ISSN: 1995-4875 CRFM Special Publication. No.14 Manual on Laboratory Testing of Fishery Products The SPS Project is funded by the European Union under the 10th Economic Development Fund and is being implemented by the Inter-American Institute for Cooperation on Agriculture (IICA) with the following regional Partners: the CARICOM Secretariat, the Caribbean Regional Fisheries Mechanism (CRFM), El Comité Nacional para la Aplicació n de Medidas Sanitarias y Fitosanitarias de la Repú blica Dominicana (CNMSF) and CARIFORUM. Manual on Laboratory Testing of Fisheries Products Copyright © 2016 by Caribbean Regional Fisheries Mechanism (CRFM) All rights reserved. Reproduction, dissemination and use of material in this publication for educational or non- commercial purposes are authorized without prior written permission of the CRFM, provided the source is fully acknowledged. No part of this publication may be reproduced, disseminated or used for any commercial purposes or resold without the prior written permission of the CRFM. Prepared by: Christine Froese, Megapesca Lda., December 2016, under contract to the Inter- American Institute for Cooperation on Agriculture (IICA), through the 10th EDF funded Sanitary and Phytosanitary Project Correct Citation: Froese, C, 2016. Manual on Laboratory Testing of Fishery Products. CRFM Special Publication. No.14. 89pp. ISSN: 1995-4875 ISBN: 978-976-8257-38-3 Cover Photo: Fishery Product Testing Laboratory, Saint Vincent Contents 1 INTRODUCTION ................................................................................................................................ 1 1.1 BACKGROUND .................................................................................................................................... 1 1.2 ABOUT THIS MANUAL ........................................................................................................................... 1 1.3 HOW TO USE THIS MANUAL ................................................................................................................... 2 2 OVERVIEW OF REQUIREMENTS ........................................................................................................ 3 2.1 OFFICIAL CONTROL LABORATORIES .......................................................................................................... 3 2.2 STAFF REQUIREMENTS .......................................................................................................................... 4 2.3 QUALITY CONTROL AND VALIDATION OF TEST METHODS .............................................................................. 5 3 MICROBIOLOGICAL PARAMETERS FOR WATER ................................................................................. 6 3.1 SAMPLING FOR WATER ANALYSIS, SAMPLE HANDLING AND SAMPLE PREPARATION ............................................ 7 3.2 MICROBIOLOGICAL METHODS ................................................................................................................ 8 4 MICROBIOLOGICAL PARAMETERS FOR FISHERY PRODUCTS ........................................................... 19 4.1 SOURCES OF DATA ............................................................................................................................. 19 4.2 SAMPLING ....................................................................................................................................... 19 4.3 MICROBIOLOGICAL METHODS .............................................................................................................. 22 5 CHEMICAL PARAMETERS ................................................................................................................ 54 5.1 INTRODUCTION ................................................................................................................................. 54 5.2 HEAVY METALS ................................................................................................................................. 54 5.3 HISTAMINE ...................................................................................................................................... 59 5.4 SULPHUR DIOXIDE AND SULPHITE RESIDUES ............................................................................................ 63 5.5 SHELLFISH POISONS (PSP, ASP, DSP) .................................................................................................. 66 5.6 CIGUATERA TOXIN ............................................................................................................................. 85 6 ADDITIONAL TESTS REQUIRED FOR PRODUCTS OF AQUACULTURE ................................................ 88 ANNEX 1: FURTHER READING.................................................................................................................. 91 List of Tables Table 1: Commercial product tests in water quality ................................................................................ 17 Table 2: Commercial product tests Colony Count .................................................................................. 24 Table 3: Typical Salmonella biochemical reactions .................................................................................... 28 Table 4: Commercial product tests for Salmonella ................................................................................... 30 Table 5: Commercial test products for Clostridium perfringens .............................................................. 34 Table 6: Commercial test products for Coliform bacteria (food and environment) ........................ 36 Table 7: Commercial product tests for Escherichia coli ............................................................................ 40 Table 8: Commercial products for Vibrio rapid detection ....................................................................... 48 Table 9: Commercial product tests for Listeria monocytogenes .............................................................. 53 Table 10: Expected sulphur dioxide (ppm) level........................................................................................ 63 Table 13: Mass of shellfish tissue on column (g) after periodate and peroxide oxidations for quantification of STX toxin ................................................................................................................... 78 Table 14: PSP concentrations at the action limit and relative toxicities to STX ............................... 79 Table 15: Minimum required performance limit for testing certain substances in food .................. 89 List of Figures Figure 1: Monier-Williams apparatus for sulphite residues ..................................................................... 66 ABBREVIATIONS µg Microgram 3-MCPD 3-monochloropropane-1,2-diol or 3-chloro-1,2-propanediol AAS Atomic Absorption Spectroscopy AFNOR Association Française de Normalisation AOAC Association of Analytical Chemists ASP Amnesic Shellfish Poisoning ASPW Alkaline saline peptone water BAM Bacteriological Analytical Manual BSI British Standards Institution BMS Bivalve Molluscan Shellfish CAs Competent Authorities CCA Chromogenic Coliform Agar Cd Cadmium CEFAS Centre for Environment, Fisheries & Aquaculture Science CEN European Committee for Standardisation Cf Correction Factor CFU Colony Forming Units CFP Ciguatera Fish Poisoning SPE COOH Solid Phase Extraction Purification of Carboxylic Acid Products CREM Ciguatoxin Rapid Extraction Method CRFM Caribbean Regional Fisheries Mechanism CTX Ciguatoxins DNA Deoxyribonucleic acid DSP Diarrheic shellfish poisoning E. Coli Escherichia coli EC European Commission EDF European Development Fund EIA Enzyme immunoassay ELISA Enzyme-linked immunosorbent assay EN English Language EPA Environmental Protection Agency (US) EU European Union FDA Food and Drug Administration GC-ECD Gas chromatography with electron capture detection GC-MS Gas Chromatography with Mass Spectrometer GLC Gas Liquid Chromatograph h Hours HABs Harmful algal blooms HACCP Hazard Analysis and Critical Control Point HCH Hexachlorcyclohexan Hg Mercury HPLC High performance liquid chromatography ICMSF International Commission on Microbiological Specifications for Foods IEC International Electrotechnical Commission ISO International Organization for Standardization IUPAC International Union of Pure and Applied Chemistry kg Kilogram LC-MS Liquid Chromatography-Mass Spectrometry LC-MS-MS Liquid Chromatography–Tandem Mass Spectroscopy MBA Mouse Bio Assay m-CP Membrane Clostridium perfringens MF Membrane Filtration MKTTn Muller-Kauffmann Tetrathionate-Novobiocin MPN Most Probable Number MRL Maximum Residue Level MRPL Minimum Required Performance Limit MU Mouse Units MUG 4-methylumbelliferyl-beta-D-glucuronide NaCl Sodium Chloride NACMCF National Advisory Committee for Microbiological Criteria for Foods NF Validation mark, European certification system by AFNOR based on EN ISO 16140 standard NSP Neurotoxic Shellfish Poisoning Pb Lead PBS Phosphate Buffered Saline PCB Polychlorinated Biphenyl PCR Polymerase Chain Reaction ppm Part per million PSP Paralytic Shellfish Poisoning PTFE Polytetrafluoroethylene RVS Rappaport-Vassiliadis Medium with Soya SO2 Sulphur Dioxide SPE Solid Phase Extraction SPS Sanitary and Phytosanitary STX Saxitoxin TCBS Thiosulphate Citrate Bile and Sucrose TSA Tryptone Soy Agar TSC Tryptose Sulfite Cycloserine TSI Triple Sugar Iron TSYEA Tryptone Soya Yeast Extract Agar TSYEB Tryptone Soya Yeast Extract Broth UKAS UK Accreditation Service VRBL Crystal Violet Neutral Red Bile