The 9th Irish Shellfish Safety Workshop was held on the 20th March, 2009, in Kenmare, County Kerry. The Workshop was co-sponsored by the Marine Institute, Bord Iascaigh Mhara, the Food Safety Authority of Ireland, and the Sea Fisheries Protection Authority, with support from IFA Aquaculture. The topics addressed at the workshop included an update on the National Biotoxin monitoring programme, and a number of research projects with Irish participation and international perspectives on toxin detection. Finding mechanisms to improve our product was a common theme with presentations on improving food safety, increasing productivity, providing easily applied test methods, and research in support of the shellfish industry. The focus of the three Workshop sessions was on a review of the year, research and legislation.

This document outlines the proceedings of the 8th Irish Shellfish Safety Scientific Workshop. This event was organised by the Marine Institute, the Food Safety Authority of Ireland and Bord Iascaigh Mhara to discuss the methods and advances of food safety with respect to shellfish health.

Water quality monitoring programmes exist in many of the Member States throughout the European Union (EU). With the implementation of the Water Framework Directive (WFD, Council Directive 2000/60/EC) all Member States must harmonise their national monitoring methods for each common metric (parameter indicative of a biological water quality element) used to determine the state of the aquatic environment to ensure consistent and comparable classification results for all biological community quality elements used (WFD Annex V, 1.4.1). The Marine Ecological Tools for Reference, Intercalibrationand Classification (METRIC) project, therefore, was designed specifically to support the Irish role in the EU Intercalibration Exercise of biological quality elements (BQEs) in order to set harmonised ecological quality criteria for the assessment of water quality in the transitional and coastal (TraC) waters of Europe. The BQEs investigated by METRIC included: Plants (phytoplankton, macroalgae andangiosperms), Benthic macroinvertebrates (soft-bottom habitat), Fish (estuarine).

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.

This paper presents a description of three of the proposed phytoplankton indices under investigation as part of a classification framework for UK and ROI marine waters. The three indices proposed for the classification process are (i) phytoplankton biomass measured as chlorophyll, (ii) the frequency of elevated phytoplankton counts measuring individual species and total cell counts and (iii) seasonal progression of phytoplankton functional groups through the year. Phytoplankton biomass is calculated by a 90th percentile measurement of chlorophyll over the growing season (April to September) compared to a predetermined reference value. Calculation of functional groups and cell counts are taken as proportional counts derived from the presence of the indicator species or group as compared to the total phytoplankton count. Initial boundary conditions for the assessment of high/good status were tested for each index. Chlorophyll reference conditions were taken from thresholds developed for previous EU directives with the setting of offshore concentrations as a reference condition. Thresholds for elevated counts of phytoplankton taxa were taken from previous EU assessments describing counts that could be impact negatively on the environment. Reference seasonal growth curves are established using phytoplankton counts from ‘‘high status’’ waterbodies. To test the preliminary boundaries for each index, a risk assessment integrating nutrient enrichment and susceptibility for coastal and transitional waters was carried out to identify WFD waterbodies in England and Wales at different levels of risk. Waterbodies assessed as having low or medium risk from nutrient enrichment were identified as type 1 and type 2 waterbodies, and waterbodies assessed as high risk were identified as type 3 waterbodies. Phytoplankton data was extracted from the risk assigned waterbodies and applied to each phytoplankton index to test the robustness of the preliminary classification ranges for each phytoplankton index.

Azaspiracids (AZAs) are a group of lipophilic polyether toxins that were discovered in shellfish from Ireland in 1995, following a food poisoning incident. Both the limited availability of pure AZAs and the co-occurrence in shellfish of other toxins in combination with AZAs have so far prevented an in-depth evaluation of the performance of the EU reference test, the mouse bioassay (MBA), for this toxin group at the regulatory limit. The present study evaluated the performance of the mouse bioassay at the example of a mussel tissue homogenate, naturally contaminated with AZAs, diluted with uncontaminated tissues to appropriate concentration levels. Concentrations were determined using liquid chromatography coupled to tandem mass spectrometry (LC-MS-MS) (7 levels ranging from levels less than the limit of quantification to a maximum of ca. 2.24 mg/kg in hepatopancreas, which corresponds to a maximum whole flesh AZA1-equivalent of ca. 0.34 mg/kg). Replicate homogenates of each concentration level were analysed by MBA on 7 independent occasions over 6 weeks. Inhomogeneity between replicate aliquot portions was evaluated using LC-MS-MS and ranged from 1.8 to 6.6% RSD for the six levels contaminated above quantification limits. This variation was similar to the variability of the LC-MS-MS method within a batch, and the difference between replicate aliquots could thus be considered negligible. Other uncertainties considered in the study included the short- and long-term variability of the LC-MS-MS method, toxic equivalence factors, relative response factors in mass spectrometric detection, additional analogues and matrix effects. A concentration-response curve was modelled as a 4-parametric logistic fit to a sigmoidal function, with an LC50 of 0.70 mg AZA1-equivalent/kg hepatopancreas tissue. Furthermore, the mathematical model of the lethality data from this study suggests that occasional negative mouse assays at high concentrations, previously observed in the Irish statutory monitoring, are at least partly due to the biological variation of mice and can be understood on a statistical basis. The mathematical model of the concentration-response curve also describes the probability of a positive mouse bioassay at the current regulatory limit of 0.16 mg/kg to be ca. 95%. Therefore, it appears that the mouse bioassay performs very well in the implementation of this limit. Hence, the present study very strongly suggests that the MBA and LC-MS-MS techniques can be considered equivalent in the implementation of the current regulatory limit of 0.16 mg/kg for Azaspiracids in shellfish.

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.

Since its initial development in the early 1970s the Irish aquaculture industry has grown to be an important contributor to the national economy. There has been a steady increase, in both output and value, as well as in job creation. The total production of farmed shellfish has increased from approximately 5,000 tonnes in 1980 to 44,678 tonnes in 2003 (Figure 1), with a first sale value of €41.8m and directly employing some 1100 people (Parsons et al, 2004). Mussels (Mytilus edulis), native oysters (Ostrea edulis), Pacific oysters (Crassostrea gigas), Clams (Tapes semidecussata) and scallops (Pecten maximus) are the main species produced. With a growing recognition and awareness internationally of the potential human health effects of the consumption of shellfish containing algal toxins, a monitoring programme was established in Ireland in the early 1980s and has continued since then. In this paper the evolution and development of the programme is described and discussed.