Department for Environment, Food and Rural Affairs

UK COMPLIANCE WITH BALLAST WATER REGULATIONS

Ref: CDEP 84/5/286

FINAL REPORT MARCH 2002

Suzanne Welch Ian Lucas

Contractor: Dr. I.A.N.Lucas School of Ocean Sciences University of Wales, Bangor Contents

List of figures and tables 3

Acknowledgements 5

Abbreviations 6

Executive summary 7

1.0 Background to the project 8

2.0 Objectives 11

3.0 Milestones 11

4.0 Methods 12 4.1 Literature review 12 4.1.1 Non-native species 12 4.1.2 Sampling methodology 13 4.2 Ports 15 4.3 Buoys 22 4.4 Database 22 4.5 General approach on port surveys 23 4.5.1 Target species 23 4.5.1.1 Target introduced pest species list for the UK 24 4.5.1.2 Marine pest species that pose a threat to the UK 24 4.5.1.3 Known or likely non-native marine species in UK waters that currently are not assigned pest species 24 4.5.1.4 Native UK marine species that appear on other port nations target species list (Australia) 25 4.6 Cysts 25

4.7 Information sources 25 4.7.1 Government agencies 25 4.7.2 Marine groups 25 4.7.3 General public 26 4.8 Sampling 26 4.9 Preliminary surveys and equipment testing 27 4.9.1 Panels 27 4.9.2 Van Veen Grab 28 4.9.3 Dredge 28 4.10 Sampling methodology 29 4.11 Preservation and curation 34 4.11.1 Cyst analysis 35 4.12 Statistical analysis 35

1

5.0 Results and observations 36

5.1 Introduction 36 5.2 Non-native species collected 37 5.3 Sampling 37 5.3.1 Plankton 37 5.3.2 Benthos 37 5.3.2.1 Mobile organisms 37 5.3.2.2 Sessile organisms 37 5.3.2.3 Infauna 38 5.3.3 Encrusting organisms 40 5.3.4 Community analysis 42

6.0 Analysis and discussion 44 6.1 Analysis of methods employed 44 6.1.1 Plankton 44 6.1.2 Dredging 45 6.1.3 Grab and Corer 45 6.1.4 Panels 45 6.1.5 Destructive sampling 46 6.1.6 Trapping 46 6.1.7 Visual search, intertidal 47 6.1.8 Visual search, subtidal 47 6.2 Time allocation for surveys 48 6.3 Analysis of results 48

7.0 Conclusions 51

8.0 Recommendations 51

References 52

Appendices i - lxxiii

A. UK maps showing the distribution of established non-native species collected during the survey i B. Port summary for Cardiff viii C. Port summary for Felixstowe xiii D. Port summary for Liverpool xix E. Port summary for Milford Haven xxvi F. Port summary for Southampton xxxvii G. Port summary for Teesside xlv H. Summary of sample methods used at each site l I. Table for dates that panels were deployed and retrieved lii J. Distribution maps for established non-native species not found liii K. Distribution maps for non-established non-native species not found lxv

2 List of Figures and Tables

Table 1: Summery of common sampling techniques. Table 2: Sampling techniques recommended by Hewitt & Martin 1996 Table 3: Habitat types at each port. Table 4: Names and ports of buoys sampled Table 5: Sampling dates for each port Table 6: Initial test sites for panels Table 7: Non-native species collected at each port. Table 8: Dredging summary for all ports and the number of species collected Table 9: Comparisons between the number of species found in Collingwood using three sample methods Table 10: Sampling methods that successfully collected non-native species, the numbers collected by wach method and the ports in which they were collected. Table 11: Total numbers of native and non-native species found at each port Table 12: Comparisons between the use of biological divers, destructive sampling and settlement panels Table 13: Suggested time allocation for surveys

Figure 1: Map of the UK showing the six ports used in this survey. Figure 2: Map showing the port of Liverpool and the sampling sites. Figure 3: Map showing the port of Cardiff and the sampling sites. Figure 4: Map showing the port of Milford Haven and the sampling sites. Figure 5: Map showing the port of Southampton and the sampling sites. Figure 6: Map showing the port of Felixstowe and the sampling sites. Figure 7: Map showing the port of Teeside and the sampling sites. Figure 8: Opening screen of database. Figure 9: Deploying the phytoplankton net with the pulley system at Teesside. Figure 10: Benthic dredge. Figure 11: Benthic dredge in use at Cardiff docks. Figure 12: Van Veen grab.

3 Figure 13: Two tier settlement panels Figure 14: Destructive scraper Figure 15: 0.25m2 quadrat used for sampling on buoys Figure 16: Base of a buoy after samples have been taken Figure 17: Shrimp and crab traps Figure 18: Underwater photograph of the non-native sea squirt, Styela clava Figure 19: Typical photograph obtained of the subtidal community in Collingwood dock, Liverpool Figure 20: Distribution and abundance of Styela clava Figure 21: Number of Carcinus maenas caught in Gladstone dock, Liverpool between March 2001 and August 2001 Figure 22: MDS plot of replicate cyst assemblages in sediment samples taken from Liverpool Figure 23: Dendrogram showing similarities between cyst assemblages in sediment samples taken from Liverpool. Figure 24: Map of Cardiff docks showing the distribution and abundance of Ficopomatus enigmaticus collected on settlement panels Figure 25: Salinity variation at Cardiff docks Figure 26: Dendrogram showing the similarity between communities at sample sites in Liverpool

4 Acknowledgements

Particular thanks to: Associated British Ports Cardiff & Barry, Associated British Ports Southampton, Mersey Docks and Harbour Company, Milford Haven Port Authority, Port of Felixstowe, Tees and Hartlepool Port Authority Ltd.

In addition, we are grateful to the following:

Esso Petroleum Company, Mersey Ferries, Merseyside Development Corporation, Neyland Yacht Club, Phillips Petroleum, Porcupine Natural History Society, Royal Maritime Auxiliary Service, Royal National Lifeboat Institute, Trinity House and the officers and crews of THV Mermaid & Patricia, Sea Angler Magazine.

John Hamer, Helen Hardy and the technical and academic staff at the School of Ocean Sciences, Menai Bridge, who helped with advice, identification and sampling

Cover page: Calcarious tubes of Ficopomatus enigmaticus, photograph by Suzanne Welch. Underwater photography by Paul Kay. Marine Wildlife Photo Agency. All other photographs by research staff working on the project.

5 Abbreviations

ABWMAC Australian Ballast Water Management Advisory Council

ACME The Advisory Committee on the Marine Environment

AQUIS Australian Quarantine and Inspection Service

CRIMP Center for Research on Introduced Pests

GEF Global Environment Facility

GESAMP The Joint Group of Experts on the Scientific Aspects of Marine Environmental Protection

ICES The International Council for the Exploration of the Sea

IMO International Maritime Organisation

IMPACT Results from the Working Group on Impacts on the Marine Environment

JAMP Joint Assessment and Monitoring Programme

JNCC Joint Nature Conservancy Council

MARPOL The International Convention for the Prevention of Pollution from Ships

MDS Non-metric multi-dimensional scaling

MEPC Marine Environmental Pollution Council

OSPAR Oslo and Paris Convention

SSSI Site of Special Scientific Interest

UNDP United Nation’s Development Programme

WGITMO Working Group on Introductions and Transfers of Marine Organisms

WHO World Health Organisation

6 Executive Summary

The aim of this project was to generate recommendations on the most appropriate monitoring programmes required to address the issue of non-native species in the ports of the UK. The continuing risk from shipborne species introductions, has prompted the IMO to produce guidelines regarding ballast water management. These encourage port states to provide the IMO with information on severe outbreaks or infestations of harmful aquatic organisms which may pose a risk. An essential prerequisite for fulfilling these guidelines/regulations on the prevention of the spread of non-native organisms is a base-line study to record those species present in the waters of docks and harbours at which ballast water is loaded.

The main objectives of the research undertaken were to: • Review survey practices and reporting within the OSPAR region and develop a port monitoring program which compliments any on going surveys and utilises existing information. • Produce and develop a baseline recording system for selected UK ports, reporting the occurrence of native species and those non-native species that already have been introduced from elsewhere. • Interpret the data collected on the distribution of non-native species. • Develop recommendations for future monitoring programmes, taking into account OSPAR and IMO requirements.

The ports and harbours within the UK are numerous and varied in their size, geography and traffic type. Bearing in mind that UK ports are generally net importers of bulk cargo and hence ships are more likely to load than to discharge ballast water, six ports for England and Wales were selected on the basis of a number of criteria. These included their biogeographical nature and the volume and type of international traffic taking place. Using a range of sampling techniques, between six and seventeen sites at each port were examined. Five of the six ports were sampled twice within a 14 month period and one, Liverpool, was sampled 8 times. Species collected were identified, catalogued and the information entered on a database.

In total, over 600 marine species were identified during the course of the project. Of the 52 established non-native marine species reported to be present in UK waters, 33 had been recorded as being present in the areas where sampling took place. Of these records, 14 were identified in the course of the collections made during routine surveys. Conclusions • This study has demonstrated that it would be possible to survey and monitor the occurrence and distribution of native and non-native marine species in the ports of England and Wales.

• Large ports, because of their continuous activity, are difficult environments in which to carry out biological surveys. UK ports alsodiffer considerably in size and in the habitats they contain. To accommodate for these variables the sampling methods employed have to be practicable and flexible.

• It has been possible to demonstrate the general distribution, at the surveyed ports, of the non-native species identified and also show a more detailed distribution of species within ports.

• There are limitations in the current methods employed for assessing the populations of planktonic organisms that require further investigation.

• A target species list has been developed, composed of those species, native and non- native, which are considered to be candidates for introduction elsewhere.

• This project represents a first step in the development of a comprehensive, detailed database on the distribution of non-native species for the UK.

7 1.0 Background

Non-native species (also termed non-indigenous, exotic, alien, invasive,) are organisms occurring outside their natural range and dispersal potential (Hopkins 2001). Whether the causes of these introductions are intentional or accidental, the occurrence of species in areas where they are not native can have far reaching and harmful effects on biodiversity and the ecosystem into which they are introduced (Carlton & Geller 1993; GESAMP 1997; Gollasch & Leppäkowski 1999). In some cases these transfers have induced and continue to produce extensive changes in the biodiversity of the region where the introduction has taken place (Carlton 1989). In other cases there have been serious social and commercial repercussions as a consequence of the establishment of the so-called non-native species (Carlton & Geller 1993). The extent of this process has only been highlighted in the scientific literature over the last twenty years or so, and the degree and significance of the phenomenon has been assessed in only a small number of bioregions.

The progressive dispersal of marine organisms is an established, natural process that normally occurs stepwise over relatively short distances and on extended time scales. Long distance transportation between oceans and continents is a much less frequent event and may be attributed to chance occurrence, for example transportation of adhering small organisms by migratory birds or on drifting debris. Other, much more long term changes in species range result from the movements of climatic or geographic barriers. Transportation and establishment of single species or groups of species by human mediation has often had its most extreme effects on the biodiversity of the receiving region as the transportation has occurred over a short period of time, has been over great distances and may have been repeated. Establishment may occur in a number of ways, for example as a result of deliberate commercial introductions, but often it is the result of an inadvertent introduction related to marine commerce. One of the most common mechanisms by which this is accomplished is related to shipping activities. In many of the shipborne introductions, organisms have travelled either as hull fouling or included in wet or dry ballast. As a consequence ports and port areas are considered to be focal points for the initial appearances of non-native species or communities. These areas will then act as sources for the wider dispersal in the newly colonised region, the secondary dispersal process may be by a slower, natural biological process or again accelerated by man’s repeated, unconscious, intervention via local shipping routes.

Although there are undoubtedly many instances when introduced species do not survive for a significant length of time in their new environment and do not cause damaging changes to local biodiversity, non-native species have the potential to cause far reaching economic and biological impacts. The following is a simple classification regarding the different ways in which a non-native introduction may effect an ecosystem: • Disrupt the existing interactions between species or food web links. • Produce hybrids with native species, resulting in changes in biological and genetic diversity • Introduce parasites and diseases. • Produce natural toxins, which may accumulate in the local food chain and be consumed by man.

Initially, Sweden acted as Lead Country on non-native species issues in OSPAR and to investigate the extent of the problem of non-native marine species, a questionnaire survey was carried out. This resulted in reports being presented to OSPAR IMPACT in 1996 and 1997 (IMPACT 96/6/1 and 97/7/1). This desk study reported that a wide range of non-native marine species had been introduced, some of which have caused serious ecological, economic and public health problems. The study recognised the urgent need for a European wide monitoring of the spread of non-native marine species and of any prevention or eradication programmes that were being developed, and suggested that the information from such programs should be recorded in an easily accessible database.

8 Within the UK, the JNCC commissioned a review of non-native marine species in British waters (Eno et al 1997). This indicated that 52 non-native marine species had become established, the report also stated that up to 55% of primary introductions were probably associated with shipping. An observation supported up by Gollasch & Leppäkowski (1999) who have produced a list of the main vectors for aquatic species introductions, ranking in importance, ship’s ballast water, hull fouling, with sediment in ballast tanks being recognised as containing the richest source of potential introductions.

The potential dangers of ballast water as a source of introduced species have been recognised by resolution 18 of the 1973 ‘International Conference on Marine Pollution’. This conference provided the basis for the establishment of the MARPOL Convention (to which all the North Sea States are now party), and Resolution 18. This called on the WHO to establish collaboration with the IMO to investigate the degree to which ballast water could act as a vector for the transmission of pathogens such as cholera. The MEPC (the senior technical body on marine pollution related matters of the IMO) adopted guidelines by resolution in 1991 and in 1993, and these were, in turn, adopted by the IMO Assembly under Resolution A.774(18) and entitled ‘International Guidelines for Preventing the Introduction of Unwanted Aquatic Organisms and Pathogens from Ships Ballast Water and Sediment Discharges’. In 1997, the IMO Assembly adopted as Resolution A.868(20) the voluntary IMO ‘Guidelines for the Control and Management of Ships Ballast Water to Minimize the Transfer of Harmful Aquatic Organisms and Pathogens (IMO 1997).

With the recognition of the need for the provision of ballast water management options, the MEPC is working on development of draft regulations for ballast water management to prevent the transfer of harmful aquatic organisms in ballast water. It is planned to hold a diplomatic conference during 2003 to adopt the new measures. The proposed instrument will be a new international convention “for the control and management of ships ballast water and sediments”. This will include requirements that would apply to all ships, including mandatory requirements for a Ballast Water and Sediments Management Plan, a Ballast Water Record Book and a requirement that new ships will carry out ballast water and sediment management procedures to a given standard or range of standards. In addition it also acknowledges that ballast exchange on the high seas is the only widely accepted technique currently available to prevent the spread of unwanted aquatic organisms in ballast water and its use should continue. However, it has been stressed that this technique has a number of limitations and it is hoped that the development of alternative treatment technologies will produce techniques that are substantially more reliable, commercially acceptable and will not infringe on the safe operation of shipping.

A range of alternative treatment options have been proposed and are currently being tested and developed, for example, heating, filtration, chemical treatment and ultra-violet irradiation of ballast water are being examined (Rigby 2001). However, the debate as to the most effective methods of ballast water treatment to minimise the risks of introducing non-native marine organisms, financially, practically and environmentally, is ongoing.

As an interim measure the IMO has also recommended port states provide information on the range of ‘local’ organisms that may be taken on board during ballast loading operations. This information should include, as well as indigenous species, ‘naturalized’ non-native species that have themselves been introduced at some time. The guidelines require that port states notify the IMO of significant occurrences of harmful organisms and pathogens (e.g. harmful algal blooms) and that this and other information (e.g. location of sewage outfalls) be supplied to ships upon request to enable ballast water management plans to be implemented.

Baseline information on the distribution and abundance of non-native marine species is also vital for the assessment of whether the regulations make a significant contribution to reducing the establishment of non-native species within UK waters. In addition, the information will provide the basis upon which the rate of spread of non-native species both within the UK and the Convention area waters can be assessed.

9 The information presented in the JNCC review (Eno et al,1997) provides a substantial basis for further direct investigations into the numbers and distribution of non-native species on and around the coasts of the UK. When this report is combined with the survey that assessed the extent of discharge of ballast water in UK ports (Laing 1995), some indication may be derived on those ports with a greater potential for containing non-native introductions. On the basis of this and other information the ports that form part of this initial survey programme have been chosen. However, it must be recognised that there are many other factors involved in the process of establishment of a non-native species and to establish a complete picture a significant number of other ports will need to be examined in the future.

10 2.0 Objectives

With reference to the UK’s compliance with ballast water regulations the aim of this project was to examine the introduction of non-native marine species within ports and harbours in England and Wales. With this in mind, the objectives are listed below.

‰ To review current survey procedures within the OSPAR region and develop a port sampling programme that will complement existing information and any on-going surveys being carried out within the convention area. ( Section 4.0)

‰ To provide a baseline record of the occurrence, distribution and abundance of non-native species in major UK ports and harbours.( Section 5.0, Appendices A-G & Database)

‰ To evaluate the present status of non-native species in UK ports and harbours.( Section 6.0)

‰ To develop recommendations for the structure of future monitoring programmes with regard to OSPAR and IMO requirements and suggest areas for future monitoring. ( Section 8.0)

3.0 Milestones

1 Contact port authorities and appoint Completed and report submitted June 2000 Research Assistant 2 Review sampling methods Completed and report submitted June 2000 3 Develop sampling protocol Completed and report submitted June 2000 4 Develop database Completed and report submitted June 2000 5 Prepare sampling equipment Completed and report submitted June 2000 6 Implement sampling regime Completed and report submitted September 2001 7 Prepare and complete distribution maps Completed and report submitted January 2002 8 DETR marine research seminar Completed and report submitted January 2002 9 Evaluate present status of alien species Completed and report submitted February 2002 10 Final report Completed and report submitted February 2002

11 4.0 Methods

4.1 Literature review

4.1.1 Non-native species

Definitions Marine species can be distinguished in a number of ways apart from native species. The main catagories and those used in this report are: - • Non-native species is “a species that has been introduced directly or indirectly by human agency (deliberately or otherwise) to an area where it has not occurred in historical times (since 5000 years BP) and which is separate from and lies outside the area where natural range extension could be expected. The species has become established in the wild and has self-maintaining populations”. (Eno et al 1997) • Established species is one occurring in a reproducing, self-sustaining population in an open ecosystem, i.e. in waters where organisms are able to migrate to other waters (Anon. 1996). • Non-established species is one unable to maintain a self-sustaining population without the deliberate intervention of man. • Cryptogenic species is one that is not demonstrably native or introduced (Carlton 1996). This category is often omitted when assigning a status to species (Eno et al 1997). As Carlton (1996) showed, in San Fransisco Bay up to 33% of species can be cryptogenic. This category must result in an underestimation of the numbers of non-native species present. • Pest species in relation to its use in this report is twofold. Used by the Australian group , these are species listed on the Australian Ballast Water Management Advisory Council’s (ABWMAC) schedule of introduced pest species. For this report a pest species is described as a marine species, native or non-native, that poses a threat to the ecology of the marine environment, health of the human population or economics of a state.

The status of any species may change within these definitions. For example, at the beginning of this report the brackish water mussel Mytilopsis leucophaeta was present in Cardiff docks. However, it was not, at that time, considered to be established. Now, since it has been shown to have a self sustaining population it is included in the list of established non-native marine species in British Waters. (See web-site: http://www.jncc.gov.uk/marine)

The UK situation The review and directory of non-native marine species in British Waters carried out for the JNCC (Eno et al, 1997) derived information from a questionnaire distributed to British marine biologists. The resulting publication provided the UK with a greater knowledge about their particular situation regarding non-native marine species than is available to many other countries, particularly those in Europe. Following this work in the UK, a number of countries have, or are still in the process of producing a desktop survey of similar format, listing non- native species. e.g. Germany: Nehring & Leuchs 1999. Holland: due 15 June 2000 (Wolff, W. J. pers. comm) and the Baltic Sea: Olenin & Lappäkoski 2000.

The focus of most studies in the literature of non-native species is often confined to a particular species (e.g. Styela clava: Coughlan 1985, Ficopomatus enigmaticus:Thorpe 1994, Sargassum muticum: Boaden 1995) There are no examples of wider sampling programmes looking for all or at sub-groupings of non-native species. Most of the reports on the occurrence and distribution of non-native marine species are compilations of opportunistic observations. Consequently, although the presence of non-natives in UK waters may be reasonably well documented the actual distribution patterns of individual species remains unclear. Some of these records may include specific reports from ports but there have been no comprehensive studies on the flora and fauna particularly related to ports and harbours.

4.1.2 Sampling methodology

12 Within the UK, or indeed the OSPAR region, there are no regulated or systematic protocols for the reporting of non-native species, nor is there any information or advice on actual survey methodology relating non-native species.

Most of the survey procedures in the UK outlined in the recent literature deal with the ongoing monitoring of community structure and changes, over time, in biotopes. (Hiscock & Hoare1975, Coulson et al 1980, Holme & McIntyre 1984, Howson et al 1994, Brazier 1998, Dalkin & Barnett 1998, Elliott et al 1998, Hartnoll 1998. Hill et al, 1998, Hiscock 1998, Hughes 1998, Murrey 1998, Worsfold 1998, Wilding et al 1999). Other survey methods are usually restricted to those used when working with an individual species, (Atkinson & Parsons1973, Thorpe 1994, Fox et al 1999).

However, it is evident that many sampling techniques are universal and exploit common methodology and equipment. Some of these methods are applicable to ports e.g. fixed settlement panels,line transects. However, some that require good visibility, no shipping or areas of little or no disturbance e.g. diving are difficult and sometimes impossible to emulate in a busy port or enclosed dock. A summary of the most frequently used methods are outlined in Table 1 below.

Table 1: Summery of common sampling techniques:

See Methods Sample area Organism types Sample techniques Section 4.10 Intertidal Hand nets, traps, Hard substrata Fish & mobile invertebrates anaesthetics, visual h & i search Surface examination. Crabs & other mobiles. Manual search i Macroalgae/invertebrates Abundance, quadrats/pin frames, weed washing Fixed settlement panels/ Encrusting / sessile invertebrates e & f destructive scraping Sediment cores (various Soft substrata Micro-flora, meio + macro infauna sizes and depths) and c & k sieves Sub-tidal Beam trawl, video, Fyke Fish & mobile invertebrates nets, traps, floorless pop h nets Crabs & other mobile Hard substrata Baited traps, beam trawl h invertebrates Collection of stones, Encrusting / sessile invertebrates bottles etc./fixed e settlement panels Sediment cores, diver operated suction cores, Van Veen or Day grabs d & k Soft substrata Micro-flora, meio + macro infauna (less compacted), Hamon or Shipek grabs (compacted sample). Plankton Water bottles Zoo & phyto plankton a & b Mesh net samplers

The European literature provides very little information on the methods suitable for port sampling. A PhD thesis has been published in the UK on mapping and monitoring in ports using restricted, but standard, techniques (Howe 1998). Worldwide, Australia and the USA have developed an awareness of the significant impacts non-native species may have on local

13 communities and economies. Much of this work dates back to Carlton, whose initial work was on the impacts of non-native species on San Francisco Bay (Carlton 1979). He has subsequently published extensively in the field and a number of his students have carried out investigations elsewhere in the USA and also in Australia. Like North America, Australia has had a number of problems with the introduction of non-native marine species that have impacted on their local environment and economy, the reasons for which have been widely discussed (Carlton 1989, Carlton & Geller 1993 etc.)

Recognising a need for research on non-native marine species, the Federal Government of Australia, through the offices of Australian Quarantine and Inspection Service (AQUIS), provided funds for the establishment of the Centre for Research on Introduced Marine Pests (CRIMP). The Centre was established in 1994 and is now located within CSIRO Marine Research at the CSIRO Marine Laboratories in Hobart, Tasmania. Their objectives are: • To develop and promote the application of techniques for earlier detection, more accurate prediction of impacts, and effective assessment of risks and costs associated with marine pest species introduced into Australian waters. • To develop new methods or improve existing measures to control the introduction and spread, and minimise the impacts of exotic marine pest species.

The Centre has produced the CRIMP port survey report series, outlining methods and results on each port surveyed (e.g. Hewit et al 1997, Hewitt et al 1999 ) and the CRIMP technical report series on particular species (e.g. Clapin and Evans 1995). The surveys are ongoing and a legal requirement for all ports in Australia handling international shipping.

The Australian project uses a target approach to their surveys.(See section 4.5 for description) Surveys are designed to maximise the likelihood that these target species will be detected by concentrated sampling on habitats and sites in the port and adjacent areas that are most likely to have been colonised by these species. Methods used ensure a comprehensive coverage of habitats, providing presence/absence information and/or semi quantitative indices of abundance.

Details of individual methods are outlined in further detail in the report (Hewitt & Martin 1996), and a summary is outlined in Table 2 below.

Table 2: Sampling techniques recommended by Hewitt & Martin1996:

HABITATS Soft Hard Seagrass/ Plankton/ Beach Sampling technique substrata substrata macroalgal nekton wrack Small cores X Large cores X X 20um mesh plankton nets X 100um mesh drop nets X Traps X X X X Qualitative visual surveys X X X X Quadrat scraping X Beam trawl / benthic sledges X X Poison stations X X X Beach seines X X X

14 4.2 Ports In choosing the ports for this survey an attempt was made to select those that were representative of the range conditions encountered found in the UK. The factors taken into account in the choices made were: • Size – physical dimensions and complexity of dock system • Positioning - estuarine or coastal • Siting – adjacent to special areas (SSSI’s, SAC’s, SPA’s etc) • Commercial type - cargo/ship type and volume of traffic • Ballast Water discharge - quantity and origin

Figure 1: Map of the UK showing the six ports used in this survey

Table 3: Habitat types at each port Port Habitat types Liverpool Enclosed docks, estuarine, high salinity Cardiff Enclosed docks, low salinity Milford Haven Open water, various tidal shore types Southampton Open water, low energy, sedimentary shores Felixstowe Estuarine Teesside Estuarine

Liverpool Liverpool port was the world's first enclosed dock system, built in 1710 on the banks of the Mersey Estuary. The Port of Liverpool, owned and operated by the Mersey Docks and Harbour Company, is Britain's most central deep water port. There are 19 docks on the north side of river and 6 on the south. Water levels are kept relatively constant by a system of reservoirs, pumps with locks to the River Mersey. The sandbanks of the surrounding estuary are highly mobile and training walls have been introduced to help channel the tidal flow. The Port of Liverpool is one of Europe’s top 10 container ports and handles nearly 30 million tons of cargo a year through a comprehensive range of terminals and facilities with shipping services linking to most areas of the world. Liverpool is the UK's major port for imports of grain and for exports of scrap metal, but is also a key port for timber and forest products, crude oil and other fossil fuels, and has a growing role in other bulk liquid and bulk solids. It also handles almost 25% of all container traffic crossing between the UK and N.America.

15 Figure 2: Map showing the port of Liverpool and the sampling sites.

16 Cardiff The Port of Cardiff is situated on the north shore of the Bristol Channel at the mouth of the River Taff. Cardiff is an artificial dock area enclosed by lock gates with limited access according to the tidal regime. The dock area is very sheltered, with freshwater input from runoffs, rivers and streams and dedicated feeders to maintain the level of water within the dock. The dock bottoms are of very fine sediment, with some subtidal and intertidal areas composed of hard substrate, such as dock walls and quays. The westerly position of the port gives sheltered accommodation suitable for vessels up to 35,000 tons dry weight

Cardiff was one of the smaller ports within the sampling programme, with three docks, 14 open storage berths and 26 hectares of storage space, it is only a tenth of that available at the Port of Liverpool. Among the port's principal traffic are various dry bulks including grain, coal, coke and sand, petroleum products, timber and steel and has regular container services between Ireland and the Mediterranean.

Figure 3: Map showing the port of Cardiff and the sampling sites.

17 Milford Haven Milford Haven is the fifth largest port in the UK, handling over 41 million gross tons of shipping and over 10,000 ship movements in 2000. It is only one of a few UK ports to report the discharge of ballast water originating from outside continental Europe with over 50,000 tonnes of ballast water discharged annually (Laing 1995).

The Port of Milford Haven is situated on the southwest coast of Wales and is enclosed by natural geographical features. It is an open water port with little freshwater input. The wide variation in the rocky coastal topography and water movements support extremely diverse, abundant and important biological communities. Habitats range from soft sediments to rocky shores. Milford Haven is one of the best examples of a drowned river valley (ria) in Britain, with large beds of eelgrass (a marine flowering plant), and sheltered reefs occupied by rich sponge communities. The area within the Milford Haven Waterway and around the Pembrokeshire Islands has been selected as a Candidate European Special Area of Conservation (under the EC Habitats and Species directive) with over 12 SSSI’s and a Special Protection Area (under the EC Birds Directive).

Figure 4: Map showing the port of Milford Haven and the sampling sites.

18 Southampton Southampton is a natural deep-water harbour with a famous double tide, providing 17 hours of rising or standing water every 24 hours. It is below the limit of the last glaciation and is therefore not troubled by large quantities of mobile sediment and consequent dredging costs. The harbour is lock-free, with a maximum tidal flow of 2 knots and a tidal range of 4.5 m, the main approach channel was recently dredged from 10.2 m to 12.6 m below chart datum.

The Port of Southampton is situated in the center of the UK's South Coast and is only 28 nautical miles from the main international shipping lanes.

Southampton is one of the major UK ports, handling 35 million tons of exports every year, which are about 7% of the UK's total. They are the UK's largest vehicle handling port, with a throughput of 516,000 vehicles in 1997, and is one of the UK's leading fruit handling ports, managing over 350,000 tons each year. Ships discharge over 50,000 tonnes of ballast water annually (Laing 1995)

Figure 5: Map showing the port of Southampton and the sampling sites.

19

Felixstowe The Port of Felixstowe is situated in the south east of the UK at the mouths of the Stour and Orwell estuaries. Felixstowe has the largest container handling facility in the UK and the longest continuous quay in the British Isles. It handles over 115,000 containers each month, a figure unsurpassed by any other UK port. The port handles worldwide traffic, importing slightly more than exporting. Total volume in 1997 was almost 30million tonnes.

The main navigation channels and berths have depths maintained for a minimal tidal influence upon its whole range of ship operations. In response to increasing numbers of the very latest deep draught vessels, Felixstowe has initiated through Harwich Haven Authority a program of further deepening of channels and berths.

Figure 6: Map showing the port of Felixstowe and the sampling sites.

20 Teesside Tees and Hartlepool Port Authority is the statutory harbour authority for the ports of Tees and Hartlepool handling over 50 million tonnes of cargo a year. The port of Teeside is situated near the North Sea coast at the mouth of the River Tees. The whole area of Teesmouth Flats and Marshes is a wetland Area of International Importance under the Ramsar Convention, and is also a ‘Special Protection Area’. It is an internationally important wildlife site with three SSSI’s within the Tees estuary.

Tees Dock is a deep-water tidal inset facility within Teesport. It is located on the south bank of the River Tees, 5km from the sea. In addition to Teesport there are numerous wharf and jetty operators on the banks of the River Tees itself. Tees dock handles almost five million tons of cargo annually. Large volumes of steel, project, bulk, and unitised traffic dominate this. However, grain, bulk minerals and chemicals are also important components. There are regular sailings to many destinations worldwide, including Europe, Scandinavia, Africa, the Far East, and Australasia. It was one of the ports in this survey to report discharge of ballast water originating from outside continental Europe, with ships discharging over 50,000 tonnes of ballast water annually (Laing 1995).

Figure 7: Map showing the port of Teeside and the sampling sites.

21

4.3 Buoys In addition to the six ports selected for this project, Trinity House was approached and gave permission to board the buoy maintenance vessels THV Mermaid and THV Patricia. Navigation buoys, after a period of up to three years at sea, were sampled immediately after they were taken on board the maintenance vessel, prior to cleaning, repair or replacement. The buoys sampled in this way (as listed in Table 4) are marked on the port maps of Liverpool (Figure 2), Milford Haven (Figure 4) and Felixstowe.(Figure 6).

Table 4: Names and ports of buoys sampled

Port Buoy Liverpool C2 Milford Haven 5A Milford Milford Shelf Esso Stack Behar Mill Bay East Chapel Rat St Annes Rows Rock Sheep Felixstowe No3 No4 Cross East Massac West Massac South Threshold

4.4 Database Information on the transfer of non-native species by all means is currently collated by the Working Group on Introductions and Transfers of Marine Organisms (WGITMO) set up by the Advisory Committee on the Marine Environment (ACME) of the International Council for the Exploration of the Sea (ICES). Within the UK, the JNCC operate a national database concerned with non-native species, but for the purposes of this project, all of the data collected has been put onto a custom made database that has been created using Microsoft Access 97 and 4.5. linked to MapInfo.

Figure 8: Opening screen of project database.

22 All data from the port surveys has been added under the relevant port, site and method of collection. This data can then be retrieved in various forms and used to produce, for example, species lists for a particular site and/or port, data to produce graphs and maps to display species distribution and abundance.

The information currently on the database has been collated in this report for each port in the following way: • Species lists of native and non-natives at that port in alphabetical order. • Maps showing presence and abundance of non-native species collected during port sampling. • Graphs of average temperature and visibility at each site within that port. These data are presented in Appendices B-G.

4.5 General approach on port surveys The main aim of the research programme was to select and test port sampling methodology and, in the process, provide a baseline listing of the species (native as well as non-native) within the study areas. Two general objectives can be defined to accomplish port surveys, the first, attempts to define the populations of plants and animals by sampling and identifying all the species encountered. These methods were employed in this study. The second method sets as its objective the search for and recognition of a restricted list of so-called ‘target’ species.

4.5.1 Target Species The application of this approach to biological surveying was proposed by Hewett & Martin (1996) and should provide a more cost effective means of data collection. To provide a comparison with the methodology applied in this study a list of target species was drawn up for the UK.

From the Non-native species list compiled by Eno et al (1997) a secondary list was produced containing those species which have been recognised as ‘pest’ species in the UK (See 4.5.1.1). A second list of those species which, on the basis of their invasive history, appear able to become established in UK waters and also have been designated ‘pest’ species in their current biological range was developed (See 4.5.1.2). Both lists were circulated to interested parties in the USA and Europe with a request for their comments and additions. The final target species list was composed of four elements • Non-native species that are present in the UK and are already classed as a ‘pest species’. • Non-native species that are major pests in overseas ports and which, on the basis of their invasive history and projected ship movements might be expected to colonise UK ports. • Known or likely non-native marine species in UK waters that currently are not assigned pest status. • Native UK species that another port state (only Australia at this time) has on its list of pest species.

The final list would need to be amended on the production of target species lists by other national port monitoring authorities.

23 4.5.2 Suggested Target Species List for the UK

4.5.2.1 Target Introduced Pest Species reported in the UK

Coscinodiscus wailesii Diatom

Gyrodinium mikimotoi Dinoflagellate Sargassum muticum Brown alga Undaria pinnatifida Brown alga Anguillicola crassus Nematode (parasitic) Marenzelleria viridis Polychaete annelid Eriocheir sinensis Decapod crustacean Crepidula fornicata Gastropod mollusc Urosalpinx cinerea Gastropod mollusc Styela clava Urochordate (sea squirt)

4.5.2.2 Marine Pest Species that pose a threat to the UK

Alexandrium catanella Dinoflagellate Gymnodinium catenatum Dinoflagellate Chattonella sp. Raphidophyte Fibrocapsa japonica Raphidophyte Mnemiopsis leydyi Ctenophore Cercopagis pengoi Branchiopod crustacean Dreissina polymorpha Bivalve mollusc

Teredo navalis Bivalve mollusc Hemigrapsus penicillatus Decapod crustacean Rapana venosa Gastropod mollusc Neogobius melanostomus Fish Ocinebrellus inornatus Gastropod mollusc

4.5.2.3 Known or likely Non-native marine species in UK waters that are not currently assigned pest status.

Thalassiosira punctigera Diatom Thalassiosira tealata Diatom Heterosigma akashiwo Raphidophyte Odontella sinensis Diatom Pleurosigma simonsenii Diatom Agardhiella subulata Red alga Antithamnionella spirographidis Red alga Antithamnionella ternifolia Red alga Asparagopsis armata Red alga Bonnemaisonia hamifera Red alga Grateloupia doryphora Red alga Grateloupia filicina var. luxurians Red alga Pikea californica Red alga Polysiphonia harveyi Red alga Solieria chordalis Red alga Colpomenia peregrina Green alga Codium fragile ssp. atlanticum Green alga Codium fragile ssp. tomentosoides Green alga

24 Spartina anglica Angiosperm Haliplanella lineata Anthozoan cniderian Gonionemus vertens Hydrozoan cniderian Clavopsella navis Hydrozoan cniderian Clymenella torquara Polychaete annelid Ficopomatus enigmaticus Polychaete annelid Goniadella gracilis Polychaete annelid Hydroides dianthus Polychaete annelid Hydroides ezoensis Polychaete annelid Janua brasiliensis Polychaete annelid Pileolaria berkeleyana Polychaete annelid Ammothea hilgendorfi Pycnogonid Eusarsiella zostericola Ostracod crustacean Acartia tonsa Copepod crustacean Balanus amphitrite Cirripede crustacean Elminius modestus Cirripede crustacean Corophium sextonae Amphipod crustacean Rhithropanopeus harrisii Decapod crustacean Aulocomya ater Bivalve mollusc Crassostrea gigas Bivalve mollusc Ensis americanus Bivalve mollusc Mercenaria mercinaria Bivalve mollusc Mytilopsis leucophaeta Bivalve mollusc Petricola pholadiformis Bivalve mollusc Mya arenaria Bivalve mollusc Tiostrea lutaria Bivalve mollusc Potamopyrgus antipodarum Gastropod mollusc

4.5.2.4 Native UK Marine species that appear on other port nations target species lists (Australia)

Sabella spallanzanii Polychaete annelid Carcinus maenas Decapod crustacean Mytilus galloprovincialis Bivalve mollusc

4.6 Cysts Coastal sediments frequently contain the resting stages of various planktonic organisms. Particular attention has been paid to the occurrence and distribution of dinoflagellate cysts, a number of which are capable of forming harmful blooms. There is evidence which suggests that the recent paralytic shellfish poisoning (PSP) outbreaks in Australia may be due to non- native dinoflagellate species introduced as cysts by ships arriving at ports in Australia (Hallegraeff, 1995). Surveys of the occurrence of dinoflagellate cysts in ships ballast tank sediments suggest that ships may transport and discharge large numbers of viable dinoflagellate cysts into UK waters (MacDonald and Davidson, 1998; Hamer et al, 2000; 2001).

4.7 Information sources 4.7.1 Government agencies Various departments were contacted (Environment Agency, JNCC, CCW), either for information and/or assistance with sampling. This was done in an attempt to pool resources and elicit the maximum amount of information to aid this project.

4.7.2 Marine groups There are various marine groups within the UK that are able to provide information, samples and assistance. The Porcupine Natural History Society was contacted to assess if they could be of assistance with information on the distribution of non-native species. Other groups and individuals working on non-natives in the UK were also alerted to our work via letter or e-mail.

25 The ‘Sea Angler’ magazine, via the internet, was also asked for information, as eel capture by the project had been unsuccessful and animals were required for analysis for the presence of the non-native parasite Anguilicola crassus.

4.7.3 General public As the general public, and in particular nature groups, field study groups and specialist species groups frequent sites on a regular basis and have extensive knowledge of local fauna and flora it was felt that any attempt to elicit their aid would be a positive one. As a result, public talks were given, in an attempt to utilize local knowledge. A small identification booklet was designed, printed and given out at such events (See Appendix L).

4.8 Sampling Carrying out biological surveys in commercial ports imposes special restrictions not encountered in surveys on non-impacted marine environments. In particular the inherent commercial activity, ship loading and discharge and ship movements cannot be hindered and also present considerable safety concerns. Shipping activities in themselves may also produce further environmental problems, for example, increased turbidity due to propeller action. Initially the experimental design, sampling procedures and data management looked to those described in the CRIMP report (Hewitt & Martin 1996). These methods are clearly defined, repeatable and have been shown to work under operational conditions. These procedures are also being used by the IMO currently carrying out The Global Ballast Water Management Programme (GloBallast) (S.Raaymakers pers comm), a three year, $10.2 million initiative under the International Waters portfolio of the Global Environment Facility (GEF). Funding from the GEF is deployed through the United Nation’s Development Programme (UNDP), to allow the IMO to assist developing countries in tackling the transfer of harmful aquatic organisms in ship’s ballast water and also to assist these countries implement their own guidelines on ballast water management. The present project has taken the methods described by Hewitt and Martin (1996) and adapted them where necessary to suit UK ports, specific site conditions, manpower and financial constraints.

Initially, each port was studied using relevant Admiralty charts and potential sampling sites that were considered to be representative of the different marine and maritime habitats within the port boundaries were highlighted for exploration. Meetings were then arranged with the relevant port authorities and individuals to allow for consultation on the nature of the survey work, access to the proposed sites, health and safety considerations and security. In some cases the Port Authority were the sole group present as all sites for sampling were within their jurisdiction (e.g. Cardiff) in other cases, (e.g. Milford Haven) both Port Authorities and individual companies operating from the port were included in the consultation process.

Each port was then visited, habitat types explored and the proposed sites viewed. Port authorities were then re-contacted to confirm permission for sampling at the specified sites or to propose and agree alternatives if the preliminary choice proved to be unsuitable. Two port authorities, Southampton and Teesside, also offered the use of their survey boats if required, these proved to be of great value, enabling visits to be made to sites not easily accessible by land. The local knowledge of the boat crews was also invaluable.

In order to attempt to cover all habitat types, the following type areas within the ports were investigated.

Commercial areas ‰ active and inactive wharves/berths & piles ‰ sediments ‰ buoys

Active non-commercial / Recreational areas ‰ marinas ‰ recreational berths & piles ‰ adjacent brackish water/estuarine areas/intertidal sites ‰ rock jetties/groynes

26 Other areas ‰ Water column

A variable number of sites at each port were sampled, each site being judged to be a representative of the whole range of habitats within that port area. Maps of each port with the sites marked have been produced and can be found in the Appendices relating to each port (Appendices B – G) Methods used at each site are in Appendix H.

All ports, with the exception of Liverpool, were fully sampled twice between March 2001 and August 2001. Liverpool was sampled each month, to give a clearer picture of the shorter time scale differences in species numbers and occurrence.

Table 5: Sampling dates for each port.

Port Date(s) sampled (2001) 17 – 19 April Teeside 4 – 6 July 30 April – 2 May Milford Haven 25 – 27 July 14 & 15 May Cardiff 12 – 13 July 29 – 31 May Southampton 1 – 3 August 26 – 28 March Felixstowe 13 June 5 & 6 March 11 & 12 April 21 & 22 May 18 & 19 June Liverpool 30 & 31 July 14th August 28 & 29 August 3rd September

4.9 Preliminary surveys and equipment testing Prior to the implementation of the full sampling regime, methods and equipment were trialled at Liverpool and Teesside, since these ports involved the shortest travelling times. Sampling methodology used is detailed in section 4.3. The equipment outlined below, when evaluated, often needed some physical modification or adjustment in the method of deployment.

Two people worked on this project during the sampling period; travelling to sites, deploying and collecting equipment and samples and identifying species. Attempts were made to ensure that all the equipment can be operated and can be deployed by one person, either from a manmade structure such as a pier or, over the side of a boat. Risk assessments required by Health & Safety legislation, however, prohibited work to be carried out alone.

4.9.1 Settlement Panels In an initial test deployment, three tier panels were put down at various sites within Liverpool and Teesside (July 2000). Each tier was positioned to lie just below the high tide mark, at mid tide and at low tide level at Teeside and at 1m, 5m and 10m below the maintained water level in the docks at Liverpool. Positions on the dock-sides were chosen which were considered to be representative of the various habitats within each port and had structures to which the panels could be attached (See table 6). Panels that were still present were collected in November 2000 (See appendix I for details on deployment and retrieval dates and of those panels lost). The panels positioned just below the high water mark at Teesside showed no signs of settlement and thus were modified to two tiers. In Liverpool, the lack of difference between panels at 5m and 10m and the abundance of

27 organisms on the panels close to the surface also led to their modification to two tiers at 1m and 3m.

Table 6: Initial test sites for panels

Port Site Attached to Habitat Liverpool Gladstone dock Dock wall Intake Dock Liverpool Pier Head Pier Estuarine Liverpool Canada Dock wall Central in the Dock system Liverpool Westfloat Dock wall Dock on the south of the river Teesside Philips Petroleum Pier Mouth of estuary Teesside Bran Sands Buoy Coastal Teesside Buoy 25 Fixed buoy structure Estuarine Teesside Billingham Wharf Upper reaches, lower salinity Teesside Tees & Hartelpool Port Jetty Central dock area, Authority estuarine Teesside Government Jetty Jetty Coastal

Two sites were not re-used. • Pier Head - Within the estuary proper at Liverpool, the current was strong and the whole structure of the pier rose and fell with the tide. The equipment was continually dragged by the current and became entangled in dangerous, inaccessible places. Were areas similar to this to be sampled in the future, fixed panels rather than suspended panels would be required. • Bran Sands - The panels were attached to the only structure at the mouth of the estuary. However, this produced too much drag, thus affecting the position of the buoy itself.

4.9.2 Van Veen Grab After testing this device in Liverpool and finding a low recovery rate, due to the small volume of sediment collected, benthic organisms were sampled by means of the dredge. The grab did, however, provide sufficient sediment for cyst analysis.

4.9.3 Dredge In order to test how best to retrieve a sediment sample using dredges a small survey boat was launched into Collingwood Dock at Liverpool. Several dredge designs were tested with different sizes of mesh used for the collection bag (See 4.10c for design used). The boat pulled the equipment initially at 2 knots across the length of the dock. This was found to be too fast, with the apparatus bouncing along over the bottom rather than digging in and collecting sediment. At a reduction to 1.4 knots all the dredges collected samples. The volume of sediment collected by each trawl averaged 10 litres. Such a volume of sediment took almost a day to analyse. A comparison of the numbers and types of organisms collected when the dredge was towed by the boat proved to be the same as those collected when the dredge as deployed from the shore.

28 4.10 Sampling methodology a. Phytoplankton & b. Zooplankton nets Both the phytoplankton net (20um mesh free-fall drop net) and the zooplankton net (100um mesh free-fall drop net) were mounted on a 50 cm ring to which the towing bridle was attached. The nets were weighted to achieve a fall rate of approx 1m per second. Each drop was made to a depth 0.5m above the bottom and a single drop was performed at each collection site. The nets were hauled up vertically through the water column at a speed of no more than 0.3m per minute. This method ensured a fully integrated sample from the whole of the water column. The water sample collected was then poured, using a funnel, through a 5mm mesh, to collect macro plankton, and then put into a labeled bottle and preserved (See 4.11 preservation techniques) for later analysis. In most cases it was necessary to deploy the nets from the side of a dock or pier. Initially there was a tendency for the user to scrape the dock wall or man made structure and collect unwanted material therefore a metal stand and pulley system that held the net 1 m away from the wall was constructed. Sampling from a boat did not require this extra equipment.

Figure 9: Deploying the phytoplankton net with the pulley system at Teeside.

c. Dredge from dock side: A hand deployed dredge was used to collect sediment. The dredge was constructed from a length of heavy gauge steel pipe 30 cm in diameter and 25 cm long. A collecting bag made of 1mm mesh was attached to one end. A central handle, protruded from the other end, to which was attached a rope, weighted at 1m and 3m. These weights ensured that the equipment settled on its side on the sediment. The dredge was pulled for 3m giving a resultant volume of sediment of approximately 2.5 liters in volume. This sediment was transferred into a container and on return to the laboratory sieved through a 0.5mm mesh. Organisms were picked by hand from the sieve surface, preserved and subsequently identified.

Figure 10: Benthic dredge

29 Figure 11: Benthic dredge in use at Cardiff docks

d. Grab: 0.25m3 Van Veen hand held grab

Three benthic samples were taken from one undredged site within a port and kept in cold storage until prepared for cyst analysis (See 4.11.1 cyst analysis).

Figure 12: Van Veen grab e. Settlement Panels: Figure 13: Two tier settlement panels

Settlement panels are clean surfaces placed into the water for a period of time, offering a site for settlement to encrusting organisms.

The slates used here were cut from Welsh roof tiles. Three 6” x 5” quadrats of slate (½” apart) were attached at each level to 6mm ply by stainless steel washers and screws. Two household bricks weighted the panels so that they would hang vertically in the water

Two sets of settlement panels were deployed at designated sites at all ports. One set was removed every 3 months. When collected, the panel was wrapped in wet newspaper and placed in a large plastic bag before being returned to the laboratory for analysis. Animals would survive for about 12 hours in these conditions and were then stored ‘live’ in running seawater prior to identification, as, in some cases e.g. anemones, delicate animals were easier to identify in life.

30 At ports where there is a tidal regime (Milford Haven, Southampton, Felixstowe and Teesside), the panels were set to hang in the water at the mid tide level and just below the low tide level. In docks where the water level remains relatively constant (Liverpool and Cardiff), panels were placed at one and three metres below the waters surface respectively. (Appendix I shows the number and dates each panel was deployed and retrieved).

In the laboratory, slates were removed individually from the wooden board and placed in a shallow tray of sea water so that the organisms could be identified under a stereomicroscope. Initially, all species were counted to provide quantitative data. However, as the number of species found increased, this protocol became too time consuming and after evaluation it was decided to note only the presence of native species but to continue to count all examples non- native species. At the Westfloat dock in Liverpool, an additional set of panels was removed and replaced each month to permit examination of the annual pattern of settlement. f. Destructive sampling

Figure 14: Scraper The destructive scraper was used to collect flora and fauna attached to hard substrates and those mobile organisms that were living amongst them.

A large net was modified by the addition of a metal scraper (arrow). The metal scraper was 50cm wide and was dragged upwards across a hard surface for 50cm. Samples were taken from an area 0.5m below the water level. Anything on the sampled surface fell into the net and was then transferred to an appropriate container and preserved for analysis in the laboratory.

All organisms collected using a destructive scraper were Trinity House allowed access to theiridentified vessels and THV counted Mermaid if they andwere THVnon-native Patricia species. and accommodated our work during their normal buoy maintenance. The buoys sampled had been in the water for up to three years. Once a buoy had been lifted onto the deck of the ship and all lifting gear had been removed, samples could be taken.

Figure 15: A 0.25m2 quadrat used on buoys

A small hand scraper was used when sampling off-shore buoys. A 0.25m2 quadrat was randomly placed against the side of the buoy and the contents scraped off and placed in a container. Three quadrats were taken from the water mark and three from the base (approx. depth ~2m). These were immediately preserved for later sorting and identification.

31 Figure16: The base of a buoy after samples have been taken

g,h. Traps

Figure 17: Shrimp and Crab trap

A. a Shrimp trap, constructed from plastic drain pipe whose ends were fitted with a fine mesh provided with an inverted cone entry port. B. the Crab trap, constructed of 25mm mesh plastic mesh, with an inverted cone entrance.

The traps were baited with mackerel, weighted and left in the water overnight. On retrieval, all animals were identified, counted and returned immediately.

A specialised, commercial, Fyke net was also deployed to trap larger fish and eels. This was set over night but unbaited. i. Visual searches. Intertidal: A transect line was laid from the high shore to the waters edge at low water. Three quadrats were thrown randomly at 5 equidistant stations along the transect and the organisms within them identified. On a rocky shore organisms were counted or percentage cover noted, at sandy and muddy sites the sediment within the quadrat was dug to a depth of 5cm and sieved through a 2mm mesh. Species were identified in the field as far as possible and left there, only those that could not be identified were preserved and returned to the laboratory . j. Visual searches. Sub-tidal: In order to compare the efficiency of destructive sampling with visual sampling by divers, Collingwood Dock in Liverpool was surveyed by SCUBA divers. The dive team consisted of four HSE qualified divers who were also experienced marine biologists and were accustomed to this type of survey work. Only one dock was surveyed in this manner because of the expense. Two survey dives were conducted with written and photographic records being made. All surveys had to be conducted with minimum disturbance to the fine sediments at the base of the dock which would have substantially reduced the visibility and adversely affected the results.

Six sample stations were established on the vertical rock habitats of the dock. Three of these sample stations were at intervals along the base of the dock wall at 4.5 m below surface level (bsl), half a meter above the bottom. Three shallower stations were also established above

32 these at 2.5 m bsl (just below the zone accessible from the surface with the sampling net). Two parallel weighted lines were suspended from the dock-side in three locations. From these lines a 3 m long survey pole was suspended horizontally at the two target depths. For widely dispersed species, counts were made in an area 0.5 m above and below the pole, ie a strip 1 x 3 m in dimensions. For more common species that would be prohibitively difficult to count in a larger area, a 0.5 x 0.5 m gridded quadrat was suspended from the pole and three replicate counts obtained. Each of the grids on the quadrat measured 0.1 x 0.1 m and for certain species at high densities, counts were made at this spatial scale, selecting squares at random from the 25 available. For some other species, counting was not possible. This was the case for Ciona intestinalis that occurred at such high densities and with a sufficient range of sizes that only percentage cover could be satisfactorily used. Indeed, for counting some of the species in quadrat sub-squares, the Ciona intestinalis had to be carefully removed first. Other species were cryptic in which case they were recorded as present or absent in quadrats.

On the second survey dive a strip of the sediment benthos was surveyed from the base of the wall towards the centre of the dock (between 5 and 6 m bsl) to survey the surface of the benthos. Here, the survey area was delineated using a modified version of a technique used by Wilson (1994) and subsequently by Sanderson (2001). A shot line was deployed from which divers reeled-out a 10 m measured line from a pole. The pole was designed to measure a fixed width and therefore, in combination with the fixed 10 m traveled, quantify the area surveyed. The pole length was chosen to be 3 m because the surveyors were advised that the visibility was unlikely to be worse than 2 m (each diver would need to be able to survey the area on one side of the pole and see one-another).

Additionally, records were made of a few species that were not encountered during actual survey work. These were scored as present and were observed whilst travelling to and from the sample station.

Figure 18: Typical photograph obtained of the subtidal community in Collingwood dock, Liverpool

33 Figure 19: Underwater photograph of the non- native sea squirt, Styela clava

k. Corer: A hand deployed microcorer (Ocean Scientific International) was used initially for the collection of replicate sediment samples for the study of dinoflagellate cysts. It was designed to be used from small boats or waterside fixtures without requiring a winch. Its total weight was 6.4 kg and it took a single core of 59mm diameter up to 150mm long. Unfortunately modifications had to be made from the outset but it subsequently proved to be unreliable. Therefore sediment for this part of the survey was collected by the hand operated grab. A corer of this type would be valuable if a historical record of cyst deposition was required. However, within a dock/port area it was rare for a site not to have been dredged or recently disturbed by ship movement, such sites were only found where commercial activity had ceased.

4.11 Preservation and Curation Wherever possible, specimens were identified in situ. Where this was not possible samples were fixed in formalin and returned to the laboratory, washed and then stored in alcohol prior to identification. Algae were stored in formalin to prevent colour loss, or pressed and dried.

All containers used for the collection of species were plastic and had fitted lids. Retained species were transferred to glass specimen tubes for long term storage.

• All Formalin used was 4% concentration. (100ml of 40% stock formalin diluted with 900ml of seawater.) • 70% Alcohol used for long term preservation of samples • Lugol’s Iodine (iodine in potassium iodide) for phytoplankton

Appropriate HSE directives were followed when using formalin, especially with respect to the washing out process of this preservative prior to long term curation.

A reference system was designed and specimens stored for quality assessment. The reference system used was: e.g. L110401ZG

L = the port name Liverpool 110401 = date 11th April 2001 Z = sample type zooplankton G = site Gladstone dock

34 4.12 General Analyses 4.12.1 Cyst analysis In developing a sampling methodology for dinoflagellate cysts in port sediments, an appreciation of small-scale variability in cyst distributions is important. Because of their nature, enclosed dock complexes such as Liverpool and Cardiff have the potential to vary significantly in species occurrence and abundance between docks. This study focused on the cyst assemblages present in the sediments of various docks within the Liverpool dock complex and aimed to determine whether assemblages vary significantly between docks and if so, what implications did this have for sampling studies.

Replicate (n = 3) sediment samples were collected using the grab from the sampling sites at Liverpool port (Fig. 2). In the laboratory, sediment samples and concentrated cyst samples were stored in the dark at 4ºC until sample analysis. Total dinoflagellate cyst concentrations for each site were determined by sieving replicate sediment samples through 100 and 20 µm mesh sieves. The material retained on the 20 µm sieve was resuspended in a known volume of filtered seawater and a sub-sample counted to determine the concentration of full cysts. The relative abundance of cyst types was estimated from concentrated cyst samples prepared using the density gradient centrifugation method of Bolch (1997). The first 300 cysts from each of three replicate samples were identified (as far as possible) and counted on an inverted microscope. Count data was square root transformed before multivariate analysis using Primer (See 4.12 for ‘Primer’).

4.12.2 Statistical analysis A number of routines in the computer software package PRIMER (Plymouth Routines in Multivariate Ecological Research) was used to examine the community structure of a selected number of ports and sites.

35 5.0 Results and Observations

5.1 Introduction The data presented here is only a sub-set of the total information collected for all the ports covered in this project. It refers predominately to the docks within the port of Liverpool. Similar analyses could be drawn up for the other five ports covered in the project. The complete data set for all ports surveyed in this project is held in the database, but the appendices contain collated information as listed in Section 4.4.

5.2 Non-native species collected Table 7 summarises the presence of non-native species collected during the sampling programme at each port. This information has been visually interpreted for each of these species in the form of an abundance map. Figure 22 is one example showing the distribution and abundance for Styela clava. All of these maps can be found in Appendix A.

Table 7: Non-native species collected at each port.

Non-native species Cardiff Felixstowe Liverpool Milford Southampton Teeside Haven Hydroides dianthus x Hydroides ezoensis x x Ficopomatus x x x x enigmaticus Elminius modestus x x x x x x Rhithroponopeus x harisii Haliplanella lineata x x Corophium sextonae x Crassostrea gigas x Styela clava x x x x Sargassum muticum x Potamopyrgus x x x x antipodarum Crepidula fornicata x x x Urosalpinx cinerea x x Odontella sinensis x x x x x

Figure 20: Distribution and abundance of Styela clava

36 5.3 Sampling 5.3.1 Plankton 30 native genera of phytoplankton and 15 of zooplankton were identified in the Liverpool plankton samples. Few of these were identified to the species level. One non-native species, Odontella sinensis, was identified and found in all ports except Cardiff. Its presence within a port was not constant or ubiquitous and it was a minor part of the plankton community (<3%). Although not in every case, many planktonic species of both plants and animals show irregular or annual patterns. In the case of phytoplankton this is related to light and nutrients, which effect division rates of these cells such that there are regular ‘blooms’ in spring and autumn. However, the dominant species in these blooms may change from year to year. Thus, there is a need for long term, regular sampling to establish the full range of organisms present at a site. The zooplankton also shows a seasonal pattern in appearance of species which may be further complicated by a tendency to local patchiness in distribution.

5.3.2 Benthos 5.3.2.1 Mobile organisms The traps used to catch mobile fauna operated successfully. However, as the results for the common shore crab, Carcinus maenas show there is a seasonal component influencing captures (See Figure 23). This demonstrates that there is a need to understand something of the biology of an organism in order to maximise sampling efficiency, or indeed make any successful captures. In this case numbers caught relate to the seasonal reproductive cycle of this species.

Figure 21: Numbers of Carcinus maenas caught in Gladstone dock, Liverpool between March 2001 and August 2001.

Total number of Carcinus maenas caught in Gladstone Dock, Liverpool

12 10 8 6

Number 4 2 0

05/03/0119/03/0102/04/0116/04/0130/04/0114/05/0128/05/0111/06/0125/06/0109/07/0123/07/0106/08/0120/08/01 Date Sampled

Fyke nets, used for catching eels and fish were also operated at Liverpool but yielded no results. However, a local fisherman with detailed local knowledge used the same equipment and collected eels on a daily basis. Eels (Anguilla anguilla) purchased from him were examined for the presence of the non-native parasitic worm Anguillicola crassus but the organism was not found.

5.3.2.2 Sessile organisms Most organisms cannot live successfully on the surface of the sediment if it is continually being disturbed. Many of the sites within ports are commercially dredged, leaving few benthic areas available for organisms to colonise. There are however, areas within some ports that are not dredged and where sessile organisms could be found e.g. approaches, open or coastal waters, intertidal or non-commercial areas. Most of the ports surveyed had an intertidal area that was then surveyed (See methods 4.10i ) but only Milford Haven.possessed easily accessible areas that were undisturbed.

37 Table 8: Dredging summary for all ports with the number of species collected.

Port Number of sites Number of times Number of species trawled trawled Milford Haven 4 2 49 Cardiff 3 2 22 Liverpool 7 6 26 Southampton 5 2 29 Teesside 4 2 6 Felixstowe 3 2 7

The dredge was successfully used to take samples of the benthos in areas where there was no commercial dredging and away from any SSSI sites. Table 8 shows that Milford Haven yielded the greatest number of species even though only four sites were included. This is probably due to the large open water areas where commercial dredging has not taken place. Liverpool had seven sites that were dredged six times and only yielded the same number of species as Southampton (another port with areas that are not dredged but are disturbed to a certain extent by the movement of shipping). Teeside and Felixstowe showed the lowest number of species overall. In these cases the low numbers may because of extensive dredging operations (Felixstowe) and a combination of dredging activity and historically high levels of industrial pollution (Teesside). In the latter example strenuous efforts to restore the river are yet to be fully rewarded because of high levels of pollutants still resident in the sediments.

5.3.2.3 Infauna The port of Milford Haven does not require deepening by dredging, consequently it has large undisturbed areas of sediment. In addition, it also has a continual input of water from the open sea. This port has the largest number of species living in the sediments. Other ports that have little opportunity for sediments to be cleaned by a through-flow of clean oxygenated water and are also continually disturbed by the movement of shipping and by commercial dredging have lower numbers of benthic species (e.g. Liverpool).

As an example of the detailed analysis that can be carried out on benthic collections, the following is a report on the types and numbers of dinoflagellate cysts found in Liverpool docks. Dinoflagellate cysts are particularly significant in that they: a) represent a past bloom of the motile phytoplankton planktonic stages, some of which produce toxins. b) produce viable cysts that have been shown to be present in large numbers in ballast tanks (Hallegraeff & Bolch 1992, Hamer 2000). c) can survive in sediments for many years.

They are therefore of considerable interest when considering potential introduced species. Fortunately, unlike the plankton they can be easily collected and identified with reasonable certainty.

Sediment samples from all the sites at Liverpool port contained a diverse assemblage of resting stages including dinoflagellate cysts, diatom spores, tintinnid (protozoa) cysts and copepod (planktonic crustaceans) eggs. A total of 14 cysts were identified to species level, ‘round brown’ cysts were grouped into a single category although the cysts found undoubtedly represented some species of the range that are known to produce these relatively uniform cyst types. A large number of dinoflagellate cysts were not ascribed to a particular species or group of species, these were primarily small (20 µm diameter), round cysts of various types. Unidentified cysts dominated the dinoflagellate cyst assemblages at all sites. Round brown, Scripsiella sp. and Pentapharsodinium dalei cysts were the most common species identified overall, followed by Protoperidinium oblongum and P. leonis. Potentially harmful cyst types were present in most samples, most notable were the cysts of Alexandrium tamarense, a species which is responsible for paralytic shellfish poisoning (PSP) and also those of the nuisance bloom forming species Scripsiella trochoidea.

38 Total cyst concentrations varied significantly between the sites (One-way ANOVA, F = 35.43; p<0001). Certain cyst types were common in samples from some sites but were not recorded in samples from other sites and in addition, the relative proportions of the various species recorded was also variable between sites. Multivariate analysis showed that cyst assemblages were significantly different between sites (One-way ANOSIM, Global-R 0.901; P<0.001). Figure 24 shows similarity in cyst assemblages found in the sediment samples. Generally, the samples collected in the main dock complex at Liverpool were more similar to each other than those found in samples from Westfloat docks on the opposite side of the river Mersey. The exception was the sample from Collingwood dock that had relatively low species diversity. The sample analysed from ‘Pierhead’ on the Mersey river itself was more similar to those from the main port than those from Westfloat dock.

Stress: 0.06

Wfs Co Al Al Wfs Al Ca Wfs Ca Ca Wfe Wfe

Wfe Gl Gl Co PH Gl Co

Figure 22: MDS plot of replicate cyst assemblages in sediment samples taken from Liverpool port. Symbols closer together are more similar than those further apart. The letters represent sampling sites (Wfe = Westfloat dock, exposed site; Wfs = Westfloat dock, sheltered site; Co = Collingwood dock; Ca = Canada dock; PH = Pier Head; Gl = Gladstone dock; Al = Albert dock). See Figure 2 for position of sampling sites.

Figure 23: Dendrogram showing similarity between cyst assemblages in sediment samples taken from Liverpool port. Symbols closer together are more similar than those further apart. The letters represent sampling sites (Wfe = Westfloat dock, exposed site; Wfs = Westfloat dock, sheltered site; Co = Collingwood dock; Ca = Canada dock; PH = Pier Head; Gl = Gladstone dock; Al = Albert dock)See Figure 2 for position of sampling sites.

39 The results for Liverpool show that dinoflagellate cysts are common in all dock sediments. Although large-scale spatial variations in cyst assemblages have been extensively reported, few studies have documented variability on a smaller scale e.g. within hundreds of metres. Dinoflagellate cyst assemblages varied significantly between the sampling sites at Liverpool. An appreciation of small-scale variability has important implications for any cyst monitoring programme, particularly one that aims to describe cyst occurrence in enclosed docks. Because cyst assemblages may vary widely between neighbouring docks, a more comprehensive sampling programme is necessary to fully describe the cyst species and numbers present throughout the dock system.

5.3.3 Encrusting organisms Three methods were used to sample for encrusting species. A summary of the numbers of species identified found can be seen in Table 9. A combination of sampling methods were employed, each of which has its advantages and disadvantages (See Section 6, Table 12), but together give a broad coverage of the communities present. The following examples/comments concern data collected from Collingwood dock at Liverpool and Cardiff docks. • Destructive sampling Destructive sampling yielded 28 species. The method was purposely limited in its application because of the time constraints. No doubt, numbers of species found would have increased if the sampled areas had been increased. However, as a quick snapshot of what can be found (to a depth of approximately 1m) it was successful and could be carried out in working docks. • Biological diving Divers were able to identify more species than was possible by destructive sampling. However, they did cover a much greater surface area and took all day to complete two dives, each of 45 minutes duration. The dock used had no commercial shipping and as such was logistically simple to access to for this method of sampling. This method would not be easily applied to active docks. • Settlement panels Panels were in the water for up to six months, numerically, this was the method that yielded the most native species but gave the lowest number of non-native species. This is a non-intrusive method suitable for most dock conditions.

Settlement panels allow a more accurate count of species present. Figure 26 shows the abundance of Ficopomatus enigmaticus on settlement panels in the Cardiff docks. The literature indicates that this species favours a relatively high temperature and water of variable salinity (Eno et al 1997). Abundance varies throughout the dock system as did some of the physical parameters (See Figure 27) Changes in salinity at various points in the system may influence the recorded distribution of this species.

Table 9: Comparison between the number of species found in Collingwood dock, Liverpool using three sampling methods

Sampling method Number of species identified Number of non-natives Destructive sampling 25 3 Biological divers 28 2 Settlement panels 32 1

40 Fig 24:Map of Cardiff docks showing the distribution and abundance of the tube worm Ficopomatus enigmaticus (See report cover), collected on settlement panels

Fig 25: Salinity variation in Cardiff Docks

Avarage salinity (ppt) at sites in Cardiff

10

5 Salinity (ppt)

0 Queen Alexander Dock Roath Basin Roath Dock Sites

41 Table 10: Sampling methods that successfully collected non-native species, the numbers collected by each method and the ports in which they were collected from. (S=Southampton, L=Liverpool, M=Milford Haven, C=Cardiff, F=Felixstowe, T=Teeside)

Sampling method used Non-native species Port found Total Settlement panels Hydroides dianthus S Hydroides ezoensis S Ficopomatus enigmaticus L,M,C,S Elminius modestus L,M,C,S,F,T Rhithroponopeus harisii C 5 Destructive sampling Haliplanella lineata L,S Hydroides ezoensis Ficopomatus enigmaticus Elminius modestus Corophium sextonae M Crassostrea gigas M Styela clava L,M,S,F 7 Intertidal surveys Sargassum muticum S Hydroides ezoensis Ficopomatus enigmaticus Elminius modestus Potamopyrgus antipodarum T,F,M Crepidula fornicata M,S,F Crassostrea gigas Urosalpinx cinerea S,F Styela clava 9 Subtidal surveys - divers Ficopomatus enigmaticus Styela clava 2 Dredges Haliplanella lineata Hydroides ezoensis Ficopomatus enigmaticus Elminius modestus Potamopyrgus antipodarum Crepidula fornicata Styela clava 7 Traps Rhithropenopeus harisii 1 Plankton nets Odontella sinensis L,M,S,F,T 1

It can been seen that intertidal surveys, dredging and destructive sampling were the most successful methods for the collection of non-native species.

Intertidal surveys in port areas, and in particular those that included rocky areas offer a variety of niches for organisms to exploit, e.g. cover for mobile organisms, hard surfaces for encrusting organisms and sediment for infauna. Therefore they represent an extremely important habitat, although often small and difficult to access, but one which should be examined as part of any survey.

5.3.4 Community analysis Although the total numbers of native species found at Liverpool is high in comparison to most of the others (n=151), it was sampled three times as often. Milford Haven in contrast not only has the highest number of native species (n=203) collected in two surveys (See Table 11) but also, as with Southampton, the greatest number of non-natives (n=10).

42 Table 11: Total numbers of native and non-native species found at each port

Port Number of native species Number of non-native species Cardiff 61 3 Felixstowe 43 6 Liverpool 151 5 Milford Haven 203 10 Southampton 67 10 Teesside 66 3

The dendrogram (Figure 28) shows similarities between sites, expressed in terms of a percentage. The data used to produce this analysis comprises of lists of species found at each site, but not their quantities.

The Pier Head site is within the estuary and hence tidal. Major differences in water flow rate, salinity and turbidity lead to the expectation that the species composition at this site would be different to that found within the enclosed docks.

With the exception of Westfloat, which is on the south side of the Mersey, the remaining docks examined are interconnected. Water levels are maintained by operation of lock gates to allow ship passage and a pumping system. There is therefore a limited flow through the system from this pump outflow to the locks. It should be borne in mind that the communities not only reflect current commercial activities but will also have been influenced by historical usage of the individual docks. This historical influence can also be seen in some of the other docks investigated e.g.Teeside.

There are differing degrees of similarity between the docks and many factors influencing them. It is not a simple matter to identify all of these factors, the influence they have on the organisms present and their relative importance. The intricacy of the relationships is illustrated as follows:

Collingwood and Nelson are physically next to each other, which would account for the species composition in both being similar (73% similarity). There is also a fresh water inflow into Collingwood that may have an impact on the types of species found there, both native and non- native, accounting to some extent for the differences with Nelson.

Alexander, which lies between Gladstone and Canada, is, however, closer in community structure to Nelson (55%), while Westfloat, which lies across the river Mersey is closer to Canada (59%) than any of the other docks. Alexander has less commercial traffic than its closest neighbours and in this respect has more in common with the quieter docks such and Collingwood and Nelson. Westfloat is a busy dock with similar traffic to Canada dock. A full analysis of these results can be found in Section 6.3 Figure 26: Dendrogram showing similarity between species communities at all sampling sites in Liverpool

43 6.0 Analysis and Discussion

6.1 Analysis of methods employed Large ports, because of their continuous activity, are difficult environments in which to carry out biological surveys. UK ports also vary considerably in terms of their location and the various habitats that they contain, so any sampling methodology used has to be both practicable and flexible. Based upon the methodology developed by the Australian Centre for Research into Introduced Marine Pests (CRIMP), the sampling methodology has been developed and tested to enable the sampling of native and non-native species resident in UK ports.

The port surveys, particularly those carried out in Liverpool, were designed to investigate the efficiency and practicality of the various methods available for the characterization, in number and type, of the organisms resident in dock systems.

Before any sampling, ports should be surveyed to identify the range of habitat types present and to select appropriate sampling sites. Sampling sites must be representative of the habitats present in a particular port but must also be realistic. This study worked with the ports sector and aimed to cause a minimum of disturbance to normal port operations.

One of the major considerations was the need to provide methods that were sufficiently sensitive to be able to demonstrate the presence within the native population of organisms, of relatively small communities of non-native species. In achieving this, the surveys have also produced a reasonably comprehensive list of resident native species. This list will also be a requirement for the successful operation of ballast water management programmes, as subsets of the species on this list may be considered as potential invasive organisms in other parts of the world.

6.1.1 Plankton sampling The technique used to take samples is tried and tested and has been shown to collect a representative sample of those organisms present in the water column. However, two problems present themselves with respect to the analysis of the plankton. Single samples represent only a very brief snap shot of the organisms that may populate a port’s waters. The phytoplankton is usually made up of a mixed assemblage of a wide range of algal species. These have the capacity to change in composition and relative numbers over a very short time scale, which is often measured in days. The zooplankton is also very variable in its composition but for different reasons. The community of animals in the plankton is made up of organisms that are permanent inhabitants but also of temporary larval stages that will change on a slightly longer, weekly time scale. Aggregation or swarming of these organisms may also produce vertical and horizontal patchiness in their distribution. Therefore, in order to gauge accurately both types of organism, samples should be taken close together, in both time and space, and over an extended period of time in order to record the annual cycling of events. Clearly such an intensive programme is beyond the capacity of any extensive port survey scheme.

The second problem that presents itself is the actual identification of the organisms sampled. Many of the organisms are small, fragile and differentiated one from another by minute changes of body structure only discernable under a scanning electron microscope. There are few taxonomists available in the UK, or indeed world wide, who would be able to deal with more than one or two groupings of these organisms, even when only required to identify native species. The problem is further complicated by the need, not only to sort and identify local species, but also to separate out and identify adult and developmental stages of non-native forms that may have originated from anywhere in the world.

Because of their presence in the water column, planktonic organisms are the easiest grouping to transport in ballast tanks and because of their high concentrations potentially the most prolific source of introduced species. Until there is a world wide use of efficient ballast water treatment systems they will continue to provoke the greatest concern. But they also, as has been noted earlier, present the greatest problems for monitoring at the donor sites of ballast water. One option is to continue to take extensive samples, particularly at ports where large

44 quantities of dock water are loaded. These samples would only be screened for so-called target species, this would still be a considerable task, particularly the maintenance of quality assurance. The use of new techniques currently under development, molecular probes for toxic algal species, for example, may offer some answers. Alternatively sampling might only be carried out in response to visible blooms in or around port areas. This strategy would not work for planktonic animals that rarely reach the concentrations of the algae. Although collected from sediment samples, the presence of the resting cysts of dinoflagellates (the algal group containing almost all the recorded toxin producing species of algae) may offer an alternative to the monitoring of phytoplankton. The presence of high cyst populations in dock sediments is indicative of past algal blooms, providing a warning that blooms may be repeated. Of even greater significance, the cysts are themselves highly resistant organisms ideally suited to transportation in ballast tanks. Management decisions would have to be made on the extent of the assessment of these types of populations of organisms.

6.1.2 Dredging The dredge was developed in response to the observation that the small hand deployed grab was found to be collecting too small and shallow sediment sample for a significant collection of organisms to be made. Testing of how best the retrieval of a sediment sample might be accomplished using dredges was carried out using different dredge types in combination with varying mesh sizes and deployment methods, see Section 4.9.3

Dredging, together with grab sampling and coring (see next section), combine to provide a suite of collecting methods which may be used as local conditions and requirements for organism groupings dictate. As the diversity of benthic flora and fauna appears to increase when the port is situated in an open coastal position or has areas that are undredged, an increase in the number of sites sampled will give a more accurate picture of the species within that portconsideration. Consideration also should be given to the use of divers to survey port areas containing sensitive sites such as SSSI’s.

6.1.3 Grab and core sampling Although the corer (65mm diameter, 300mm length) was the most expensive and complex piece of equipment this model proved to be unreliable. Even after modifications were made it persistently malfunctioned. Consequently the small hand operated Van Veen grab, which was easy and simple to use from the dockside or boat deck was substituted to collect small volumes of sediment, usually triplicate samples were taken from each site. If a vertical profile of the sediment were required to provide, for example, a time series on cyst deposition, then coring, either by a remote device or deployed by divers, would be necessary. It should be borne in mind that sediment samples/cores should be taken from ports at areas where deposition and undisturbed accumulation of uncompacted fine sediment to a depth of 20-30cm is likely to occur. Therefore, decisions on sampling sites can only be made after an examination of the commercial dredging history within the survey areas.

Resting stages in dock sediments represent a potentially useful tool for monitoring of planktonic assemblages, particularly because many of the harmful bloom causing species of dinoflagellates produce cysts that are relatively easily recognisable. CRIMP have suggested a methodology for the sampling of dinoflagellate cysts but to date there are few accounts of the occurrence and distribution of dinoflagellate resting cysts in port sediments.

6.1.4 Settlement panels The benefit of using this system to study sessile flora and fauna is that the panels can be removed, quickly transported to the laboratory and the organisms examined alive. Table 9 also shows that this method has the potential to collect the greatest number of species. Many of the panels at Felixstowe were lost (Appendix I) and this could have led to the low numbers of species found at that port. Initially, three tier panels were deployed at chosen sites in Liverpool and Teesside (see Section 4.9.1) The settlement patterns on these panels were found to show significant differences between the subtidal and intertidal panels only. Therefore, subsequent panel deployments were modified to lie at these two levels, as seen in Figure15.

45 Some panels were broken or removed from the water, a certain amount of loss must be expected, but can be minimized by placement at inaccessible and/or unobtrusive sites and tagging with contact details. Data loss can be kept to a minimum by frequent checking and replacement.

The panels used in this survey were left in position for varying lengths of time. It was found that checking on the rate and degree of colonization was to be recommended, too short a deployment resulting in negligible settlement but too long a period in the water produced overcrowding with consequent problems in identification. But with an ability to predict the likely populations types at a particular port, which should be possible from the visual observation of dock areas and a knowledge of the seasonal abundance of these types of organism, sampling intervals should be relatively easy to determine. At Liverpool docks, to ascertain the optimum time for the panels to be exposed, panels left out for more than one month during the period between June – November became very heavily colonized and difficult to examine. Those put out between November and May could reasonably be left for 3 – 6 months.

6.1.5 Destructive sampling While panel deployment can give an impression of the short term progressive settlement on clean surfaces, sampling of naturally occurring surfaces was necessary to give an indication of the population structure of these long term stabilized surfaces. Destructive sampling was a quick and easy method of sampling any hard substrate. The only disadvantages were that the maximum depth that it could be used at was 0.75m below the surface and it sometimes destroyed friable barnacles and tube worms beyond recognition.

This method of sampling, although time consuming to analyse was probably one of the most quantitative in the number of species collected (See Table 9) in comparison with others used for encrusting species. Destructive sampling also collected more non-natives in Collingwood dock at Liverpool than either divers or settlement panels. However, since this comparison has only been completed for one dock, the results are not conclusive.

The presence of seasonal patterns in, for example the macroalgae, suggests that a sample taken on a single visit may not reveal the full range of plants and animals which may colonize any one surface at different times of the year. However, unlike the plankton, the number of visits needed to produce a complete species list would be small. The weakness of this method was the inability to sample a natural vertical surface from the water surface to the bottom. Such a survey method could be carried out using divers to collect samples, as was the case in the CRIMP methodology, but would have the same considerable financial implications that apply to visual surveys using divers.

An additional source of information on surface living organisms explored was the fouling communities of navigation buoys. These structures are maintained by harbour boards or, in many cases, by Trinity House and serviced regularly. Thus it is possible to determine the period over which settlement has taken place on surfaces which may offer organisms a slightly different set of environmental conditions from those present in ports themselves. Such sites may favour introduced species at an early stage in their process of establishment. An example is the first record of the brown alga Undaria pinnatifida on mainland Britain from a floating pontoon in Southampton Water. In the samples examined in this work it was apparent that the submerged surfaces of buoys support communities similar to but not identical with those on shore. Within Milford Haven it is worth noting that seven of the ten non-native species found there were collected from these buoys.

6.1.6 Trapping The crab traps worked well for common species of crabs and the occasional small fish. The shrimp traps deployed with them were generally unsuccessful, even when modified. Fyke nets, which were deployed at Liverpool docks in an attempt to catch eels were also unproductive. Commercial fishermen were operating, but the survey team was unsuccessful, lacking, it was suspected, specialist local knowledge on the deployment of this type of net.

46 6.1.7 Visual search, intertidal Intertidal searches proved valuable, with the proviso that the full tidal range must be investigated. Intertidal areas within ports were not common and those surveyed were small and narrow. However, these biotopes should be investigated wherever possible. Shores may require investigation when they lie in close proximity to ports. On rocky shores many common species were identified in situ with the few the few problematic organisms returned to the laboratory for further investigation. The investigation of sandy and muddy substrates were more time consuming since they required the digging and sieving of known volumes of sediment. Additional information on the organisms from these types of shore could be gained from the collection and identification of dead, stranded organisms or parts of organisms i.e. mollusc shells whose populations may be sublittoral.

Where docks and harbours are closely associated with sites of special scientific interest. as for example, Milford Haven, it should be possible to obtain detailed species lists from appropriate agencies.

6.1.8 Visual search, subtidal Visual subtidal surveys have been used extensively as part of the CRIMP surveys, to sample settled surfaces such as pier pilings and dock walls. However, there are a number of implications over and above considerations of the scientific value of such methods. There must be some concern in terms of the scientific return from the considerable financial costs involved. Such a burden might be tolerable for large ports, but might be considered excessive for smaller facilities. But perhaps of more significance are the safety aspects, diving is a high risk activity, as is recognised by the HSE. While scientific survey diving may be acceptable in ‘quiet’ docks with little or no commercial activity, surveying of any sort, let alone diving, in, for example, Felixstowe, which may have thirty vessels arriving and departing each day, would be an extremely high risk activity. In practice the port authorities are unlikely to authorise such activities.

The financial implications of using divers to survey ports and harbours restricted their use in this study, but it was considered necessary to carry out comparisons of diving driven surveys with shore based surveys. There were pros and cons to this method as compared with destructive sampling and settlement panels.

Table 12: Comparisons between the use of biological divers, destructive sampling and settlement panels.

Biological divers Destructive sampling Settlement panels

Pros Cons Pros Cons Pros Cons Saw and identified Cost £1000+ per Generally will collect Can destroy some Panels can be Surface area marginally more day all that is present if species beyond placed at any depth available is very than destructive (minimum for enough samples are identification for any length of small in samples one day) taken. time comparison to the size of a dock Can survey more Unable to Inexpensive Inexpensive Panels left too than 1m below identify small Preserving/washin long will become surface mobile fauna or g/ identification is to encrusted to those organisms time consuming distinguish all that need (cost implications species that have internal itself) grown on them examination. Identification in Will collect smaller Will collect smaller situ therefore time organisms for later organisms for later saving and identification identification protective

Within a dock system divers may be an unnecessary expense, our data suggests that the visual surveys by divers are no more efficient than the destructive sampling method. . They must be able to identify species present and such biological knowledge comes with an added cost. Their major attribute would be being able to collect, for subsequent identification, over the whole vertical profile to the bottom and therefore potentially collecting species that are only found lower down on the profile. Alternatively, surveys could use commercial divers following a strict protocol. They would make a video record of specific areas designated by the biological

47 survey and collect samples at specified depths for later identification. Both the video record and samples would then have to be processed. In open water ports however, as has been indicated earlier, such activity may not be possible.

6.2 Time allocation for surveys As a preliminary investigation this project was restricted by time and manpower, and in practice a port state would need working practices approaching the scale of the investigations being done in Australia if a comprehensive baseline assessment were to be carried out.

In order to draw up a realistic timetable for any future survey, the following information has been provided as a guide. Table 13 identifies approximate timings for each of the activities described that relate to the survey itself, but no time is included for preparation, making or repairing equipment, data entry or report writing.

Table 13: Suggested time allocation for surveys

Method Collection time Identification time (hrs) Temp, salinity, turbidity ½ hr nil etc. (abiotic data) Phytoplankton ¼ hr 6 Zooplankton ¼ hr 6 Drag 1 hr 7 Destructive sampling ½ hr 48 Panels (1 month) ½ hr 48 Cores/grabs ½ hr 48 Traps (overnight) ½ hr ½ hr Intertidal survey 4 hrs 4 hrs Subtidal survey 1 day nil 3 days 10 days maximum mininum*

* this estimate assumes no taxonomic difficulties or problems are encountered.

This represents a simple survey, sampling one site only. If preparatory work, data handling and report writing is added, the time requirement increases to a minimum of 3 weeks per site. If plankton samples and panels were to be collected and identified each month, an additional week per month would be required.This estimate assumes at least two full time workers would be involved in the work.

6.3 Analysis of results The choice of analytical methods used to compare species distributions has been limited by the lack of a complete set of quantitative data. Counting all the species found proved to be too costly in terms of time, especially for native species. Therefore, similarity matrices and various data plots were employed to give an overall comparison of docks or ports. From these, general patterns could be described and preliminary conclusions drawn. For example, from Figure 28, showing the similarity between the sites used in Liverpool, it can be concluded that docks within a port may appear to be superficially similar, but are not identical. It also highlights that it would be possible, in the case of the Liverpool dock system, to use either Collingwood or Nelson docks as sampling sites, as they are probably sufficiently similar to preclude the need to sample both. A similar assumption may also be made with the pairing of Canada and Gladstone docks.

A judgement, then, has to be made on the degree of similarity or difference that can be considered acceptable. The omission of one site may preclude the discovery of an important species, whether native or non-native. There is, therefore, a circular argument that says that all sites in all docks should be surveyed in order to recognise those that are so similar that they need not be surveyed. It clearly is not possible to carry out such intensive survey

48 programmes, but equally a single site at each port is unlikely to give an adequate representation of its biological diversity.

The crux of the process in deciding the sampling strategy at a specific port is therefore the preliminary port survey. The collection and analysis of a broad selection of physical, chemical and biological data on a port should be able to highlight those areas which can be taken as represent the whole spectrum of habitats which are available for colonisation by organisms, whether they be native or non-native species. The associations of such factors as water movement, tidal range, turbidity, salinity, temperature, bottom type, vertical surfaces type, and adjacent sources of pollution as well as a generalised appraisal of the biological communities would form the basis on which decisions would be made. Other special factors may need to be taken into account, for example docking patterns of ships and their ports of origin may influence the possibility that non-native species are present. Not only current patterns are important, historic shipping patterns at a port or dock may be reflected in the spectrum of organisms that are present. Any subsequent sampling programme should also be able to take into account effects produced by change in usage. The timing of the sampling regime also needs to be considered. Seasonal variations are evident and need to be taken into account.

Despite focussed efforts, no previously unrecorded non-native specie were found during this study, suggesting that in the UK, our knowledge of non-native marine species is relatively comprehensive. However, various non-native species were recorded from ports at which they had not previously been recorded. It is not clear whether this is due to recent spread of species or because of the limited knowledge of the fauna and flora of UK ports.

Approximately half of the established non-native species recorded in the literature were collected within the sampling areas of this survey. A number of species were not collected (See Appendix J) and it is not clear whether these have not survived at that site, have a restricted range at that site which was not sampled in this survey, or were not recognized. The list prepared by Eno (1997) although of great value as a starting point for this survey is of restricted value for comparative purposes. It was compiled from individual records rather than as a product of a systematic survey and contains a number of older records, sometimes with limited detailed information on distributions. There is no monitoring system available to confirm whether a species has survived to the present and no reporting system which actively looks for newly arrived species and follows their spread. Thus the actual number of non-native species resident in the UK may well be greater than current estimates.

Nevertheless it would appear that non-native marine species within the UK do not represent a large proportion of the marine flora and fauna. Only 4% of the species collected and counted on this survey were non-native. Had all the species identified also been counted, this proportion would have been much lower.

Direct numerical comparison with other bioregions is probably not appropriate, the situation in Australia, for example, has been accentuated by a number of factors related to its geographical isolation. The extent of endemicity of the UK and European flora and fauna is also difficult to define. However, further investigative work is necessary if a full picture of the present status of non-native and native species is to be accomplished.

Ports, as the sites of most ship borne introductions, can be considered as the ‘hot spots’ for introduced non-native species. Monitoring of ports would therefore also provide an initial warning of the appearance of a non-native species. It would then be possible to make decisions as to whether any attempts were to be made to eradicate the introduction or institute management practices to limit further local spread of the species concerned.

Although it is clear that it is possible to establish protocols to monitor the marine species present in ports, there are a number of further points that may also require consideration. These are listed without detailed comment as they fall outside the specific remit of this project.

• As ballast water management and treatment legislation has yet to be instituted, there is the possibility that re-surveying of ports may be necessary to take account of subsequent introductions.

49 • This project has only attempted to identify the physically larger end of the spectrum of micro-organisms, but there is some evidence of the transportation of human pathogens in ships’ ballast water. • If any sort of national port survey were to be instituted, consideration would be necessary with respect to the general management structure, a detailed cost analysis and the setting up and maintenance of a database accessible to mariners.

Without comprehensive baseline information on the occurrence and distribution of non-native marine species, the effectiveness of various policies aimed at minimising further introductions and reducing the spread of established species cannot be measured. This project represents a first step in this process.

50 7.0 Conclusions

• This study has demonstrated that it would be possible to survey and monitor the occurrence and distribution of native and non-native marine species in the ports of England and Wales.

• Large ports, because of their continuous activity, are difficult environments in which to carry out biological surveys. UK ports also differ considerably in extent and the habitats they contain. To accommodate these variables the sampling methods employed have to be practicable and flexible.

• It has been possible to demonstrate the general distribution, at the surveyed ports, of the non-native species identified and also show a more detailed distribution of all species within ports.

• There are limitations in the current methods employed for assessing the populations of planktonic organisms which require further investigation.

• This project represents a first step in the development of a comprehensive database on the distribution of non-native species for the UK.

• A target species list has been developed, composed of those species, native and non- native, which are considered to be candidates for introduction elsewhere.

8.0 Recommendations

Consideration should be given to the establishment of a national programme designed to survey the ports of the UK for that group of marine organisms (‘target species’) considered most likely to be introduced to other maritime nations via shipping.

Consultations should be initiated towards the formulation of an internationally acceptable target species list.

Ports be encouraged to collect and make available information on the occurrence and distribution of native and non-native marine species on a voluntary basis.

Further investigations be carried out into the problems of assessing the populations of planktonic organisms in ports.

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