Scottish Marine and Freshwater Science Volume 4 Number 4: Measurement of Contaminants and their Effects in Environmental Samples - Proposal for the Revision of the Sampling Programme

EC Directives

Water Framework Directive ( WFD)

The Water Framework Directive (2000/60/ EC) was adopted by the European Parliament and the Council of the European Union on 22 December 2000. The Directive became law in Scotland during 2003 through the Water Environment and Water Services (Scotland) Act 2003 (the WEWS Act) which sets out the new arrangements for the protection of the water environment in Scotland. The monitoring programme became operational in Scotland in 2007. The aim of the WFD is to protect all transitional ^[2] and coastal waters ^[3] , prevent deterioration and enhance the status of aquatic ecosystems. Three groups of priority substances ( Appendix 1) were identified by the WFD: Priority Hazardous Substances ( PHS), Priority Substances ( PS) and Priority Substances subject to review to Priority Hazardous Substances ( PSR). In addition, Member States are required to list, control and monitor Specific Pollutants that are of concern to the river basin. Originally thirty-three substances or groups of substances were classed as PHS and PS (13 PHSs and 20 PSs), including metals, herbicides, insecticides, fungicides, biocides, volatile organic compounds, alkylphenols, PAHs and phthalates. The European Commission is required to review the list of PSs every four years and identify, where appropriate, new PSs or PHSs and any need to revise the Environmental Quality Standards ( EQSs) or the status of existing PSs (2000/60/ EC, Article 16). The outcome of the first review was reported in 2011 (( COM(2011)876)). As a result of this review the following additional substances were proposed as priority substances: aclonifen, bifenox, cybutryne, cypermethrin, dichlorvos, terbutryn, 17-alpha-ethinylestradiol, 17-beta-estradiol and diclofenac, and the following as priority hazardous substances: dicofol, PFOS and its derivatives, quinoxyfen, dioxins and dioxin-like compounds, HBCDD and heptachlor/heptachlor epoxide. More recently, the new Priority Substances Directive (2013/39/ EU) was published with the revised list of PSs/ PHSs, together with their EQS values for freshwater, saltwater and, where applicable, biota; the pharmaceutical compounds (17-alpha-ethinylestradiol, 17-beta-estradiol and diclofenac) have not been included as PSs, and are likely to be included within the next review of PSs to be completed by 2017. Biological effects measurements are not required for WFD although their role in providing information on substances not known a priori to be present in the environment, and on the effects of mixtures of substances has been recognised by the EU. The Scottish River Basin District Directions sets out the environmental standards to be applied by SEPA ⁷.

The objective for PHSs is for the cessation of discharges, emissions and losses within 20 years of appropriate measures being introduced, with the "ultimate aim of achieving concentrations in the marine environment near background values for naturally occurring substances and close to zero for man-made synthetic substances" ( WFD, Article 1); close-to-background/zero concentrations are required in order to achieve "High Ecological Status" ( WFD Annex V, 1.2). For PS the objective is for progressive reduction of discharges, emissions and losses. The WFD requires that the chemical status of water bodies to be categorised as 'Good' or 'Failing to achieve good'. Chemical status applies to territorial waters (out to 12 miles), whilst ecological status applies to coastal waters (out to 3 nautical miles for Scotland). The over-all objective of the WFD is to achieve a good chemical status, in addition to a good ecological status, by 2015.

The majority of monitoring for WFD is undertaken in Scotland by SEPA, with contaminant monitoring occurring in transitional and coastal waters at surveillance sites and operational sites. Operational sites are related to identified pressures whereas surveillance sites are not related to pressures and contaminant concentrations found at these sites should be representative of background concentrations. Concentrations of trace organic substances monitored in water samples from transitional and coastal waters in Scotland are well below EQS and frequently below the limit of detection of the analyses; consequently monitoring in water was stopped when there was sufficient evidence to show there was no risk of failing EQS. SEPA has continued monitoring trace metals in waters at selected sites where concentrations are greater than the limit of detection of the analyses.

Dangerous Substances Directive ( DSD) and the EC Shellfish Waters Directive ( SWD)

SEPA undertakes the analysis of organic compounds and trace metals to meet the requirements of the EC Dangerous Substances Directive ( DSD) (76/464/ EEC) and the EC Shellfish Waters Directive ( SWD) (79/923/ EEC, updated in 2006 (2006/113/ EC). The DSD requires environmental monitoring wherever there is a consented discharge of a specified dangerous substance to determine if EQSs are exceeded and the analysis of either sediment or biota annually to determine if the substances are accumulating. The Directive lists 11 metals and 52 organic compounds, however, only those with known discharges are monitored. Concentrations are compared to EQSs set by the DSD. EQSs are concentrations below which a substance is not believed to be detrimental to aquatic life and were derived using acute toxicity testing and have only been established for water. The WFD will take over the provisions of the DSD, which will be repealed 22 December 2013 (13 years after the date of entry into force of the WFD).

The SWD requires the analysis of water and shellfish biannually. The Directive states that the concentration of the substance in the water column or shellfish flesh should not harm the shellfish. The Directive prescribes the minimum quality criteria which must be met by shellfish waters, and guideline values which Member States must endeavour to observe. Analysis for SWD is undertaken by SEPA for Scotland, concentrations in shellfish and water samples are compared to EC Directive Quality Standards for Shellfish Growing Waters. The SWD allows reduced frequency or no monitoring where there is no risk of failing the Directive standards. In practice the concentration of hazardous substances in water samples from shellfish waters are low (below SWD quality standard) and frequently below the limit of detection of the analysis so monitoring in water samples was discontinued. However, contaminants (trace metals, PCBs and PAHs) continue to be monitored in shellfish flesh by SEPA. Only metals and organochlorine pesticide data are required by the SWD, but PCBs and PAHs are also measured by SEPA to provide data for the OSPAR CEMP. The SWD will be repealed in 22 December 2013, after which monitoring for contaminants in shellfish flesh will be covered by the existing WFD requirements.

Contaminant and Biological Effects Monitoring Programmes Currently Undertaken in Scotland

Biota

Biota contaminant monitoring programmes currently undertaken in Scotland are described in the Marine Scotland Science Descriptor 9 (contaminants in food) report ⁸. SEPA monitor contaminants (trace metals, PCBs and PAHs) in native mussels collected from the intertidal zone from 56 sites across Scotland between January and March each year ⁸ (Fig. 1). SEPA also measure PBDEs in mussels from the Firth of Clyde and Firth of Forth. Samples are collected to assess compliance with the standstill clause in the DSD, to assess compliance with the SWD, and for OSPAR CEMP, so as to assess progress in respect of the OSPAR Hazardous Substances Strategy. This data is submitted to the UK Marine Environment Monitoring and Assessment National ( MERMAN) database. Contaminants have also been measured in mussels (2001-2012) as part of MSS's Long Term Hazardous Substances Monitoring Programme and submitted to MERMAN. Farmed, rope grown mussels (Loch Etive and Loch Ewe) and wild mussels (Straad on the west coast, Shell Bay and Aberdeen Breakwater on the east coast) ⁸ (Fig. 1) have been analysed for PAHs, PCBs and PBDEs, although this programme has now been concluded. MSS sample fish for contaminants as part of their annual CSEMP and Clyde trend monitoring cruises (Fig. 2). The OSPAR CEMP programme requires the determination of lead, cadmium, PCB, and PBDE concentrations in fish liver, and mercury in fish muscle tissue. For CSEMP, MSS monitor contaminants in plaice, dab or flounder from 10 sites annually. The use of modern analytical methodologies means that data for the three specified metals (and several others) are routinely obtained for both liver and muscle of fish. CSEMP contaminant data for fish are submitted to MERMAN and from there to international databases ( ICES).

Figure 1 Scottish shellfish sites, including MSS's Long Term Hazardous Substances Monitoring Programme ( LTHSMP) 2005 - 2012 sites, MSS integrated assessment sites and SEPA's Shellfish Water and DSD sites. MSS sites include 2 rope grown sites (Loch Etive and Loch Ewe) and 3 wild mussel sites (Straad on the west coast and Shell Bay and Aberdeen Breakwater, this sampling programme has now stopped). SEPA's sites are all wild mussel sites. Grey lines show the Scottish sea areas ².

Figure 2 Scottish fish monitoring sites sampled annually for contaminant analysis by MSS. The grey lines show the Scottish sea areas ².

In addition, a range of biological effects are or have been measured for CSEMP and other monitoring and research programmes. MSS conduct annual determinations of hepatic CYP1A enzyme activity ( EROD) in flatfish collected from specific CSEMP locations and from some Clyde trend monitoring sites. Since 2007, biliary PAH metabolites have also been measured in flatfish from these programmes. Additionally, annual fish disease surveys of dab were undertaken from North Sea sites between 2002 and 2008 (reported to MERMAN for 2004-2008) and EROD samples were analysed from these surveys between 2002 and 2008.

MSS began a 3 year research programme in 2009/10 intended to align MSS biological effects monitoring with the scheme for integrated assessment of contaminants and their effects ^9,10, although several of the effects measurements are not currently required for the OSPAR CEMP. This scheme is designed to address the MSFD requirement for determining whether the presence of contaminants causes environmental harm. During this project, MSS has developed expertise in internationally recognised biological effects techniques and obtained data on the concentrations and effects of contaminants in offshore fish (dab; 6 sites annually), inshore fish (flounder; 5 sites annually), and inshore mussels (4 sites in the Clyde in 2010; 4 sites in the Forth in 2011; and four sites on the Scottish east coast in 2012). The mussels monitoring programme continued to include sampling at 4 sites in the Clyde and one in the Solway in 2013.

As part of CSEMP, SEPA and MSS have undertaken a triennial monitoring programme for imposex in dogwhelks from around the Scottish coastline. The most recent survey was in 2010. Whilst a UK survey was due in 2013, it has been deferred to 2014 when it will fall within the MSFD monitoring cycle. In addition, MSS monitor imposex in Sullom Voe every two years on behalf of the Shetland Oil Terminal Environmental Advisory Group, most recently in 2013 (data not yet available).

Table 1 Mandatory and Voluntary CEMP determinands (contaminants and biological effects) monitored in biota annually by SEPA and MSS Y, currently monitored annually; N, not currently monitored

	SEPA (mussels)	MSS (fish and mussels)
Mandatory ( CEMP)
Cadmium	Y (1999 - Present)	Y (1999 - Present)
Mercury	Y (1999 - Present)	Y (1999 - Present)
Lead	Y (1999 - Present)	Y (1999 - Present)
CB28, 52, 101, 118,138, 153, and 180.	Y (1999 - Present)	Y (1999 - Present)
PAHs: anthracene, benz[ a]anthracene, benzo[ ghi]perylene, benzo[ a]pyrene, chrysene, fluoranthene, ideno[1,2,3-cd]pyrene, pyrene and phenanthrene	Y (1999 - Present)	Y (1999 - Present, mussels only)
BDE28, 47, 66, 85, 99, 100, 153, 154, 183 and 209	Y (2011- Present, Clyde and Forth only)	Y (2005 - Present)
Hexabromocyclododecane ( HBCDD)	N	N (Limited fish data, mainly from Clyde)
Imposex in dogwhelks	N UK triennial monitoring programme, 1998-2010; no longer have the capability to determine imposex.	Y UK triennial monitoring programme, 1992-present Biennial monitoring on behalf of Sullom Voe Oil Terminal Environmental Advisory Group from 1987 to 2013
Voluntary (pre- CEMP)
alkylated PAHs: C1-, C2-, and C3-naphthalenes, C1-, C2- and C3-phenanthrenes, and C1-, C2- and C3-dibenzothiophenes and the parent compound dibenzothiophene	Y (2010 - Present)	Y (1999 - Present, mussels only)
PFOS	N	N (Method under development)
Polychlorinated dibenzodioxins and furans	N, (One off survey in Clyde and Forth in 2009.	N
TBT in biota	N	N
CYP1A enzyme activity ( EROD) in flatfish	N	Y (1999-present)
Biliary PAH metabolites in flatfish	N	Y (2007-present)
Liver histopathology in flatfish	N	Y (2002-2008, 2010-present)
Liver macroscopic neoplasms in flatfish	N	Y (2002-2008, 2010-present)
Non- CEMP, recommended as part of ICES integrated assessment scheme
Copper	Y (1999-present)	Y (1999-present)
Zinc	Y (1999-present)	Y (1999-present)
External disease in flatfish	N	Y (2002-2008, 2010-present)
Plasma vitellogenin in flatfish	N	N (Outsourced in 2010)
DNA damage in flatfish haemocytes (micronucleus)	N	N (Outsourced in 2010-2011)
Lysosomal Membrane Stability ( LMS) in flatfish liver	N	Y (2010-present)
Lysosomal Membrane Stability ( LMS) in mussel haemocytes	N	Y (2009-present)
DNA damage in mussel haemocytes (Comet assay)	N	Y (2009-present)
Mussel histopathology	N	Y (2009-present)
Stress-on-stress in mussels	N	Y (2009-present)

Sediments

Sediment samples are collected and analysed for contaminants and benthic invertebrates ^[4] as part of UK CSEMP. This sediment sampling programme was modified for Scotland in 2005 following a review of the statistical power of the programme to detect a trend. Previously, five replicate samples were collected from each fixed CSEMP site and the results used to determine trends in contaminants and the benthic community. Sediments are not homogeneous so this approach provided low within-year variability but large between-year variability, which limited the power of the programme to detect a trend. It also led to data from a small area being used to represent a wide sea area. For these reasons, collecting five replicate samples at a fixed site was replaced by collecting 3-10 (depending on the size of the geographical area, or strata) more widely dispersed samples from within the same strata (based on WFD water bodies, if present), with sediment samples being collected in areas of fine sediment only. British Geological Survey maps showing sediment type were used to identify suitable areas. Most of the east coast consists of shell and gravel and, therefore, was unsuitable for contaminant analysis. Dispersing the sampling over a wider area increased within-year variability but reduce between-year variability so increasing the power of the programme to detect trends. Furthermore, the sampling programme will also be more representative of the wider sea area. This approach is similar to the approach recommended by the ICES Working Group on Marine Sediments ( WGMS) in 2013. WGMS were asked by OSPAR to produce a sampling strategy for spatial monitoring of sediments on a sub-regional scale. It recommended that for a sub-region ( e.g. southern North Sea), the areas of fine sediment be identified and, depending upon local hydrography and sediment transport patterns, grouped to form a number of coherent strata (which may transcend national boundaries). A statistically defined number of samples would then be randomly collected from each of the strata; the number of samples collected in each stratum is defined in order to provide sufficient power to determine whether a concentration exceeds the EAC/ EQS and depends upon the variability of any data already available for that strata.

Figure 3 Locations of SEPA and MSS sediment sites sampled as part of the UK CSEMP. An example of MSS stratified random ( SR) sampling sites from 2009 are shown along with the MSS fixed (F) locations, sampled annually since 1999. SEPA use a stratified fixed ( SF) sampling protocol and these sites have been sampled annually since 2005.

For CSEMP sites, MSS adopted a random stratified approach to sediment sampling whereby samples are collected from 5-10 random sites in a strata (based on WFD water bodies, where present) every year, although samples are only collected from fine sediments (Fig. 3). Sampling at the six fixed sites sampled since 1999 is also undertaken annually (Fig. 3), however, since 2005 only one sample is collected (1999 - 2005 five replicate samples were collected). Sediment samples are collected for both hazardous substances and benthic invertebrate analysis.

In 2005, SEPA adopted a stratified fixed sampling approach where five fixed locations in each selected water body (eight water bodies, 40 samples) are sampled (Fig. 3). SEPA made further modifications to the sediment programme in 2010 when a new 3 yearly monitoring programme was introduced for benthic invertebrates ¹¹. The 3 yearly monitoring programme is based on 3 regions, west, east and north. For contaminants annual sampling has been retained in the Forth (3 strata) and Clyde (2 strata) with triennial sampling in Minches and Malin Sea (1 stratum) and Moray Firth (1 stratum).

Table 2 Mandatory and voluntary OSPAR CEMP determinands monitored in sediments by SEPA and MSS.

Y, currently monitored annually; N, not currently monitored

	SEPA*	MSS
Mandatory ( CEMP)
Cadmium	Y (1999 - Present)	Y (1999 - Present)
Mercury	Y (1999 - Present)	Y (1999 - Present)
Lead	Y (1999 - Present)	Y (1999 - Present)
Tributyltin	N	N (Monitored historically, but stopped as below detection limits)
CB28, 52, 101, 118, 138, 153 and 180.	Y (1999 - Present)	Y (1999 - Present)
PAHs: anthracene, benz[ a]anthracene, benzo[ ghi]perylene, benzo[ a]pyrene, chrysene, fluoranthene, ideno[1,2,3- cd]pyrene, pyrene and phenanthrene	Y (1999 - Present)	Y (1999 - Present)
BDE 28, 47, 66, 85, 99, 100, 153, 154, 183 and 209	N	Y (2005 - Present)
HBCDD	N	N (Limited monitoring, in the Clyde in 2009
Cofactors (particle size, organic carbon, lithium and aluminium)	Y	Y
Voluntary (pre- CEMP)
alkylated PAHs C1-, C2-, and C3-naphthalenes, C1-, C2- and C3-phenanthrenes, and C1-, C2- and C3-dibenzothiophenes and the parent compound dibenzothiophene	Y (2010 - Present)	Y (1999 - Present)
PFOS	N	N (Method under development)
Polychlorinated dibenzodioxins and furans	N	N
General biological effects	N (Sediment bioassays discontinued in 2011 due to resourcing)	N

*Since 2010 sediment sites are generally visited on a three yearly cycle. The exception is the Forth and Clyde.

Data Assessment

Contaminant and biological effects ( EROD, bile metabolites, fish disease) data collected for the above sediment and biota monitoring programmes are submitted annually (by 1 ^st June) to the UK MERMAN database; this data is subsequently submitted to ICES (September of the same year). The MERMAN and ICES databases are being updated to allow data to be submitted for effects in mussels ( e.g. LMS, stress-on-stress, comet assay) and for additional effects in fish ( e.g. LMS, micronucleus). Currently the majority of contaminant and effects data collected in Scotland is held in the MERMAN database and is readily available for Descriptor 8 data assessments. Data collected as part of one-off surveys/research projects and some of SEPA's WFD data is not submitted to MERMAN, but are held on other databases ( e.g. MSS contaminants database). Station dictionary names will be obtained for any new sites proposed for Descriptor 8 monitoring and this data will be submitted to MERMAN. Contaminant data from the Food Standards Agency in Scotland ( FSAS) that may be useful for D8 assessments ( e.g. dioxins in shellfish) are not currently submitted to MERMAN, although FSAS have agreed to try and do this. However, there may be resourcing issues for the addition of new contaminants/effects to MERMAN, as this will require changes to the database structure.

Contaminants and effects data have been assessed for the OSPAR Quality Status report (1998 - 2008) ¹², UK Charting Progress 2 (1999 - 2007) ¹ and Scotland's Marine Atlas (1999 - 2009) ². Charting Progress 2 and Scotland's Marine Atlas were used for the UK's initial assessment for the MSFD. MSS have also assessed data on persistent organic pollutants from 1999 up to 2010; PCB, PBDE and PAH concentrations in sediment and biota from Scottish sea areas were assessed ¹³. Temporal and spatial trends were investigated and the concentrations assessed against internationally agreed assessment criteria. Rules were developed for the aggregation of the contaminant data across each sea area. An overall assessment for each sea area was then assigned, looking at the frequency of sites or strata within each sea area that were above or below the relevant assessment criteria.

MSS have also been influential in the development of guidelines by ICES for the integrated monitoring and assessment of contaminants and their effects ¹⁴. These guidelines have been accepted by OSPAR for a three-year trial (2012-2015) and rely upon a coherent approach to sample collection, sample analysis, data handling, and data assessment for both contaminants and biological effects, and associated supporting data.

Suitability of the Existing Scottish Monitoring Programmes to Meet MSFD Descriptor 8 Requirements

Current contaminant and biological effects monitoring undertaken as part of Scottish monitoring programmes, including UK CSEMP, will help fulfil the requirement of MSFD Descriptor 8. However, there are limitations, particularly in the spatial coverage of the datasets and the range of contaminants monitored.

Spatial Coverage for MSFD

The MSFD (Article 4) lists 10 sub-regions for monitoring purposes, the relevant sub-regions for the UK are:

• The Greater North Sea, including the Kattegat and the English Channel ( OSPAR Region II)
• The Celtic Seas ( OSPAR Region III)

Scottish waters fall within OSPAR Region II (includes the Scottish sea areas east Shetland, Fladen, Moray Firth, east Scotland coast, Forties, and Forth) and OSPAR Region III (includes the Scottish sea areas Solway Firth and North Channel, Clyde, Minches and Malin Sea, Hebrides, Rockall, Bailey, north Scotland coast, Faroe-Shetland, west Shetland).

Under the UK CSEMP, SEPA sample annually shellfish (mussels) from all Scottish sea areas with a coastline and SEPA/ MSS sample dogwhelks from around Scotland on a triennial basis. Sediment samples are collected annually in the Solway Firth and North Channel, Clyde, Minches and Malin Sea, east Shetland and west Shetland, Fladen, Moray Firth, east Scotland and Forth, which covers both OSPAR Region II and III. Samples are not collected in all Scottish sea areas e.g. Forties and Hebrides.

For CSEMP, fish samples are collected annually in the same sea areas as sediments (but not West Shetland), although the sites in the Forth sea area are within the estuary and therefore not suitable for use under the MSFD which is not applicable to transitional waters. The Hebrides, Rockall, Bailey, north Scotland coast, Faroe-Shetland and Forties sea areas are not routinely sampled for contaminants or effects monitoring for either biota or sediment, although fish were sampled from Rockall, Bailey and Hebrides (2006-2012) and the north Scotland coast (2012) as part of research projects on deep sea fish or MSFD Descriptor 9.

The main source of contaminants in the offshore areas will be from diffuse impacts (although there may be some licensed inputs as a result of petroleum-related activities in some of these areas) from atmospheric deposition or shipping and contaminant concentrations are likely to be low, such as seen in the Fladen for PBDEs and PCBs. It will not be necessary to monitor every Scottish sea area on an annual basis, particularly if concentrations are below the EQS/ EAC (or equivalent assessment criteria) (See Section - "Proposed MSFD Descriptor 8 Monitoring for Scotland").

The available sediment type may not be suitable for contaminant monitoring in all Scottish sea areas. Furthermore, obtaining undisturbed sediment samples in deep water sea areas (Rockall and Bailey) can be difficult; although information is not currently available on the sediment type in these sea areas, there are likely to be areas of fine sediment. Sediment in the east Scotland coast, north Scotland coast, west Shetland and Hebrides sea areas are mainly sand, gravel or rock, with limited areas of fine sediment, and therefore, it may be difficult to find suitable sediment for contaminant monitoring in these areas (Fig. 4). Small areas are currently monitored in the east Scotland coast and west Shetland regions, however, collecting a suitable sample for contaminant monitoring in these area can be difficult due to a lack of suitable sediment. Sampling in the east of Shetland is also undertaken in a small area of fine sediment close to Shetland, sampling here should change to focus on the larger area of fine sediment in the east Shetland area (Fig. 3), using stratified random sampling. Previous sediment sampling in this area has been undertaken with samples being analysed for PAHs ¹⁵; similar to the Fladen area concentrations were found to be close to background and therefore monitoring every 6 years in this area may be sufficient.

Figure 4 Map showing the Sediment type of offshore sediment around Scotland from the British Geological Survey ( BGS)

Although fish are sampled from most of the Scottish sea areas with a coastline (not Hebrides or west Shetland), existing fish sampling is focussed on the Clyde and Forth sea areas (Fig. 2), with 3 CSEMP sites (2 estuarine and one coastal), five Clyde trend monitoring stations and one estuarine research site. This is because the Clyde and the Forth are subject to the greatest inputs.

The MSFD covers the UK Exclusive Economic Zone, including coastal waters, but not transitional waters ( i.e. estuaries). WFD also includes coastal waters, which for chemical status includes territorial waters to 12 nautical miles. The MSFD applies in coastal waters only in so far as particular aspects of the environmental status of the marine environment are not already addressed through the WFD or other Community legislation. Hence, contaminant monitoring in coastal waters for WFD purposes may also be used for Descriptor 8 assessments. However, WFD has a requirement for sediment and biota trend monitoring of some Priority Substances (see Appendix 1), this is covered by CSEMP for those contaminants considered to be an issue in Scottish waters ¹⁶. The majority of WFD monitoring in water samples from transitional and coastal waters has shown concentrations are below limits of detection and EQS. Therefore, monitoring of organic contaminants in water samples for WFD has stopped and only trace metals are monitored at sites where concentrations are above the limit of detection. MSFD also has additional requirements not covered by WFD, such as biological effects measurements. Therefore, effects measurement are required in coastal waters for Descriptor 8 monitoring. Monitoring of contaminants and effects measurements in offshore, coastal and transitional waters is undertaken as part of UK CSEMP for OSPAR purposes and will be used for Descriptor 8 assessments; following the integrated assessment research project, an additional shellfish biological effects monitoring has been established.

Shellfish are monitored from around the Scottish coastline (Fig. 1); this is done primarily by SEPA, although MSS undertakes a limited biological effects monitoring programme. Monitoring uses mussels for contaminants and effects, with dogwhelks used for assessing the impacts of TBT (imposex). The mussel contaminants and imposex programmes are sufficient for MSFD purposes; the mussels biological effects programme covers each of the coastal Scottish sea areas on a rolling basis; as with the fish sampling, the numbers of sites in each varies depending upon the perceived environmental hazards, thus there are more sites in the Forth and Clyde than on the northwest coast. The Moray Firth, north and southwest coasts have few sites where mussels can be obtained.

Determinands Required for MSFD

Descriptor 8 covers those substances which are classed as priority substances for WFD ( Appendix 1). This list covers a much wider range of contaminants than are currently monitored in marine waters as part of the OSPAR CEMP. Many of these substances will not be relevant for marine waters as they are from land-based sources and concentrations are low (< EQS) in water samples collected from coastal and transitional waters. In 2012 the OSPAR Working Group on Monitoring and on Trends and Effects of Substances in the Marine Environment ( MIME) reviewed the contaminants and effects that member states were planning on monitoring for Descriptor 8 and developed a list of common indicators for use in the next JAMP and in assessment of GES for Descriptor 8. This list was reviewed and amended by OSPAR in June 2013 ( OSPAR 13/21/1, Annex 4), to produce a minimum set of determinands that should be monitored in all north east Atlantic MSFD sub-regions (for the UK, OSPAR regions II and III, see previous section) in order to allow comparisons to be made across regions (from Directive 2008/56/ EC these are north east Atlantic). For contaminants, the list of common indicators includes the OSPAR CEMP determinands: Hg, Cd, Pb, PCBs and PBDEs in sediments and biota, and PAHs and organotins in sediments. Imposex/intersex in gastropod molluscs was the only biological effect agreed as an OSPAR common indicator.

Currently, as part of the UK CSEMP, sediment and biota from some Scottish sea areas are monitored for all the MIME common indicators, except TBT in sediment. TBT is no longer monitored in sediment for CSEMP as TBT concentrations are low and usually below detection limits, although TBT concentrations continue to be monitored at dredged spoil disposal sites as part of post-disposal monitoring undertaken by MS Licensing Operations Team. Of these, almost all disposal site sediments were assessed as being unlikely to have acute exposure risks and TBT was undetectable in many ¹⁷. Female gastropod molluscs exposed to TBT develop male sexual characteristics, a condition known as imposex and monitoring for imposex is mandatory under the CEMP; within the UK this has taken place on a triennial basis (the last survey was in 2010). The occurrence and severity of imposex in gastropods collected from the Scottish coastline has decreased markedly since the ban on the use of TBT as an antifoulant was introduced. The most recent available imposex data (2009-2011) indicate 11 Scottish sites where the EAC is exceeded. Five of these sites are from Sullom Voe and are being sampled in 2013. Defra are proposing that a UK-wide imposex survey take place in 2014 of any sites which were not classified as consistent with GES in both 2007 and 2010. In Scotland this will require 7 sites to be sampled, six sites that exceed the EAC in 2010 and one which was borderline. SEPA no longer have the capability to determine imposex, hence they will collect the samples and MSS will analyse them.

OSPAR also produced a list of "candidate indicators", that require further development before being considered for adoption as "common indicators". This list included four contaminants and five biological effects, although none of these were prioritised for urgent development and inclusion in the OSPAR 2017 Intermediate Assessment. The contaminant candidate indicators are PAHs and TBT in biota and hexachlorobenzene ( HCB) and hexachlorobutadiene ( HCBD) in sediment and biota. The biological effects candidate indicators are externally visible fish diseases, EROD activity, biliary PAH metabolites, lysosomal stability and micronuclei.

Of the OSPAR contaminant candidate indicators, PAHs in biota are already monitored in Scotland through SEPA's mussel monitoring, whereas TBT, HCB and HCBD in biota and, HCB and HCBD in sediment are no longer monitored. HCB and HCBD are classed as WFD PHSs, but are not included on the OSPAR CEMP or pre- CEMP. Hazardous substances included on the OSPAR CEMP and/or classed as priority substances for WFD were reviewed and prioritised in 2010 for Scotland ¹⁶. This report concluded that the occurrence of HCB and HCBD in the Scottish marine environment was low with concentrations generally below detection limits, therefore, monitoring of these contaminants was a low priority and is not required under WFD. Ni and its compounds is a WFD PS, but not listed under the OSPAR CEMP/pre- CEMP and was not suggested as a common or candidate indicator, whilst trend analysis is not required (it is not hydrophobic), both MSS and SEPA routinely report Ni as part of their CEMP trace metals analyses in sediments and biota.

Of the biological effects measurements listed as OSPAR candidate indicators, externally visible fish disease, EROD and bile metabolites are routinely monitored in fish samples by MSS as part of UK CSEMP and reported to national ( MERMAN) and international ( ICES) databases. Lysosomal stability and micronuclei have also been measured in Scottish waters as part of a MSS integrated contaminants and biological effects research project. MSS have now established the lysosomal stability techniques, however, the micronuclei analyses were out-sourced and there may be a need to develop this expertise in Scotland; MERMAN and ICES databases are being adapted to accept these data.

Assessment Criteria Currently Available for Contaminants and Biological Effects

Criteria have been developed for the assessment of data collected for WFD and OSPAR purposes and which will be used for Descriptor 8 assessments. For WFD, Environmental Quality Standards ( EQS) are required to enable assessments of the chemical status of a water body to be made. For consistency between Directives, EQSs are also to be used in assessments for the MSFD, although EQSs have not been developed for sediments. The recently published Priority Substances Directive (2013/39/ EU) specifies freshwater and saltwater EQS values for 25 priority substances ( PSs) and 9 priority hazardous substances ( PHSs) and for 6, non-priority, substances, together with biota EQS values for 1 PS and 10 PHSs (see Appendix 1). EQSs are the boundary between "Good" and "Failing to achieve good" status. Concentrations of each contaminant must be below EQS before the water body is classified as having "Good" chemical status. The EU Technical Guidance Document on how to define EQS values specifies that Quality Standards are produced for several different matrices (freshwater, saltwater, biota, sediment, human health, etc.) and that it is the most protective of these that is chosen as the EQS value. For each PS/ PHS, the EC produced a dossier including additional Quality Standards for each different matrix; for hydrophobic contaminants these could be converted to provide sediment or biota assessment criteria by the use of equilibrium partitioning theory.

Within OSPAR, assessment criteria have been developed for contaminants in sediment and biota so as to permit environmental status assessments to be made. Background Concentrations and Background Assessment Concentrations ( BACs) have been developed for OSPAR CEMP determinands in biota and sediment to allow assessment of whether concentrations are at the OSPAR target of close-to-background; Background Concentrations ( BCs) determined as freely dissolved concentrations (C _free) in saltwater by passive sampling are available for some CEMP substances ( e.g. PCBs, PAHs). Environmental Assessment Criteria ( EACs) are concentration thresholds designed to be protective of the most sensitive species and have been developed for most CEMP determinands. Where it has not been possible to develop EACs, other assessment criteria were used in the OSPAR Quality Status report 2010, including EC Food Regulation limits and the Effects Range-Low ( ERL) ¹² of the United States National Oceanic and Atmospheric Administration were used ( NOAA, 1999). However, it is recognised that there is a need to further develop assessment criteria for both existing CEMP determinands and the pre- CEMP determinands. OSPAR also have a number of Ecological Quality Objectives (EcoQOs) which are environmental indicators stating aspirations for a healthy North Sea as part of the ecosystem approach; one of these is for imposex in gastropod molluscs. Assessment criteria have been developed for a suite of effects measurements, though currently not approved for use. The ICES/ OSPAR Study Group on Integrated Monitoring of Contaminants and Biological Effects ( SGIMC) developed assessment criteria analogous to contaminant BACs and EACs for biological effects determinants. Background Assessment Criteria are available for all biological effects common and candidate indicators and Environmental Assessment Criteria are available for four (imposex, fish disease, LMS and PAH metabolites) ¹⁸.

The Assessment Criteria to be used for MSFD assessments are WFD EQSs or, where these are not available ( e.g. for sediments), OSPAR EACs. The EC WFD Working Group E (Chemicals) is expected to develop additional EQSs, nonetheless, if EQSs/ EACs do not become available, sediment ERLs will be used. For OSPAR assessments, BACs are additionally available for sediments, biota and (for some hydrophobic compounds) freely-dissolved water concentrations (determined by passive sampling). Currently available assessment criteria for CEMP determinands and the OSPAR common and candidate indicators are listed below.

Metals (Hg, Cd, Pb, Ni): An EQS is available for Hg in biota (whole fish), but not for sediments or mussels. For sediment, OSPAR BACs are available, but there are no EACs; ERLs were used to assess sediments for the OSPAR Quality Status Report ( QSR) 2010. OSPAR BACs are available for mussels and fish, but there are no EACs; EC food regulations were used for the OSPAR QSR 2010. Although not required for MSFD, ERLs are available for Ni in sediment, but no assessment criteria are available for Ni in biota. Saltwater EQSs exist for dissolved Ni, Cd and Pb and are used by SEPA for assessing transitional waters.

PCBs: OSPAR BACs and EACs are available for the ICES7 PCBs (ortho- PCBs, non dioxin-like) for use in sediment, but no EQSs are available. For fish liver and mussels BACs are available; BCs are available for saltwater C _free. For the OSPAR QSR 2010, an EAC was calculated using biota sediment accumulation factor ( EAC ^passive). For dioxin-like PCBs there is a WFD EQS for whole fish.

PAHs: BACs are available for parent PAHs in sediment but there are no EACs or EQSs available and ERLs were used for the OSPAR QSR 2010. For PAHs in mussels (candidate indicator), BACs and EACs are available for parent PAHs and EQSs exist for fluoranthene and benzo[ a]pyrene. EQSs exist for naphthalene and anthracene in total water and PAH BCs exist for saltwater C _free. Alkylated PAHs are also on the pre- CEMP; there are no BACs or EACs available, but sediment BCs and ERLs are available for some e.g. 2-methyl naphthalene.

PBDEs: BACs and EACs are not available for sediment or biota. There is an EQS for biota (whole fish) however, this value, 0.0085 µg kg ^-1 wet weight, is several orders of magnitude below current achievable detection limits.

TBT : OSPAR BAC and EAC are not available for sediment (common indicator) and the EQS is for water. BAC and EAC are available for shellfish. The Priority Substance Directive ( PSD) dossier on TBT includes quality standards for sediments and biota, although these are several orders of magnitude below the detection limits currently achievable by Competent Monitoring Authorities. Within OSPAR, the significance of TBT is assessed using its biological effect (imposex) on gastropods; this will also be used for MSFD.

HCB : OSPAR BACs are available for HCB in mussels and fish, but there are no EACs and no BACs or EACs for sediment. There is a BC for saltwater C _free and a WFD EQS for whole fish and ERLs for sediment.

HCBD : There are no BACs or EACs for sediment or biota. However, there is an EQS for whole fish.

Imposex in gastropods: Common indicator. OSPAR have a six class assessment scheme from which an OSPAR EcoQO and ICES biological effect BAC/ EACs have been developed.

Externally visible fish diseases: Candidate indicator. ICES BACs and EACs are available for dab, cod and flounder.

Liver macroscopic neoplasms: Included on the OSPAR pre- CEMP, but not a candidate indicator. ICES BAC and EAC values are available for dab.

Liver histopathology (contaminant specific): Included on the pre- CEMP, but not a candidate indicator. ICES BAC and EAC values are available for dab.

Bile metabolites of PAH: Included on the pre- CEMP and a candidate indicator. ICES BAC and EAC values are available for dab.

Lysosomal stability: Included on the pre- CEMP and a candidate indicator. ICES BAC and EAC values are available for fish and mussels.

Micronuclei: Biomarker of genotoxicity, included on the pre- CEMP and as a candidate indicator. ICES BAC values are available for dab, flounder, cod, viviparous blenny and mussels.

EROD : Biomarker of exposure to planar organic compounds ( e.g. PAH, dioxins). Included on the pre- CEMP and as a candidate indicator. ICES BAC values are available for dab, flounder, plaice, and cod.

DNA Adducts: Biomarker of effects due to PAHs. Included on the pre- CEMP, but not a candidate indicator. ICES BAC and EAC values available for dab, flounder, haddock and cod.

Sediment Bioassays: Included on the pre- CEMP, but not a candidate indicator. ICES BAC and EAC values available for Corophium and Arenicola bioassays.

Water bioassays: Included on the pre- CEMP, but not a candidate indicator. ICES BAC and EAC values available for copepod, sea urchin embryo, oyster embryo and mussel embryo bioassays.