• The role of Azadinium spinosum (Dinophyceae) in the production of azaspiracid shellfish poisoning in mussels

      Salas, Rafael; Tillmann, Urban; John, Uwe; Kilcoyne, Jane; Burson, Amanda; Cantwell, Caoimhe; Hess, Philipp; Jauffrais, Thierry; Silke, Joe (Elsevier, 2011)
      Azaspiracids (AZAs) are a group of lipophilic polyether compounds first detected in Ireland which have been implicated in shellfish poisoning incidents around Europe. These toxins regularly effect shellfish mariculture operations including protracted closures of shellfish harvesting areas for human consumption. The armoured dinoflagellate Azadinium spinosum Elbrächter et Tillmann gen. et sp. nov. (Dinophyceae) has been described as the de novo azaspiracid toxin producer; nonetheless the link between this organism and AZA toxin accumulation in shellfish has not yet been established. In August 2009, shellfish samples of blue mussel (Mytilus edulis) from the Southwest of Ireland were analysed using liquid chromatography–tandem-mass spectrometry (LC–MS/MS) and were found to be above the regulatory limit (0.16 μg g−1 AZA-equiv.) for AZAs. Water samples from this area were collected and one algal isolate was identified as A. spinosum and was shown to produce azaspiracid toxins. This is the first strain of A. spinosum isolated from Irish waters. The Irish A. spinosum is identical with the other two available A. spinosum strains from Scotland (3D9) and from Denmark (UTHE2) in its sequence of the D1–D2 regions of the LSU rDNA. A 24 h feeding trial of blue mussels (M. edulis) using an algal suspension of the Irish A. spinosum culture at different cell densities demonstrated that A. spinosum is filtered, consumed and digested directly by mussels. Also, LC–MS/MS analysis had shown that AZAs were accumulating in the shellfish hepatopancreas. The toxins AZA1 and -2 were detected in the shellfish together with the AZA analogues AZA3, AZA6, AZA17 and -19 suggesting that AZA1 and -2 are metabolised in the shellfish within the first 24 h after ingestion of the algae. The levels of AZA17 detected in the shellfish hepatopancreas (HP) were equivalent to the levels of AZA1 but in the remainder tissues the levels of AZA17 were four to five times higher than that of AZA1, only small quantities of AZA3 and -19 were present with negligible amounts of AZA6 detected after the 24 h period. This could have implications in the future monitoring of these toxins given that at present according to EU legislation only AZA1–AZA3 is regulated for. This is the first report of blue mussels’ (M. edulis) feeding on the azaspiracid producing algae A. spinosum from Irish waters.
    • Salmon Mortalities at Inver Bay and McSwyne’s Bay Finfish farms, County Donegal, Ireland, during 2003

      Cronin, M.; Cusack, C.; Geoghegan, F.; Jackson, D.; McGovern, E.; McMahon, T.; O'Beirn, F.X.; Ó Cinneide, M.; Silke, J. (Marine Institute, 2004)
      This report details the investigations into a major mortality of farmed salmon at Inver Bay and McSwyne’s Bay, Co. Donegal in July 2003. Previous reports were provided on 29th July 2003 and on 11th August 2003. The information is based upon analysis and research by MI scientists, a review of environmental data, survey reports by external consultants, inputs from veterinary practitioners who visited the site, reports from DCMNR staff in Killybegs, and site visits made by DCMNR / MI inspectors. Following a review meeting of the principal investigators on the 9th October, 2003, MI proceeded to carry out further scientific investigations. DCMNR also commissioned Kirk McClure Morton Consulting Engineers (KMM) to carry out a parallel investigation of the mortalities at Inver Bay and McSwynes Bay salmon farms. MI provided support as required to the KMM study, the report for which was furnished to DCMNR and MI on 11 February 2004. (KMM, 2004) MI wishes to acknowledge the high level of co-operation and assistance that it received from the owners and staff of Creevin Fish Farm Ltd, Eany Fish Products Ltd and Ocean Farms Ltd. It also wishes to acknowledge the assistance of veterinary practitioners, DCMNR staff and others in the course of this investigation.
    • A sensitive liquid chromatography/tandem mass spectrometry method for the determination of natural and synthetic steroid estrogens in seawater and marine biota, with a focus on proposed Water Framework Directive Environmental Quality Standards

      Ronan, J.; McHugh, B. (Wiley, 2013)
      RATIONALE: Trace levels of natural and synthetic steroid estrogens estrone (E1), 17b-estradiol (E2) and 17a-ethynyl estradiol (EE2) have been demonstrated to exert adverse effects in exposed organisms. E2 and EE2 have been proposed for inclusion in the Water Framework Directive (WFD) list of priority pollutants; however, the detection and accurate quantification of these compounds provide significant challenges, due to the low detection limits required. METHODS: A sensitive method combining ultrasonication, solid-phase extraction (SPE) and liquid chromatography/tandem mass spectrometry, with electrospray ionisation in negative mode (LC/ESI-MS/MS), capable of determining E1, E2 and EE2 at concentrations between 0.07 and 60 ng/L for seawater and between 0.4 and 200 ng/g wet weight in Mytilus spp. is reported. Recoveries at the limit of quantification (LOQ) ranged from 95 to 102% and 88 to 100% for water and tissue, respectively. Salinity (12 to 35%) and typical marine particulate matter loadings (between 10 and 100 mg/L) were not found to affect analyte recoveries. RESULTS: The first detection of E1 by LC/MS/MS in Irish marine waters (Dublin Bay, at 0.76 ng/L) is reported. Steroids were not detected in Galway Bay, or in any mussel samples from Dublin, Galway and Clare. The level of E2 detected in the dissolved water phase was below the proposed WFD Environmental Quality Standard (EQS) in other surface waters. CONCLUSIONS: The proposed method is suitable for the detection of E1, E2 and EE2 at biologically relevant concentrations and, due to the specificity offered, is not subject to potential interferences from endogenous E1 and E2 which often complicate the interpretation of estrogenic biomarker assays.
    • Simulated sunlight inactivation of norovirus and FRNA bacteriophage in seawater

      Flannery, J.; Rajko-Nenow, P.; Keaveney, S.; O'Flaherty, V.; Doré, W. (Wiley, 2013)
      Aims: To investigate norovirus (NoV) and F-specific RNA (FRNA) bacteriophage inactivation in seawater under simulated sunlight and temperature conditions representative of summer (235 W m−2; 17°C) and winter (56 W m−2; 10°C) conditions in Ireland. Methods and Results: Inactivation experiments were carried out using a collimated beam of simulated sunlight and 100 ml of filtered seawater seeded with virus under controlled temperature conditions. NoV concentrations were determined using RT-qPCR, and FRNA bacteriophage concentrations were determined using RT-qPCR and by plaque assay. For all virus types, the fluence required to achieve a 90% reduction in detectable viruses (S90 value) using RT-qPCR was not significantly different between summer and winter conditions. S90 values for FRNA bacteriophage determined by plaque assay were significantly less than those determined by RT-qPCR. Unlike S90 values determined by RT-qPCR, a significant difference existed between summer and winter S90 values for infectious FRNA bacteriophage. Conclusions: This study demonstrated that RT-qPCR significantly overestimates the survival of infectious virus and is therefore unsuitable for determining the inactivation rates of viruses in seawater. Significance and Impact of the Study: Results from this study provide initial data on the inactivation of NoV and FRNA bacteriophage in seawater under representative summer and winter conditions and will be of interest to shellfish and water management agencies alike.
    • Small intestinal injuries in mice caused by a new toxin, Azaspiracid, isolated from Irish mussels

      Ito, E.; Satake, M.; Ofuji, K.; McMahon, T.; Silke, J.; James, K.; Yasumoto, T. (UNESCO, 2001)
      Pathological changes of the small intestine caused by a new toxin, azaspiracid, from Irish mussels were studied. Human poisoning cases included both diarrhetic shellfish and paralitic shellfish poisoning symptoms. The present paper focused on the former. Injuries were observed in the Upper part of the small intestine, where lamina propria in the villi became atrophied at the initial stage, followed by desquamation of epithelial cells and shortening of villi. The injuries were different from the DSP toxin okadaic acid; 1) they developed very slowly after a lag time of about 3 hr, 2) recovery was very late, 3) initial target and process were different.
    • Stable isotope analysis of baleen reveals resource partitioning among sympatric rorquals and population structure in fin whales

      Ryan, C.; McHugh, B.; Trueman, C.N.; Sabin, R.; Deaville, R.; Harrod, C.; Berrow, S.D.; O'Connor, I. (Inter-Research, 2013)
      Stable isotope analysis is a useful tool for investigating diet, migrations and niche in ecological communities by tracing energy through food-webs. In this study, the stable isotopic composition of carbon and nitrogen in keratin was measured at growth increments of baleen plates from 3 sympatric species of rorquals (Balaenoptera acutrostrata, B. physalus and Megaptera novaeangliae), which died between 1985 and 2010 in Irish and contiguous waters. Bivariate ellipses were used to plot isotopic niches and standard ellipse area parameters were estimated via Bayesian inference using the SIBER routine in the SIAR package in R. Evidence of resource partitioning was thus found among fin, humpback and minke whales using isotopic niches. Highest δ15N values were found in minke whales followed by humpback, and fin whales. Comparison between Northeast Atlantic (Irish/UK and Biscayan) and Mediterranean fin whale isotopic niches support the current International Whaling Commission stock assessment of an isolated Mediterranean population. Significantly larger niche area and higher overall δ 15N and δ 13C values found in fin whales from Irish/UK waters compared to those sampled in adjacent regions (Bay of Biscay and Mediterranean) suggest inshore foraging that may be unique to fin whales in Ireland and the UK. Isotopic profiles support spatial overlap but different foraging strategies between fin whales sampled in Ireland/UK and the Bay of Biscay. Stable isotope analysis of baleen could provide an additional means for identifying ecological units, thus supporting more effective management for the conservation of baleen whales.
    • Summary Report on 2015 Residue Monitoring of Irish Farmed Finfish and 2015 Border Inspection Post Fishery and Fishery Product Sample Testing

      Residues Monitoring Programme, Chemistry Section, Marine Environmental Food Safety Services (Marine Institute, 2017)
      On behalf of the Department of Agriculture, Food and Marine (DAFM), the Marine Institute carries out monitoring of chemical residues in finfish for aquaculture sector. This monitoring is set out in the annual National Residue Control Plan, which is approved by the European Commission, and is an important component of the DAFM food safety controls and is implemented under a service contract with the Food Safety Authority of Ireland. Since 1999, the Marine Institute has implemented the National Residues Monitoring Programme for aquaculture. This is carried out on behalf of the Sea Fisheries Protection Authority, which is the responsible organisation for residue controls on farmed finfish. The outcome for residues levels in farmed finfish during 2015 remains one of consistently low occurrence. In 2015, in excess of 676 tests and a total of 1,845 measurements were carried out on 128 samples (i.e. 124 target samples & 4 suspect samples) of farmed finfish for a range of chemical substances, including banned and unauthorised substances, various authorised veterinary treatments and environmental contaminants.
    • Summary Report on 2016 Residue Monitoring of Irish Farmed Finfish and 2016 Border Inspection Post Fishery Product Testing undertaken at the Marine Institute

      Marine Institute (Marine Institute, 2018)
      On behalf of the Department of Agriculture, Food and Marine (DAFM), the Marine Institute carries out monitoring of chemical residues in finfish for aquaculture sector. This monitoring is set out in the annual National Residue Control Plan, which is approved by the European Commission, and is an important component of the DAFM food safety controls and is implemented under a service contract with the Food Safety Authority of Ireland. Since 1999, the Marine Institute has implemented the National Residues Monitoring Programme for aquaculture. This is carried out on behalf of the Sea Fisheries Protection Authority, which is the responsible organisation for residue controls on farmed finfish. The outcome for residues levels in farmed finfish during 2016 remains one of consistently low occurrence. In 2016, in excess of 691 tests and a total of 1,933 measurements were carried out on 136 samples (i.e. 126 target samples & 10 suspect samples) of farmed finfish for a range of chemical substances, including banned and unauthorised substances, various authorised veterinary treatments and environmental contaminants.
    • Summary Report on 2017 Residue Monitoring of Irish Farmed Finfish and 2017 Border Inspection Post Fishery Product Testing undertaken at the Marine Institute

      Marine Institute (Marine Institute, 2018-11-27)
      On behalf of the Department of Agriculture, Food and Marine (DAFM), the Marine Institute carries out monitoring of chemical residues in finfish for aquaculture sector. This monitoring is set out in the annual National Residue Control Plan, which is approved by the European Commission, and is an important component of the DAFM food safety controls and is implemented under a service contract with the Food Safety Authority of Ireland. Since 1999, the Marine Institute has implemented the National Residues Monitoring Programme for aquaculture. This is carried out on behalf of the Sea Fisheries Protection Authority, which is the responsible organisation for residue controls on farmed finfish. The outcome for residues levels in farmed finfish during 2017 remains one of consistently low occurrence. In 2017, in excess of 775 tests and a total of 2,250 measurements were carried out on 141 samples of farmed finfish for a range of chemical substances, including banned and unauthorised substances, various authorised veterinary treatments and environmental contaminants.
    • Summary Report on 2018 Residue Monitoring of Irish Farmed Finfish & 2018 Border Inspection Post Fishery Product Testing undertaken at the Marine Institute

      Glynn, D.; McGovern, E.; Kelly, C.; Moffat, R.; Farragher, E. (Marine Institute, 2019)
      On behalf of the Department of Agriculture, Food and Marine (DAFM), the Marine Institute carries out monitoring of chemical residues in finfish for aquaculture sector. This monitoring is set out in the annual National Residue Control Plan, which is approved by the European Commission, and is an important component of the DAFM food safety controls and is implemented under a service contract with the Food Safety Authority of Ireland. Since 1999, the Marine Institute has implemented the National Residues Monitoring Programme for aquaculture. This is carried out on behalf of the Sea Fisheries Protection Authority, which is the responsible organisation for residue controls on farmed finfish. In 2018, in excess of 920 tests and a total of 2,611 measurements were carried out on 171 samples of farmed finfish for a range of residues. Implementation of the Aquaculture 2018 Plan involves taking samples at both farm and processing plant: • 123 target samples taken at harvest: 110 farmed salmon and 13 freshwater trout. • 48 target samples were taken at other stages of production: 40 salmon smolts and 8 freshwater trout. All 2018 samples were compliant. For target sampling of farmed fish, a summary table of the residue results from 2005 - 2018 is outlined in Table 1. Overall, the outcome for aquaculture remains one of consistently low occurrence of residues in farmed finfish, with no non-compliant target residues results for the period 2006-2014, 0.11% and 0.10% non-compliant target residues results in 2015 and 2016 respectively and no non-compliant target results in 2017 and 2018.
    • Survey of toxaphene concentrations in fish from European waters

      McHugh, B.; Nixon, E.; Klungsoyr, J.; Besselink, H.; Brouwer, A.; Rimkus, G.; Leonards, P.; de Boer, J. (2000)
      Toxaphene, a suspected carcinogen, is a broad spectrum chlorinated pesticide. The objective of this study was to provide information on the toxicological risks posed by toxaphene to the consumer of fish from European waters. The levels of 3 toxaphene congeners in various fish species from different geographical locations were determined. These data were then used to provide information on the exposure of toxaphene to the consumer of fish.
    • Survival and growth of juvenile freshwater mussels (Unionidae) in a recirculating aquaculture system

      O'Beirn, F.X.; Neves, R.J.; Steg, M.B. (American Malacological Union, 1998)
      An indoor recirculating aquaculture system was constructed to provide suitable conditions to culture juvenile freshwater mussels. In the first of three growth trials, Villosa iris (I. Lea, 1829) juveniles were cultured for 22 wk, and grew from an initial mean length of 0.4 mm to 2.7 mm. Survival was 26.8% overall. In the second trial, growth and survival were compared between juveniles of V. iris held in sediment and without sediment. The initial mean length of both groups was 2.7 mm, and this experiment ran for 17 wk. The juvenile mussels in sediment grew to a mean length of 5.7 mm with 85% survival, significantly greater (p < 0.01) than juveniles held without sediment (4.5 mm, 74% survival). In the third trial, two cohorts of juvenile Lampsilis fasciola Rafinesque, 1815, increased in length from I.I mm and 1.4 mm to 3.3 mm and 4.1 mm, respectively, with comparable survival (78.7% versus 64.5%). Results of these trials demonstrate that juvenile mussels can be reared successfully within recirculating systems. One of the factors deemed important in successful culture is continuous feeding of an appropriate food source. In this study, a unialgal culture of Neochloris oleabundans Chantanachat and Bold, 1962, was used throughout. Regular cleaning of the system and water replacement also was important. Finally, the culture of juveniles in sediment appears to be an important factor in ensuring good growth and survival. This phenomenon could be related to pedal feeding behavior, proper orientation of the mussels for filtering efficiency, or stability from physical disturbance.
    • A test battery approach to the ecotoxicological evaluation of cadmium and copper employing a battery of marine bioassays

      Macken, A; Giltrap, M; Ryall, K; Foley, B; McGovern, E; McHugh, B; Davoren, M (Springer, 2009)
      Heavy metals are ubiquitous contaminants of the marine environment and can accumulate and persist in sediments. The toxicity of metal contaminants in sediments to organisms is dependent on the bioavailability of the metals in both the water and sediment phases and the sensitivity of the organism to the metal exposure. This study investigated the effects of two metal contaminants of concern (CdCl2 and CuCl2) on a battery of marine bioassays employed for sediment assessment. Cadmium, a known carcinogen and widespread marine pollutant, was found to be the least toxic of the two assayed metals in all in vivo tests. However CdCl2 was found to be more toxic to the fish cell lines PLHC-1 and RTG-2 than CuCl2. Tisbe battagliai was the most sensitive species to both metals and the Microtox® and cell lines were the least sensitive (cadmium was found to be three orders of magnitude less toxic to Vibrio fischeri than to T. battagliai). The sensitivity of Tetraselmis suecica to the two metals varied greatly. Marine microalgae are among the organisms that can tolerate higher levels of cadmium. This hypothesis is demonstrated in this study where it was not possible to derive an EC50 value for CdCl2 and the marine prasinophyte, T. suecica. Conversely, CuCl2 was observed to be highly toxic to the marine alga, EC50 of 1.19 mg l-1. The genotoxic effect of Cu on the marine phytoplankton was evaluated using the Comet assay. Copper concentrations ranging from 0.25 to 2.50 mg l-1 were used to evaluate the effects. DNA damage was measured as percent number of comets and normal cells. There was no significant DNA damage observed at any concentration of CuCl2 tested and no correlation with growth inhibition and genetic damage was found.
    • Toward design criteria in constructed oyster reefs: oyster recruitment as a function of substrate type and tidal height

      O'Beirn, F.X.; Luchenbach, M.W.; Nestlerode, J.A.; Coates, G.M. (National Shellfisheries Association, 2000)
      Restoration of degraded oyster reef habitat generally begins with the addition of substrate that serves as a reef base and site for oyster spat attachment. Remarkably, little is known about how substrate type and reef morphology affect the development of oyster populations on restored reefs. Three-dimensional, intertidal reefs were constructed near Fisherman's Island, Virginia: two reefs in 1995 using surfclam (Spisula solidissima) shell and six reefs in 1996 using surfclam shell, oyster shell, and stabilized coal ash. We have monitored oyster recruitment and growth quarterly at three tidal heights (intertidal, mean low water, and subtidal) on each reef type since their construction. Oyster recruitment in 1995 exceeded that observed in the two subsequent years. High initial densities on the 1995 reefs decreased and stabilized at a mean of 418 oyster/m2. Oyster settlement occurred on all reef types and tidal heights in 1996; however, postsettlement mortality on the surfclam shell and coal ash reefs exceeded that on the oyster shell reefs, which remained relatively constant throughout the year (mean = 935 oysters/m2). Field observations suggest that predation accounts for most of the observed mortality and that the clam shell and coal ash reefs, which have little interstitial space, suffer greater predation. Oyster abundance was consistently greatest higher in the intertidal zone on all reefs in each year studied. The patterns observed here lead to the preliminary conclusion that the provision of spatial refugia (both intertidal and interstitial) from predation is an essential feature of successful oyster reef restoration in this region. In addition, high levels of recruitment can provide a numerical refuge, whereby the oysters themselves will provide structure and increase the probability of an oyster population establishing successfully on the reef.
    • Toxic phytoplankton in Irish waters

      Silke, J.; McMahon, T.; Nolan, A. (1995)
      The subject of harmful and toxic marine algae has recently gained a growing public and scientific interest both in Ireland and abroad because of the occurrence of these toxins in shellfish.
    • Toxicological risks to humans of toxaphene residues in fish

      Leonards, P.E.G.; Besselink, H.; Klungsøyr, J.; McHugh, B.; Nixon, E.; Rimkus, G.G.; Brouwer, A.; de Boer, J. (Wiley, 2011)
      A revised risk assessment for toxaphene was developed, based on the assumption that fish consumers are only exposed to toxaphene residues that differ substantially from technical toxaphene due to environmental degradation and metabolism. In vitro studies confirmed that both technical toxaphene and degraded toxaphene inhibit gap junctional intercellular communication that correlates with the mechanistic potential to cause tumour promotion. In vivo rat studies established the NOAEL for degraded and technical toxaphene at the highest dose tested in the bioassay. Toxaphene residue intakes from European fishery products were estimated and compared to the provisional tolerable daily intakes (TDIs) from various regulatory agencies including Canada, the United States, Germany. The estimated intake was also compared to a new calculated provisional MATT pTDI. The MATT pTDI is based upon new toxicological information (in vivo rat studies) developed on a model for environmental toxaphene residues rather than technical toxaphene. A MATT pTDI (1.08 mg total toxaphene for a person of 60 kg) for tumour promotion potency was adopted for use in Europe and is hitherto referred to as the MATT pTDI. These new data result in a better estimate of safety and a higher TDI than previously used. Based on realistic fish consumption data and recent baseline concentration data of toxaphene in European fishery products the toxaphene intake for the consumers of Germany, Ireland, Norway and The Netherlands was estimated. For an average adult fish consumer the average daily intake of toxaphene was estimated to be 1.2 µg, and 0.4, 0.5, and 0.2 µg for the consumers of Norway, Germany, Ireland, and The Netherlands, respectively. The toxaphene intake of these average fish consumers was far below the MATT pTDI of 1.08 mg/60 kg body weight. In conclusion, based on the most relevant toxicological studies and the most realistic estimates of fish consumption and recent concentrations of toxaphene in European fishery products, adverse health effects are unlikely for the average European consumer of fishery products. In no case is the MATT pTDI exceeded.
    • The use of immunoassay technology in the monitoring of algal biotoxins in farmed shellfish

      Wilson, A.; Keady, E.; Silke, J.; Raine, R. (International Society for the Study of Harmful Algae and Intergovernmental Oceanographic Commission of UNESCO, 2013)
      The use of immunoassay technology as an adjunct method for monitoring biotoxins in shellfish was investigated at aquaculture sites in Killary Harbour, Ireland, during summer 2009. Sub-samples of mussels (Mytilus edulis) were taken from batches collected as part of the Irish National Phytoplankton and Biotoxin Monitoring Programme (NMP). Samples were analysed for Diarrhetic Shellfish Poisoning (DSP) toxins using a commercially available ELISA immunoassay kit. The results were compared with those obtained by chemical (liquid chromatography with mass spectrometry, LC-MS) and biological (mouse bioassay, MBA) methods from the monitoring programme. DSP levels increased in late June 2009 over the European Union maximum permitted level of 0.16 μg g-1 and positive MBA results led to harvest closures. This event was reflected in both the chemical and immunoassay results, where a positive relationship between them was found.
    • Use of LC-MS testing to identify lipophilic toxins, to establish local trends and interspecies differences and to test the comparability of LC-MS testing with the mouse bioassay: an example from the Irish biotoxin monitoring programme 2001

      Hess, P.; McMahon, T.; Slattery, D.; Swords, D.; Dowling, G.; McCarron, M.; Clarke, D.; Gobbons, W.; Silke, J.; O'Cinneide, M. (Conselleria de Pesca e Asuntos Maritimos da Xunta de Galicia and Intergovernmental Oceanographic Commission of UNESCO, 2003)
      During 2001, the Marine Institute has extended its range of chemical tests to include the analysis of DSP toxins by Liquid Chromatography coupled to Mass Spectrometry (LC-MS). Thus the range of compounds determined extends from domoic acid over DSP compounds (okadaic acid and DTXs) to azaspiracids (AZAs). These tests complement the mouse bioassay, which is the current reference method for lipophilic toxins within the European Community. The development and performance characteristics of the LC-MS method are discussed. Isomer patterns and interspecies differences are discussed as well as local trends in time and variability at one production site at a given time. Comparison of the LC-MS results with the results from the mouse bioassay showed good agreement (93%), and a small but significant number of discrepancies (7%). Overall, the chemical testing has proven to be an invaluable tool in the assessment of shellfish toxicity in Ireland. Lacks of standards and reference materials are discussed as well as the need for further research into the equivalence of methods.