations were performed.

2.8.2 Multivariate Analysis

Multi-Dimensional (MDS) plots were generated in PRIMER 6 to identify if there was any grouping

between sites in metal profiles during each sampling event. Ordination of metal concentrations was

performed using MDS scaling in PRIMER 6, based on ranked matrices of dissimilarities between

samples, employing the square root transformation Euclidean distance, as a measure of dissimilarity.

Goodness of fit (stress) was assessed using Kruskal‟s stress formula and compared to maximum

values recommended by Sturrock and Rocha (2000).

Analysis of similarities (ANOSIM) was undertaken in PRIMER 6 to assess if there was a significant

difference in metal profiles. If a significant difference was found, pairwise comparisons were

performed to assess which pairwise site comparisons were contributing to the difference.

Power analysis was undertaken on the first round of data in order to help design and modify, where

applicable, future oyster bioaccumulation studies. A Type I error rate of 5% (0.05) was adopted here,

and a Type II error rate of 20% (0.2, power 80%) was considered acceptable. A 50% effect size was

used. A 50% effect size has been considered suitable in other studies that have used S. glomerata

as a biomonitor (Robinson et al. 2005).

HUNTER WATER


BURWOOD BEACH WWTW


3 RESULTS

3.1 Organic Chemicals

Analysis of organic chemicals included a suite of organochlorine (OC) and organophosphate (OP)

pesticides, polychlorinated biphenyls (PCBs) congeners and total PCBs (summation of PCB

congeners). Following the January - April 2012 deployment, OC pesticides were detected at Outfall B

and 2000 S. At Outfall B, four chemicals were detected at or above the LOR including heptachlor,

trans-chlordane, cis-chlordane and dieldrin (with respective averages of 0.009 mg/kg, 0.028 mg/kg,

0.007 mg/kg and 0.007 mg/kg). Trans-chlordane was detected in oysters from 2000 S at an average

concentration of 0.085 mg/kg. These concentrations are below the ANZFA MRLs (ANZFA 2011) of

0.05 mg/kg for heptachlor, trans-chlordane and cis-chlordane and 0.1 mg/kg for dieldrin. All other

organic chemicals were below the LOR.

For the May - July 2012 deployment and the March - May 2013 deployment, all OCs, OPs, PCB

congeners and total PCBs were lower than the LOR (0.01 mg/kg).

A summary table of organic concentrations detected in oyster tissue, with comparison to ANZFA

MRLs for molluscs (ANZFA 2011) is provided in Appendix 1. As the majority of organics were below

the LOR, statistical comparisons were not carried out.

3.2 Metals / Metalloids

Metals/metalloids in oyster tissue, including arsenic, cadmium, chromium, cobalt, copper, iron, lead,

manganese, nickel, selenium, silver and zinc are presented in Figures 3.1 - 3.13. These graphs also

include comparisons to the available ANZFA MRLs (ANZFA 2011), concentrations of

metals/metalloids that have been previously detected in NSW estuarine background locations and

concentrations in time zero samples (i.e. oysters before each deployment). The full results of

metals/metalloids that were detected in oyster tissue, along with comparison to the ANFZA MRLs,

concentrations of metals/metalloids in S. glomerata from background locations in NSW (Scanes and

Roach 1999) and time zero concentrations (i.e. before deployment) is also provided in Appendix 2.

Where arsenic exceeded 1 mg/kg, these oysters were tested for inorganic arsenic. Inorganic arsenic

is the portion of total arsenic that is inorganic and includes the most toxic forms, arsenite (As3+

) and

arsenate (As5+

). All samples tested for inorganic arsenic were below the LOR of 0.05 mg/kg and the

ANZFA MRL of 1 mg/kg. ANFZA MRLs are also available for cadmium (2 mg/kg), lead (2 mg/kg),

mercury (0.5 mg/kg), selenium (1 mg/kg) and zinc (1000 mg/kg). All oyster tissue samples were well

below these limits.

Most metal/metalloid concentrations were lower than or similar to those reported by Scanes and

Roach (1999) for oysters from NSW background estuarine locations. Concentrations of copper,

selenium and zinc were higher across most sites and of a similar magnitude spatially.

HUNTER WATER


BURWOOD BEACH WWTW


Prior to deployment, oysters were tested for metals/metalloids (time zero), to compare whether

concentrations changed after each deployment period. It was found that following each deployment

most metals/metalloids (including arsenic, cadmium, chromium, copper, lead, mercury, nickel,

selenium, silver and zinc) had higher values in field deployed oysters in comparison to the respective

time zero oysters for that deployment.

3.3 Univariate Analysis of Metals / Metalloids

General linear model ANOVAs or univariate PERMANOVAs were used to assess for significant

differences in oyster tissue metal/metalloid concentrations among times and sites to determine if

there were significant interactions between time and site. The results of these ANOVA analyses are

provided in Table 3.1.

Overall, there were no patterns to show consistent significant spatial variability of any metals along a

distance gradient or between sites (i.e. there were no metals that were consistently elevated over the

three sampling events at outfall sites or that decreased with distance from the outfall).

For many metals, including arsenic, cadmium, cobalt, iron, manganese, silver and zinc, there were

significant interactions found between time and site indicating that the patterns among sites were

inconsistent between deployment periods. Patterns in arsenic, cobalt, iron and silver were

inconclusive and there was high temporal variability and spatial variability between sites. The

patterns for cadmium and zinc suggest that values were different across most sites following one

deployment, in comparison to the other two sampling events. Cadmium concentrations were lower

during May - July 2012 with the exception of samples collected at 2000 S. Concentrations of zinc

were higher during March - May 2013, except at outfall B. For manganese and lead, during March -

May 2013 concentrations were different at both outfall sites in comparison to other sites. During

March - May 2013, manganese concentrations were significantly elevated at the outfall sites in

comparison to the 500 m and 2000 m sites, but lead concentrations were significantly lower at the

outfall sites in comparison to all other sites. A significant main effect was found for the factor of time

for nickel and selenium, whereby concentrations were higher during March - May 2013 in comparison

to January - April 2012 and May - July 2012.

Linear regressions were used to determine if there were any significant relationships between metal

concentrations and distance from the outfall, for each deployment period. Cadmium was found to

significantly decrease with distance from the outfall during January - April 2012 but significantly

increased with distance during March - May 2013. Iron significantly decreased with distance from the

outfall during May - July 2012 and March - May 2013. Manganese was also found to significantly

decrease with distance from the outfall during March - May 2013.

HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

2

1

0

Ars

en

ic (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

2

1

0

Ars

en

ic (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall B

Outfall A100m S

500m S2000m S

TIME ZERONSW BG

2

1

0

Site (distance from outfall)

Ars

en

ic (

mg

/k

g)

LOR

LOR = 0.02 (mg/kg)

January - April 2012

May - July 2012

March - May 2013

Figure 3.1 Concentrations of arsenic in S. glomerata tissue (mg/kg, wet weight) following eight

weeks offshore deployment during January - April 2012, May - July 2012 and March - May 2013

(mean ± SE). For each site, N = 2 - 3 replicate samples with 10 composite oysters / replicate.

LOR = Limit of Reporting. Colours indicate different distances from the outfall.

HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.50

0.25

0.00

Ca

dm

ium

(m

g/

kg

)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.50

0.25

0.00

Ca

dm

ium

(m

g/

kg

)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.50

0.25

0.00


Ca

dm

ium

(m

g/k

g)

LOR

MRL = 2.0 (mg/kg) LOR = 0.01 (mg/kg)


May - July 2012

March - May 2013

Figure 3.2 Concentrations of cadmium in S. glomerata tissue (mg/kg, wet weight) following

eight weeks offshore deployment during January - April 2012, May - July 2012 and March - May

2013 (mean ± SE). For each site, N = 2 - 3 replicate samples with 10 composite oysters /

replicate. LOR = Limit of Reporting. Colours indicate different distances from the outfall.

HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.2

0.1

0.0

Ch

rom

ium

(m

g/

kg

)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.2

0.1

0.0

Ch

rom

ium

(m

g/

kg

)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.2

0.1

0.0


Ch

rom

ium

(m

g/

kg

)

LOR


May - July 2012

March - May 2013

LOR = 0.05 (mg/kg)

Figure 3.3 Concentrations of chromium in S. glomerata tissue (mg/kg, wet weight) following




HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.08

0.04

0.00

Co

ba

lt (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.08

0.04

0.00

Co

ba

lt (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.08

0.04

0.00


Co

ba

lt (

mg

/k

g)

LOR

LOR = 0.01 (mg/kg)


May - July 2012

March - May 2013

Figure 3.4 Concentrations of cobalt in S. glomerata tissue (mg/kg, wet weight) following eight




HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

30

15

0

Co

pp

er

(mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

30

15

0

Co

pp

er

(mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

30

15

0


Co

pp

er

(mg

/k

g)

LOR

May - July 2012

LOR = 0.01 (mg/kg)

March - May 2013


Figure 3.5 Concentrations of copper in S. glomerata tissue (mg/kg, wet weight) following eight




HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

30

15

0Iro

n (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

30

15

0Iro

n (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

30

15

0


Iro

n (

mg

/k

g)

LOR

LOR = 0.01 (mg/kg)


May - July 2012

March - May 2013

Figure 3.6 Concentrations of iron in S. glomerata tissue (mg/kg, wet weight) following eight




HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.08

0.04

0.00Lea

d (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.08

0.04

0.00Lea

d (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.08

0.04

0.00


Lea

d (

mg

/k

g)

LOR

MRL = 2.0 (mg/kg) LOR = 0.01 (mg/kg)


May - July 2012

March - May 2013

Figure 3.7 Concentrations of lead in S. glomerata tissue (mg/kg, wet weight) following eight




HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

2

1

0

Ma

ng

an

ese

(m

g/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

2

1

0

Ma

ng

an

ese

(m

g/

kg

)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

2

1

0


Ma

ng

an

ese

(m

g/

kg

)

LOR

LOR = 0.01 (mg/kg)


May - July 2012

March - May 2013

Figure 3.8 Concentrations of manganese in S. glomerata tissue (mg/kg, wet weight) following




HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.010

0.005

0.000

Merc

ury

(m

g/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.010

0.005

0.000

Merc

ury

(m

g/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.010

0.005

0.000


Merc

ury

(m

g/k

g)

LOR

MRL = 0.5 (mg/kg) LOR = 0.01 (mg/kg)


May - July 2012

March - May 2013

Figure 3.9 Concentrations of mercury in S. glomerata tissue (mg/kg, wet weight) following




HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.30

0.15

0.00Nic

ke

l (m

g/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.30

0.15

0.00Nic

ke

l (m

g/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.30

0.15

0.00


Nic

kel (m

g/k

g)

LOR

LOR = 0.01 (mg/kg)


May - July 2012

March - May 2013

Figure 3.10 Concentrations of nickel in S. glomerata tissue (mg/kg, wet weight) following eight




HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.50

0.25

0.00

Sele

niu

m (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.50

0.25

0.00

Sele

niu

m (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.50

0.25

0.00


Sele

niu

m (

mg

/k

g)

LOR

MRL = 1.0 (mg/kg) LOR = 0.01 (mg/kg)


May - July 2012

March - May 2013

Figure 3.11 Concentrations of selenium in S. glomerata tissue (mg/kg, wet weight) following




HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.30

0.15

0.00Silver

(mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.30

0.15

0.00Silver

(mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

0.30

0.15

0.00


Silver

(mg

/k

g)

LOR

LOR = 0.02 (mg/kg)


May - July 2012

March - May 2013

Figure 3.12 Concentrations of silver in S. glomerata tissue (mg/kg, wet weight) following eight




HUNTER WATER


BURWOOD BEACH WWTW


2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

500

250

0Zin

c (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

500

250

0Zin

c (

mg

/k

g)

LOR

2000m N

500m N B

500m N A100m N

Outfall BOutfall A

100m S500m S

2000m S

TIME ZERONSW BG

500

250

0


Zin

c (

mg

/k

g)

LOR

MRL = 1000 (mg/kg) LOR = 0.01 (mg/kg)


May - July 2012

March - May 2013

Figure 3.13 Concentrations of zinc in S. glomerata tissue (mg/kg, wet weight) following eight




HUNTER WATER


BURWOOD BEACH WWTW


Table 3.1 Factorial GLM ANOVAs on metal/metalloids (mg/kg, wet weight) concentrations in

oyster tissue deployed during January - April 2012, May - July 2012 and March - May 2013.

N = 2 - 3 replicates (of a composite sample of 10 oysters) per site.

Arsenic Cadmium

Source DF MS F p MS F p

Time 2 0.05 2.96 0.09 0.08 19.67 0.00**

Site 7 0.02 1.15 0.39 0.00 0.45 0.85

Time * Site 13 0.02 3.36 0.00** 0.00 3.73 0.00**

Error 41 0.00 0.00

Transformation ln (x+1) n/a

Cobalt Copper

Source DF MS F pa MS F p

Time 2 0.00 16.00 0.00** 0.36 9.17 0.00**

Site 7 0.00 3.02 0.02* 0.09 2.15 0.11

Time * Site 13 0.00 3.15 0.00** 0.04 1.66 0.11

Error 41 0.00 0.02

Transformation n/a ln (x+1)

Iron Lead


a

Time 2 42.75 3.62 0.04* 0.00 30.53 0.00**

Site 7 35.41 3.00 0.01* 0.00 2.66 0.03*

Time * Site 13 34.68 2.94 0.00** 0.00 1.11 0.39

Error 41 11.81 0.00

Transformation n/a n/a

Manganese Nickel


a

Time 2 3.74 37.51 0.00** 0.01 8.53 0.00**

Site 7 0.36 3.58 0.00** 0.00 2.38 0.05

Time * Site 13 0.67 6.68 0.00** 0.00 1.74 0.10

Error 41 0.10 0.00

Transformation n/a n/a

a = randomization p-value, * = < 0.05, ** = < 0.01.

HUNTER WATER


BURWOOD BEACH WWTW


Table 3.1 (continued) Factorial GLM ANOVAs on metal/metalloids (mg/kg, wet weight)

concentrations in oyster tissue deployed during January - April 2012, May - July 2012 and

March - May 2013. N = 2 - 3 replicates (of a composite sample of 10 oysters) per site.

Selenium Silver

Source DF MS F p MS F pa

Time 2 0.04 11.88 0.00** 0.01 3.62 0.03*

Site 7 0.01 3.05 0.04* 0.00 2.65 0.02**

Time * Site 13 0.00 1.47 0.17 0.00 2.58 0.02**

Error 41 0.00 0.00

Transformation ln (x+1) n/a

Zinc

Source DF MS F p

Time 2 96775.76 17.12 0.00**

Site 7 4949.69 0.88 0.55

Time * Site 13 5744.73 2.41 0.02*

Error 41 2386.59

Transformation n/a

a = randomization p-value, * = < 0.05, ** = < 0.01.

HUNTER WATER


BURWOOD BEACH WWTW


Table 3.2 Regressions of oyster tissue metal/metalloids concentration with distance from the

outfall during each sampling event. N = 2 - 3 replicates (of a composite sample of ten oysters)

per site.


Metals DF R2 F Standard p-value

Permutation T

Randomisation p-value Slope

Arsenic 1, 22 0.438 17.211 0.000**

+ve

Cadmium 1, 22 0.295 9.219 0.006**

-ve

Cobalt 1, 22 0.014 0.318 0.578 Copper 1, 22 0.002 0.0330 0.857 Iron 1, 22 0.126 3.0166 0.089 Lead 1, 22 0.157

-2.030 0.011* -ve

Manganese 1, 22 0.649 40.624 0.000**

+ve

Nickel 1, 22 0.041

-0.960 0.262 Selenium 1, 22 0.103 2.537 0.125

Silver 1, 22 0.100 2.468 0.130 Zinc 1, 22 0.148 3.851 0.062

May - July 2012

Arsenic 1,23 0.136 3.609 0.070 Cadmium 1,23 0.156

2.060 0.033* +ve

Cobalt 1,23 0.027 0.629 0.436 Copper 1,23 0.017 0.403 0.531 Iron 1,23 0.003 0.701 0.793

-ve

Lead 1,23 0.012 0.283 0.599 Manganese 1,23 0.046 1.104 0.304 Nickel 1,23 0.016 0.377 0.545 Selenium 1,23 0.111 2.867 0.104 Silver 1,23 0.028 0.665 0.423 Zinc 1,23 0.038 0.918 0.348

March - May 2013

Arsenic 1,13 0.112

-1.281 0.262 Cadmium 1,13 0.298 5.532 0.035*

+ve

Cobalt 1,13 0.010 0.137 0.717 Copper 1,13 0.006 0.076 0.786 Iron 1,13 0.289 5.297 0.038*

-ve

Lead 1,13 0.014

-0.432 1.000 Manganese 1,13 0.200

-1.810 0.031* -ve

Nickel 1,13 0.065 0.904 0.358 Selenium 1,13 0.062 0.869 0.368 Silver 1,13 0.041 0.562 0.467 Zinc 1,13 0.009 0.118 0.737

*= < 0.05, **= < 0.01

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3.4 Multivariate Analysis of Metal Profiles

3.4.1 January - April 2012

A non-metric MDS plot was generated in PRIMER 6 to compare similarities of metal/metalloid profiles

among sites for the January - April 2012 oyster deployment. Time zero samples were also included

in this analysis to determine how the multivariate metal profile changed post deployment. Chromium

and mercury were excluded due to all values being less than the LOR. The MDS plot is presented in

Figure 3.14.

The plot indicates that there is a weak gradient with distance from the outfall. The outfall sites and, to

a lesser extent 100 m sites, are clustered together although within site variability is high as samples

are not closely clustered. The time zero samples are however, quite segregated from the post-

deployment samples. There is little difference between the 500 and 2,000 m sites with overlapping of

these samples. Vectors on the plot suggest that higher concentrations of cadmium, cobalt, copper,

iron, manganese and zinc in two 100 m replicates (located to the left of the MDS plot) are driving the

separation of these samples on the plot.

Transform: Square root

Resemblance: D1 Euclidean distance

DistanceTimeZero

Outfall

100

500

2000

TimeZeroTimeZero

TimeZero

Outfall

OutfallOutfallOutfall

Outfall

Outfall North

North

North

South

South

South North

NorthNorth

South South

South North

North

North

South

South

South

Cadmium

Cobalt

Copper

Iron

Manganese

Silver

Zinc

2D Stress: 0.02

Figure 3.14 MDS plot of multivariate suite of metals/metalloids for each sample from the

January – April 2012 deployment. Symbols indicate the direction of each sample. N = 3

replicates (of a composite sample of 10 oysters) per site.

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3.4.2 May - July 2012

The MDS plot for May - July 2012 is presented in Figure 3.15. Time zero samples were also included

in this analysis to determine how the multivariate metal profile changed post deployment. Chromium

and mercury were excluded due to all values being less than the LOR. One sample taken at time

zero is clustered separately from all other samples showing that this sample had a different metal

profile. Vectors on the plot suggest that cadmium, copper, cobalt, iron, lead, nickel, manganese,

silver and zinc are responsible for this separation. The plot shows that there is little difference

between the metal profiles of sites with overlapping of most samples / distances.



DistanceTimeZero

Outfall

100

500

2000

TimeZero

TimeZero

TimeZero

Outfall

OutfallOutfall

Outfall Outfall

Outfall

SouthSouthSouth

North

NorthNorth

North

NorthNorthNorthNorth

North

South

South

South

North

NorthSouth

South

Cadmium

Cobalt

Copper

Iron

Lead

Manganese

Nickel

Selenium

SilverZinc

2D Stress: 0.01

Figure 3.15 MDS plot of multivariate suite of metals/metalloids for each sample from the May -

July 2012 deployment. Symbols indicate the direction of each sample. N = 2 - 3 replicates (of a

composite sample of 10 oysters) per site.

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3.4.3 March - May 2013

The MDS plot for March - May 2013 is presented in Figure 3.16. Time zero samples were also

included in this analysis to determine how the multivariate metal profile changed post deployment.

Chromium and mercury were excluded due to all values being less than the LOR. Time zero samples

and two samples from the outfall are clustered separately from all other samples. Vectors on the plot

suggest that the metals arsenic, cadmium, copper, cobalt, iron, selenium and zinc are responsible for

this separation. No strong gradient of impact with distance from the outfall can be detected here.



DistanceTimeZero

Outfall

100

500

2000

TimeZero

TimeZeroTimeZero

OutfallOutfall

OutfallOutfall

North

NorthNorth

SouthSouthNorth

North

South

SouthNorth North

Arsenic

Cadmium

Cobalt

Copper

Iron Nickel

Selenium

Zinc

2D Stress: 0.01

Figure 3.16 MDS plot of multivariate suite of metals/metalloids for each sample from the March

- May 2013 deployment. Symbols indicate the direction of each sample. N = 2 - 3 replicates (of

a composite sample of 10 oysters) per site.

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3.4.4 Overall

MDS plots were generated to compare metal/metalloids profiles over the three deployment periods to

determine overall patterns in the results. These are provided in Figures 3.17 - 3.19. The overall

MDS plots do not show any distinct patterns between distances or direction from the outfall. The

MDS plot by deployment periods shows that March - May 2013 was different and clustered separately

from January - April 2012 and May - July 2012. The vectors suggest that this was due to

concentrations of copper, cobalt, iron and zinc.

A PERMANOVA was undertaken in PRIMER 6 to determine if there were significant differences

among time or site in the suite of metals tested (Table 3.3). The analysis found that there was a

significant interaction between time and site suggesting that the patterns were inconsistent across

deployment periods (which reflects the univariate ANOVAs on individual metals as many also found

an interaction between time and site).

During January - April 2012, there were a number of differences in the suite of metals/metalloids

between sites. The 500 N site was different to Outfall A, 500 S and 2000 N. The 100 S site was also

different to Outfall A and 2000 N. In May - July 2012, the 500 S was different to 2000 N and Outfall A.

Following March - May 2013, the Outfall B site was different to 100 N and 500 S.

Table 3.3 PERMANOVA analysis of a suite of metals/metalloids in the S. glomerata tissue

following three deployment periods.

Factor DF MS Pseudo F Ratio p-value Permutations

Time 2 64.901 13.581 0.001** 999

Site 7 4.518 2.315 0.046* 999

Time * Site 13 4.857 2.489 0.007** 998

Error 41 1.952

* = <0.05, ** = < 0.01.

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DistanceOutfall

100

500

2000

Cobalt

Copper

Iron

Zinc

2D Stress: 0.03

Figure 3.17 MDS plot of multivariate suite of metals/metalloids by distance from the outfall,

pooled over deployment period. N = 3 replicates (of a composite sample of ten oysters) per

site.



TimeJanuary- April 2012

May- July 2012

March- May 2013

Cobalt

Copper

Iron

Zinc

2D Stress: 0.03

Figure 3.18 MDS plot of multivariate suite of metals/metalloids by deployment period, pooled

over distance from the outfall. N = 3 replicates (of a composite sample of ten oysters) per site.

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DirectionOutfall

North

South

Cobalt

Copper

Iron

Zinc

2D Stress: 0.03

Figure 3.19 MDS plot of multivariate suite of metals/metalloids by direction from the outfall,

pooled over deployment period. N = 3 replicates (of a composite sample of ten oysters) per

site.

3.5 Power Analysis

A power analysis was carried out on the January - April 2012 data to estimate which sample sizes

would be required to detect a significant difference between sites for all metals/metalloids. A Type I

error rate of 5% (0.05) was used and a Type II error rate of 20% (0.2, power 80%) was considered

acceptable and a 50% effect size was used.

The power analysis estimated the amount of replication required to detect a significant difference

(p < 0.05) with a 50% effect size (Appendix 1) and is presented in Table 3.4. The estimate of

required sample size was three replicates per site for most metals. The low estimate of sample size

is likely due to the fact that replicate samples were a composite of ten oysters, reducing the variability.

Copper and cobalt had an estimate of four replicates. Nickel had an estimate of twelve replicates,

which was higher due to the higher variation between samples.

Overall, the variability within sites in oyster metal concentrations was low and the adopted sampling

size of three replicates (of a composite sample of ten oysters) per site should be sufficient to detect

differences in most metal concentrations with the exception of cobalt, copper and nickel.

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Table 3.4 Estimates of sample sizes required to detect a significant difference between

metal/metalloids in oysters based on power analysis of January - April 2012 sampling data.

Metal/Metalloid Sample Size Estimate

Arsenic 3

Inorganic Arsenic n/e

Cadmium 4

Chromium n/e

Cobalt 3

Copper 4

Iron 3

Lead n/e

Manganese 3

Mercury n/e

Nickel 12

Selenium 3

Silver 3

zinc 3

n/e = not estimable due to no variation between samples (all or majority were less than the LOR).

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4 DISCUSSION


and biosolids discharges to lead to bioaccumulation of chemicals, over a range of spatial scales,

using Sydney rock oysters, S. glomerata, as a biomonitor. A specific requirement of the study was to

establish the zone in which there is a detectable increase in the concentration of chemicals in oysters

that is related to the outfall. Concentrations of a suite of organic and metal/metalloid chemicals were

measured in oyster tissue following four deployments of eight weeks in the receiving waters of the

Burwood Beach WWTW.

Concentrations of organics and metals/metalloids in oysters were compared to the ANZFA MRLs

(ANZFA 2011). These were used as a guide only (in the absence of other available guidelines) rather

than for assessing health risks for oyster consumption, as there are no commercially grown oysters

within the boundaries of this study.

4.1 Organics

Organic chemical levels in S. glomerata tissue were consistently lower than available ANZFA Food

Standard MRLs for Molluscs (ANZFA 2011). For the majority of measurements, organic chemicals

were lower than the LOR, but there were some exceptions.

In assessment of a suite of OC and OP pesticides, there were four detections of heptachlor, trans-

chlordane, cis-chlordane and dieldrin at Outfall B during January - April 2012. As they were detected

very close to the outfall, it is likely that the source of OC contamination is likely to be Burwood Beach

WWTW. The concentrations were lower than the ANZFA MRLs (2011). These pesticides have

sometimes been detected in routine analysis of effluent and biosolids which has been undertaken by

Hunter Water during 2006 – 2013, but at low concentrations. In effluent, mean concentrations of <

0.005 µg/L for heptachlor, 0.001 µg/L for chlordane and 0.000 for dieldrin have been measured. In

biosolids, mean concentrations of 0.006 µg/L have been measured for dieldrin (heptachlor and

chlordane have not been measured in biosolids). Previous measurements of organic chemicals in

oysters deployed in the receiving waters of Burwood Beach WWTW have not detected any OC or OP

pesticides (Ajani et al. 1999; NSW EPA 1996, reviewed in further detail in the introduction). During

January - April 2012, cis-chlordane was also detected in oysters from 2000 S. This site is located

near Redhead. The study of NSW EPA 1996; Ajani et al. 1999 also used Redhead as a reference

location and DDD was the only organic chemical that exceeded the LOR of 0.02 mg/kg.

Concentrations of PCB congeners and total PCB concentrations were lower than the LOR following

all deployments. No PCBs congeners were detected following all deployments (i.e. in January - April

2012, May- July 2012 and March - May 2013). This is similar to the findings of Ajani et al. 1999 and

NSW EPA 1996, who did not detect PCBs in oysters using the same LOR of 0.01 mg/kg.

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4.2 Metals

Assessment of metal/metalloid concentrations in S. glomerata tissue demonstrated that most metals

were at low concentrations. No metals/metalloids were found to exceed the available ANZFA MRLs

(ANZFA 2011). There were no consistent significant differences in the spatial patterns of

metals/metalloids to suggest that oysters deployed closer to the Burwood Beach WWTW had

accumulated higher concentrations of metals/metalloids than those located further away from the

outfall.

Concentrations of total arsenic in oysters exceeded 1 mg/kg during all sampling events. There is no

ANZFA MRL for total arsenic but the limit for inorganic arsenic is 1 mg/kg. Inorganic arsenic is the

portion of total arsenic that is inorganic and includes the most toxic forms, arsenite (As3+

) and

arsenate (As5+

) which have the capacity to inhibit enzymes and disrupt metabolic activities in marine

invertebrates (Cox 1995). All samples that exceeded 1 mg/kg for total arsenic were tested to

determine the proportion of inorganic arsenic and were all below the LOR of 0.05 mg/kg.

The majority of metals and metalloids in S. glomerata during this study were lower or at similar

concentrations to those that were reported by Scanes and Roach (1999). The exception was

selenium which was higher in most samples in comparison to the background level reported by

Scanes and Roach (1999). This was also reported by Andrew-Priestley (2011), who similarly

deployed S. glomerata in the receiving waters of Burwood Beach WWTW and at reference locations,

Redhead and Fingal Island (described in further detail in introduction). It was found that oysters at all

locations had higher levels of selenium in comparison to those reported by Scanes and Roach (1999).

There are likely to be differences between background metal concentrations between oysters in

estuarine locations and offshore environments but it does provide a point of comparison in the

absence of studies on oysters grown offshore. In general concentrations of metals and metalloids in

oysters living in NW estuaries would be expected to have higher concentrations as higher levels are

found in sediments of NSW estuaries in comparison to the continental shelf; Birch 2000).

Background levels of metals and metalloids may have also been higher during the 1990‟s in

comparison to the present day due to differences in environmental management.

Some metals were found to significantly decrease (p < 0.05, linear regression) with distance from the

outfall during one or two sampling events including cadmium during January - April 2012, iron during

May - July 2012 and March - May 2013 and manganese during March - May 2013. This result could

indicate that Burwood Beach WWTW is an occasional source of these metals into the marine

environment but as this result was not consistent across all sampling events this assumption should

be viewed with caution. Cadmium and manganese have been measured in the final treated effluent

and biosolids from Burwood Beach WWTW during 2006 - 2013 with higher concentrations detected in

the biosolids. Cadmium has been measured in biosolids at concentrations 0.5 - 128 µg/L with an

average of 5.93 µg/L while manganese has been measured in biosolids at concentrations 33 -

1270 µg/L with an average of 360 µg/L (N = 152; Hunter Water 2013).

Measurements of nickel in oysters may suggest an occasional impact. During January - April 2012,

nickel was elevated at 100 m N in comparison to the other sites, this pattern was not seen during the

following two deployment periods. This result could indicate that Burwood Beach WWTW is an

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BURWOOD BEACH WWTW


occasional source of nickel in the marine environment close to the diffuser but again this result was

not consistent across sampling events so should be viewed with caution. Nickel has been measured

in biosolids from Burwood Beach WWTW, from 2006 - 2013, at concentrations 30 - 180 µg/L, with an

average of 47.21 µg/L (N = 152; Hunter Water 2013). Nickel is mainly used as alloys in an extensive

range of everyday domestic and industrial uses including building materials, batteries, mobile phones,

medical equipment, transport and power generation.

Oysters were tested for metals/metalloids prior to each deployment and most metals/metalloids were

found to increase following deployments in Burwood Beach WWTW receiving waters, including

arsenic, cadmium, copper, lead, mercury, nickel, selenium, silver and zinc. Increases in

metal/metalloid concentrations relative to time zero samples do not appear to be related to the outfall

as there were no patterns between elevated concentrations and sites or distance from the outfall.

This suggests that background concentrations of metals/metalloids are higher in the offshore waters

off Newcastle. The oysters deployed were depurated in clean waters for two weeks prior to

deployments (which may have reduced the concentrations of metals and metalloids). However, the

concentrations of metals/metalloids in oysters measured in this study are higher in comparison to

those measured in the concurrent study (with the same design) undertaken at Boulder Bay. Higher

concentrations in oysters deployed in Newcastle waters, in comparison to time zero oysters, may be

due to proximity to the Port of Newcastle which includes three coal export terminals and the historic

industrial background of the city which included a major steelworks (BHP).

Concentrations of mercury, selenium and zinc were higher in oysters deployed in March - May 2013.

For selenium, this was likely because concentrations in the time zero oysters were also higher relative

to the time zero oysters that were sampled in January - April 2012 and May - July 2012. For mercury

and zinc, it is unknown why there were temporal differences. Seasonal environmental changes such

as salinity, temperature and other water quality changes, as well as differences in the reproductive

status, weight and health of oysters can all affect the bioaccumulation of metals in oysters (Phillips

1980). Ajani et al. (1999) and NSW EPA (1996) also found differences between metal/metalloid

concentrations between deployments; concentrations were higher in oysters collected in August

1992, in comparison to February deployments in 1992 - 1996 and respective August deployments in

1993 - 1996.

The findings of this study show that overall metal/metalloids concentrations in oysters following

deployments were low and there was no evidence to suggest an impact from the Burwood Beach

WWTW. There are two possible scenarios for these findings:

1. The first scenario is that metal concentrations were not elevated in Burwood Beach WWTW

receiving waters and the WWTW was not a significant source during the deployments. Hence no

significant differences could be detected in the temporal patterns of metals and metalloids.

2. The second scenario is that the deployment period of eight weeks was not sufficient for S.

glomerata to equilibrate some of the metals/metalloids within their tissue. The rates of uptake

and equilibration in oysters has not been studied extensively or established for all metals,

however eight weeks exposure has been demonstrated as sufficient time of laboratory exposure

of non-essential metalloid/metals to accumulate to concentrations which are significantly different

from controls (Watling 1983; Boisson et al. 2003; Spooner et al. 2003) or to equilibrate (i.e. reach

HUNTER WATER


BURWOOD BEACH WWTW


a plateau) in tissue (Scanes and Roach 1999). A longer exposure time may be required for

essential metals, such as, zinc and copper, as while oysters have been demonstrated to

bioaccumulate essential metals (George et al. 1978; Phillips and Yim 1981; Brown and

McPherson 1992; Phillips and Rainbow 1993; Phillips 1995; Spooner et al. 2003), it has been

suggested that molluscs may exhibit a degree of homeostasis for these metals depending on the

exposure concentration. Essential metals have a minimum metabolic requirement and as

suggested by Langston et al. 1998 (pg. 233) “copper concentrations may be buffered according to

the requirements for the copper containing pigment, haemocyanin”.

Importantly, no metals/metalloids consistently exhibited differences among sites or with distance from

the outfall. The multivariate profiles also suggest that there are no differences in the suite of metals

between sites.

It would be expected that with increases in future discharges of effluent and biosolids at Burwood

Beach WWTW that higher concentrations of organic chemicals and metal/metalloids would be found

via oyster biomonitoring studies.

HUNTER WATER


BURWOOD BEACH WWTW


5 CONCLUSIONS


and biosolids discharges to lead to bioaccumulation of chemicals, over a range of spatial scales,

using Sydney rock oysters, S. glomerata, as a biomonitor. A specific requirement of the study was to

establish the zone in which there is a detectable increase in the concentration of chemicals in oysters

that is related to the outfall. Concentrations of a suite of organic and metal/metalloid chemicals were

measured in oyster tissue following four deployments of eight weeks in the receiving waters of the

Burwood Beach WWTW.

Concentrations of organics and metals/metalloids in oysters were compared to the ANZFA MRLs

(ANZFA 2011). These were used as a guide only (in the absence of other available guidelines) rather

than for assessing health risks for oyster consumption, as there are no commercially grown oysters

within the boundaries of this study.

Organic chemical levels in oyster tissue at all sites, including the outfall site, were consistently

lower than available ANZFA Food Standard MRLs for Molluscs (ANZFA 2011). For the May -

July 2012 and the March - May 2013 deployments, all OCs, OPs, PCB congeners and total PCBs

were lower than the LOR (0.01 mg/kg). As the majority of organics were below the LOR,

statistical comparisons were not carried out. However in the January - April 2012 deployment,

some OC pesticides (i.e. heptachlor, trans-chlordane, cis-chlordane and dieldrin) were detected,

which does indicate their presence in the environment. The Burwood Beach WWTW discharge is

likely to be a source of these chemicals and should be continued to be monitored.

Most metals were at low concentrations in oysters following deployment. No metals/metalloids

were found to exceed the available ANZFA MRLs (ANZFA 2011). There were no consistent

significant differences in the spatial patterns of metals/metalloids to suggest that oysters deployed

closer to the Burwood Beach WWTW had accumulated higher concentrations of

metals/metalloids.

Oysters were tested for metals/metalloids prior to each deployment and most metals/metalloids

were found to increase following deployments in Burwood Beach WWTW receiving waters,

including arsenic, cadmium, copper, lead, mercury, nickel, selenium, silver and zinc. Increases in

metal/metalloid concentrations relative to time zero samples do not appear to be related to the

outfall as there were no patterns between elevated concentrations and sites or distance from the

outfall.

Most metal/metalloid concentrations were lower than or similar to those reported by Scanes and

Roach (1999) for oysters from NSW background estuarine locations. Concentrations of copper,

selenium and zinc were higher across most sites and of a similar magnitude spatially.

It would be expected that with increases in future discharges of effluent and biosolids at Burwood

Beach WWTW that higher concentrations of organic chemicals and metal/metalloids would be

found via oyster biomonitoring studies.

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BURWOOD BEACH WWTW


6 ACKNOWLEDGEMENTS

We would like to thank those that assisted with the design and implementation of this study.

Consulting Environmental Engineers, Hunter Water, NSW EPA, NSW Marine Parks and NSW DPI

Fisheries assisted with the design of the sampling program and methodology. XL Oysters (Lemon

Tree Passage) provided all the oysters for the study. McLennan‟s Dive Services undertook

deployment and retrieval of oyster moorings and oyster bags. The National Measurement Institute

undertook all laboratory analyses in oyster tissue. All surveys were undertaken under NSW Fisheries

Permit # P110051-1.2 and NSW Marine Parks Permit #2011/046.

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BURWOOD BEACH WWTW


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BURWOOD BEACH WWTW


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BURWOOD BEACH WWTW


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HUNTER WATER


BURWOOD BEACH WWTW


Appendix 1 – Organic Chemical Concentrations in Oysters

HUNTER WATER


BURWOOD BEACH WWTW


Summary of average trace organic concentrations (wet weight, mg/kg) measured in oysters following eight weeks offshore deployment for each

deployment period. For each site, N = three replicate samples with 10 composite oysters / replicate. Missing sites are indicated.


Class

Chemical

Comparison Criteria Average concentration (mg/kg, wet weight) with Distance from Outfall

ANZFAa LOR Outfall Mixing Zone Mixing Zone Reference Time Zero

mg/kg mg/kg A B 100 N 100 S 500 N A 500 N B

500 S 2000 N 2000 S TIME ZERO

n= 3 n=3 n= 3 n= 3 n= 3 n= 0 n= 3 n= 3 n= 3 n= 3

Organochlorine (OC) Pesticides

HCB 0.1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

Heptachlor 0.05 0.01 <0.01 0.0086 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

Heptachlor epoxide 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

Aldrin 0.1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

gamma-BHC (Lindane) 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

alpha-BHC 0.01 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

beta-BHC 0.01 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

delta-BHC 0.01 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

trans-Chlordane 0.05 0.01 <0.01 0.028 <0.01 <0.01 <0.01 n/a <0.01 <0.01 0.0085 <0.01

cis-Chlordane 0.05 0.01 <0.01 0.007 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

Oxychlordane 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

Dieldrin 0.1 0.01 <0.01 0.007 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

pp-DDE 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

pp-DDD 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

pp-DDT 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

HUNTER WATER


BURWOOD BEACH WWTW



Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 3 n=3 n= 3 n= 3 n= 3 n= 0 n= 3 n= 3 n= 3 n= 3

Endrin - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

Endrin Aldehyde - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

Endrin Ketone - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

alpha-Endosulfan - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

beta-Endosulfan - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

Endosulfan Sulfate - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

Methoxychlor - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <0.01

PCB Congeners PCB # 8 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 18 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 28 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 44 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 52 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 66 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 77 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 101 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 105 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 118 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 126 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 128 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

HUNTER WATER


BURWOOD BEACH WWTW



Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 3 n=3 n= 3 n= 3 n= 3 n= 0 n= 3 n= 3 n= 3 n= 3

PCB # 138 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 153 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 169 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 170 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 180 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 187 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 195 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 206 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

PCB # 209 - 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

Total PCB's < 0.5 0.01 <10 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 <0.01 <10

Organophosphate (OP) Pesticides

Dichlorvos - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Demeton-S-Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Diazinon - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Dimethoate - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Chlorpyrifos - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Chlorpyrifos Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Malathion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Fenthion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Ethion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

HUNTER WATER


BURWOOD BEACH WWTW



Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 3 n=3 n= 3 n= 3 n= 3 n= 0 n= 3 n= 3 n= 3 n= 3

Fenitrothion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Chlorfenvinphos (E) - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Chlorfenvinphos (Z) - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Parathion (Ethyl) - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Parathion Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Pirimiphos Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Pirimiphos Ethyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Azinphos Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

Azinphos Ethyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 <0.02 <0.02

HUNTER WATER


BURWOOD BEACH WWTW


May - July 2012

Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 3 n=3 n= 3 n= 3 n= 3 n= 0 n= 3 n= 2 n= 2 n= 3


HCB 0.1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Heptachlor 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Heptachlor epoxide 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Aldrin 0.1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

gamma-BHC (Lindane) 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

alpha-BHC 0.01 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

beta-BHC 0.01 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

delta-BHC 0.01 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

trans-Chlordane 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

cis-Chlordane 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Oxychlordane 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Dieldrin 0.1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

pp-DDE 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

pp-DDD 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

pp-DDT 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Endrin - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Endrin Aldehyde - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Endrin Ketone - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

HUNTER WATER


BURWOOD BEACH WWTW


May - July 2012

Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 3 n=3 n= 3 n= 3 n= 3 n= 0 n= 3 n= 2 n= 2 n= 3

alpha-Endosulfan - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

beta-Endosulfan - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Endosulfan Sulfate - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

Methoxychlor - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01

PCB Congeners PCB # 8 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 18 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 28 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 44 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 52 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 66 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 77 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 101 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 105 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 118 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 126 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 128 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 138 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 153 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 169 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

HUNTER WATER


BURWOOD BEACH WWTW


May - July 2012

Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 3 n=3 n= 3 n= 3 n= 3 n= 0 n= 3 n= 2 n= 2 n= 3

PCB # 170 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 180 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 187 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 195 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 206 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

PCB # 209 - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10

Total PCB's < 0.5 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <10


Dichlorvos - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Demeton-S-Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Diazinon - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Dimethoate - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Chlorpyrifos - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Chlorpyrifos Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Malathion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Fenthion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Ethion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Fenitrothion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Chlorfenvinphos (E) - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Chlorfenvinphos (Z) - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

HUNTER WATER


BURWOOD BEACH WWTW


May - July 2012

Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 3 n=3 n= 3 n= 3 n= 3 n= 0 n= 3 n= 2 n= 2 n= 3

Parathion (Ethyl) - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Parathion Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Pirimiphos Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Pirimiphos Ethyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Azinphos Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

Azinphos Ethyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02 <0.02

HUNTER WATER


BURWOOD BEACH WWTW


October - December 2012

Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 2 n=2 n= 3 n= 2 n= 3 n= 0 n= 2 n= 2 n= 0 n= 3


HCB 0.1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

Heptachlor 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

Heptachlor epoxide 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

Aldrin 0.1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

gamma-BHC (Lindane) 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

alpha-BHC 0.01 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

beta-BHC 0.01 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

delta-BHC 0.01 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

trans-Chlordane 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

cis-Chlordane 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

Oxychlordane 0.05 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

Dieldrin 0.1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

pp-DDE 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

pp-DDD 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

pp-DDT 1 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

Endrin - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

Endrin Aldehyde - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

Endrin Ketone - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

alpha-Endosulfan - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

HUNTER WATER


BURWOOD BEACH WWTW



Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 2 n=2 n= 3 n= 2 n= 3 n= 0 n= 2 n= 2 n= 0 n= 3

beta-Endosulfan - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

Endosulfan Sulfate - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

Methoxychlor - 0.01 <0.01 <0.01 <0.01 <0.01 <0.01 n/a <0.01 <0.01 n/a <0.01

PCB Congeners PCB # 8 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 18 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 28 - 0.002 <0.002 0.0016 <0.002 0.0018 0.0015 n/a 0.0013 <0.002 n/a <0.002

PCB # 44 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 52 - 0.002 <0.002 <0.002 0.00155 0.00155 0.00155 n/a 0.0014 <0.002 n/a <0.002

PCB # 66 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 77 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 101 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 105 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 118 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 126 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 128 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 138 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 153 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 169 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 170 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

HUNTER WATER


BURWOOD BEACH WWTW



Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 2 n=2 n= 3 n= 2 n= 3 n= 0 n= 2 n= 2 n= 0 n= 3

PCB # 180 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 187 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 195 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 206 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

PCB # 209 - 0.002 <0.002 <0.002 <0.002 <0.002 <0.002 n/a <0.002 <0.002 n/a <0.002

Total PCB's < 0.5 0.002 <0.002 0.0016 <0.002 0.00285 0.002 n/a 0.0020 <0.002 n/a <0.002


Dichlorvos - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Demeton-S-Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Diazinon - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Dimethoate - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Chlorpyrifos - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Chlorpyrifos Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Malathion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Fenthion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Ethion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Fenitrothion - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Chlorfenvinphos (E) - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Chlorfenvinphos (Z) - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Parathion (Ethyl) - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

HUNTER WATER


BURWOOD BEACH WWTW



Class

Chemical




500 S 2000 N 2000 S TIME ZERO

n= 2 n=2 n= 3 n= 2 n= 3 n= 0 n= 2 n= 2 n= 0 n= 3

Parathion Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Pirimiphos Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Pirimiphos Ethyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Azinphos Methyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

Azinphos Ethyl - 0.02 <0.02 <0.02 <0.02 <0.02 <0.02 n/a <0.02 <0.02 n/a <0.02

HUNTER WATER


BURWOOD BEACH WWTW


Appendix 2 – Metal/Metalloid Concentrations in Oysters

HUNTER WATER


BURWOOD BEACH WWTW


Arsenic Inorganic As Cadmium Chromium Cobalt Copper Iron Lead

NT2_46 NT2_56 NT2_46 NT2_46 NT2_46 NT2_46 NT2_46 NT2_46

Deployment Period Site and Rep mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg

Time Zero (January- April 2012) Time Zero 1 0.76 0.14 <0.05 0.07 18 21 0.02



January- April 2012 Outfall A 1 0.85 0.42 <0.05 0.04 14 9.6 0.02

January- April 2012 Outfall A 2 0.68 0.28 <0.05 0.03 11 9.4 0.02

January- April 2012 Outfall A 3 0.84 0.32 <0.05 0.04 13 10 0.02

January- April 2012 Outfall B 1 0.74 0.28 <0.05 0.04 10 9.1 0.02

January- April 2012 Outfall B 2 0.63 0.31 0.08 0.03 14 9.5 0.02

January- April 2012 Outfall B 3 0.79 0.34 <0.05 0.04 20 13 0.03

January- April 2012 100 N 1 0.83 0.33 0.05 0.04 18 12 0.02

January- April 2012 100 N 2 0.68 0.29 <0.05 0.04 17 14 0.05

January- April 2012 100 N 3 0.63 0.33 <0.05 0.05 23 20 0.04

January- April 2012 100 S 1 0.84 0.37 <0.05 0.05 19 17 0.03

January- April 2012 100 S 2 0.95 0.3 0.06 0.05 18 21 0.03

January- April 2012 100 S 3 0.92 0.34 <0.05 0.06 24 22 0.04

January- April 2012 500 N 1 0.86 0.3 <0.05 0.04 17 18 0.03

January- April 2012 500 N 2 0.67 0.36 <0.05 0.06 24 26 0.03

January- April 2012 500 N 3 0.77 0.34 <0.05 0.06 18 28 0.03

January- April 2012 500 S 1 0.79 0.29 <0.05 0.04 17 15 0.02

January- April 2012 500 S 2 0.92 0.28 <0.05 0.04 13 14 0.03

January- April 2012 500 S 3 1.1 < 0.05 0.3 <0.05 0.05 17 18 0.02

HUNTER WATER


BURWOOD BEACH WWTW





January- April 2012 2000 N 1 1.1 < 0.05 0.27 <0.05 0.05 14 18 0.02

January- April 2012 2000 N 2 1.1 < 0.05 0.3 <0.05 0.05 14 16 0.02

January- April 2012 2000 N 3 0.95 0.24 <0.05 0.05 14 19 0.02

January- April 2012 2000 S 1 1.2 < 0.05 0.3 <0.05 0.04 22 23 0.02

January- April 2012 2001 S 2 0.91 0.28 <0.05 0.04 17 13 0.02

January- April 2012 2002 S 3 0.92 0.26 <0.05 0.04 15 19 0.02

Time Zero (May- July 2012) Time Zero 1 0.71 0.17 <0.05 0.07 20 26 0.03

Time Zero (May- July 2012) Time Zero 2 1.2 <0.05 0.08 <0.05 0.04 8 13 0.02

Time Zero (May- July 2012) Time Zero 3 1.5 <0.05 0.15 0.08 0.08 13 92 0.06

May- July 2012 Outfall A 1 0.73 0.18 <0.05 0.04 16 16 0.04



May- July 2012 Outfall B 1 0.78 0.18 <0.05 0.05 17 15 0.03

May- July 2012 Outfall B 2 0.76 0.19 <0.05 0.05 19 16 0.04

May- July 2012 Outfall B 3 1 0.21 0.05 0.06 21 30 0.06

May- July 2012 100 S 1 1.1 <0.05 0.22 <0.05 0.05 19 20 0.05

May- July 2012 100 S 2 0.75 0.17 <0.05 0.05 17 21 0.04

May- July 2012 100 S 3 0.8 0.17 0.07 0.04 16 19 0.04

May- July 2012 100 N 1 1.2 <0.05 0.27 <0.05 0.07 23 26 0.07

May- July 2012 100 N 2 1.1 <0.05 0.16 <0.05 0.05 16 18 0.04

May- July 2012 100 N 3 1.2 <0.05 0.24 <0.05 0.06 22 20 0.04

HUNTER WATER


BURWOOD BEACH WWTW





May- July 2012 500 N A 1 1.1 <0.05 0.18 0.06 0.04 13 16 0.04

May- July 2012 500 N A 2 1.1 <0.05 0.26 0.05 0.05 22 22 0.04

May- July 2012 500 N A 3 1.2 <0.05 0.21 <0.05 0.05 19 20 0.04

May- July 2012 500 N B 1 0.84 0.22 0.05 0.06 20 18 0.05

May- July 2012 500 N B 2 0.94 0.22 0.05 0.05 21 18 0.04

May- July 2012 500 N B 3 0.73 0.21 <0.05 0.05 21 14 0.03

May- July 2012 500 S 1 1.2 <0.05 0.21 0.06 0.06 20 24 0.06

May- July 2012 500 S 2 1 <0.05 0.19 <0.05 0.05 21 19 0.05

May- July 2012 500 S 3 1.5 <0.05 0.25 <0.05 0.06 20 23 0.05

May- July 2012 2000 N 1 0.93 0.18 0.09 0.04 13 14 0.03

May- July 2012 2000 N 2 1.2 <0.05 0.2 <0.05 0.05 15 17 0.04

May- July 2012 2000 S 1 1.1 <0.05 0.31 <0.05 0.07 26 24 0.06

May- July 2012 2001 S 2 0.82 0.25 <0.05 0.05 19 18 0.04

Time Zero (March- May 2013) Time Zero 1 0.68 0.16 <0.05 0.07 14 35 0.04



March- May 2013 Outfall A 1 1.2 <0.05 0.31 <0.05 0.06 27 20 0.03

March- May 2013 Outfall A 2 1.2 <0.05 0.29 <0.05 0.06 20 18 0.03

March- May 2013 Outfall B 1 0.77 0.24 <0.05 0.04 18 13 0.02

March- May 2013 Outfall B 2 0.73 0.26 <0.05 0.05 19 17 0.03

March- May 2013 100 N 1 1.2 <0.05 0.3 <0.05 0.06 25 19 0.03

HUNTER WATER


BURWOOD BEACH WWTW





March- May 2013 100 N 2 1.3 <0.05 0.41 <0.05 0.07 25 22 0.04

March- May 2013 100 N 3 0.9 0.34 <0.05 0.07 20 22 0.04

March- May 2013 100 S 1 1.2 <0.05 0.28 <0.05 0.06 21 17 0.03

March- May 2013 100 S 2 1.1 <0.05 0.31 <0.05 0.05 20 19 0.04

March- May 2013 500 N A 1 1.2 <0.05 0.28 <0.05 0.06 21 18 0.03

March- May 2013 500 N A 2 1.1 <0.05 0.29 <0.05 0.06 21 15 0.03

March- May 2013 500 S 1 1.2 <0.05 0.34 <0.05 0.06 26 19 0.03

March- May 2013 500 S 2 1 <0.05 0.36 <0.05 0.04 22 12 0.03

March- May 2013 2000 N 1 0.82 0.36 <0.05 0.06 24 14 0.03

March- May 2013 2000 N 2 0.91 0.4 <0.05 0.06 20 13 0.03

HUNTER WATER


BURWOOD BEACH WWTW


Manganese Mercury Nickel Selenium Silver Zinc

NT2_46 NT2_56 NT2_46 NT2_46 NT2_46 NT2_46

Deployment Period Site and Rep mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg

Time Zero (August- September 2012) Time Zero 1 1.8 <0.01 0.12 0.32 0.18 270



August- September 2012 Outfall A 1 0.57 <0.01 0.11 0.35 0.21 300



August- September 2012 Outfall B 1 0.7 <0.01 0.1 0.28 0.18 330



August- September 2012 100 N 1 0.81 0.01 0.11 0.34 0.24 370


August- September 2012 100 N 3 1 0.01 0.24 0.44 0.28 430

August- September 2012 100 S 1 1.1 <0.01 0.15 0.47 0.32 400


August- September 2012 100 S 3 1 <0.01 0.16 0.47 0.31 510





August- September 2012 500 S 2 1.5 0.01 0.12 0.49 0.17 280

HUNTER WATER


BURWOOD BEACH WWTW



NT2_46 NT2_56 NT2_46 NT2_46 NT2_46 NT2_46



August- September 2012 2000 N 1 2.7 <0.01 0.12 0.51 0.18 310






Time Zero (May- July 2012) Time Zero 1 1 0.01 0.08 0.36 0.25 330

Time Zero (May- July 2012) Time Zero 2 3.6 <0.01 0.05 0.42 0.1 130

Time Zero (May- July 2012) Time Zero 3 5.1 <0.01 0.11 0.66 0.13 230

May- July 2012 Outfall A 1 0.51 <0.01 0.09 0.32 0.15 310



May- July 2012 Outfall B 1 0.39 <0.01 0.11 0.33 0.19 310



May- July 2012 100 S 1 0.41 <0.01 0.11 0.43 0.2 370

May- July 2012 100 S 2 0.44 <0.01 0.12 0.33 0.14 290

May- July 2012 100 S 3 0.4 <0.01 0.11 0.36 0.17 290

May- July 2012 100 N 1 0.83 <0.01 0.14 0.51 0.37 400

May- July 2012 100 N 2 0.87 <0.01 0.11 0.41 0.17 300

HUNTER WATER


BURWOOD BEACH WWTW



NT2_46 NT2_56 NT2_46 NT2_46 NT2_46 NT2_46


May- July 2012 100 N 3 0.69 <0.01 0.1 0.46 0.25 390

May- July 2012 500 N A 1 0.84 <0.01 0.09 0.46 0.15 250

May- July 2012 500 N A 2 0.54 <0.01 0.12 0.42 0.2 410

May- July 2012 500 N A 3 0.41 <0.01 0.11 0.49 0.24 330

May- July 2012 500 N B 1 0.83 <0.01 0.18 0.34 0.21 350

May- July 2012 500 N B 2 0.36 <0.01 0.12 0.35 0.26 370

May- July 2012 500 N B 3 0.3 <0.01 0.11 0.33 0.22 340

May- July 2012 500 S 1 0.64 <0.01 0.13 0.46 0.25 330

May- July 2012 500 S 2 0.6 <0.01 0.1 0.42 0.21 390

May- July 2012 500 S 3 1.1 <0.01 0.11 0.58 0.23 410

May- July 2012 2000 N 1 0.52 <0.01 0.11 0.33 0.15 250

May- July 2012 2000 N 2 1 <0.01 0.08 0.44 0.17 280

May- July 2012 2000 S 1 0.41 <0.01 0.13 0.46 0.32 430

May- July 2012 2001 S 2 0.37 <0.01 0.1 0.38 0.22 350

Time Zero (March- May 2013) Time Zero 1 1.7 <0.01 0.1 0.4 0.17 290



March- May 2013 Outfall A 1 1.3 0.01 0.1 0.58 0.21 550

March- May 2013 Outfall A 2 2.2 <0.01 0.09 0.6 0.19 430

March- May 2013 Outfall B 1 0.71 <0.01 0.08 0.42 0.17 390

March- May 2013 Outfall B 2 0.44 <0.01 0.07 0.42 0.18 350

HUNTER WATER


BURWOOD BEACH WWTW



NT2_46 NT2_56 NT2_46 NT2_46 NT2_46 NT2_46


March- May 2013 100 S 1 0.93 0.01 0.09 0.53 0.25 510

March- May 2013 100 S 2 1.2 0.01 0.11 0.67 0.21 540

March- May 2013 100 S 3 0.89 0.01 0.09 0.46 0.19 450

March- May 2013 100 N 1 1.4 0.01 0.1 0.57 0.18 510

March- May 2013 100 N 2 0.94 <0.01 0.08 0.5 0.19 450

March- May 2013 500 N A 1 1.1 <0.01 0.09 0.52 0.2 450

March- May 2013 500 N A 2 1.2 <0.01 0.09 0.58 0.19 480

March- May 2013 500 S 1 0.66 0.01 0.1 0.63 0.26 590

March- May 2013 500 S 2 1.3 0.01 0.06 0.49 0.13 470

March- May 2013 2000 N 1 0.49 <0.01 0.08 0.46 0.23 450

March- May 2013 2000 N 2 0.55 0.01 0.08 0.47 0.2 490

HUNTER WATER


BURWOOD BEACH WWTW


Appendix 3: Power Analysis

HUNTER WATER


BURWOOD BEACH WWTW


Power analyses to determine the suitability of the sample size used in

sampling period 1.

Power Analysis determined that a sample size of 3 (based on 50% effect size using reference

distance) should be sufficient to detect a significant difference in for most metal concentrations among

sites. The exceptions were cobalt, copper and nickel (which had respective estimates of 4, 4 and 12).

Arsenic

0.50.40.30.20.10.0

1.0

0.8

0.6

0.4

0.2

0.0

Maximum Difference

Po

we

r

A lpha 0.05

StDev 0.11

# Lev els 2

A ssumptions

3

Size

Sample

Power Curve for One-way ANOVA

HUNTER WATER


BURWOOD BEACH WWTW


Cadmium

0.140.120.100.080.060.040.020.00

1.0

0.8

0.6

0.4

0.2

0.0

Maximum Difference

Po

we

r

A lpha 0.05

StDev 0.03

# Lev els 2

A ssumptions

3

Size

Sample


Cobalt

0.0250.0200.0150.0100.0050.000

1.0

0.8

0.6

0.4

0.2

0.0

Maximum Difference

Po

we

r

A lpha 0.05

StDev 0.008

# Lev els 2

A ssumptions

4

Size

Sample


HUNTER WATER


BURWOOD BEACH WWTW


Copper

1086420

1.0

0.8

0.6

0.4

0.2

0.0

Maximum Difference

Po

we

r

A lpha 0.05

StDev 2.84

# Lev els 2

A ssumptions

4

Size

Sample


Iron

14121086420

1.0

0.8

0.6

0.4

0.2

0.0

Maximum Difference

Po

we

r

A lpha 0.05

StDev 2.9

# Lev els 2

A ssumptions

3

Size

Sample


HUNTER WATER


BURWOOD BEACH WWTW


Manganese

1.41.21.00.80.60.40.20.0

1.0

0.8

0.6

0.4

0.2

0.0

Maximum Difference

Po

we

r

A lpha 0.05

StDev 0.32

# Lev els 2

A ssumptions

3

Size

Sample


Nickel

0.090.080.070.060.050.040.030.020.010.00

1.0

0.8

0.6

0.4

0.2

0.0

Maximum Difference

Po

we

r

A lpha 0.05

StDev 0.05

# Lev els 2

A ssumptions

12

Size

Sample


HUNTER WATER


BURWOOD BEACH WWTW


Selenium

0.250.200.150.100.050.00

1.0

0.8

0.6

0.4

0.2

0.0

Maximum Difference

Po

we

r

A lpha 0.05

StDev 0.03

# Lev els 2

A ssumptions

2

Size

Sample


Silver

0.100.080.060.040.020.00

1.0

0.8

0.6

0.4

0.2

0.0

Maximum Difference

Po

we

r

A lpha 0.05

StDev 0.02

# Lev els 2

A ssumptions

3

Size

Sample


HUNTER WATER


BURWOOD BEACH WWTW


Zinc

200150100500

1.0

0.8

0.6

0.4

0.2

0.0

Maximum Difference

Po

we

r

A lpha 0.05

StDev 41.58

# Lev els 2

A ssumptions

3

Size

Sample


HUNTER WATER


BURWOOD BEACH WWTW


Appendix 4: NMI QA/QC Reports

Page 1 of 2

QUALITY ASSURANCE REPORT

Client: WORLEYPARSONS SERVICES PTY LTDNMI QA Report No: WORL23/120411 Sample Matrix: Biota

Analyte Method LOR Blank Sample DuplicatesSample Duplicate RPD LCS Matrix Spike

mg/kg mg/kg mg/kg mg/kg % % %Organics Section

OC Pesticides N12/09540 N12/009540HCB NR19 0.01 <0.01 <0.01 <0.01 - - -Heptachlor NR19 0.01 <0.01 <0.01 <0.01 - 124 115Heptachlor epoxide NR19 0.01 <0.01 <0.01 <0.01 - - -Aldrin NR19 0.01 <0.01 <0.01 <0.01 - 78 109gamma-BHC (Lindane) NR19 0.01 <0.01 <0.01 <0.01 - 77 115alpha-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -beta-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -delta-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -trans-Chlordane NR19 0.01 <0.01 <0.01 <0.01 - - -cis-Chlordane NR19 0.01 <0.01 <0.01 <0.01 - - -Oxychlordane NR19 0.01 <0.01 <0.01 <0.01 - - -Dieldrin NR19 0.01 <0.01 <0.01 <0.01 - 103 132pp-DDE NR19 0.01 <0.01 <0.01 <0.01 - 90 130pp-DDD NR19 0.01 <0.01 <0.01 <0.01 - 95 134pp-DDT NR19 0.01 <0.01 <0.01 <0.01 - 76 91Endrin NR19 0.01 <0.01 <0.01 <0.01 - 80 133Endrin Aldehyde NR19 0.01 <0.01 <0.01 <0.01 - - -Endrin Ketone NR19 0.01 <0.01 <0.01 <0.01 - - -alpha-Endosulfan NR19 0.01 <0.01 <0.01 <0.01 - - -beta-Endosulfan NR19 0.01 <0.01 <0.01 <0.01 - - -Endosulfan Sulfate NR19 0.01 <0.01 <0.01 <0.01 - - -Methoxychlor NR19 0.01 <0.01 <0.01 <0.01 - - -Surrogate : DF-DDE NR19 - - 97 78 22 89 131

OP Pesticides N12/009563 N12/009540Dichlorvos NR19 0.01 <0.01 <0.01 <0.01 - - -Demeton-S-Methy NR19 0.01 <0.01 <0.01 <0.01 - - -Diazinon NR19 0.01 <0.01 <0.01 <0.01 - 107 100Dimethoate NR19 0.01 <0.01 <0.01 <0.01 - - -Chlorpyrifos NR19 0.01 <0.01 <0.01 <0.01 - 100 85Chlorpyrifos Methy NR19 0.01 <0.01 <0.01 <0.01 - - -Malathion (Maldison NR19 0.01 <0.01 <0.01 <0.01 - - -Fenthion NR19 0.01 <0.01 <0.01 <0.01 - - -Ethion NR19 0.01 <0.01 <0.01 <0.01 - 70 132Fenitrothion NR19 0.01 <0.01 <0.01 <0.01 - - -Chlorfenvinphos (E) NR19 0.01 <0.01 <0.01 <0.01 - - -Chlorfenvinphos (Z) NR19 0.01 <0.01 <0.01 <0.01 - - -Parathion (Ethyl) NR19 0.01 <0.01 <0.01 <0.01 - 87 65Parathion Methy NR19 0.01 <0.01 <0.01 <0.01 - - -Pirimiphos Ethy NR19 0.01 <0.01 <0.01 <0.01 - - -Pirimiphos Methy NR19 0.01 <0.01 <0.01 <0.01 - - -Azinphos Methy NR19 0.01 <0.01 <0.01 <0.01 - - -Azinphos Ethyl NR19 0.01 <0.01 <0.01 <0.01 - - -Surrogate : TPP - - 127 137 7.6 93 84

Results expressed in percentage (%) or mg/kg wherever appropriateAcceptable Spike recovery is 50-150%Acceptable RPDs on spikes and duplicates is 40%RPD= Relative Percentage DifferenceThis report shall not be reproduced except in ful

Signed:Danny SleeOrganics Manager, NMI-Pymble

Date: 1/05/2012

Recoveries

Australian GovernmentNational Measurement Institute

1 Suakin Street, Pymble NSW 2073 Tel: +61 2 9449 0111 Fax: +61 2 9449 1653 www.measurement.gov.au

National Measurement Institute

Page 2 of 2




ug/kg ug/kg ug/kg ug/kg % % %Organics SectionPCB Congeners N12/009540 N12/009540

#8 NR_19 10 <10 <10 <10 - - -#18 NR_19 10 <10 <10 <10 - - -#28 NR_19 10 <10 <10 <10 - - -#44 NR_19 10 <10 <10 <10 - - -#52 NR_19 10 <10 <10 <10 - 66 139#66 NR_19 10 <10 <10 <10 - - -#77 NR_19 10 <10 <10 <10 - - -#101 NR_19 10 <10 <10 <10 - - -#105 NR_19 10 <10 <10 <10 - - -#118 NR_19 10 <10 <10 <10 - 78 125#126 NR_19 10 <10 <10 <10 - - -#128 NR_19 10 <10 <10 <10 - - -#138 NR_19 10 <10 <10 <10 - 81 135#153 NR_19 10 <10 <10 <10 - - -#169 NR_19 10 <10 <10 <10 - - -#170 NR_19 10 <10 <10 <10 - - -#180 NR_19 10 <10 <10 <10 - 82 134#187 NR_19 10 <10 <10 <10 - - -#195 NR_19 10 <10 <10 <10 - - -#206 NR_19 10 <10 <10 <10 - - -#209 NR_19 10 <10 <10 <10 - - -

Results expressed in percentage (%) or ug/kg wherever appropriateAcceptable Spike recovery is 50-150% (PCB)Acceptable RPDs on spikes and duplicates is 40%RPD= Relative Percentage Difference.This report shall not be reproduced except in full


Date: 1/05/2012

Recoveries




Page 1 of 1

Client: WORLEYPARSONS SERVICES PTY LTD NMI QA Report No: WORL23/120926T1 Sample Matrix: OYSTER

Analyte Method LOR Blank DuplicatesSample Duplicate RPD LCS Matrix Spike

mg/kg mg/kg mg/kg mg/kg % %Inorganics Section N12/025858 N12/025858

Arsenic NT2.46 0.05 <0.05 1.5 1.5 0 105 100Cadmium NT2.46 0.01 <0.01 0.16 0.15 6.5 99 100Chromium NT2.46 0.05 <0.05 0.07 0.09 24 106 97Cobalt NT2.46 0.01 <0.01 0.08 0.08 0 96 100Copper NT2.46 0.01 <0.01 13 14 7.4 102 98Iron NT2.46 0.5 <0.5 88 96 8.7 103 101Lead NT2.46 0.01 <0.01 0.06 0.06 0 90 100Manganese NT2.46 0.01 <0.01 5.1 5.1 0 101 96Mercury NT2.46 0.01 <0.01 <0.01 <0.01 ND 102 110Nickel NT2.46 0.01 <0.01 0.12 0.1 18 78 96Selenium NT2.46 0.05 <0.05 0.66 0.66 0 108 95Silver NT2.46 0.02 <0.02 0.12 0.14 15 109 99Zinc NT2.46 0.01 <0.01 210 240 13 106 86 Filename = K:\Inorganics\Quality System\QA Reports\TE\QAR2012\Food and Misc\Legend:Acceptable recovery is 75-120%.Acceptable RPDs on duplicates is 44% at concentrations >5 times LOR. Greater RPD may be expected at <5 times LOR.LOR = Limit Of Reporting ND = Not DeterminedRPD = Relative Percent Difference NA = Not ApplicableLCS = Laboratory Control Sample.#: Spike level is less than 50% of the sample's concentration, hence the recovery data is not reliable.**: reference value not available

Comments:Results greater than ten times LOR have been rounded to two significant figures.This report shall not be reproduced except in full.

Signed:

Dr Michael WuInorganics Manager, NMI-North Ryde

Date: 5/10/2012


Recoveries


105 Delhi Road, North Ryde, 2113. Tel: +61 2 9449 0111 Fax: +61 2 9449 1653 www.measurement.gov.au


Page 1 of 1


Client: WORLEY PARSONS SERVICES PTY LTDNMI QA Report No: WORL23/120926 Sample Matrix: Biota



OC PesticidesHCB NR19 0.01 <0.01 NA NA NA - NAHeptachlor NR19 0.01 <0.01 NA NA NA 112 NAHeptachlor epoxide NR19 0.01 <0.01 NA NA NA - NAAldrin NR19 0.01 <0.01 NA NA NA 125 NAgamma-BHC (Lindane) NR19 0.01 <0.01 NA NA NA 101 NAalpha-BHC NR19 0.01 <0.01 NA NA NA - NAbeta-BHC NR19 0.01 <0.01 NA NA NA - NAdelta-BHC NR19 0.01 <0.01 NA NA NA - NAtrans-Chlordane NR19 0.01 <0.01 NA NA NA - NAcis-Chlordane NR19 0.01 <0.01 NA NA NA - NAOxychlordane NR19 0.01 <0.01 NA NA NA - NADieldrin NR19 0.01 <0.01 NA NA NA 127 NApp-DDE NR19 0.01 <0.01 NA NA NA 120 NApp-DDD NR19 0.01 <0.01 NA NA NA 118 NApp-DDT NR19 0.01 <0.01 NA NA NA 111 NAEndrin NR19 0.01 <0.01 NA NA NA 117 NAEndrin Aldehyde NR19 0.01 <0.01 NA NA NA - NAEndrin Ketone NR19 0.01 <0.01 NA NA NA - NAalpha-Endosulfan NR19 0.01 <0.01 NA NA NA - NAbeta-Endosulfan NR19 0.01 <0.01 NA NA NA - NAEndosulfan Sulfate NR19 0.01 <0.01 NA NA NA - NAMethoxychlor NR19 0.01 <0.01 NA NA NA - NASurrogate : DF-DDE NR19 - - NA NA NA 89 NA

OP PesticidesDichlorvos NR19 0.01 <0.01 NA NA NA - NADemeton-S-Methyl NR19 0.01 <0.01 NA NA NA - NADiazinon NR19 0.01 <0.01 NA NA NA 102 NADimethoate NR19 0.01 <0.01 NA NA NA - NAChlorpyrifos NR19 0.01 <0.01 NA NA NA 100 NAChlorpyrifos Methyl NR19 0.01 <0.01 NA NA NA - NAMalathion (Maldison) NR19 0.01 <0.01 NA NA NA - NAFenthion NR19 0.01 <0.01 NA NA NA - NAEthion NR19 0.01 <0.01 NA NA NA 112 NAFenitrothion NR19 0.01 <0.01 NA NA NA - NAChlorfenvinphos (E) NR19 0.01 <0.01 NA NA NA - NAChlorfenvinphos (Z) NR19 0.01 <0.01 NA NA NA - NAParathion (Ethyl) NR19 0.01 <0.01 NA NA NA 101 NAParathion Methyl NR19 0.01 <0.01 NA NA NA - NAPirimiphos Ethyl NR19 0.01 <0.01 NA NA NA - NAPirimiphos Methyl NR19 0.01 <0.01 NA NA NA - NAAzinphos Methyl NR19 0.01 <0.01 NA NA NA - NAAzinphos Ethyl NR19 0.01 <0.01 NA NA NA - NASurrogate : TPP - - NA NA NA 102 NA

Results expressed in percentage (%) or mg/kg wherever appropriate.Acceptable Spike recovery is 50-150%Acceptable RPDs on spikes and duplicates is 40%.RPD= Relative Percentage Difference.This report shall not be reproduced except in full.

Signed:Danny SleeOrganics Manager, NMI-North Ryde

Date: 10/10/2012

Recoveries


105 Delhi Road, North Ryde NSW 2113 Tel: +61 2 9449 0111 www.measurement.gov.au


Page 1 of 4





OC Pesticides N12/019846 N12/019846HCB NR19 0.01 <0.01 <0.01 <0.01 - - -Heptachlor NR19 0.01 <0.01 <0.01 <0.01 - 124 126Heptachlor epoxide NR19 0.01 <0.01 <0.01 <0.01 - - -Aldrin NR19 0.01 <0.01 <0.01 <0.01 - 98 101gamma-BHC (Lindane) NR19 0.01 <0.01 <0.01 <0.01 - 119 119alpha-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -beta-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -delta-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -trans-Chlordane NR19 0.01 <0.01 <0.01 <0.01 - - -cis-Chlordane NR19 0.01 <0.01 <0.01 <0.01 - - -Oxychlordane NR19 0.01 <0.01 <0.01 <0.01 - - -Dieldrin NR19 0.01 <0.01 <0.01 <0.01 - 121 121pp-DDE NR19 0.01 <0.01 <0.01 <0.01 - 105 104pp-DDD NR19 0.01 <0.01 <0.01 <0.01 - 123 112pp-DDT NR19 0.01 <0.01 <0.01 <0.01 - 124 134Endrin NR19 0.01 <0.01 <0.01 <0.01 - 120 116Endrin Aldehyde NR19 0.01 <0.01 <0.01 <0.01 - - -Endrin Ketone NR19 0.01 <0.01 <0.01 <0.01 - - -alpha-Endosulfan NR19 0.01 <0.01 <0.01 <0.01 - - -beta-Endosulfan NR19 0.01 <0.01 <0.01 <0.01 - - -Endosulfan Sulfate NR19 0.01 <0.01 <0.01 <0.01 - - -Methoxychlor NR19 0.01 <0.01 <0.01 <0.01 - - -Surrogate : DF-DDE NR19 - - 97 97 0.0 96 96

OP Pesticides N12/019846 N12/019846Dichlorvos NR19 0.01 <0.01 <0.02 <0.02 - - -Demeton-S-Methyl NR19 0.01 <0.01 <0.02 <0.02 - - -Diazinon NR19 0.01 <0.01 <0.02 <0.02 - 120 129Dimethoate NR19 0.01 <0.01 <0.02 <0.02 - - -Chlorpyrifos NR19 0.01 <0.01 <0.02 <0.02 - 111 120Chlorpyrifos Methyl NR19 0.01 <0.01 <0.02 <0.02 - - -Malathion (Maldison) NR19 0.01 <0.01 <0.02 <0.02 - - -Fenthion NR19 0.01 <0.01 <0.02 <0.02 - - -Ethion NR19 0.01 <0.01 <0.02 <0.02 - 125 129Fenitrothion NR19 0.01 <0.01 <0.02 <0.02 - - -Chlorfenvinphos (E) NR19 0.01 <0.01 <0.02 <0.02 - - -Chlorfenvinphos (Z) NR19 0.01 <0.01 <0.02 <0.02 - - -Parathion (Ethyl) NR19 0.01 <0.01 <0.02 <0.02 - 126 146Parathion Methyl NR19 0.01 <0.01 <0.02 <0.02 - - -Pirimiphos Ethyl NR19 0.01 <0.01 <0.02 <0.02 - - -Pirimiphos Methyl NR19 0.01 <0.01 <0.02 <0.02 - - -Azinphos Methyl NR19 0.01 <0.01 <0.02 <0.02 - - -Azinphos Ethyl NR19 0.01 <0.01 <0.02 <0.02 - - -Surrogate : TPP - - 62 56 10 56 100



Date: 16/08/2012

Recoveries




Page 2 of 4








Date: 16/08/2012

Recoveries




Page 3 of 4





OC Pesticides N12/019864 N12/019865HCB NR19 0.01 <0.01 <0.01 <0.01 - - -Heptachlor NR19 0.01 <0.01 <0.01 <0.01 - 124 118Heptachlor epoxide NR19 0.01 <0.01 <0.01 <0.01 - - -Aldrin NR19 0.01 <0.01 <0.01 <0.01 - 98 96gamma-BHC (Lindane) NR19 0.01 <0.01 <0.01 <0.01 - 119 106alpha-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -beta-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -delta-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -trans-Chlordane NR19 0.01 <0.01 <0.01 <0.01 - - -cis-Chlordane NR19 0.01 <0.01 <0.01 <0.01 - - -Oxychlordane NR19 0.01 <0.01 <0.01 <0.01 - - -Dieldrin NR19 0.01 <0.01 <0.01 <0.01 - 121 111pp-DDE NR19 0.01 <0.01 <0.01 <0.01 - 105 100pp-DDD NR19 0.01 <0.01 <0.01 <0.01 - 123 104pp-DDT NR19 0.01 <0.01 <0.01 <0.01 - 124 123Endrin NR19 0.01 <0.01 <0.01 <0.01 - 120 118Endrin Aldehyde NR19 0.01 <0.01 <0.01 <0.01 - - -Endrin Ketone NR19 0.01 <0.01 <0.01 <0.01 - - -alpha-Endosulfan NR19 0.01 <0.01 <0.01 <0.01 - - -beta-Endosulfan NR19 0.01 <0.01 <0.01 <0.01 - - -Endosulfan Sulfate NR19 0.01 <0.01 <0.01 <0.01 - - -Methoxychlor NR19 0.01 <0.01 <0.01 <0.01 - - -Surrogate : DF-DDE NR19 - - 99 122 21 96 89

OP Pesticides N12/019864 N12/019865Dichlorvos NR19 0.01 <0.01 <0.02 <0.02 - - -Demeton-S-Methyl NR19 0.01 <0.01 <0.02 <0.02 - - -Diazinon NR19 0.01 <0.01 <0.02 <0.02 - 120 119Dimethoate NR19 0.01 <0.01 <0.02 <0.02 - - -Chlorpyrifos NR19 0.01 <0.01 <0.02 <0.02 - 111 112Chlorpyrifos Methyl NR19 0.01 <0.01 <0.02 <0.02 - - -Malathion (Maldison) NR19 0.01 <0.01 <0.02 <0.02 - - -Fenthion NR19 0.01 <0.01 <0.02 <0.02 - - -Ethion NR19 0.01 <0.01 <0.02 <0.02 - 125 123Fenitrothion NR19 0.01 <0.01 <0.02 <0.02 - - -Chlorfenvinphos (E) NR19 0.01 <0.01 <0.02 <0.02 - - -Chlorfenvinphos (Z) NR19 0.01 <0.01 <0.02 <0.02 - - -Parathion (Ethyl) NR19 0.01 <0.01 <0.02 <0.02 - 126 135Parathion Methyl NR19 0.01 <0.01 <0.02 <0.02 - - -Pirimiphos Ethyl NR19 0.01 <0.01 <0.02 <0.02 - - -Pirimiphos Methyl NR19 0.01 <0.01 <0.02 <0.02 - - -Azinphos Methyl NR19 0.01 <0.01 <0.02 <0.02 - - -Azinphos Ethyl NR19 0.01 <0.01 <0.02 <0.02 - - -Surrogate : TPP - - 68 54 23 56 92



Date: 16/08/2012

Recoveries




Page 4 of 4








Date: 16/08/2012

Recoveries




Page 1 of 1

Client: WORLEYPARSONS SERVICES PTY LTD NMI QA Report No: WORL23/120926T2 Sample Matrix: OYSTER



Arsenic inorganic NT2.56 0.05 <0.05 <0.05 <0.05 ND 98 100 Filename = K:\Inorganics\Quality System\QA Reports\TE\QAR2012\Food and Misc\Legend:Acceptable recovery is 75-120%.Acceptable RPDs on duplicates is 44% at concentrations >5 times LOR. Greater RPD may be expected at <5 times LOR.LOR = Limit Of Reporting ND = Not DeterminedRPD = Relative Percent Difference NA = Not ApplicableLCS = Laboratory Control Sample.#: Spike level is less than 50% of the sample's concentration, hence the recovery data is not reliable.**: reference value not available


Signed:


Date: 12/10/2012


Recoveries




Page 1 of 1

Client: WORLEYPARSONS SERVICES PTY LTD NMI QA Report No: WORL23/130531T1 Sample Matrix: SEAFOOD

WORL23/130531/1T1



Arsenic NT2.46 0.05 <0.05 1.7 1.7 0 97 99Arsenic Inorganic NT2.56 0.05 <0.05 <0.05 <0.05 ND 106 86Cadmium NT2.46 0.01 <0.01 0.35 0.37 5.6 96 100Chromium NT2.46 0.05 <0.05 <0.05 <0.05 ND 100 98Cobalt NT2.46 0.01 <0.01 0.06 0.06 5.1 96 100Copper NT2.46 0.01 <0.01 17 18 5.7 101 100Iron NT2.46 0.5 <0.5 26 26 0 89 96Lead NT2.46 0.01 <0.01 0.04 0.04 0 87 100Manganese NT2.46 0.01 <0.01 4.0 4.1 2.5 102 100Mercury NT2.46 0.01 <0.01 <0.01 <0.01 ND 101 99Nickel NT2.46 0.01 <0.01 0.10 0.10 0 92 100Selenium NT2.46 0.05 <0.05 0.68 0.74 8.5 101 100Silver NT2.46 0.02 <0.02 0.2 0.21 4.9 98 100Zinc NT2.46 0.01 <0.01 330 330 0 105 92

Filename = K:\Inorganics\Quality System\QA Reports\TE\QAR2013\Food & Misc\Legend:Acceptable recovery is 75-120%.Acceptable RPDs on duplicates is 44% at concentrations >5 times LOR. Greater RPD may be expected at <5 times LOR.LOR = Limit Of Reporting ND = Not DeterminedRPD = Relative Percent Difference NA = Not ApplicableLCS = Laboratory Control Sample.#: Spike level is less than 50% of the sample's concentration, hence the recovery data is not reliable.**: reference value not available


Signed:


Date: 20/06/2013


Recoveries




Page 1 of 4


Client: WORLEYPARSONS SERVICES PTY LTD

NMI QA Report No: WORL23/130531 Sample Matrix: Biota

Analyte Method LOR Blank Sample Duplicates

Sample Duplicate RPD LCS Matrix Spike

mg/kg mg/kg mg/kg mg/kg % % %

Organics SectionOC Pesticides N13/014608 N13/014608

HCB NR19 0.01 <0.01 <0.01 <0.01 - - -

Heptachlor NR19 0.01 <0.01 <0.01 <0.01 - 91 103

Heptachlor epoxide NR19 0.01 <0.01 <0.01 <0.01 - - -

Aldrin NR19 0.01 <0.01 <0.01 <0.01 - 91 98

gamma-BHC (Lindane) NR19 0.01 <0.01 <0.01 <0.01 - 78 103

alpha-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -

beta-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -

delta-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -

trans-Chlordane NR19 0.01 <0.01 <0.01 <0.01 - - -

cis-Chlordane NR19 0.01 <0.01 <0.01 <0.01 - - -

Oxychlordane NR19 0.01 <0.01 <0.01 <0.01 - - -

Dieldrin NR19 0.01 <0.01 <0.01 <0.01 - 93 113

pp-DDE NR19 0.01 <0.01 <0.01 <0.01 - 83 79

pp-DDD NR19 0.01 <0.01 <0.01 <0.01 - 97 92

pp-DDT NR19 0.01 <0.01 <0.01 <0.01 - 74 62

Endrin NR19 0.01 <0.01 <0.01 <0.01 - 94 110

Endrin Aldehyde NR19 0.01 <0.01 <0.01 <0.01 - - -

Endrin Ketone NR19 0.01 <0.01 <0.01 <0.01 - - -

alpha-Endosulfan NR19 0.01 <0.01 <0.01 <0.01 - - -

beta-Endosulfan NR19 0.01 <0.01 <0.01 <0.01 - - -

Endosulfan Sulfate NR19 0.01 <0.01 <0.01 <0.01 - - -

Methoxychlor NR19 0.01 <0.01 <0.01 <0.01 - - -

Surrogate : DF-DDE NR19 - - 73 72 1.4 63 70

OP Pesticides N13/014608 N13/014608

Dichlorvos NR19 0.01 <0.01 <0.01 <0.01 - - -

Demeton-S-Methyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Diazinon NR19 0.01 <0.01 <0.01 <0.01 - 106 127

Dimethoate NR19 0.01 <0.01 <0.01 <0.01 - - -

Chlorpyrifos NR19 0.01 <0.01 <0.01 <0.01 - 117 124

Chlorpyrifos Methyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Malathion (Maldison) NR19 0.01 <0.01 <0.01 <0.01 - - -

Fenthion NR19 0.01 <0.01 <0.01 <0.01 - - -

Ethion NR19 0.01 <0.01 <0.01 <0.01 - 110 100

Fenitrothion NR19 0.01 <0.01 <0.01 <0.01 - - -

Chlorfenvinphos (E) NR19 0.01 <0.01 <0.01 <0.01 - - -

Chlorfenvinphos (Z) NR19 0.01 <0.01 <0.01 <0.01 - - -

Parathion (Ethyl) NR19 0.01 <0.01 <0.01 <0.01 - 116 112

Parathion Methyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Pirimiphos Ethyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Pirimiphos Methyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Azinphos Methyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Azinphos Ethyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Surrogate : TPP - - 95 96 1.0 57 89

Results expressed in percentage (%) or mg/kg wherever appropriate.Acceptable Spike recovery is 50-150%Acceptable RPDs on spikes and duplicates is 40%.

RPD= Relative Percentage Difference.

This report shall not be reproduced except in full.


Date: 26/06/2013

Recoveries

Australian Government




Page 2 of 4






mg/kg mg/kg mg/kg mg/kg % % %

Organics SectionOC Pesticides N13/014618 N13/014618

HCB NR19 0.01 <0.01 <0.01 <0.01 - - -

Heptachlor NR19 0.01 <0.01 <0.01 <0.01 - 91 100

Heptachlor epoxide NR19 0.01 <0.01 <0.01 <0.01 - - -

Aldrin NR19 0.01 <0.01 <0.01 <0.01 - 91 92

gamma-BHC (Lindane) NR19 0.01 <0.01 <0.01 <0.01 - 78 103

alpha-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -

beta-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -

delta-BHC NR19 0.01 <0.01 <0.01 <0.01 - - -

trans-Chlordane NR19 0.01 <0.01 <0.01 <0.01 - - -

cis-Chlordane NR19 0.01 <0.01 <0.01 <0.01 - - -

Oxychlordane NR19 0.01 <0.01 <0.01 <0.01 - - -

Dieldrin NR19 0.01 <0.01 <0.01 <0.01 - 93 112

pp-DDE NR19 0.01 <0.01 <0.01 <0.01 - 83 80

pp-DDD NR19 0.01 <0.01 <0.01 <0.01 - 97 92

pp-DDT NR19 0.01 <0.01 <0.01 <0.01 - 74 62

Endrin NR19 0.01 <0.01 <0.01 <0.01 - 94 114

Endrin Aldehyde NR19 0.01 <0.01 <0.01 <0.01 - - -

Endrin Ketone NR19 0.01 <0.01 <0.01 <0.01 - - -

alpha-Endosulfan NR19 0.01 <0.01 <0.01 <0.01 - - -

beta-Endosulfan NR19 0.01 <0.01 <0.01 <0.01 - - -

Endosulfan Sulfate NR19 0.01 <0.01 <0.01 <0.01 - - -

Methoxychlor NR19 0.01 <0.01 <0.01 <0.01 - - -

Surrogate : DF-DDE NR19 - - 78 69 12 63 70

OP Pesticides N13/014618 N13/014618

Dichlorvos NR19 0.01 <0.01 <0.01 <0.01 - - -

Demeton-S-Methyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Diazinon NR19 0.01 <0.01 <0.01 <0.01 - 106 127

Dimethoate NR19 0.01 <0.01 <0.01 <0.01 - - -

Chlorpyrifos NR19 0.01 <0.01 <0.01 <0.01 - 117 119

Chlorpyrifos Methyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Malathion (Maldison) NR19 0.01 <0.01 <0.01 <0.01 - - -

Fenthion NR19 0.01 <0.01 <0.01 <0.01 - - -

Ethion NR19 0.01 <0.01 <0.01 <0.01 - 110 82

Fenitrothion NR19 0.01 <0.01 <0.01 <0.01 - - -

Chlorfenvinphos (E) NR19 0.01 <0.01 <0.01 <0.01 - - -

Chlorfenvinphos (Z) NR19 0.01 <0.01 <0.01 <0.01 - - -

Parathion (Ethyl) NR19 0.01 <0.01 <0.01 <0.01 - 116 115

Parathion Methyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Pirimiphos Ethyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Pirimiphos Methyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Azinphos Methyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Azinphos Ethyl NR19 0.01 <0.01 <0.01 <0.01 - - -

Surrogate : TPP - - 84 82 2.4 57 86

Results expressed in percentage (%) or mg/kg wherever appropriate.Acceptable Spike recovery is 50-150%Acceptable RPDs on spikes and duplicates is 40%.


This report shall not be reproduced except in full.


Date: 26/06/2013

Recoveries





Page 3 of 4






ug/kg ug/kg ug/kg ug/kg % % %

Organics Section

PCB Congeners N13/014608 N13/014608

#8 NR_19 2 <2 <2 <2 - - -

#18 NR_19 2 <2 <2 <2 - - -

#28 NR_19 2 <2 <2 <2 - - -

#44 NR_19 2 <2 <2 <2 - - -

#52 NR_19 2 <2 <2 <2 - 89 117

#66 NR_19 2 <2 <2 <2 - - -

#77 NR_19 2 <2 <2 <2 - - -

#101 NR_19 2 <2 <2 <2 - - -

#105 NR_19 2 <2 <2 <2 - - -

#118 NR_19 2 <2 <2 <2 - 87 124

#126 NR_19 2 <2 <2 <2 - - -

#128 NR_19 2 <2 <2 <2 - - -

#138 NR_19 2 <2 <2 <2 - 99 115

#153 NR_19 2 <2 <2 <2 - - -

#169 NR_19 2 <2 <2 <2 - - -

#170 NR_19 2 <2 <2 <2 - - -

#180 NR_19 2 <2 <2 <2 - 93 115

#187 NR_19 2 <2 <2 <2 - - -

#195 NR_19 2 <2 <2 <2 - - -

#206 NR_19 2 <2 <2 <2 - - -

#209 NR_19 2 <2 <2 <2 - - -

Results expressed in percentage (%) or ug/kg wherever appropriate.

Acceptable Spike recovery is 50-150% (PCB)

Acceptable RPDs on spikes and duplicates is 40%.


This report shall not be reproduced except in full.Signed:

Danny SleeDate: Organics Manager, NMI-North Ryde

26/06/2013

Recoveries





Page 4 of 4






ug/kg ug/kg ug/kg ug/kg % % %

Organics Section

PCB Congeners N13/014618 N13/014618

#8 NR_19 2 <2 <2 <2 - - -

#18 NR_19 2 <2 <2 <2 - - -

#28 NR_19 2 <2 <2 <2 - - -

#44 NR_19 2 <2 <2 <2 - - -

#52 NR_19 2 <2 <2 <2 - 89 94

#66 NR_19 2 <2 <2 <2 - - -

#77 NR_19 2 <2 <2 <2 - - -

#101 NR_19 2 <2 <2 <2 - - -

#105 NR_19 2 <2 <2 <2 - - -

#118 NR_19 2 <2 <2 <2 - 87 94

#126 NR_19 2 <2 <2 <2 - - -

#128 NR_19 2 <2 <2 <2 - - -

#138 NR_19 2 <2 <2 <2 - 99 117

#153 NR_19 2 <2 <2 <2 - - -

#169 NR_19 2 <2 <2 <2 - - -

#170 NR_19 2 <2 <2 <2 - - -

#180 NR_19 2 <2 <2 <2 - 93 107

#187 NR_19 2 <2 <2 <2 - - -

#195 NR_19 2 <2 <2 <2 - - -

#206 NR_19 2 <2 <2 <2 - - -

#209 NR_19 2 <2 <2 <2 - - -

Results expressed in percentage (%) or ug/kg wherever appropriate.

Acceptable Spike recovery is 50-150% (PCB)

Acceptable RPDs on spikes and duplicates is 40%.


This report shall not be reproduced except in full.Signed:

Danny SleeDate: Organics Manager, NMI-North Ryde

26/06/2013

Recoveries





Documents

Oyster Biomonitoring Study€¦ · oysters each were deployed at eight locations with increasing distance from the Burwood Beach outfall. Oysters were deployed in approximate north-eastern