Aquaculture research, tập 42, số 1, 2011

Aquaculture Research, 2011, 42, 1^20 doi:10.1111/j.1365-2109.2010.02666.x Quantitative microbial source apportionment as a tool in aiding the identification of microbial risk factors in shellfish harvesting waters: the Loch Etive case study Carl M Stapleton1, David Kay1, Shona H Magill2, Mark D Wyer1, Cheryl Davies3, John Watkins3, Chris Kay1, Adrian T McDonald4 & John Crowther5 CREH, Catchment and Coastal Research Centre, River Basin Dynamics and Hydrology Research Group, IGES, University of Aberystwyth, Ceredigion, UK Scottish Association for Marine Science, Dunsta¡nage Marine Laboratory, Oban, UK CREH Analytical, Hoyland House, Leeds, UK Faculty of the Environment, University of Leeds, Leeds, UK CREH, University of Wales, Lampeter, Ceredigion, UK Correspondence: D Kay, CREH, Catchment and Coastal Research Centre, River Basin Dynamics and Hydrology Research Group, IGES, University of Aberystwyth, Ceredigion SY23 3DB, UK E-mail: dave@crehkay.demon.co.uk Abstract Sanitary surveys of shell¢sh harvesting waters are now a routine component of regulatory monitoring These provide a qualitative appraisal of potential pollutant sources impacting on shell¢sh microbial quality The information provided by this type of screening level appraisal is very useful, but does not a¡ord quantitative assessment of the di¡erent pollution sources and their complex dynamic relationships which result in a highly episodic £ux of microbial parameters into shell¢sh harvesting waters The potential £uxes derive from treated sewage and industrial e¥uents, intermittent discharges from the sewerage system and di¡use sources of pollution, principally from livestock farming areas, but also from urban surface water drainage None of these sources are routinely monitored for the faecal indicator parameters used as compliance measures by regulators worldwide and almost no high-£ow information is available with which to construct any quantitative £ux estimates to provide a credible evidence base for the design of remediation strategies where there is a need to improve water quality within a harvesting area This study was conducted at Loch Etive, near Oban, Scotland, UK It applies an approach to Quantitative Microbial Source Apportionment developed to inform management and remediation of water quality at bathing water locations The results suggested that, in this case study r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd location, di¡use sources of microbial indicator organisms derived from livestock farming activities in catchments draining to the loch were the dominant high-£ow contribution of bacterial loadings This ¢nding was unexpected by local managers who had perceived ‘environmental’ water quality as ‘high quality’ in this traditionally pristine area of west Scotland The ¢ndings led to a series of recommendations for future management of Scottish shell¢sh harvesting waters directed at appropriate data acquisition, through a detailed sampling programme design, to acquire microbial £ux data from all sources, particularly during high-£ow event conditions It was recommended that such data acquisition was essential to the design of any remediation strategies that need a credible evidence base directing appropriate investment in interventions designed to attenuate microbial £ux from either the sewerage infrastructure and/or adjacent farming activities The utility of this study could be further enhanced through microbial tracer studies to establish connectivity between the key hydrological inputs (both those studied here and potential sources outside of the lower basin) and the shell¢sh beds Keywords: faecal indicator organisms, Quantitative Microbial Source Apportionment (QMSA), bathing, shell¢sh, Water Framework Directive, Clean Water Act, source characterization Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al Introduction In the European Community (EC), shell¢sh growing waters (SGW) are identi¢ed as a ‘protected area’ by the Water Framework Directive (WFD) (see Annex IV: Section1(ii); Anonymous 2000) Article 4c of this Directive places a legal responsibility on EC member states to ensure that these protected areas achieve compliance with ‘any standards and objectives’ de¢ned in the WFD or its daughter directives The principal water quality standards in nearshore marine and estuarine waters are de¢ned in terms of microbiological parameters, which indicate pollution by faecal wastes from human and/or animal sources These indicator bacteria include the coliforms and enterococci In the case of shell¢sh waters and shell¢sh £esh, the coliform bacteria and, the intestinally derived component of this group, i.e Escherichia coli, are the principal compliance parameters in current standard systems de¢ned in the Shell¢sh Water Directive (Anonymous 1979, 1997, 2006a) and the Shell¢sh Hygiene Directive (Anonymous 1991), now replaced by EC Regulation No 854/2004 (Anonymous 2004) Parallel and complementary EC Directives covering coastal bathing waters in the United Kingdom also de¢ne coliform standards (Anonymous 1976, 2006b) as well as marine water quality criteria based on epidemiological evidence which use the enterococci group of indicator bacteria (WHO 2003; Kay, Bartram, Pruss, Ashbolt, Wyer, Fleisher, Fewtrell, Rogers & Rees 2004; Kay, Ashbolt, Wyer, Fleisher, Fewtrell, Rogers & Rees 2006) Regulation of the microbial quality of shell¢sh £esh, which is a food product, is the responsibility of the Food Standards Agencies in the United Kingdom This is responsible for the classi¢cation of SGW into four categories (de¢ned as A^D) determined by E coli standards assessed by pre-determined sampling of shell¢sh £esh However, the environmental regulators in the United Kingdom, i.e the Environment Agency (EA) in England and Wales and the Scottish Environmental Protection Agency in Scotland, are responsible for the implementation of the WFD and consequential compliance with daughter directives, i.e they must so regulate the environment to result in a required food hygiene criteria The EA in England and Wales is charged to ensure that ‘bacteriological standards should be achieved which allow designated waters to achieve at least category B standard under the system applied by the Food Standard Agency to classify shell¢sh harvesting areas Aquaculture Research, 2011, 42, 1^20 for food safety purposes’ (EA 2003a) The WFD provides the mechanism for environmental regulators to achieve this statutory requirement This is outlined in Article11of theWFD that required that Member States produce a‘Programme of Measures’ (POM) designed to achieve required standards in each river basin This is to include integrated management and control of both ‘point source’ discharges [Article 11.3(g)] and ‘di¡use’ pollution [Article 11.3(h)] These POMs contribute to resultant basin and sub-basin management plans required under Article 13.5 The principal sources of the bacterial compliance parameters for near-shore waters used for shell¢sh harvesting and/or bathing is faecal material derived from human and/or animal populations In the United Kingdom, the human input is characterized by ‘point source’ discharges of treated e¥uents from sewage works and/or intermittent discharges from the sewerage system during times of high rainfall when combined sewage over£ows (CSOs), and/or storm tank over£ows (SSOs), operate to allow excess volumes of untreated sewage e¥uent, diluted by surface water, to exit the sewerage system once its capacity is exceeded (Kay, Crowther, Stapleton, Wyer, Fewtrell, Edwards, Francis, McDonald, Watkins & Wilkinson 2008; Kay, Kershaw, Lee, Wyer, Watkins & Francis 2008) The predominant source of di¡use microbial pollution is the livestock farming sector Livestock pollution of streams impacting on coastal protected areas can be associated with animals voiding faeces directly to the land surface or into streams and onto riparian areas (Kay, Edwards, Ferrier, Francis, Kay, Rushby,Watkins, McDonald,Wyer, Crowther & Wilkinson 2007; Kay, Crowther, Stapleton, Wyer, Fewtrell, Anthony, Bradford, Edwards, Francis, Hopkins, Kay, McDonald, Watkins & Wilkinson 2008) Other signi¢cant sources include farmyards where diary cattle concentrate for milking, leaving faeces on hard standing areas, which create signi¢cant ‘dirty water’ problems as rainfall mobilizes the faeces from concrete surfaces (Edwards & Kay 2008) Indeed, accepted animal waste handling and disposal practices also contribute to the di¡use microbial loading through land applications and slurry spreading activities In addition, practices designed to concentrate livestock for stock management and husbandry, such as woodchip corrals, can produce signi¢cant source areas of faecal indicator £ux from catchment systems (McDonald, McDonald, Kay & Watkins 2008) By comparison with other water quality parameters such as the nutrients (principally compounds r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Aquaculture Research, 2011, 42, 1^20 Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al Causes of Impairment for 303(d) Listed Waters Cause of Impairment Group Name Number of Causes of Impairment Reported Pathogens 10481 Mercury 8855 Metals (other than mercury) 7364 Nutrients 6861 Sediment 6251 Organic Enrichment/Oxygen Depletion 6233 Polychlorinated Biphyneyls (PCBs) 6179 pH/Acidity/Caustic conditions 3860 Cause Unknown - Impaired Biota Turbidity 3199 3055 Figure Reasons for impairment in all USAwaters covered by the CleanWater Act on 17 September 2009 The microbial parameters implied by the word ‘Pathogens’are coliforms and enterococci (redrawn from http://iaspub.epa.gov/waters10/ attains_nation_cy.control?p_report_type=T#causes_303d) National Cumulative TMDLs by Pollutant This chart includes TMDLs since October 1, 1995 Pollutant Group Number of Causes of Impairment Addressed Number of TMDLs Pathogens 6953 Mercury 6671 Metals (other than mercury) 6443 Nutrients 4275 Sediment Organic Enrichment/Oxygen Depletion 3080 1774 7177 6704 6565 5107 3569 1849 Temperature 1536 1572 Salinity/Total Dissolved Solids/Chlorides/Sulfates 1523 1530 pH/Acidity/Caustic conditions 1507 1548 Ammonia 1028 1079 Figure Total maximum daily load investigations completed by the USEPA to17 September 2009 under the terms of the Clean Water Act since 1995 The microbial parameters implied by the word ‘Pathogens’are coliforms and enterococci (redrawn from http://iaspub.epa.gov/waters10/attains_nation_cy.control?p_report_type=T#tmdl_by_pollutant) of nitrogen and phosphorus), pesticides, sediment and oxygen demand, the microbial parameters have received very little scienti¢c and/or research attention that can be used by the regulatory community to design ‘evidence-based’ POMs at the catchment scale to achieve water quality standards guaranteed to ensure appropriate food hygiene standards de¢ned by E coli concentrations in shell¢sh £esh (Kay, McDonald, Stapleton,Wyer & Fewtrell 2006) This is potentially signi¢cant, given the experience in the United States, where legislation very similar to the EC WFD is seen in the Clean Water Act (CWA; Anonymous 2002) The United States Environmental Protection Agency (USEPA) now publishes running totals of the reasons for non-compliance of all waters covered under the CWA (USEPA 2009) This is termed ‘impairment’ under the US legislation Figure1shows the top 10 reasons for impairments to date under the CWA The US equivalent for the EC POMs is a Total Maximum Daily Load (TMDL) study to de¢ne remedial measures needed to end the impairment Figure shows the top 10 US TMDL pollutant groups from studies completed since 1995 Implementation of the CWA is some 20 years ahead of the WFD and the signi¢cance of microbial parameters is clear from Figs and If this experience is repeated in the comparable developed countries of the EC, then the comparative lack of scienti¢c attention to r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al Aquaculture Research, 2011, 42, 1^20 Materials and methods The Loch Etive study Figure Location of quantitative microbial source apportionment studies in the United Kingdom 1996^2007 the microbial parameters in catchment science and modelling to date may yet prove unfortunate The foundation of any POM or TMDL is a quanti¢cation of the microbial £uxes from each contributing source impacting, or causing impairment of, a shell¢sh harvesting water There are very few such studies, to date, reporting empirically derived data from ¢eld survey and water quality analyses of shell¢sh harvesting waters However, Quantitative Microbial Source Apportionment (QMSA) studies, at UK bathing waters, have been conducted for each of the sites outlined in Fig and the approach developed has received peer review scrutiny in a series of international journal papers (Wyer, Kay, Dawson, Jackson, Jones,Yeo & Whittle 1996; Wyer, Kay, Crowther,Whittle, Spence, Huen, Wilson & Carbo 1998; Kay, Wyer, Crowther, Stapleton, Wilkinson & Glass 2005; Kay, Wyer, Crowther, Stapleton, Bradford, McDonald, Greaves, Francis & Watkins 2005; Wither, Greaves, Dunhill, Wyer, Stapleton, Kay, Humphrey, Watkins, Francis, McDonald & Crowther 2005; Stapleton, Wyer, Crowther, McDonald, Kay, Greaves, Wither, Watkins, Francis, Humphrey & Bradford 2008) Loch Etive is a large sealoch on the west coast of Scotland (Fig 4) and is divided into two main basins, upper and lower (Magill, Black, Kay, Stapleton, Kershaw, Lees, Lowther, Francis, Watkins & Davies 2008) The loch has a large catchment area (approximately1350 km2) with a number of freshwater inputs to the system The largest of the inputs to the lower basin is the River Awe, which enters the loch to the east of Taynuilt A hydroelectric power scheme (HEP) is located on this river, thus regulating the £ow The immediate catchment area of the study has a low human population density (approximately 2500 concentrated along the shoreline) with a further 1000 (approximately) within km of the mouth of the loch In common with many sealoch coastlines of the west coast of Scotland, the catchment consists of a number of small villages and many rural dwellings Approximately 53% of the immediate population is served by public network sewage facilities Accurate detailed information could not be gathered on all private domestic sewage systems in the catchment, but it is estimated that a large number of dwellings are served by septic tank systems However, in a number of areas, shoreline dwellings discharge raw sewage into the loch, both outside and within the SGW One wastewater treatment works (WwTW; activated sludge secondary treatment, population equivalent design capacity 51400) operates within the village of Taynuilt, serving around 60% (approximately 700) of the local population in that area The treated e¥uent and intermittent discharges of untreated storm e¥uent are discharged into the River Nant downstream of the tidal limit Three public network septic systems operate within the area The two largest systems discharge to marine outfalls outwith the main lower basin of the loch There are CSO and Emergency Over£ows within the loch itself, but are outside the SGW A small network in the village of Bonawe discharges raw sewage directly into the loch within the SGW This network is small in capacity (23 dwellings), but is in close proximity to the shell¢sh production sites Agriculture in the catchment is dominated by livestock grazing for sheep and cattle In excess of 5000 ewes and 500 cattle graze within the immediate catchment Sheep numbers during the summer months, when lambs are present, may be approximately double this estimate Much of the grazing areas are in close proximity to the shoreline Livestock census data for the agricultural parishes in the r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Aquaculture Research, 2011, 42, 1^20 Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al Figure Riverine and sewage sample points monitored during the study (seeTable1for sample point details) Rain gauges were located at sites 202, 204, 207 and 302 Stage recording monitors were located at sites 202, 204, 207, 208, 209 and 210 catchment area indicate that sheep numbers have decreased by approximately 10% from 2000 to 2005 Examination of available historical classi¢cation data collected by Fisheries Research Services (1999^ 2000), and Food Standards Agency Scotland (FSAS; 2000^2006) indicated variation in E coli levels between the production sites in Loch Etive lower basin, which houses the sampling sites It should be noted that the historical data used in this sanitary survey were collected before the implementation of more stringent sampling protocols by FSAS in 2007 These protocols required sampling to be taken from ¢xed monitoring points by designated Environmental Health O⁄cers (who are local government employees in the United Kingdom) Potential variations in sampling points used before 2007 means that the interpretation of historical data within the sanitary survey must be viewed with caution Study design A QMSA study was implemented to quantify faecal indicator organism (FIO) budgets from sewage and riverine sources draining to the lower basin of Loch Etive, Scotland, above the Falls of Lora road crossing The budgets were constructed to provide an indication of the relative contribution of 26 out of 34 identi¢ed sewage and stream inputs This did not include study of the fate and transport of faecal indicators within the waters of the loch itself that might infer a link from any discharge to compliance monitoring points Other inputs, for example from avian populations direct to the loch, were also not quanti¢ed It should be noted that this study considered a summer condition when other investigations have suggested that agricultural sources of faecal indicators in Scottish streams are at their maximum (Rodgers, Soulsby, Hunter & Petry 2003; Kay, Aitken, Crowther, Dickson, Edwards, Francis, Hopkins, Je¡rey, Kay, McDonald, McDonald, Stapleton, Watkins, Wilkinson & Wyer 2007) Similar previous studies have largely been driven by bathing water compliance considerations and have, thus, centred on summer £uxes Empirical data were acquired in two ¢eld campaigns spanning the 2006 and 2007 summer periods Water quality, stream and sewage e¥uent £ow data were collected during 2006 while further water quality and river stage data were collected during 2007 River and sewage discharge, stream level and rainfall Flow data for the River Awe at the HEP barrage, approximately km upstream of the sampling point (Fig 4), were supplied by the operators of the HEP, Scottish Southern Energy Flow throughout the r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al Aquaculture Research, 2011, 42, 1^20 Table Riverine and sewage e¥uent sample sites visited during the 2006 and 2007 surveys Site code Location River and stream sample points 201 Abhainn Achnacree 202 Inion Farm streamÃw 203 R Esragan 204 Blacreen BurnÃw 205 Kenmore Bay stream 206 Lusragan Burn 207 Culnadalloch streamà 208 Allt Nathaisw 209 R Luachraganw 210 Allt na h-Airdew 211 R Nant 212 R Awe 213 Abhainn Achnacree 214 Allt nam Ban 215 Inion Farm stream 216 Kenmore stream 220 Achnacloich Plantation Burn 221 Mussel Farm Beck 222 Stream to Rubha nan Carn 223 Stream to Rubha Ban 224 Allt Tig Dhonnchaidh 225 Allt an t-Siomain 226 Allt Ardachy 227 Allt Dail a Mhuilinn 228 Eas Mhaodain 229 Stream near ‘Sheep Wash’ 230 Un-named Stream 231 Stream north of Bonawe 232 Allt Garbh Sewage effluent sample points 301 Taynuilt WwTW FE 302 Taynuilt WwTW CSO 303 Taynuilt WwTW FE1CSO 304 Connel CSOz Location/description Year sampled Catchment area (km2) Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit ptc R Luachragan Tidal limit Tidal limit Tidal limit Tidal limit ptc Allt nam Ban ptc Abhainn Achnacree Upstream of Inion Farm Upstream Ardchattan School Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit Tidal limit 2006 2006 and 2007 2006 2006 2006 2006 2006 2006 2006 2006 2006 and 2007 2006 and 2007 2006 2006 2006 2006 2007 2007 2007 2007 2007 2007 2007 2007 2007 2007 2007 2007 2007 7.0 0.7 15.6 5.4 0.2 21.8 1.8 10.3 18.2 4.7 45.0 828.6 — — — — 0.2 0.2 0.1 0.3 1.6 2.4 1.6 0.3 0.6 0.5 0.1 0.1 0.8 Secondary treated FE CSO effluent Combined FE and CSO effluent CSO effluent 2006 2006 2006 2006 — — — — and and and and 2007 2007 2007 2007 ÃRain gauge locations wRiver level and £ow gauging locations zDid not operate during 2006 sampling ptc, prior to con£uence; FE, ¢nal e¥uent; CSO, combined sewage over£ow 2006 study period was constant at 14.2 m3 s À 1, with the exception of ‘freshets’ of three 12-h periods over the weekends of 8/9, 16/17 and 22/23 July 2006, when the £ow was 23.7 m3 s À For the remaining period of the ¢eld study, planned freshets were not released due to low levels in Loch Awe Five catchment outlet sites were monitored for stage through installation of manometric-level recorders and open-channel £ow gauging (EA 2003b) These data were used to derive stage-discharge relationships with r2-values (coe⁄cient of determination) ranging between 98.3% (site 209) and 99.2% (sites 204 and 210) Rainfall input (mm) was monitored at four tip- ping bucket rain gauges, installed for the duration of the study across the catchment (Fig 4,Table 1) The hourly discharge records for £ow monitoring stations were split into two components: (i) base-£ow and (ii) high-£ow event response to rainfall This was achieved using a combination of computer programmes (PASCAL) and visual inspection of individual events by detailed hydrograph analysis) Given the regulated £ow of the RiverAwe, it was considered inappropriate to separate the £ow into base £ow and high £ow because this was unlikely to be driven by rainfall However, to estimate a representative £ow budget for Loch Etive, discharge from the River Awe should be r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Aquaculture Research, 2011, 42, 1^20 Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al included within both the base-£ow and the high-£ow components To achieve this, £ow in the River Awe was considered to be ‘high £ow’ during the periods of high £ow in the River Luachragan, this being the largest and most proximal of the gauged catchments Discharge from catchments from which no £ow data were available were based on catchment area and rainfall input using 2006 rainfall data from the nearest available CREH rain gauge The proportion of rainfall contributing to total £ow and the base-£ow index (proportion of total £ow contributing to base£ow volumes) was taken from the nearest most similar monitored catchment Flow data for Taynuilt WwTW [inlet, CSO and combined ¢nal e¥uent (FE) and CSO £ow] were available only for part of the 2006 survey period (i.e 456 h between July 2006 and 26 July 2006) from the sewerage undertaker The FE £ow was calculated as the di¡erence of the combined FE1CSO and CSO data In the absence of a time-series of £ow for the complete study period (i.e July 2006 to 10 August 2006; 34 days), it was felt that the most robust method for estimating the £ow would be to scale the £ow for the 28-day period between 28 June 2006 and 26 July 2006 by a factor of 1.214 (i.e 34/28) While this method would provide a reasonable estimation of the total £ow over the study period, it does not provide an hourly time-series for the period of the study after the sewage £ow monitors were removed Consequently, while overall budget estimates of £ow and FIO delivery can be made, a full timeseries of £ow and FIO delivery can only be provided up to the end of the available WwTW £ow data The hourly 2006 discharge record for the FE at Taynuilt WwTW was split into base-£ow and high£ow event components in response to rainfall This was achieved through detailed inspection of the £ow record and comparison with a typical daily dry weather £ow pattern derived using data from eight dry days and rainfall records Flows that deviated signi¢cantly from the typical daily dry weather £ow pattern corresponding with rainfall were categorized as high £ows All discharges from the CSO at Taynuilt WwTW were categorized as high £ows The other sewage e¥uent sample site, a CSO in Connel, did not have any £ow measurement equipment installed Riverine freshwater and sewage e¥uent sampling Sampling was carried out between 14 July and August 2006 and between August and 14 September 2007 Samples were collected from 29 river and stream sites (16 sites during 2006 with a further 13 additional sites sampled in 2007) (Fig 4,Table1) During 2006,11sites were selected to allow budget calculations, while four sites were added to the sampling regime after initial results indicated particularly high base-£ow FIO concentrations at three catchment outlets: two were located within the Abhainn Achnacree catchment, one on the main river (site 213) and one on a tributary, Allt nam Ban (site 214), both upstream of their con£uence; one site was added to Inion Farm stream, upstream of Inion Farm (site 215), and one was located upstream of Ardchattan school on the Kenmore stream (site 216) The 2007 survey included three sites sampled during 2006: Inion Farm stream (site 202), the River Nant (site 211) and the River Awe (site 212) plus a further 13 new catchment outlets (Fig 4, Table 1) These new streams were all relatively small, with the largest catchment area being that of Allt an t-Siomain at 2.4 km2 Four sewage sample sites were sampled during both the 2006 and the 2007 surveys (Table 1) Three sites were located at Taynuilt WwTW and included the continuous treated FE discharge (site 301), the intermittent untreated CSO e¥uent at the works (site 302) and the combined e¥uent sampled from their common outfall (site 303) The forth sample point was of the storm over£ow e¥uent from the CSO in Connel, although this did not spill during the 2006 study period Samples were collected during the 2007 survey, although the lack of any £ow data precludes its inclusion in the budget estimates Three sampling runs were scheduled during each week of the respective study periods This was amended in response to rainfall, in order to target sampling during hydrograph events and obtain samples from intermittent sewage e¥uent discharges from storm tanks and CSOs Multiple samples were often collected during such events Samples were obtained either directly into 150 mL sterile disposable plastic containers (Sureseal SelectTM, SLS, Nottingham, UK) using a laboratory clamp and telescopic landing rod or by lowering a clean stainless-steel can into the £owing water/e¥uent If the sampling can was used, this was lowered into the £ow three times, on the ¢rst two occasions, the can was rinsed and the water/e¥uent was discarded After the third collection, a sample was obtained by pouring into a 150 mL sterile plastic container On return to the sampling vehicle, the inside of the sampling can was immediately dried with absorbent paper towel and then wiped clean with an r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al alcohol-impregnated cloth (Azo wipeTM, Synergy Healthcare, Swindon, UK), allowing time for the alcohol to evaporate on the journey to the next sampling location Samples were stored in the dark inside a cool box during transport to the laboratory Laboratory methods, data analysis and £ux estimation Indicator organism enumerations [colony-forming units (CFU) 100 mL À 1] followed Standing Committee of Analysts Blue Book methods based on membrane ¢ltration (Standing Committee of Analysts (SCA) 2000, 2002, 2006) Sample dilutions were determined from the initial sampling run and faecal coliforms (FC) and intestinal enterococci (EN) enumerations were performed at two or three sample dilutions A complete duplicate analysis was carried out on at least one sample collected during each sampling run for quality control purposes All samples were analysed within 24 h, in accordance with the Blue Book methods (SCA 2000, 2002, 2006) A total of 469 samples were collected during the 2006 ¢eld study period, with a further 563 samples collected during 2007 Results from each river and stream sampling site were classi¢ed into two categories, base £ow and high £ow, according to £ow conditions For temporary stations monitored during 2006, the £ow separated discharge record was used as a basis for this classi¢cation.Where continuous records were unavailable, the record from the nearest neighbouring site was used Samples collected during the 2007 ¢eld survey were separated using stage records from temporary monitors, stage board readings at each individual site and ¢eld team notes As the £ow of the River Awe was regulated, separation of the microbiological data into base-£ow and rainfall-impacted high£ow categories is not applicable Therefore, all data from each survey period for site 212 were used to calculate a geometric mean (GM) for each FIO, which was used to characterize water quality during the periods assigned to both base £ow and high £ow For the purposes of statistical analyses, samples where no organisms were detected were recorded as the detection limit value The distribution of microbial concentrations found in stream and sewage e¥uent samples, taken under base-£ow and high-£ow conditions, showed a closer approximation to normality when log10 transformed All microbial concentration data were, therefore, log10 transformed before statistical analysis The SPSS statistical compu- Aquaculture Research, 2011, 42, 1^20 ter software package (SPSS 2002) was used for statistical analyses Descriptive statistics were used to characterize the distribution of bacterial concentrations at each sampling location These statistics include the GM, calculated as the antilog of the mean of log10 -transformed concentrations, the standard deviation of log10 -transformed concentrations, the 95% con¢dence interval for the mean and the range of values at each site The signi¢cance of di¡erences between GM concentrations was examined using Student’s t-test to compare the means of log10 -transformed concentrations The methodology included Levene’s test for homogeneity of variances to determine whether or not to use a t-test assuming equal or unequal variances in the two groups compared with the hypothesis of equal variance being rejected at Po0.05 All statistical tests were assessed at a 0.05 (i.e 95% con¢dence level or 5% signi¢cance level) Faecal indicator organism £ux under base-£ow and high-£ow conditions was estimated as the product of discharge and GM faecal indicator concentration for each site, i.e the base-£ow and high-£ow GM faecal indicator concentration for each source was multiplied by the appropriate base-£ow and high£ow discharge volume for the budget period (de¢ned as the 816-h period between July 2006 and 10 August 2006) The total load for each budget was calculated as the sum of the loads from each source contributing to the lower basin of Loch Etive Di¡erent budget scenarios were constructed, including estimates of the situation during the ¢eld study periods plus a hypothetical scenario to investigate the potential impact of remediation measures within catchments identi¢ed as contributing a disproportionate £ux of FIOs Because of the large amount of information generated by this project, only FC results are presented throughout, given that coliforms are the compliance parameter for shell¢sh waters Where variations in EN results were present, these are described in the text A full description of the results from this study is available in Magill et al 2008 Results River and sewage e¥uent discharge A total volume of 4.9  107 m3 was discharged during the 2006 study period into the lower basin of Loch Etive from the studied catchments and sewage sources (Table 2) Di¡use catchment sources (i.e the rivers) accounted for virtually the entire discharge budget (99.99%) Of this, 78% was discharged during r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Aquaculture Research, 2011, 42, 1^20 Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al Table Estimated discharge budget (m3) and percentage contribution to Loch Etive from sampled catchments for the period 7/7/06 and 10/8/06 Discharge volume (m3) Base flow 201 Abhainn Achnacreeà 202 Inion Farm stream 203 River Esraganw 204 Blacreen Burn 205 Kenmore Bay streamz 206 Lusragan Burn‰ 207 Culnadalloch streamz 209 River Luachragan 210 Allt na h-Airde 211 River Nantk 212 River Awe 220 Achnacloich Plant Burnz 221 Mussel Farm Beckz 222 Str to Rubha nan Carnz 223 Stream to Rubha Banz 224 Allt Tig DhonnchaidhÃà 225 Allt an t-SiomainÃà 226 Allt ArdachyÃà 227 Allt Dail a MhuilinnÃà 228 Eas MhaodainÃà 229 Str passing Sheep WashÃà 230 Un-named Streamz 231 Stream north of Bonawez 232 Allt Garbhz 301 Taynuilt WwTW FE 302 Taynuilt WwTW CSO Rivers Total Sewage Total Total (all sources) 2.63 2.57 6.02 2.07 6.72 2.39 4.33 2.41 1.38 5.95 3.54 5.86 5.55 3.20 8.33 6.13 9.13 6.02 1.21 2.43 1.78 4.63 5.65 3.09 3.34 — 3.81 3.34 3.81                          105 104 105 105 103 105 104 105 105 105 107 103 103 103 103 104 104 104 104 104 104 103 103 104 103  107  103  107 % Contribution to discharge budget High flow 3.19 2.30 7.30 2.50 6.03 2.82 3.33 2.84 1.06 7.00 7.53 4.51 4.27 2.46 6.41 5.50 8.18 5.40 1.09 2.18 1.60 4.15 5.07 2.77 1.16 7.99 1.06 1.24 1.06                              105 104 105 105 103 105 104 105 105 105 106 103 103 103 103 104 104 104 104 104 104 103 103 104 103 101 107 103 107 Total flow 5.81 4.87 1.33 4.57 1.28 5.21 7.66 5.25 2.44 1.29 4.29 1.04 9.82 5.66 1.47 1.16 1.73 1.14 2.30 4.61 3.38 8.78 1.07 5.86 4.51 7.99 4.87 4.59 4.87                              105 104 106 105 104 105 104 105 105 106 107 104 103 103 104 105 105 105 104 104 104 103 104 104 103 101 107 103 107 Base flow High flow Total flow 0.54 0.05 1.24 0.42 0.02 0.49 0.09 0.50 0.28 1.22 72.76 0.01 0.01 0.01 0.02 0.13 0.19 0.12 0.02 0.05 0.04 0.01 0.01 0.06 0.01 0.00 78.29 0.01 78.30 0.65 0.05 1.50 0.51 0.02 0.58 0.07 0.58 0.22 1.44 15.48 0.01 0.01 0.01 0.01 0.11 0.17 0.11 0.02 0.04 0.03 0.01 0.01 0.06 o0.01 o0.01 21.70 o0.01 21.70 1.19 0.10 2.74 0.94 0.03 1.07 0.16 1.08 0.50 2.66 88.24 0.02 0.02 0.01 0.03 0.24 0.36 0.23 0.05 0.09 0.07 0.02 0.02 0.12 0.01 0.00 99.99 0.01 100.00 Flow estimate based on: ÃInion Farm rain and Blacreen Burn £ow wBlacreen Burn rain and £ow zBlacreen Burn rain and Inion Farm stream £ow ‰Culnadalloch rain and R Luachragan £ow zCulnadalloch rain and Allt na h-Airde £ow kTaynuilt WwTW rain and R Luachragan £ow ÃÃInion Farm rain and £ow base-£ow events, although this distribution was skewed somewhat by the regulated £ow from the River Awe (site 212), which contributed 88% of the total £ow discharged during the study period (4.3  107 m3, of which 83% designated as base £ow; Table 2) and accounted for 93% of the base-£ow discharge volume (Fig.5) During high-£ow conditions, the River Awe accounted for a lower proportion of the £ow (71%), with the other rivers contributing a larger proportion of the freshwater/e¥uent input to the loch than they did during base-£ow conditions (Table 2, Fig 5) The next largest inputs were the River Esragan (site 203) and the River Nant (site 211), both of which accounted for approximately 2.7% of the £ow into the loch (Table 2, Fig.5).The smallest £ow was from Rubha nan Carn (site 222), which discharged and estimated 3.2  103 m3 during the study period Only two of the gauged rivers discharged a greater proportion of their £ow during high-£ow conditions: Blacreen Burn (site 204; 55% during high-£ow events, over a period of 125 h ^ 15% of the study period) and the River Luachragan (site 209; 54% during high-£ow events, over a period of 140 h ^ 17% of the study period) The remaining three monitored rivers discharged a greater volume during base-£ow conditions (Table 2) Consequently, only those rivers whose r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Aquaculture Research, 2011, 42, 142^147 Consumer behaviour and environmental preferences D Whitmarsh & M G Palmieri Table Variables used in the logistic regression model Variable Type Measure Definition BUY AGECLASS JOBGRADE REGION ENVIRONMENT Dependent Independent Independent Independent Independent Dichotomous Dichotomous Dichotomous Dichotomous Continuous for respondents who buy weekly or monthly, otherwise for respondents aged 30 years or below, otherwise for respondents in managerial or professional posts, otherwise for respondents resident in Orkney, Shetland or the Western Isles, otherwise Priority weight attached to minimizing environmental impacts of salmon farming (calculated using analytic hierarchy process) involving the calculation of a consistency ratio (CR) Higher CR levels indicate greater inconsistency in responses, and following the normal convention (Saaty & Vargas 2000), it was decided to exclude cases where the CR was 410% This had the e¡ect of reducing the number of observations used in the model to 256 The logistic regression results are given in Table All the independent variables were statistically signi¢cant, and the sign on the B-coe⁄cient in each instance (positive or negative) indicated that the relationship with the dependent variable was as expected To summarize, respondents were found to be more likely to buy salmon on a regular basis if they were aged over 30, were employed in managerial or professional posts (a proxy for higher income) and were resident in the mainland regions The crucial result for our purposes, however, relates to environmental preferences Here, the sign is negative, implying that the higher the priority attached to minimizing the environmental impact of salmon farming, the lower the likelihood that salmon would be purchased on a regular basis Implications The results of the survey indicate that public attitudes towards the environmental performance of Scottish salmon farming translate into purchasing behaviour This ¢nding is consistent with other studies showing the emergence of ‘green’ values among consumers, including concern over whether ¢sh originates from sustainable sources (Ja¡ry, Pickering, Ghulam, Whitmarsh & Wattage 2004; Muir 2005) There is a debate to be had over whether there are a particular set of issues that underpin these values, and how far concern over sustainability encompasses other areas such as animal welfare In our survey, the environmental performance of salmon farming was de¢ned in terms of three speci¢c Table Results of the logistic regression Variable B SE Significance Odds ratio Constant AGECLASS JOBGRADE REGION ENVIRONMENT 1.525 À 1.210 1.149 À 0.933 À 0.017 0.462 0.392 0.290 0.292 0.006 0.001 0.002 0.000 0.001 0.004 4.594 0.298 3.156 0.393 0.983 Chi square 48.5 (d.f 54, sig 50.000) Nagelkerke pseudo-R2 50.234 N 256 impacts, but it is quite possible that some respondents at least may have held a rather broader conception of the problem and why they might consider ¢sh farming to be ‘unsustainable’ The main implication of this study is that the future development of salmon markets will be shaped by the information that consumers receive ^ both positive and negative ^ about the aquaculture industry and its environmental record It is instructive to compare our results with those of a survey undertaken in Valencia (Spain) by Honkanen and Olsen (2009), which showed that consumer knowledge and understanding about aquaculture was quite limited, with some people holding ambivalent attitudes towards farmed ¢sh This in turn, the authors suggest, may potentially lead to lower ¢sh consumption (p 306) One of the lessons from the Valencia study would appear to be that consumers who hold ambivalent views require more information about the practice of ¢sh farming so that environmental issues can be sensibly evaluated A similar point can be made with regard to the results reported here, although the ¢ndings suggest that the explanation for infrequent consumption of salmon is due to more than simply lack of knowledge or ambivalence on the part of consumers The key point to come out of our research is that the pattern of consumption is determined at least in part by people’s values, with environmental preferences (‘green’ values) being a factor a¡ecting the propensity r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 142^147 145 Consumer behaviour and environmental preferences D Whitmarsh & M G Palmieri to purchase The challenge for the aquaculture industry certainly involves increasing public understanding of ¢sh farming practice, but arguably, it also requires a strategy of positioning products in the market so that people who are currently disinclined to buy salmon on account of environmental concerns are persuaded otherwise Acknowledgments Research for this paper was undertaken as part of the European Commission project ECASA (Ecosystem Approach for Sustainable Aquaculture), Contract No 006540 References Al-Mazrooei N., Chomo G.V & Omezzine A (2003) Purchase behaviour of consumers for seafood products Agricultural and Marine Sciences 8,1^10 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(2008) Aquaculture in the Ecosystem SpringerVerlag, NewYork, NY, USA Honkanen P & Olsen S.V (2009) Environmental and animal welfare issues in food choice: the case of farmed ¢sh British Food Journal 111, 293^309 Ja¡ry S., Pickering H., GhulamY.,Whitmarsh D & Wattage P (2004) Consumer choices for quality and sustainability labelled seafood products in the UK Food Policy 29, 215^228 Kangas J (1995) Supporting the choice of the sports ¢shing site Journal of Environmental Management 43, 219^231 Lane D.E (2007) Planning in ¢sheries-related systems In: Handbook of Operations Research in Natural Resources (ed by A.Weintraub, C Romero,T Bjorndal & R Epstein), pp 237^271 Springer Science and Business Media, NewYork, NY, USA Leung P., Muraoka J., Nakamoto S.T & Pooley S (1998) Evaluating ¢sheries management options in Hawaii using analytic hierarchy process (AHP) Fisheries Research 36, 171^183 Mardle S & Pascoe S (1999) A review of applications of multiple criteria decision making techniques to ¢sheries Marine Resource Economics 14, 41^63 Mardle S & Pascoe S (eds.) (2003) Multiple objectives in the management of EU ¢sheries: preference elicitation CEMARE Report 64, University of Portsmouth, UK Mardle S., Pascoe S & Herrero I (2004) Management objective importance in ¢sheries: an evaluation using the analytic hierarchy process Environmental Management 33, 1^11 Muir J (2005) Managing to harvest? 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Trade-o¡ analysis of sustainability attributes Marine Policy 32, 465^474 Verbeke W., Vanhonacker F., Sioen I., Van Camp J & De Henauw S (2007) Perceived importance of sustainability and ethics related to ¢sh: a consumer behaviour perspective Ambio 36, 580^585 Whitmarsh D & Palmieri M.G (2009) Social acceptability of marine aquaculture: the use of survey-based methods for eliciting public and stakeholder preferences Marine Policy 33, 452^457 Whitmarsh D & Wattage P (2006) Public attitudes towards the environmental impact of salmon aquaculture in Scotland European Environment 16,108^121 World Bank (2006) Aquaculture: changing the face of the waters Meeting the promise and challenge of sustainable aquaculture Report 36622 ^ GLB Young J.A, Brugere C & Muir J.F (1999) Green grow the ¢shesoh? Environmental attributes in marketing aquaculture products Aquaculture Economics and Management 3,7^17 r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 142^147 147 Aquaculture Research, 2011, 42, 148^159 doi:10.1111/j.1365-2109.2010.02764.x Abstracts Seasonal availability of food for primary production and mussel growth at Killary Harbour, Ireland A Smith, R.R Cave and the UISCE Project Team Dept of Earth and Ocean Sciences, National University of Ireland, Galway, Ireland Correspondence: a.smith11@nuigalway.ie Tel: 1353 863166540 Killary is a ¢ord-like estuary on the West coast of Ireland, economically important for farming of long line blue mussels (Mytilus edulis) Local growers report a decrease in annual tonnage and stock size in recent years The UISCE project studied three shell¢sh aquaculture sites around Ireland, including Killary, in order to gain a better understanding of the processes in£uencing shell¢sh growth Water quality sampling was carried out at 12 locations in Killary, monthly in winter and bi-weekly in spring/summer, from June 2007 to May 2008 Samples taken at all 12 locations were analysed for particulate matter, organic carbon/nitrogen, chlorophyll-A, phytoplankton and macro-nutrients Hydrodynamic data were acquired to complement the water quality data set Seasonal variations in food availability for mussel growth and primary productivity are reported here It was found that food availability at Killary is primarily in£uenced by wind and tidal movement Sites located toward the middle of the estuary where lower growth has been reported were found to be nutrient limited A range of husbandry practice changes may increase production at these slow growing sites in the Estuary Development of a vaccine against Rainbow Trout Fry Syndrome Alison L Morgan, Remi L.M Gratacap, Niall A Auchinachie, Kim D Thompson and Alexandra Adams Institute of Aquaculture, University of Stirling, Stirling, Scotland, FK9 4LA Correspondence: alm2@stir.ac.uk; Tel: 144 (0) 1786 467904; Fax: 144 (0) 1786 472133 Flavobacterium psychrophilum is the causative agent of Rainbow Trout Fry Syndrome Primarily thought 148 to be a disease of salmonids, there is increasing evidence that it a¡ects a variety of other freshwater ¢sh The disease occurs in most areas of the world including USA, Canada, Chile, Australia, Japan, Korea and several European countries, causing serious mortalities and severe economic losses in hatcheries and farms It is also no longer a disease exclusively of fry but is now regularly reported in adult ¢sh There is no commercial vaccine available for this disease and the aim of this project was to develop a vaccine that could be administered to both fry and adult ¢sh Through this study a new culture media has been created, a new serotyping system has been developed and different formulations of vaccine have been tested under laboratory conditions The most successful formulation is a whole cell (formalin killed) vaccine containing isolates representative of the three most common serotypes, all of which are typical to European and USA outbreaks of the disease Field trials are planned for the Spring/Summer and the vaccine will be administered via immersion, injection and using a novel technology for oral presentation Holding crustaceans for the live market: understanding recovery of trawled Norway Lobsters (Nephrops Norvegicus) Simon Sinclair, Amaya Albalat and Douglas Neil Faculty of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, UK The Norway lobster (Nephrops norvegicus) is the target of the most valuable ¢shery in the UK There is an increasing interest for the export of live animals In Scotland, the most common method of capture is trawl (about 95%) although some animals are also caught by creels Traditionally, creel-caught animals are exported to the live market although recently an increasing number of Nephrops have been coming from trawl capture Therefore, the aim of the present work was to understand how Nephrops recover after trawling and to optimise a holding protocol for Nephrops to target the live market To this end, Nephrops were caught by otter trawl in the Clyde sea area and recovered on board using r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd Aquaculture Research, 2011, 42, 148^159 tanks with running seawater Once at port, animals were transported in air to the commercial holding facilities and left overnight Mortality was scored, arginine phosphate, haemolymph and muscle Llactate were analysed as measures of stress and ATP and its breakdown products were measured in tail muscle in order to calculate the adenylate energy charge The results show the recovery potential of this species after trawl and indicate the importance of optimal early holding conditions if the live market is to be targeted This work was supported by a grant from a Knowledge Transfer Partnership (KTP) Award to the University of Glasgow Committee generated environmental management solutions for aquaculture in British Columbia B Taekema1, S Dedominicis2, K Hoyseth3, K Maier4, M Parker5, P Galloway6 British Columbia Ministry of Environment, 2Marine Harvest Canada, 3Department of Fisheries and Oceans Canada, 4Mainstream Canada, 5Grieg Seafood, 6BC Salmon Farmers Association Correspondence: Bernie Taekema RP Bio, BC Ministry of Environment, 2080-A Labieux Rd., Nanaimo BC CanadaV9T 6J9 Tel: 250 751-3175 Email: Bernie.Taekema@gov.bc.ca British Columbia’s (BC) Fin¢sh Aquaculture Waste Management Technical Committee is comprised of representatives from several of the BC salmon production companies, two provincial government ministries (Environment and Agriculture & Lands) and the federal Department of Fisheries and Oceans Canada It is a solutions-based committee with clear, strong government leadership that develops and ensures strong environmental management and performance as well as long-term sustainability This committee is di¡erentiated by; 1) it is ongoing, not ad hoc requiring full participation by all members, 2) the use of independent science to support evidence-based decisions through-out a given process and 3) the committee’s work has evolving objectives as science and circumstance change The committee’s work identi¢es new research directions and utilizes outside expertise to ensure that its decisions are based on sound, relevant science By e¡ective and meaningful engagement of all interests of its members and by allowing public interest to Abstracts guide direction, the committee has fostered an e⁄cient business-like transparent process that has resulted in several successes, including: the development of ecological threshold parameters and the creation of new regulations for hard bottom sites Committee recommendations focus on harmonization of agency objectives and needs and recommendations are often incorporated in updated government regulations and/or associated protocols Environmental sustainability of mariculture in Scottish waters: development of environmental sustainability indicators and exploring their possible responses to climate change Clare Greathead FRS Marine Laboratory, 375 Victoria Road, Aberdeen, AB119DB Correspondence: C Greathead, FRS Marine Laboratory, 375 Victoria Road, Aberdeen, AB11 9DB; Tel: 01224 295526; Email: greatheadc@marlab.ac.uk The demand for ¢sh is growing but the amount of ¢sh acquired through ¢sheries is decreasing, therefore aquaculture will either need to expand to ¢ll the gap or the demand for ¢sh will need to reduce To ensure that any growth in the aquaculture industry is acceptable it will need to be shown to be sustainable; however, clear indicators of environmental sustainability at the industry or regional scale have not been applied speci¢cally in Scotland In this project three Indicators have been developed so far within a ‘‘Pressure-State-Response’’ framework; these are for benthic enrichment, nutrient release and the occupation of space These indicators have used data from both Fisheries Research Services and the Scottish Environment Protection Agency to produce scales of impact and have used existing thresholds to de¢ne sustainable levels of impact Further indicators will include another for benthic impact, the ecosystem e¡ects of nutrient release, disease control, and escapes These indicators will provide all interested parties with the tools to assess the sustainability of the aquaculture industry and plan ahead for potential impacts from any potential expansion of the aquaculture industry They will also improve our understanding of where the actual environmental limitations are and where further expansion of aquaculture would be environmentally unsustainable r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 149 Abstracts Aquaculture Research, 2011, 42, 148^159 Gadoid antimicrobial peptides: sequencing, characterization and expression analysis Dawn Shewring, Jun Zou,Yolanda CorripioMiyar, and Chris Secombesà Scottish Fish Immunology Research Centre, University of Aberdeen, Aberdeen AB24 2TZ, UK ÃAuthor for correspondence Tel: 01224 272857; email: c.secombes@abdn.ac.uk Approaches to controlling infection in the culture of marine ¢sh species have attracted widespread interest in recent years Antimicrobial peptides (AMPs) have been extensively studied in mammals and these molecules play a major role in innate immune responses Adaptive immune systems in ¢sh are very slow to respond and it is believed they rely heavily on their innate system Cathelicidins are a family of AMPs which are manufactured as pre-pro-peptides, and stored in an inactive form Upon infection the peptide is cleaved to release the active mature form responsible for microbicidal activity Within teleosts, cathelicidins have so far been identi¢ed in salmonids and Atlantic cod In this study a recently published cod cathelicidin sequence was used to design primers for use in PCR with haddock (Melanogrammus aegle¢nus) tissue samples Cloning and sequencing of PCR products revealed two haddock cathelicidin genes with di¡ering lengths of their mature peptides Expression studies using qPCR are being undertaken to compare expression levels in various tissues of these Gadoid genes in control and immunostimulated ¢sh It is believed that if these Gadoid cathelicidin genes are inducible by immunostimulation, they may o¡er some protection against infection for stocks bred in the aquaculture industry An ecosystem approach to aquaculture management Culhane, F.Ã; Whyte, C.Ã; Fernandes,T.Ã; Potts, T.,wTett, P.*w ÃEdinburgh Napier University,10 Colinton Road, Edinburgh, EH10 5DT, Scotland and wScottish Association for Marine Science, Dunsta¡nage Marine Laboratory, Oban, Argyll, PA371QA, Scotland Corresponding Author: Fiona Culhane, School of Life Sciences, Edinburgh Napier University, 10 Colinton Road, Edinburgh, EH10 5DT, Telephone 0044-1314552350, Email f.culhane@napier.ac.uk 150 The ecosystem approach to management aims to integrate the ecological, economic and social aspects of ecosystems in order to achieve sustainable development An EU FP6 project, SPICOSA (Science and Policy Integration for Coastal System Assessment), applies scienti¢c knowledge through modelling to facilitate integrated management of coastal zones A systems approach framework (SAF), which follows a DPSIR (Drivers-Pressure-State-Impact-Response) cause-&-e¡ect chain, is followed in this project to examine the interactions between components of the ecosystem and external drivers The aim is to allow outcomes of prospective policy decisions to be demonstrated to stakeholders SPICOSA demonstrates the SAF at 18 study sites across Europe One of these sites is the fjordic sea-loch, Fyne, in western Scotland Interactions between mussel aquaculture and leisure boating in Loch Fyne were modelled using Extend software This approach illustrates potential, indirect negative impacts on mussel aquaculture As a result of increased concentrations of antifouling paints in the Loch due to simulated increased numbers of leisure boats, the abundance of planktonic organisms may decrease, limiting food required for mussel growth This could have economic implications for aquaculture and tourism, as well as, to the wider society Effect of diets containing soybean trypsin inhibitor on growth performance, digestive proteases and intestinal structure in juvenile seabream (Sparus aurata L.) E Santigosa2, M.A SaŁenz de RodrigaŁnìez1, A Rodiles3, F.J Alarco¤n1 Department of Applied Biology, University of Almer|¤ a, 04120 Almer|¤ a, Spain.2Department of Physiology, University of Barcelona, 08028, Barcelona, Spain.3IFAPA Aguas del Pino, Cartaya-Punta Umbr|¤ a, 21459, Huelva, Spain Correspondence: F J Alarco¤n, Department of Applied Biology, University of Almer|¤ a, 04120 Almer|¤ a, Spain Phone: 134 950 015 954; Fax: 134 950 015 476 (e-mail: falarcon@ual.es) Juvenile seabream were fed diets containing 0.0, 2.0 or 4.0 g kg À of soybean trypsin inhibitor for 30 days Growth performance, digestive protease activity and intestinal histology were studied at 0, 15 and 30 days No signi¢cant di¡erences with respect to ¢sh r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 Aquaculture Research, 2011, 42, 148^159 fed free-inhibitor diet were found in weight gain, speci¢c growth and feed conversion when fed ¢sh with inhibitor-supplemented diets Only speci¢c growth rate at day 15 decreased signi¢cantly with inhibitor inclusion, though this e¡ect reverted at day 30 Zymograms of intestinal extracts showed that soybean inhibitor diminished the intensity of some higher molecular weight alkaline proteolytic fractions at day 15 in distal intestine However, at day 30 the inhibition e¡ect on these active bands was not found and total protease activity was similar to ¢sh fed a diet without inhibitor Histological examination revealed no noticeable di¡erences of the intestinal structure between any of the diet groups Inhibitor administration did not a¡ect seabream growth performance or intestine histology after 30 days, and only a decrease in total alkaline protease activity was found after15 days of administration of inhibitor supplemented diets Results obtained con¢rm that juvenile seabream have capacity for compensating dietary soybean trypsin inhibitor in a mid-term period Dietary administration of a probiotic strain in juvenile Solea senegalensis: effect on growth performance and intestine functionality Alarco¤n, F.J.1, D|¤ az-Rosales, P.2, Chabrillo¤n, M.2, Arijo, S.2, Leo¤n-Rubio, J.M.2, SaŁenz de RodrigaŁnìez, M.A.1, Balebona, M.C.2, Morinìigo, M.A.2 Department of Applied Biology, University of Almer|¤ a, 04120 Almer|¤ a, Spain.2Dept of Microbiology Fac of Sciences, University of MaŁlaga Campus Teatinos 29071-MaŁlaga Spain Correspondence: F J Alarco¤n, Department of Applied Biology, University of Almer|¤ a, 04120 Almer|¤ a, Spain Phone: 134 950 015 954; Fax: 134 950 015 476 (e-mail: falarcon@ual.es) The e¡ects of the dietary administration of a probiotic from the Shewanella genus was assessed by measuring growth and feed e⁄ciency, intestinal activity and structure of the intestine of juvenile Solea senegalensis A bacterial preparation was sprayed on the feed (9 cfu g À 1) and ¢sh were fed twice a day with experimental diets for 60 days After the trial, signi¢cantly better results were obtained for all the zootechnical parameters in ¢sh fed on the diet including probiotic when compared to the control The activity of the intestinal brush border membrane Abstracts showed no signi¢cant di¡erences in leucine aminopeptidase activity when comparing the control group and the group fed with probiotic However, mean values of leucine aminopeptidase activity were higher in the posterior intestine than in the anterior intestine The di¡erences were statistically signi¢cant for the ¢sh fed probiotic-supplemented diet (Po0.05) Microscopic examination revealed accumulation of lipid droplets in the enterocytes of ¢sh receiving the control diet, but not in those fed on probiotic These results suggest that the absorptive capacity of the brush border membrane was more e⁄cient in ¢sh fed probiotic supplemented diet Results obtained showed that this probiotic strain enhances growth performance and feed utilisation of juvenile Senegalese sole A systemic approach to Integrated Coastal Management in the aquaculturedominated Pampanga delta, Philippines Franc°ois MialheÃa,Yanni Gunnellb, Catherine Meringa, Lionel Dabbadiec a Department of Geography, CNRS UMR 8586, Universite¤ Paris-Diderot, case courrier 7001, place Jussieu, 75251 Paris Cedex 05, France, bDepartment of Geography, CNRS UMR 8591, Universite¤ Paris-Diderot, case courrier 7001, place Jussieu, 75251 Paris Cedex 05, France, cAquaculture Research Unit, Centre de Coope¤ ration Internationale en Recherche Agronomique pour le De¤ veloppement (CIRAD), BP 5095, 34033 Montpellier cedex 1, France ÃCorresponding author: (10033) 0628071466, framialhe@hotmail.fr Territory refers to a geographic area in which land use and resource management involve interconnected ecological, historical, demographic, social, cultural and economic dimensions Territories can be approached in a systemic manner in which components and their attributes are characterized and mapped over time so that changes in system state can be revealed and analyzed Integrated Coastal Management (ICM), which aims to promote sustainable development through territorial planning, is an iterative process that operates in a context of exposure to recurring hazards, to social con£ict, to scarcity, and to the degradation of natural capital Here we showcase a synthesis of remote sensing and Geographical Information System and ¢eld analysis carried out in the Pampanga river delta, a coastal area of the Philippines that has become an r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 151 Abstracts Aquaculture Research, 2011, 42, 148^159 extensive farming territory dominated by aquaculture Scaling up from the individual homestead to the global market, the spatial organisation of the system and driving factors of change are presented as a nested analysis of existing subsystems The methodology proposed is both replicable and adaptive, providing a template for rational planning, policy making and management activities in other settings SALMOTRIP: A feasibility study of commercial triploid Atlantic salmon production Migaud, H.1,Taylor J.1, Hansen T.2, Kole A.3, Guy, D.4, Storset, A.5, Breck, O.6, Hunter, D.7 and Moss G.8 University of Stirling, Stirling, Scotland; 2Institute of Marine Research, Bergen Norway; 3Agrotechnology and Food Innovations,Wageningen, Netherlands; 4Landcatch Natural Selection Ltd, Alloa, Scotland; 5Aqua Gen AS, Trondheim, Norway; 6Centre for Aquaculture Competence, Hjelmeland, Norway; 7Marine Harvest Ltd., Fort William, Scotland; 8SARL Salmo Cherbourg, France Correspondence: H Migaud, Institute of Aquaculture, University of Stirling, Stirling, Scotland, FK9 4LA Correspondence: herve.migaud@stir.ac.uk; Tel: 144 (0) 1786 467886 A trans-national EU collaboration between key salmon industry companies and research centres was established to explore the feasibility of commercial triploid salmon production Interest arises from growing pressure on the industry to minimise environmental pollution due to escapees interacting with wild populations Furthermore, greater public awareness and the need to protect natural resources require development and implementation of new environmental regulations and operating standards Triploidy is not a new concept, originally used to avoid maturation However, poor performance, higher mortalities/deformities saw triploidy abandoned in favour of photoperiod control of pre-harvest maturation Although successful, farmed stocks remain reproductively competent and threat of escapes persists Triploid induction is the only method that can produce sterile ¢sh Other bene¢ts include: avoiding sexual maturation reduces disease risk; widens harvest windows; reduces running costs/visual impact in sea cages as photoperiod control is not 152 needed; and allow breeding companies to protect IPR on selected strains The project focuses on the optimisation of induction protocols; identi¢cation of physiological requirements and sensitivities; development of out-of-season smolt regimes; explores selective breeding programs speci¢c to triploid; commercial ¢eld trials; and examines public perception, marketing and consumer acceptance Only when this knowledge is available can the potential for triploidy be realised Data generated will also aid legislative decision making regarding future aquaculture policies Protective effects of Spirulina platensis supplementation against liver inflammation during exercise training in humans Chi-cheng Wang1, Shien-ming Yang2,Yu-ywan Chang1,Yu-kwan Yang3, & Hsueh-kuan Lu3à Department and Graduate School of Physical Education, National Taiwan Sport University, Taichung 404, Taiwan, ROC, 2Athletics Department & Graduate school, National Taiwan Sport University,Taichung 404,Taiwan, ROC, 3Sport Science Research Center, National Taiwan Sport University,Taichung 404,Taiwan, ROC Correspondence: H-K Lu, Sport Science Research Center, National Taiwan Sport University,Taichung 404,Taiwan, ROC E-mail: hklu@ntcpe.edu.tw To evaluate the e¡ects of supplementation of aquaculture blue-green algae, Spirulina platensis, against liver in£ammation during exercise training, sixteen National Taiwan Sport University student volunteers were recruited Before and after a three week period of exercise training with spirulina or methyl cellulose (control) dietary supplementation, blood samples of subjects were dtaken The results showed that there were no signi¢cant di¡erences between the treatement and controls for plasma concentrations of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidaes (GPx) However, signi¢cant di¡erences (po.05) between spirulina and control supplementation were found for lactate dehydrogenase (LDH) and glutamicoxaloacet transaminase (GOT) In conclusion, these results suggested that the dietary supplementation of spirulina exerts protective e¡ects against liver in£ammation after the exercise training, and that this was not related to antioxidation enzyme system r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 Aquaculture Research, 2011, 42, 148^159 Male and female gonad fatty acids of the commercial bivalve Donax trunculus I Mart|¤ nez-Pita, I Hachero-Cruzado, E Prieto, C SaŁnchez-Lazo and O Moreno IFAPA CentroAgua del Pino Consejer|¤ a deAgricultura y Pesca Ctra El Rompido-Punta Umbr|¤ a 3.8 Km Huelva, Spain Correspondence: I Hachero-Cruzado, IFAPA Centro Agua del Pino Consejer|¤ a de Agricultura y Pesca Ctra El Rompido-Punta Umbr|¤ a 3.8 Km, Huelva, Spain Tfno: 0034959024950, fax: 034959024929, Email: ismael hachero@juntadeandalucia.es The aim of this study was to analyze the male and female gonad fatty acid composition of a commercial bivalve species Donax trunculus, from the south coast of Spain Male and female fatty acid composition shows high percentages of 14:0, 16:0, 18:0, 20:5n-3 and 22:6n-3 Two plasmologen derivatives have been detected 17DMA and 18DMA and found to be present in a relative high proportion Signi¢cant di¡erences between both sexes have been found in the fatty acid pro¢le The saturates16:0 and18:0 have higher values in males than in females while 14:0 level is lower in males The most important di¡erences in the monounsaturates is observed in 16:1n-7, 18:1n-7 and 20:1n-7, which are higher in females than in males The two major polyunsaturates 22:5n-3 and 22:6n-3, are lower in females than in males However, minor polyunsaturates as 18:2n-4, 18:3n-4, 20:4n-6 and 22:5n-6 have higher levels in females than in males These di¡erences between sexes probably indicate di¡erent requirements of males and females during gametogenesis although the presence of a large number of gametes in the mature gonad must be also considered Growth, feed efficiency, and fatty acids composition of juvenile Senegalese sole (Solea senegalensis) fed diets containing different levels of protein and lipid Rodiles, A.1, Hachero-Cruzado, I.1, Herrera, M.1, Olmo, P.1 and Alarco¤n, F.J.2 IFAPA‘‘Agua del Pino’’ Ctra Cartaya- Punta Umbr|¤ a, s/ n 21459 Huelva Spain.2Department of Applied Biology, University of Almer|¤ a, 04120 Almer|¤ a, Spain Correspondence: I Hachero, IFAPA ‘‘Agua del Pino’’ Crtra Cartaya- Punta Umbr|¤ a, s/n 21459 Huelva Spain Phone: 134959024943; Fax: 134959024929 (e-mail: ismael.hachero@juntadeandalucia.es) Abstracts Senegalese sole (25 g) were fed 45/12, 45/17, 45/22, 50/12, 50/17 and 55/12 protein/lipid diets for weeks Final body weights did not show signi¢cant di¡erences between treatments, with the lowest and highest mean values for 45/22 (47.6 g) and 50/ 17 (54.7 g) diets, respectively However, the lipid level in diets showed a signi¢cant e¡ect on zootechnical indexes, sharing highest mean values for ¢sh fed diets containing 12 and 17% lipid The e¡ect of protein level was only signi¢cant for the hepatosomatic index The ¢sh muscle fatty acid (FA) composition did not show any di¡erences among treatments Intestine and muscle showed a FA pro¢le very close in comparison to the liver The lipid level of diets had signi¢cant e¡ects on the FA composition of the intestine (18:2n-6), liver (18:3n-3, 20:4n-6, 22:5n-3, 22:6n-3), and muscle (18:2n-6, 18:3n3) The protein level in the diets a¡ected the FA pro¢le of intestine (18:1n-9), and liver (20:4n-6) Senegalese sole present high PUFA levels in their tissues independently of the treatment Results suggest that a maximum of 17% of crude lipid, and a minimum of 45% of crude protein could be used in the feed without detrimental e¡ect on growth of juvenile S senegalensis The impact of dietary supplementation with astaxanthin on egg quality in Atlantic cod broodstock (Gadus morhua, L.) J Sawanboonchun,W.J Roy, D A Robertson, G Bell Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, Scotland, UK Correspondence: J Sawanboonchun, Westhaugh Road, Stirling, UK, FK9 5GF Tel 144 7790744273 Email: jarin.sawanboonchun@stir.ac.uk This study investigated the e¡ect on egg quality of dietary supplementation of Atlantic cod broodstock with the carotenoid astaxanthin (ASTA) Duplicate groups of farm-reared Atlantic cod broodstock were fed either a control diet with no added ASTA, or an ASTA supplemented diet for months prior to peak spawning The results indicated that ASTA uptake into eggs from the broodstock diet was highly e⁄cient A correlation between the egg ASTA content and fertilisation success of individual batches was identi¢ed These results clearly demonstrate signi¢cant bene¢ts of ASTA supplementation of cod brood- r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 153 Abstracts Aquaculture Research, 2011, 42, 148^159 stock feeds in terms of improved egg quality and larval production Use of 3D hydrodynamics and particle tracking models for sustainable marine cage culture in Scotland J Moreno Navas (1) L Ross (2) and T Telfer (3) (1) PhD Student Institute of Aquaculture University of Stirling FK94LA Stirling.UK (jmn1@stir.ac.uk) (2) Professor, Institute of Aquaculture University of Stirling FK94LA Stirling.UK (l.g.ross@stir.ac.uk) (3) Lect, Institute of Aquaculture University of Stirling FK94LA Stirling UK (tct1@stir.ac.uk) Tel: 01786767882 Fax: 01786472133 A three-dimensional hydrodynamics and Particle Tracking models would be useful for a wide range of research and management applications for sustainable aquaculture cage culture in Scotland A 3D hydrodynamic model, coupled to a particle tracking model, has been applied to study the circulation patterns, mean current speeds, and the quiescent period in an Irish fjord; an area with restricted exchange Although the whole area is within a proposed Special Area of Conservation (SAC) and is host to many important aquaculture activities with up to16 operators currently licensed for mussel, oyster, clams, scallops, abalone and salmon production A particle tracking model simulates the instantaneous massive release of 21000 particles representing discharge from ¢n-¢sh cages and allows investigation of the circulation, exchange and retention characteristics the water body from and between salmon production areas and enables the analysis of residence time and the ocean-fjord exchange characteristics This information is important for understanding of nutrient and chemical discharges, transmission of diseases and sea lice and for carrying capacity modelling This modelled information has been integrated into a 3D GIS, using Arc View 3.2, thus providing a user interface for visualization interrogation of results and as an input into other spatial modeling projects Marine Laboratory, 375 Victoria Road, Aberdeen, 3University of Aberdeen, Ocean Laboratory, Newburgh Correspondence: Karen Rodgers, University of Aberdeen, Cruickshank Building, Aberdeen, AB243UU Email: k.rodgers@abdn.ac.uk Phone: 07706387572 Although copper is an essential micronutrient, it can be toxic at high concentrations The use of antifouling coatings containing copper has raised concern related to their potential environmental impact This study has focused on copper from ¢sh farms using combined surveys of water, sediment and biota with experiments to investigate its environmental signi¢cance Copper concentrations in seawater samples from around ¢sh cages were found to be above the environmental quality standard (5 mg/l) for UK waters When these and higher concentrations were used to assess the impact of copper on phytoplankton and macroalgae, only concentrations of 40 mg/l were found to have a measurable impact It is therefore unlikely that copper concentrations in seawater are a¡ecting microalgal growth Copper concentrations in sediment were found to be extremely variable but were often high When a range of sediment copper concentrations were used in a toxicity test on a marine amphipod, higher concentrations were found to cause accumulations of copper in the animals, but not mortality The high copper concentrations found in some sediments close to ¢sh farm cages may lead to benthic community function being compromised This possibility is currently under investigation Reducing the impact of sea lice on fish health and welfare through selective breeding Environmental impact of copper based antifoulants at fish farms K Gharbi1, J E Bron2, L Matthews1, R J Roberts3 & M J Stear1 Institute of Comparative Medicine, University of Glasgow, United Kingdom, 2Institute of Aquaculture, University of Stirling, United Kingdom, 3Landcatch Natural Selection Ltd, United Kingdom Correspondence: Karim Gharbi, Institute of Comparative Medicine, Faculty of Veterinary Medicine, University of Glasgow, Glasgow, Scotland, UK G61 1QH, email: k.gharbi@vet.gla.ac.uk K Rodgers1, I M Davies2, G.I Paton1, C Robinson2, M Solan3, D Standing1 University of Aberdeen, Institute of Biological and Environmental Sciences, Aberdeen, 2Fisheries Research Service Serious outbreaks of sea lice in recent months have highlighted the limitations of traditional control methods available to salmon farmers Here we discuss breeding approaches for increasing resistance 154 r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 Aquaculture Research, 2011, 42, 148^159 to Lepeophtheirus salmonis in commercial salmon strains Pilot trials under experimental conditions indicate that susceptibility to infection is under relatively strong genetic control, with resistant families showing a 40% reduction in chalimus stages relative to susceptible families (h2 50.28 Æ 0.08, after correction for ¢sh weight) Genetic markers have also been identi¢ed that are associated with resistance to infection under farm conditions To quantify potential gains from traditional and molecular breeding, we are also developing mathematical models for predicting the response to selection and its e¡ect on parasite transmission within individual cages Selective breeding will not necessarily eliminate the need for other approaches to control sea lice However, genetic gains are permanent and cumulative, thus providing the industry with a sustainable alternative to over-reliance on chemotherapeutants, and an additional tool for developing integrated pest management strategies A decision support system for site selection and sustainable development of marine fish farming in Indonesia K R Niederndorfer1, S A van der Wulp1, R Mayerle1, F Foders2, K Sugama3, A Hana¢4, K-H Runte1, K-J Hesse1 Christian-Albrechts-University Kiel, Research and Technology Center Westcoast (FTZ) in Buesum, Germany, 2Christian-Albrechts University Kiel, Institute of World Economics (IfW), Germany, 3Department of Marine A¡airs and Fisheries (DKP), Indonesia, 4Gondol Research Institute for Mariculture (GRIM), Indonesia Correspondence: M.Sc Katharina R Niederndorfer, Christian-Albrechts-University Kiel, FTZ-Corelab, OttoHahn-Platz 3, 24118 Kiel, Germany, ph 0049 (0)431 880 3730, fax 0049 (0)431 880 7303, e-mail: niederndorfer@corelab.uni-kiel.de This presentation summarizes advancements in the development of an existing Decision Support System (DSS) for the sustainable environmental and socioeconomic management of coastal ¢sh farms worldwide The investigations have been carried out at selected coastal areas in Indonesia within the framework of the research project Science for the Protection of Indonesian Coastal Ecosystems funded by the German Ministry of Education and Research and by the Indonesian Ministry of Fisheries and Marine A¡airs from 2007 until 2010 The decision making process draws primarily upon data bases, ¢eld Abstracts measurements and models of ¢sh farm emissions, hydrodynamics, sediment and water quality and socio-economics The major components of the DSS are explained, including suitability analysis, site de¢nition, production planning, nutrient £ux and impact assessment The signi¢cance of the carrying capacity in terms of water and sediment quality is underlined as essential parameter for the sustainable operation of the ¢sh farms Special attention is given to a hydrodynamic index of suitable location for estimation of the maximum sustainable production rates of ¢sh farms Stock management strategies to optimise growth potential in on-growing of marine fish M Cowan, H Migaud, A Davie, D Penman Correspondence: H Migaud, Institute of Aquaculture, University of Stirling, Stirling, Scotland, FK9 4LA Correspondence: herve.migaud@stir.ac.uk; Tel: 144 (0) 1786 467886 Management strategies developed to combat the problem of early maturation in marine aquaculture are species speci¢c Regarding Atlantic cod, photoperiod manipulation is the preferred management approach however there are welfare concerns as to its application in commercial conditions In this project, work has investigated the impact of traditional (metal halide) and novel (green cathode) lighting systems on welfare and light sensitivity in the species Results have revealed that the application of such lighting does not induce a signi¢cant chronic stress response (plasma cortisol and glucose analysis) or non speci¢c immune response (lyzozyme activity), suppress feeding behaviour or severely damage the retina Melatonin analysis did however indicate that even under constant lighting (LL), cod perceived the overlying ambient photoperiod Consequently, a longer term outdoor study is currently being conducted to investigate the perception of day/night light intensity ratio in cod under LL in combination with shade netting to suppress the intensity of ambient illumination Overall this project is intended to improve the competitiveness and sustainability of the marine aquaculture industry within the UK by re¢ning photoperiod manipulation in Atlantic cod farming and enhancing their growth potential This work is supported by SARF, the British Marine Fin¢sh Association and Intravision Aqua AS r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 155 Abstracts Aquaculture Research, 2011, 42, 148^159 Effects of sudden salinity changes on the oxygen consumption and osmoregulatory parameters in the Senegalese sole (Solea senegalensis Kaup, 1858.) M HerreraÃ1, C Aragaìo2, I Hachero1, I Ru|¤ z-Jarabo3, L.Vargas-Chaco¡3, J M Mancera3, L Conceic°aìo2 IFAPA Agua del Pino Ctra Cartaya-Punta Umbr|¤ a 21459 Cartaya (Spain), 2CCMAR Universidade Algarve (Campus de Gambelas) P - 8005-139 Faro (Portugal), 3Departamento de Biolog|¤ a Facultad de Ciencias del Mar yAmbientales Universidad de CaŁdiz.11510 Puerto Real (Spain) ÃCorrespondence author: Marcelino Herrera Rodr|¤ guez IFAPA Agua del Pino Ctra Cartaya-Punta Umbr|¤ a 21459 Cartaya (Spain) Ph.: 0034 959024942 Fax: 0034 959024929 marcelino.herrera@juntadeandalucia.es Senegalese sole juveniles were held in £ow-through respirometers and submitted to a sudden salinity change Fish with a body weight of 40^50g were held in chambers with initial salinity of 37.5 ppt (the same as in the culture tanks) and, after an acclimation of hours, the salinity was changed to or 55 ppt A control group did not experience any salinity alteration Oxygen consumption was registered every 14 and ¢sh were sampled (gill biopsy and plasma) at di¡erent times (at t 4.5 h and t h in order to determine gill Na1,K1ATPase activity and plasma cortisol concentration At t 4.5 h, oxygen consumption showed an inverse relationship with salinity, while gill Na1,K1- ATPase activity increased Plasma cortisol levels presented a ‘‘U-shaped’’ relationship with salinity At h, oxygen consumption was higher in the control group, while cortisol and gill Na1,K1- ATPase activity varied in parallel to salinity Our results suggest that sudden salinity changes evoked an acute stress situation (cortisol increased up to 15  over the basal level) which was slightly attenuate after h Fish quickly adapted their respiration rate to the new environmental conditions and this rate returned progressively to its normal values after the shock Regarding enzyme activity, ¢sh ATPase activity increased (55 ppt) or decreased (5 ppt) depending on the salinity, as described in other studies Strategic waste management and minimisation in aquaculture M Taylor and R Kelly Correspondence: Thistle Environmental Partnership, Shandon Crescent, Edinburgh, EH111QE Tel: 0131 228 156 1579, Mob: 07773 429 408; Email: mark@thistle-env co.uk This study identi¢ed waste arisings and disposal routes for Scottish aquaculture (excluding e¥uent), with a focus on salmon farming, and identi¢ed opportunities and barriers to improve waste management and waste minimisation practices Of the estimated annual waste arisings of 9,300 tonnes, approximately 59% was land¢lled, 18% recycled (or reused),19% incinerated (18% with energy recovery) and 2% disposed of to land Plastics and ¢sh mortalities were the most dominant wastes, followed by timber, paper and cardboard, steel, domestic wastes and special waste There was concern regarding the availability of facilities for handling ¢sh mortalities in Scotland, both routine and event related ^ approximately half of which are sent to Widnes in Cheshire for incineration (although a new plant was planned for set up in Brechin) An assessment of ¢sh mortality provision in Scotland and contingency planning was recommended and, depending upon the ¢ndings, research into alternative treatment methods should be undertaken Although an infrastructure for recycling feed bags has developed, there is opportunity to improve the sustainability of waste management and implement waste minimisation initiatives, including improved reusue and recycling of redundant plant and equipment and feed bags Speci¢c measures and further research was recommended Consumer attitudes to feed sustainability M Taylor and R Kelly Correspondence: Thistle Environmental Partnership, Shandon Crescent, Edinburgh, EH111QE Tel: 0131 228 1579, Mob: 07773 429 408; Email: mark@thistle-env co.uk This study aimed to identify possible barriers to the increased use of plant materials as a partial substitute to ¢shmeal and ¢sh oil in feed for Scottish farmed salmon A consumer attitudes study found that consumer attitudes are not a barrier to substitution providing that the health bene¢ts from eating salmon and product quality are una¡ected Feed manufacturers con¢rmed they could produce feed at an increased level of substitution that allows farmed ¢sh to meet omega health claims Nevertheless, some supermarkets remained opposed to in- r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 Aquaculture Research, 2011, 42, 148^159 creased substitution as were a minority of ¢sh farmers Some product standards are also a barrier It was also noted that plant substituted diets not appear to compromise £esh quality, ¢sh health, welfare and performance, although some elements of the supply chain have concerns in this area To facilitate wider acceptance of increased substitution the feed industry should present data on the use of such diets in commercial aquaculture production and con¢rm these meet relevant health claims A supply chain forum to present and discuss data was recommended The environmental sustainability of using plant derived meals and oils was considered and the positive steps taken by feed manufacturers to address sustainability issues was highlighted Serum protein changes in injection of killed Aeromonas hydrophila in common carp M DadarÃ, R Peyghan; G.H Khajeh University of Shahid Chamran, Ahwaz, Iran ÃCorrespondence: No 12- Mozafari st Behbehan ^ Iran Tel: 0098 916 6715209 Email: mdadar77@yahoo.com Aeromonas hydrophila is one of the most important pathogenic bacteria of ¢sh, causing bacterial septicemia In this research, serum protein changes of common carp were considered after the injection of formalin killed Aeromonas hydrophila The bacteria were injected into 60 ¢sh by intraperitoneal (30 ¢sh) and intramuscular (30 ¢sh) methods As controls, 40 ¢sh were injected with normal saline intramuscularly and intraperitonealy Blood was taken from all ¢sh 15 days after injection Blood protein serum was studied using the gel electrophoresis method to assess the in£uence on levels of pre-albumin, albumin, a1-globulin, a2-globulin, b-globulin, g1-globulin and g2-globulin The results showed that in intraperitoneal injection of killed bacteria, total albumin as well as the ratio of albumin/globulin (A/G) was signi¢cantly greater than the control group The other fractions showed no signi¢cant di¡erences In intramuscular normal saline injected control group, the amount of a1-globulin was signi¢cantly higher than the bacteria injected group (po0.05) For the other fractions there was no signi¢cant di¡erence between the two groups After 15 days, the total globulin in the intraperitoneal control injection group was signi¢cantly higher than bacteria injected group Abstracts The importance of lipid reserves in the transmission of Gyrodactylus spp in 3-spine sticklebacks (Gasterosteus aculeatus L.) M Grano-MaldonadoÃ1, J Bron1, S Irving2, M Longshaw2 & A Shinn1 Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, UK, 2Cefas Weymouth Lab, Barrack Road, Weymouth, DT4 8UB, UK Correspondence: M Grano-Maldonado, Parasitology Group, Institute of Aquaculture, Pathfoot Building, University of Stirling, FK9 4LA, Scotland, UK Tel: 144 (0) 1786 467883; Fax: 144 (0) 1786 472133 ÃEmail: m.i.granomaldonado@stir.ac.uk Non-feeding life-cycle stages, such as dispersal stages of parasites, are dependant for survival upon ¢nite energy reserves gathered during feeding phases The likelihood of death thus increases as the organism ages, generating an age-dependant mortality curve In many such stages, lipids represent the principal form of stored energy reserves, often being stored as large droplets Confocal laser scanning microscopy has been used to investigate the number, size and distribution of lipid droplets in Gyrodactylus, using a working hypothesis that these droplets function as a proxy for nutritional state The work demonstrated that the droplets were particularly associated with the gut and that individuals are extremely variable in the amount of stored lipid that they carry All stages, including individuals just released from the birth pore, carried lipid droplets TEM imaging con¢rmed the presence of lipid droplets It is likely that transferring worms require a bu¡er of stored reserves to protect them against failure to establish contact with a host, this allowing survival o¡ a host for several days G salaris, for instance, has been reported to survive o¡ a host for up to six days and G gasterostei survives up to days Ongoing work is investigating the in£uence of environmental variables upon the decline of lipid storage products and will try to relate the measured lipid levels with the survival of the parasite The effectiveness of formulated feed supplemented with Catharanthus roseus leaf juice on Penaeus monodon culture N Aye Winn1, M Too Nyi Bu2, A Kyi3 Aquaculture Department, Department of Biotechnology, Yangon Technological University, Myanmar, 2Ani- r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 157 Abstracts Aquaculture Research, 2011, 42, 148^159 mal Biotechnology, Yangon Technological University, Myanmar, 3Ministery of Science and Technology, Yangon, Myanmar Correspondence: Nandar Aye Winn, Aquaculture Department, Department of Biotechnology, Yangon Technological University, Myanmar E.mail: nandar.¢sh@ gmail.com The inhibition e¡ect on pathogenic bacteria of Catharanthus roseus (L.) crude water leaf juice through feed on growth and survival rate were investigated in improved extensive culture system of black tiger shrimp (Penaeus monodon) Larval shrimp of mean body weight 0.02 g at PL(20^25) were cultured in four plots in duplicate for ¢ve months at a stocking density of 500 fry per plot of (0.01) hectare pond During the experiment the control plot ponds E1(Ia,Ib) were fed with normal diet feed (F-1)and the treated plot ponds E2(IIa, IIb), E3 (IIIa, IIIb,) E4 (IVa, IVb) were fed with leaf extract coated formulated feed at the respective concentrations of 50 ml (F-2), 80 ml (F-3), 100 ml (F-4), per kilogram of feed The survival, growth and health of the shrimp was checked by cast net samples and the physio-chemical parameters of culture plot were monitored once each week The primary objective of this study was to improve farm-made feed to increase survival rates and reduce water pollution The antimicrobial properties of four di¡erent concentrations of crude leaf juice extract were used to test antimicrobial activity against the isolated pathogenic bacteria of Vibrio hervyi The results suggested that the crude leaf juice was the most e¡ective treatment forV hervyi After the experiment, mean survival and total harvest weight of duplicate plots in treated groups E2 (71% and 20000 g) and E3 (74% and 21000 g) were showed that the higher than treated group E1 (67% and18500 g) and control (65% and 17800 g) However, there was signi¢cant di¡erence (Po0.05) in body weight between control and E3 and E4 experiments The results showed that the F4 diet was the best amongst three to support good growth rate and higher survival followed by diet F-3 Water quality data showed that water quality remained within optimal ranges for all the experimental plots Based on these results, it may concluded that di¡erent concentrations of C roseus crude leaf extract coated commercial diet feed may enhance the immune system and may improve resistance to pathogenic bacteria, increasing survival of black tiger shrimp in grow-out production Moreover, the crude leaf extract seemed to reduce the e¡ects of environmental stress and may improve the environmental conditions in the black tiger shrimp culture system 158 Study of Aeromonas, Vibrio and Mycobacterium infection in some of aquarium fishes of Ahvaz-Iran Peyghan, Rahim.Ã, Ghorbanpor, Masaod., Blooki, Adele Veterinary Faculty, Shahid Chamran University ^ P.O Box: 61335-145, Ahvaz - Iran Email: rpeyghan@yahoo.com There were no reports of Aeromonas,Vibrio and Mycobacterium infection in aquarium ¢shes of Ahvaz This study was carried out to determine the health status of four important species of aquarium ¢shes (Poecilia reticulate, Cheirodon axeliodi, Poecilia sp and Carassius auratus) in Ahvaz Over a period of a year,120 ¢sh (30 specimens of each species) were transferred to the laboratory and examined The skin and gills were examined by routine methods Bacterial infections were studied by sampling the liver and kidney and culturing samples on TSA, TCBS and Lowenstein ^ Jensen media The media were incubated at 25 1C and appropriate diagnostic and biochemical tests conducted on pure grown colonies 20% of Cardinal tetra were infected with Aeromonas spp (4 Aeromonas hydrophila and Aeromonas sobria) 30% of Carassius auratus were infected with Aeromonas spp (3 Aeromonas hydrophila and Aeromonas sobria) 33% of Poecilia sp were infected to Aeromonas spp (2 Aeromonas cavia, Aeromonas sobria and Aeromonas hydrophila) 20% of Poecilia reticulate were infected with Aeromonas spp (5 Aeromonas hydrophila and Aeromonas sobria) NeitherVibrio spp nor Mycobactrium spp were detected or isolated in the ¢sh species sampled In conclusion, it seems aquarium ¢shes of Ahvaz were not infected with serious pathogens such as Mycobacteria and Vibrio However 25.8% of ¢sh cases showed diseases related to genus Aeromonas Apoptosis in the pathogenesis of Salmon pancreas disease T Herath, K D Thompson, A Adams, J E Bron and R H Richards Institute of Aquaculture, University of Stirling, Stirling, UK, FK9 4LA Correspondence: Tharangani Herath, Institute of Aquaculture, University of Stirling, Stirling, UK E mail: tkh1@stir.ac.uk Salmon pancreas disease (SPD) has recently become a threat to the Scottish salmon industry This severe, contagious disease is characterised by pathology in r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 Aquaculture Research, 2011, 42, 148^159 the pancreas, heart and skeletal muscle Pathogenesis of the disease has not been fully elucidated Here we report the results of an in-vitro experiment that describes possible cell death mechanisms of this disease An Irish isolate of SAV-1, that causes SPD, was used to infect CHSE-214 cells and the cells were examined at 1h, h, h, and 1, 3, and days post-infection (p.i.) Electron microscopy revealed chromatin marginalisation at h and 24 h p.i and chromatin condensation at 24 h p.i Formation of electron dense multiple micronuclei, with loss of cytoplamic characteristics, were evident at 48 h p.i Nuclear fragmentation was also noticed under confocal microscopy with a £uorescent dye (Hochest 33258) in infected cells Further, these cells gave positive signals with ‘terminal deoxynucleotidyl transferasemediated dUTP nick-end labelling’ (TUNEL) The DNA laddering started to appear from 48 h p.i in infected cells Cytoplasmic caspase activation was also found in infected cells, stained with anticaspase antibody This data suggests progressive apoptosis associated cell death in SAV-1 infection and may represent the major mechanism of cellular pathogenesis in this disease Abstracts upon the cytokines they release Whilst it is not known if Th exist in lower vertebrates, the molecules that characterise them are known and many of the cytokines they release have been identi¢ed in ¢sh In this study we have cloned the IL-22 gene from the gadoid haddock (Melanogrammus aegle¢nus), to assess whether it is a useful marker of protective responses following vaccination with the Gram negative pathogenVibrio anguillarum Haddock were vaccinated with 109 cfu/ml of formalin-killed bacteria (V anguillarum, MT2582 serotype O2) and a control group was injected with an equivalent volume of saline Two months post vaccination ¢sh were challenged by immersion where vaccinated ¢sh reached a relative percentage of survival (RSP) of 100% Next, comparisons were made in terms of immune gene expression analysed by real-time PCR in samples at four time points, weeks post-immunisation and 24, 48 and 72 h post-challenge Head kidney, a major systemic lymphoid tissue, and gill, a mucosal tissue, were chosen for the transcript analysis Our results show that ¢sh vaccinated with an e⁄cacious vaccine can mount a rapid response resulting in IL-22 expression at mucosal sites when encountering the homologous pathogen and this may contribute to the protection seen IL-22 upregulation in the gills of haddock (Melanogrammus aeglefinus) after Vibrio anguillarum vaccination Y Corripio-Miyar, J Zou and C J Secombes Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Scotland, UK y.corripio@abdn.ac.uk Following vaccination, T helper (Th) cells drive immune responses in di¡erent directions, dependent r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 148^159 159 [...]... Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Aquaculture Research, 20 11, 42, 21^27 doi:10.1111/j.1365-2109.2010.02667.x Applied epidemiology with examples from UK aquaculture James F Turnbull1, Iain K Berrill1, Darren M Green1, Ryan Kaye1, David Morris1, Alexander G Murray2, Jorge del-Pozo3 & Andrew Shinn1 1 Institute of Aquaculture, University of Stirling, Stirling,... Aquaculture 136,195^207 r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 21^27 27 Aquaculture Research, 20 11, 42, 28^34 doi:10.1111/j.1365-2109.2010.02726.x What has been done to minimize the use of antibacterial and antiparasitic drugs in Norwegian aquaculture? Paul J Midtlyng1, Kari Grave2 & Tor Einar Horsberg2 1 LaboratoryAnimal Unit, Norwegian School of Veterinary... accomplished r 2011 Blackwell Publishing Ltd, Aquaculture Research, 42, 28^34 Aquaculture Research, 20 11, 42, 28^34 Minimize the use of antibacterial and antiparasitic drugs P J Midtlyng et al within a few months It appears that the situation in other countries, for example in Scotland or Chile, is quite di¡erent and, in most cases, proceeds more slowly In the Norwegian system, marine aquaculture licenses... evaluation of vaccine prototypes by the National Veterinary Institute In return, veterinary pharmaceutical companies were o¡ered free inclusion of vaccine r 2011 Blackwell Publishing Ltd, Aquaculture Research, 42, 28^34 Aquaculture Research, 20 11, 42, 28^34 Minimize the use of antibacterial and antiparasitic drugs P J Midtlyng et al (a) # of months 0 3 Oct Jan SiteA x x 6 9 12 15 18 21 24 27 30 33 36... Djupvik HO., Willumsen FV., Tveit AM & Tangen K (1994) Infectious salmon anaemia (ISA) epidemiology in Norway Preventive Veterinary Medicine 19, 277^290 r 2011 Blackwell Publishing Ltd, Aquaculture Research, 42, 28^34 Aquaculture Research, 20 11, 42, 35^40 doi:10.1111/j.1365-2109.2010.02675.x A characterization and sensitivity analysis of the benthic biotopes around Scottish salmon farms with a focus... drainage ditches, streams, etc., maintenance of bu¡er strips along river corridors and avoidance of r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Aquaculture Research, 20 11, 42, 1^20 (a) Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al 2006 total faecal coliform budget showing contribution of base flow and high flow sources... animal origin intended for human consumption O⁄cial Journal of the European Communities L226, 83^127 r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Aquaculture Research, 20 11, 42, 1^20 Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al Anonymous (2006a) Council Directive 2006/113/EC of 12 December 2006 on the quality required... conditions (Fig 6) Similar elevations in GM faecal indicator concentrations during high-£ow conditions r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1^20 Aquaculture Research, 20 11, 42, 1^20 Microbial risk factors in Loch Etive shell¢sh waters C M Stapleton et al 8 log10 concentration (cfu 100 ml–1) 7 6 5 4 3 2 1 0 Figure 6 Mean, range and 95% con¢dence... confounders, or the association is said to be confounded An example is the signi¢cant statistical r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 21^27 Aquaculture Research, 20 11, 42, 21^27 association between drowning and the application of sunblock In this case the confounding e¡ect is obvious: people that swim outdoors are both more likely to wear... salmonid movements between registered Scottish sites in 2003 Arrows indicate the direction of movement 24 r 2010 The Authors Journal Compilation r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 21^27 Aquaculture Research, 20 11, 42, 21^27 scienti¢c disciplines Aquatic animal health problems are most e¡ectively addressed by selecting the correct tools or techniques for the study, not looking for studies