Leatherhead Food International Leatherhead Food International

Edited by
Rhea Fernandes

This edition first published 2009 by Leatherhead Publishing, a division of Leatherhead Food International Ltd Randalls Road, Leatherhead, Surrey KT22 7RY, UK URL: http://www.leatherheadfood.com and Royal Society of Chemistry Thomas Graham House, Science Park, Milton Road, Cambridge, CB4 0WF, UK URL: http://www.rsc.org Regstered Charity No. 207890

ISBN: 978-1-905224-76-0

A catalogue record of this book is available from the British Library

© 2009 Leatherhead Food International Ltd

The contents of this publication are copyright and reproduction in whole, or in part, is not permitted without the written consent of the Chief Executive of Leatherhead International Limited. Leatherhead International Limited uses every possible care in compiling, preparing and issuing the information herein given but accepts no liability whatsoever in connection with it.

All rights reserved Apart from any fair dealing for the purposes of research or private study, or criticism or review as permitted under the terms of the UK Copyright, Designs and Patents Act, 1988, this publication may not be reproduced, stored or transmitted, in any form or by any means, without the prior permission in writing of the Chief Executive of Leatherhead International Ltd, or in the case of reprographic reproduction only in accordance with the terms of the licences issued by the Copyright Licencing Agency in the UK, or in accordance with the terms of the licences issued by the appropriate Reproduction Rights Organization outside the UK. Enquiries concerning reproduction outside the terms stated here should be sent to Leatherhead International Ltd at the address printed on this page.

Printed and bound by Biddles Ltd., King’s Lynn

Associate Prof. Covadonga Arias Microbial Genomics Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America Dr. Simon Derrick Special Projects The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom Carlos Abeyta, Jr. Supervisory Microbiologist Pacific Regional Laboratory Northwest U.S. Food and Drug Administration 22201 23rd Dr. S.E. P. O. Box 3012 Bothell WA 98021-4421 United States of America Linda Nicolaides, M.Ph., FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom

Prof. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom Jeffrey L.C. Wright C.M., Ph.D., FCIC Carl B. Brown Distinguished Professor of Marine Science UNCW Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America Rhea Fernandes and Dr. Peter Wareing Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom Eugenia Choi and Dr. Jenny Pfleger Regulatory Advisors Leatherhead Food International Randall Road Leatherhead Surrey KT22 7RY United Kingdom


which has been the chief factor in ensuring the completion of this book. I thank the contributing authors for their collaboration. for his scientific input and Victoria Emerton. Alison Turner. Technical Team Leader for her valuable editorial contribution. and growth characteristics for key pathogens associated with the product. All handbooks also contain a review of the related legislation in Europe and UK. and Ann Pernet for the work put into typesetting and indexing. both in the UK and elsewhere. thank you for all the support. I dedicate this book to my husband . Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product. and a detailed list of contacts for various food authorities. Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged. The books in the series are designed as easy-to-use guides to the microorganisms found in foods.FOREWORD The Microbiology Handbook series includes Dairy Products. I also acknowledge and thank Jackie Apps. My gratitude is extended to Dr Peter Wareing. potential hazards associated with the consumption of a range of products. Fish and Seafood. The books are intended as a source of information for microbiologists and food scientists working in the food industry and responsible for food safety. guides to HACCP. published by Leatherhead Food International and RSC Publishing. Rhea Fernandes Leatherhead Food International v .Goldwyn. Training Manager. and Meat Products.

vi .

CHILLED AND FROZEN RAW FISH 1.1 Introduction 2.6 Published Microbiological Criteria 4.7 References 3.7 Published Microbiological Criteria 1.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.4 Spoilage 4.3 Initial Microflora 2.8 References MOLLUSCAN SHELLFISH 3.7 Published Microbiological Criteria 2.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.8 Further Reading CRUSTACEAN SHELLFISH 4.4 Spoilage 3. vii .2 Initial Microflora 4.5 Spoilage 2.4 Spoilage 1.3 Processing and its Effects on the Microflora 1.7 References 4.1 Definitions 1.8 References 1. 3.3 Processing and its Effects on the Microflora 4.5 Pathogens: Growth and Survival 4.4 Processing and its Effects on the Microflora 2.2 Initial Microflora 3. 4.2 Initial Microflora 1.5 Factors Affecting Fresh Fish Spoilage 1.3 Processing and its Effects on the Microflora 3.6 Published Microbiological Criteria 3.2 Definitions 2.6 Pathogens: Growth and Survival 2.6 Pathogens: Growth and Survival 1.1 Definitions 4.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.5 Pathogens: Growth and Survival 3.1 Definitions 3.

854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption 212 9. 8.3 Stages of a HACCP Study 8.8 Ciguatoxins 7.4 Regulation (EC) No.5.3 Regulation (EC) No. CURED.7 Food Hygiene (England) Regulations 2006.7 Further Reading FISH AND SHELLFISH TOXINS 7.2 Legislative Structure 190 9.2 Initial Microflora 6.8 Further Reading FERMENTED FISH 6.2 Initial Microflora 5.4 Implementation and Review of the HACCP Plan 8. S.2 Definitions 8.6 Lipophilic Shellfish Toxins (LST) 7.I.5 Pathogens: Growth and Survival 6.6 Regulation (EC) No.1 Introduction 189 9.1 Introduction 8.5 Pathogens: Growth and Survival 5. 852/2004 on the General Hygiene of Foodstuffs 191 9.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.8 Guidance 222 9.7 References 5.3 Processing and its Effects on the Microflora 5.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. EU FOOD HYGIENE LEGISLATION 189 9.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.4 Spoilage 6.3 Processing and its Effects on the Microflora 6. 7.11 Conclusion 7.6 Published Microbiological Criteria 5.5 Regulation (EC) No. 14 (Hygiene requirements specific to the UK) 221 9.3 Tetrodotoxin 7. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.1 Introduction 7. SMOKED AND DRIED FISH 5.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.6 References 6.1 Definitions 6. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.9 Azaspiracids 7. 9.1 Definitions 5.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.10 Further Reading 223 viii .10 Cyclic Imines 7.4 Spoilage 5. 2006 No.9 Other Relevant Legislation 223 9.5 References 93 93 95 96 108 113 116 117 120 123 123 126 127 135 136 137 140 141 141 142 144 146 148 151 153 155 158 160 165 166 175 175 176 177 184 186 6.

7 Staphylococcus aureus 10. 10. 10.4 Listeria spp.8 Vibrio cholerae 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .10 Vibrio vulnificus 10.9 Vibrio parahaemolyticus 10.1 Aeromonas spp. 10.5 Plesiomonas 10.10 PATHOGEN PROFILES 10.6 Salmonella spp.3 Clostridium perfringens 10.2 Clostridium botulinum 10.


.B. Saebo A. Geographical region. and fish and shellfish toxins. Wageningen.A. The Microbiology Handbook. Luten J. smoked and dried fish. Wageningen Academic Publishers. Nollet L. Bekaert K.INTRODUCTION Fish and seafood are a main source of animal protein in the diet. crustacean shellfish. Cross N.M. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs. Milk. The second edition of this handbook is a review of the entire book for currency of information. 2006. Because of their health advantages over red meats. Seafood processing: basic sanitation practices.H. 94767.. Poultry. Wiley. xi . and. Chandan R. chilled and frozen prepared fish. Catches can be gathered from seas. in Handbook of Food Products Manufacturing: Health. Jacobsen C. Hui Y.. season.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish.. whether they are pelagic (surface to mid-water) or demersal (bottom) feeders will influence the numbers and types of microorganisms present on freshly caught seafood. 2007. fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins. Safety and Processing of Wild and Farmed Fish. for fish. fermented fish. Eds. the consumption of fish and seafood has increased. cured. Often contamination is from human and animal sources. rivers and lakes whose water can range from pristine to contaminated. Clark S. thus. Seafood Research from Fish to Dish: Quality. Meat. Oehlenschlager J. Seafood and Vegetables. Further Reading General Fish/Seafood Microbiology Cross N. molluscan shellfish.. Hoboken....

H. 87121. Davies A.A.H. 200-23.H. 2004. basic sanitation practices. Modified-atmosphere packaging of fish and fish products. Cambridge. Control of indigenous pathogenic bacteria in seafood. Proceedings of the International Seafood Conference. Gram L. Martin R. Dalgaard P. Bengtsson N. Noordwijkerhout. Nip W-K.. Cambridge. 8 (2). 431-43. Microbiological spoilage of fish and fish products..Y. in Microorganisms in Foods 6: Microbial Ecology of Food Commodities. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products.. The fish industry.. Ashie I. Ventresca P. 2003. Simpson B. Bruinsma B. Amsterdam. New York.D.P.R.. Heinitz M. Noordwijkerhout.. London.L. Smith J. 1997. Smith J. Dalgaard P.. Ohlsson T. Taylor S.. and Clostridium botulinum in smoked fish and shellfish.L. Spoilage and shelf-life extension of fresh fish and shellfish.S. in Food Plant Sanitation. November 1995.S. 543-61. 121-37. in Minimal Processing Technologies in the Food Industry. Tong P.H. 245-66 Bremner H. Salmonella spp. Elsevier. in Seafood from Producer to Consumer: Integrated Approach to Quality. 174-249.B. 33 (1). Johnson J. Eds. in Seafood from Producer to Consumer: Integrated Approach to Quality. 2004. 1996. Food Control. Woodhead Publishing Ltd. Huss H. 318-23. Ed. 2002. 372-6. 459-72.. Fish and fish products (Microorganisms in fish products). Huss H. Oxford.. Microbiology of fish and fish products. Luten J. 1997.L. Journal of Food Protection. Gorham J. 2002.M. 167-77. CRC Critical Reviews in Food Science and Nutrition. 60 (4). Stanfield P.J. in Advances in Food and Nutrition Research.A.N. November 1995.. Rawles D. Blech Z. International Commission on Microbiological Specifications for Foods. Marcel Dekker. Flick G. Minimal processing in practice: seafood. The incidence of Listeria spp. Safety and Quality Issues in Fish Processing... Blackie.H. Seafood: processing. Luten J. Gram L. 1997.International Commission on Microbiological Specifications for Foods.E. Predictive microbiological modelling and seafood quality.K. McCarthy S. 61 (3). 36 (1-2). Amsterdam. Ed.M. 1996. Hall G. Huss H.A. Elsevier. 413-30. Ed. in Food Processing: Principles and Applications. 1997. Kluwer Academic / Plenum Publishers.. Gudmundsson M.Y. Woodhead Publishing Ltd. Blackwell Publishing. Eds.. Blech Z. Ed. Ed. New York.. 91-8. Seafood processing: basic sanitation practices. 2005. 1996. in Fish Processing Technology. Eds. Stanfield P. 1997. London.B. 1998. Academic Press.. Oxford. Biogenic amines in fish and shellfish. Hui Y. Journal of Food Protection. International Journal of Food Microbiology. in Kosher Food Production.. Proceedings of the International Seafood Conference. xii . Vo1ume 39. 329-65. Ed.R. Hafsteinsson H. Hui Y. Blackwell Publishing.

McMeekin T.H. 5 (3). Sofos J. 1994. Food Technology. London. Bonnell A. 269-82. Amsterdam. Berlin. Jakobsen M. in Fisheries Processing: Biotechnological Applications. International Commission on Microbiological Specifications for Foods.. Evaluation of the bacteriological quality of seafood. 2257. Copenhagen. 305-17. Martin A. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. A new look at microbiological standards. Liston J. Qualitative and quantitative characterization of spoilage bacteria from packed fish. Huss H. International Journal of Food Microbiology. PHLS Microbiology Digest. The microbiology of seafood products. Volume 5: Microbiological Specifications of Food Pathogens. 1990. Pearson A. in Food and Beverage Technology International: USA. 1991. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. poultry and fish. Twigg B. 11 (2). Jakobsen M. Microbial growth and its control in meat. Garrett E.H. 1992. Amsterdam. Microbial hazards of seafood consumption.S. Blackie. Olley J.H. Hobbs G. Fish spoilage and safety. 5662. Ed. 16 (1). Amsterdam. Ross T. August 1991. Institut fur Veterinarmedizin. Bernoth E-M.International Commission on Microbiological Specifications for Foods. 1996. 319-33.N. Glasgow.A. 1994. Dutson T.D. Microbiological quality of fishery products. in Quality Attributes and Their Measurement in Meat. Elsevier. 1995. Elsevier. Eds. Ward D. August 1991. 90-106. 459-78. 118-20. Blackie.M.M.J. Ed. Bacterial spoilage of seafood. Copenhagen. in Public Health Aspects of Seafood-borne Zoonotic Diseases. Food Science and Technology Today. Seafood toxins of microbiological origin. Dalgaard P. Chapman & Hall. Chapman and Hall. Gram L.. Liston J. Activities Report of the R&D Associates.. Eds. Liston J. 26 (3). Ed. Evaluation of the bacteriological quality of seafood. 1994. Sterling Publications Ltd. Eds. Copenhagen. Elsevier.. guidelines. Eds.. xiii . Gibson D. Microbiological indicators and seafood processing. Liston J. 166-73. and specifications for seafood products. Ward D. 46 (1). in Microorganisms in Foods.R. 1-17.R. Poultry and Fish Products. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. 1994. 1989. 1995. Jakobsen M. 54-61. 1990. Liston J. Modelling of microbial activity and prediction of shelf-life for packed fish. London. 93-105. Ed. 1992. Fish: microbiological spoilage and safety. London. 25-39.D.M.A. 265-79.. in Quality Assurance in Seafood Processing: A Practical Guide. 1992. 44 (12). Ed. 26 (3). Application of predictive microbiology to assure the quality and safety of fish and fish products. International Journal of Food Microbiology. Gilbert R.. London. 1994. Dalgaard P. International Journal of Food Microbiology. Huss H. 1992.R. 121-8. Bacterial pathogens transmitted by seafood. Bonnell A. Huss H.. August 1991. 359-403.. Gram L.

Microbiology of frozen fish and related products. Hobbs G. Ed. Applied Science Publishers. Hackney C. Simmonds C. 1987.K. London.R.. Microbiological hazards associated with fishery products.spoilage of fish. 1985.J. Davies R. 103. CSIRO Food Research Quarterly. Hobbs G. 42 (3). 42 (3). Elsevier Applied Science Publishers.. 8-16. 1988. Microbiology of fish. The bacteriology of fish handling and processing.R. Part I . in Microbiology of Frozen Foods. Wiley. and specifications and inspection of seafood products. Lamprecht E. 90-3.S. 46 (1). Ed. Journal of Food Quality. Robinson R. 199-226.M. Volume 1. London. guidelines. 71-117. in Essays in Agricultural and Food Microbiology. 169-208.Pedrosa-Menabrito A. Shelf-life extension of fresh fish . 1988. 1982. Hodgkiss W. 117-27. 104-9. Garrett E. xiv . Norris J. Food Technology. Regenstein J. Microbiological standards. in Developments in Food Microbiology. 11 (2)..a review. Chichester. Food Technology. Ed.K. 1988. Eyles M. Dicharry A.. 1986.C. Seafood-borne bacterial pathogens of marine origin.

but typically present a low fat percent when compared to other meats. However. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. which can be up to 67%. and is mostly interspersed between the muscle fibres.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. it shows a typical season pattern especially in migratory species such as herring or mackerel. lean fish are those fish that store most of their fat in the liver. Fish fillet composition can vary significantly within the same species due to feed intake.2 . and spawning season. giving it its characteristically flaky structure. 1 . In addition. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. In this chapter. fins and a streamline body.5% (1).20%. typically 3 and 15% of total weight. most fish contain omega 3-fatty acids and other essential nutrients. Muscle composition and structure of fish also differ from those found in other meat. The lipid fraction is the component showing the greatest variation. and excludes molluscs and Crustacea. respectively (1). typically fluctuates between 0. The lipid content. while fatty fish have fat cells distributed along their bodies. The connective tissue content of fish is also lower than that found in meat. and other forms of aquatic animal as part of their ‘fish’ definition. ‘fish’ also refers to the flesh of such animals used as food. migratory patterns.21%. Fish flesh is dominated by the abundance of white muscle in relatively short segments. Fish are an important part of a healthy diet since they contain high quality protein. Protein content in fish fillet varies typically from 16 . some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. In addition.1. crustaceans. offering less than 0. Fish can be divided into fatty and lean fish. This super class of vertebrates includes all the bony and cartilaginous finfish. Fish fillets are a poor source of carbohydrates. “fish” will be used for fresh and seawater finfish. CHILLED AND FROZEN RAW FISH Associate Prof.

DMA is dimethyl amine. frozen or preserved. hence. 2 . and maintained at or below -2 °C (normally below -12 °C) during storage. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. transportation and sale. spoilage microflora describes an association of microorganisms that. Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases. odour and texture. transportation and sale. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. This technique is primarily used for bulk storage. transportation and sale. and flavour based on the maturity. and texture of the fish. Quality refers to palatability and organoleptic characteristics such as tenderness. Controlled-atmosphere packaging (CAP) refers to packaging in an atmosphere where the composition of the gases is continuously controlled during storage. and maintained at or below 7 °C. usually carbon dioxide (CO2). Evisceration is the removal of the viscera from a fish. during storage. but no further control is exercised during storage. at -2 to -4 °C. juiciness. but not below 3 °C during storage. Spoilage describes changes that render fish objectionable to consumers. firmness. Superchilled fish is fish that has been cooled to. nitrogen (N2) and oxygen (O2). through their development on fish. marbling. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. Fresh fish is raw fish that has not been processed. colour. Organoleptic refers to qualities such as appearance. The proportion of each component is fixed when the mixture is introduced. Frozen fish is fish that has been cooled to. colour. and maintained at temperatures just below the freezing point.FISH AND SEAFOOD Chilled fish is fish that has been cooled to. renders that fish objectionable to consumers.

as with other vertebrates. In general. It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish. Shewanella and Pseudomonas. Moraxella. standard culture-dependent methods can only recover between 1 to 10% of total bacteria present in any given sample. potential human pathogens may be present in both scenarios. 1. some species (most Vibrios) require sodium chloride for growth. However.) are also common aquatic bacteria. For example. The number and diversity of microbes associated with fish depend on the geographical location. whenever possible. posing a concern for seafoodborne illnesses (2). More accurate. are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest. Gram-positive organisms such as Bacillus. Flavobacterium. These pathogens can be divided into two groups: organisms naturally present on fish (Table 1.2 Initial Microflora The subsurface flesh of live. The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. gills and the gastrointestinal tract of fish. healthy fish is considered sterile and should not present any bacteria or other microorganisms. and those that although not autochthonous to the aquatic environment. Lactobacillus and coryneforms can also be found in varying proportions (1). Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp.I). In fact. and typical of the fish flora. Human pathogenic bacteria can be part of the initial microflora of fish. the natural fish microflora tends to reflect the microbial communities of the surrounding waters. since they heavily depend on the type of sample analysed and the protocol used for isolation. molecular-based methods have not yet been used to address this issue. It is difficult to estimate how many microorganisms are typically associated with fish. microorganisms colonise the skin. Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters.CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed. It must be noted that mesophilic bacteria can rapidly overgrow psychrotropic organisms. Clostridium. Micrococcus.II) (3). the season and the method of harvest. several culture media containing sodium chloride and more than one incubation temperature should be used. processing or storage (Table 1. Gastrointestinal tracts and gills typically yield high bacteria numbers. On the contrary. 3 . although these are influenced by water quality and feed.

and mud Ubiquitous. Toxin levels 0. sewage.109 cells/g) * Some sources considered L.>106 Salmonella spp.0. In addition. 4 . Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp.1 .I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. monocytogenes as part of the natural aquatic flora It is apparent from the above that there is potentially a very diverse range of organisms present on fish.FISH AND SEAFOOD TABLE 1. estuarine environments. vulnificus) 5 .103 High (107 . during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria. birds.37 °C Unknown Unknown TABLE 1.37 °C High for most species.106 cells/g (exception: V.14 . and risk associated with the consumption of seafood is low (2.102 101 .26 °C 10 . stream water. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. 105 . numbers of pathogenic bacteria in raw fish tend to be low.II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.106 cells/g. Plesiomonas shigelloides 3 . Escherichia coli Yersinia enterocolitica 101 .35 °C 8 . human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . 4). However.19 µg/kg bodyweight from < 102 .1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.

and especially when fish is stored on board for longer periods. The length of time that set nets have been in the water or the time trawlers’ nets are towed. offering better control of fish health prior to. boxes and/or baskets. In eviscerated fish. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea. and conditions of storage. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. with very little control over the condition of the fish at the time of death or the duration of the killing process. cortisone levels increase during prolonged stress and can alter the fillet quality. the cuttings provide areas of exposed flesh that are open to microbial attack. with fish such as tuna and cod being more commonly eviscerated than sardines. aquaculture practices have been expanding worldwide. If evisceration is carried out at sea. Fish may be eviscerated prior to storage at sea . During capture and storage. or traps. and spoilage. in recent decades. However. and the killing process is designed to minimise stress. in which the bacterial load 5 . In addition. This contrasts greatly with the meat industry. icing or freezing. hook and line. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria.3. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating.3 1. Increasingly. potential contaminants added during handling and processing. After capture. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. ropes.a practice that may have both advantages and disadvantages. and their re-use will increase the crosscontamination of other fish with such microorganisms. Physical damage such as loss of scales. finfish will almost invariably come into contact with nets. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating. handling and processing Wild finfish are usually caught by net. with the production of off-odours. birds and soil. however. digestion and off-flavours in uneviscerated fish. mackerel or herring. decks.1 Processing and its Effects on the Microflora Capture. 1. The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. and during harvest. human hands and clothing. in which the health of each animal can be assessed prior to slaughter. chilled brine or refrigerated seawater at -2 °C. These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans. has an effect on the amount of stress and physical damage that the fish will suffer during capture.

Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. if there is some oxygen demand from the product. These containers are also used for displaying the fish during auction at the quayside. These changes can be deleterious. Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. As with all foods. 1. will result in fully anoxic conditions. Good Manufacturing Practice (GMP) and control of the sanitary conditions of the transport and processing environments are essential to limit additional risk of disease caused by fish consumption (6. careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. nitrogen. but if applied properly should extend the life of the product. allowing more growth of spoilage organisms. a reduction in the oxygen tension is achieved which.3. however. and minimising handling will all reduce microbial cross-contamination. not part of the normal flora of the meat animals (8). 7). they are usually process contaminants. fish have a higher post mortem pH. Physical barriers not only protect from physical damage. for example. oxygen etc. partial fermentation of sugars occur leading to lower pH.). Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. and of the quality of the water used for ice and to wash fish. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. ferric ion (Fe3+)). and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO). Anaerobic atmospheres have less effect on fresh fish shelf life. often in the absence of adequate refrigeration.FISH AND SEAFOOD can be substantial. from water activity (aw) through to physical damage. cleaning of the work environment. by actively altering the composition of the 6 . What is more. The atmospheric conditions surrounding a product may be passively or actively altered. Packaging changes the intrinsic and extrinsic parameters affecting a product.2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. in time. In the absence of oxygen. and to limit the loss of quality (5). but isolate the food in an environment different from the bulk atmosphere. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. By vacuum packing foods. In MAP. However. use of effective detergents and disinfectants.

Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains. The microflora of meat is not the same as that of whole. VP is one of the oldest forms of altering the interior gaseous environment of a pack. the pathogens that may be able to grow before spoilage occurs. the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . but can dissolve into fish liquids and deform packages. after additive application. slower-growing flora (11). which are able to grow rapidly at low temperature. Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). gutted or filleted fish. and the promotion of a predominantly Gram-positive. although less information on their effectiveness is available. Modified-atmosphere preservation of fish was first reported in the 1930s. and the problems of muscle structure damage by the modified-atmosphere (11). the inhibition of common 'spoilage' bacteria (Pseudomonas. and increase in-pack drip (12). Flavobacterium. Replacement of oxygen with nitrogen. but only in recent years has it seen a marked expansion in use and market share. does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). High concentrations of carbon dioxide have the most pronounced microbial effect. Micrococcus and Moraxella). an inert and odourless gas. The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). Sulphur dioxide. The products. including salt. and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. may not be considered ‘fresh’ fish. The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. the higher pH and reduction potential (Eh) of the fish muscle. In the United States. MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). phosphates. a modified-atmosphere may contain any gas necessary for the desired effect.3 °C and produce toxins. without overt signs of spoilage. discolour pigmented fish (11). botulinum. sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). Many additives have been tried in conjunction with changed atmosphere. This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. as they are able to grow at temperatures as low as 3. Gases used in MAP of fish most commonly include carbon dioxide and nitrogen.CHILLED AND FROZEN RAW FISH surrounding gas.

The addition of high levels of carbon dioxide to these fish. These conditions include storage of the product at ≤ 3.g. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). For other less fatty marine fish. The replacement gas should contain at least 40% carbon dioxide. 4. but is recognised as a drawback of carbon dioxide application. Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met.g. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. Subsequent storage must be at or below 3 °C. from both a microbiological and a chemical standpoint. use of high-quality raw fish. as already stated. The problem of water loss in MAP fish is common. Only the highest-quality fish. adequate shelf life and cooking requirements. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13). and the use of a HACCP plan. TMAO) that can be reduced to produce 8 . Although all fish species need to be treated separately. Another problem facing the application of MAP to marine fish is the existence of carbon dioxide-resistant bacteria that can rapidly produce off-odours (e. In the United Kingdom. greater advantages are seen. at all times. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. 5.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). owing to the dissolution of cellular structures. For fatty fish. should be used. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1. this problem occurs to a lesser extent with meat. 3. however. however. adequate product labelling in respect of storage temperature. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. The growth of these bacteria is reduced by high carbon dioxide concentrations but. botulinum. trimethylamine and sulfides).3 °C at all points from packaging onwards. Storage studies have shown that the extension in shelf life of MAP cod fillets was proportional to carbon dioxide concentration up to around 50% (V/V). may result in unacceptable colour and textural change and hence little shelf life extension (11). It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. 2.

1. with no smell. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. reactions caused by activities of the fish’s own enzymes. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . and the flesh turns from bright and elastic to dull and soft (1).4. When working with fillets. A. C (C=reject) Significantly. 18) and are probably the most widely used.CHILLED AND FROZEN RAW FISH off-odours. Loss of freshness and spoilage cannot be separated as processes. Over time.III. so lower carbon dioxide concentrations may have better preservative effects. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids). The eyes turn from convex and clear to concave and opaque. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. texture. and this pattern is the basis of freshness grading schemes. Taste panel evaluation of cooked fish may be used to score or grade colour. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). TABLE 1. the skin turns from iridescent to dull and bleached with bacterial slime. Degradation of whole fresh fish stored in ice generally follows a set pattern. Various methods for whole fish freshness evaluation by a trained panel have been proposed. B.III Fish-quality scales Name of scale Torry Scale Quality Index Method European Grading Scheme Scoring system 10 to 0 (≤ 4 reject) 0 to 24 (arbitrary reject) Extra. as traditional spoilage patterns of the external organs are typically very distinct. smell and 9 .1 Spoilage Fresh fish spoilage and methods of evaluation Food spoilage can be considered as any change that renders the product unacceptable for human consumption (10). 17. these are summarised in Table 1. to brown and slimy with an intense off-odour.4 1.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. the gills from pink and shiny. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. and metabolic activities of microorganisms present in the fish. or for that matter evaluation of the taste of the fish.

ii) Neutral taste. the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). Because fish tend to have relatively little residual glycogen compared with mammals. Generally. an intense odour compound. rigor mortis is the first noticeable change in the fish. spoiled and putrid. ammoniacal. an anti-freeze. AMP. By measuring the concentrations of the six components. cabbage-like. chemical. an osmoregulator. IMP. However. the muscles relax again and the fish returns to a flaccid state. The process of ATP breakdown is used as an indicator of fish freshness (24). iii) Traces of sour. the pH drop of the muscle is correspondingly less. %). is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. the Torrymeter. a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. As the fish degrades.g. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. or simply a waste product present due to bioaccumulation (22). sulphurous and/or rancid smells. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical.5 (18. measurements of the falling skin resistance may be made and compared with a calibration curve to estimate the time that the fresh fish has been stored in ice. From being flaccid. which increases from 0% towards 100% with time. texture becomes soft and watery or hard and dry. fruity and/or bitter off-flavours. reduction in ATP and increase 10 . development of sickly sweet. iv) Enhancement of the spoilage characteristic of phase iii. HxR and Hx) gives a quantity (K-value. After death. To a varying extent. After some time. the Eh of fish muscle remains relatively high (20). or its remaining shelf life (1). as little endogenous reduction of TMAO occurs (18). ADP. sea-weedy taste. all marine fish use/have TMAO (21).8 to 6. biochemical and microbiological actions. the conductance generally increases. After death. 19). 23). rigor resolves. little flavour. the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. Changes in the resistance of the fish skin after death are used as the basis for tests that employ an electrode measurement of skin resistance (e. These processes are strongly influenced by the physical conditions of storage. bacterial reduction of TMAO to TMA. TMAO permits a high Eh to remain in the muscle tissue. this will generally correlate with the length and severity of rigor. thus. The pH of the muscle will drop after death depending upon the amount of residual glucose or glycogen that is reduced anaerobically to lactate with the coproduction of ATP. possibly slightly metallic.FISH AND SEAFOOD taste of cooked fish. post-rigor values are typically in the range of pH 5. During the initial storage. RT Freshmeter or Fishtester).

also termed TVN). Neither does the change in pH during storage correlate well with the production of TVB-N. Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. 27) produced by aerobic respiration and other forms of metal ion reduction (28). not to describe freshness. However. the importance of microbial action increases with time (5. hypoxanthine. Comparing results for different fish species. The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. For some fish species. using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. The variation in post mortem fish pH may likewise influence the amount of bases being liberated to the air and consequently affect the odour characteristics of the fish. 7). for example the TMA fraction. Lipid oxidation and other oxidative changes lead to oxidative rancidity. 26). a correlation can be made between the spoilage of the fresh fish and the production of TVB-N. however. however. Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. 11 . so the measurement is effective only during the initial loss of freshness. which may also be formed by bacteria (25). as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. Chemical degradation continues after the initial post mortem phase. but rejection of whole fresh fish by sensory methods such as the EC grading scheme (32) is based upon non-specific odour detection and physical appearance. 18). does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection. as microbial spoilage is limited by the low storage temperature (1. or a specific fraction of the volatile bases. Quality indices based upon the products of microbial metabolism do not explain changes in quality until microbial growth produces measurable changes in the fish. Evaluation of Total Volatile Base-Nitrogen (TVB-N. and are especially important in the spoilage of frozen fish. the majority of volatile bases are produced from the soluble non-protein nitrogen of the fish (free amino acids and other low-molecular-weight nitrogenous compounds). these measurements are usually used to quantify the amount of spoilage. colour changes. This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. and a segregated weak acid to absorb them. the residual acid is then titrated. 31). Oxidative rancidity is known to reduce the quality of fatty fish in particular (19.CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). is later significantly reduced by bacterial action. The Conway method (and variations of it) uses a strong inorganic base to volatilise the bases in the fish sample. therefore. During spoilage.

All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent.5 1. yoghurt spoilage by yeasts and clostridial spoilage of cheese (5) are examples where it has been recognised for many years that a particular minority of the microbial flora present in the product is responsible for its spoilage. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. a minority are often associated with the majority of the spoilage. careful temperature control during storage is not only important in terms of quality loss. the results of altering one parameter may have far-reaching consequences on others. biochemical and microbiological reactions taking place within and on the fish. The concept of specific spoilage organisms (SSOs) is not new. 5).FISH AND SEAFOOD 1. 1. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. but also crucial for the assurance of consumer safety. The factors are interactive and cannot be completely isolated. For example. Fish spoil as a result of the chemical. 1. Also. the slower the spoilage processes proceed (within limits).and temperate-water fish spoilage rates may be many. The sum of these changes may result in a different spoilage profile. 34). realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 . The reasons for the differences between tropical. For fresh fish. by altering the gas atmosphere (extrinsic).5.1 Factors Affecting Fresh Fish Spoilage Temperature By far the most effective method of reducing the rate of whole fresh fish spoilage is temperature control (1. For fresh fish iced immediately after capture and continually stored in ice.5.3 Recognised specific spoilage organisms (SSOs) The degree of spoilage leading to sensory rejection of fish is partly dependent on the perception of the consumer. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH.2 Fish-spoilage bacteria The parameters affecting the multiplication of microorganisms in foods have been categorised into two general groups: intrinsic (inherent qualities of the food) and extrinsic (qualities of the food environment) (35).5. the lower the temperature of storage.

1. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. it is often isolated as about 1 . It is also present in fresh water.g. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s. 13 . i. therefore. Members of the genus are able to produce pigments.1 Pseudomonas spp. As previously noted. plus other characteristic compounds of the species responsible for spoilage. the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. the spoilage of a product may be strongly influenced by the conditions under which the product is held. The importance of Sh. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. These organisms are described as potential spoilers. but only if or when they reach numbers capable of producing sufficient spoilage compounds to effect rejection do they become the SSOs of the product (9. 35).2 Shewanella putrefaciens Sh. its ability to reduce TMAO to TMA. which produced the characteristic spoilage compounds of the fish (30. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms.e. ‘oniony’ and ‘faecal’ odours from the production of ketones. It is able to grow as fast as. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp.3. putrefaciens spoilage of fish is due to its biochemical action on muscle. Pseudomonas spp.5. (9) and they are considered SSO of iced freshwater fish (35). or faster than the rest of the flora of ice-stored fresh marine fish (9). 35. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine. produce hydrogen sulphide (H2S) from cysteine.5. frozen) fish products. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. on fish are diverse and in many cases species-specific. They are generally characterised as Gram-negative rods. more especially.3. 36). and may play some role in the spoilage of freshwater fish. putrefaciens growth may be used as indicators of spoilage (30). although only since the late 1960s has it been realised that specific metabolites of Sh. mediated spoilage is characterised by ‘fruity’. processed (e. obligate respiratory bacteria. 37). aldehydes. catalase-positive. oxidase-positive. motile with polar flagella. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice.10% of the total flora of fresh fish from temperate marine waters. During the 1970s.CHILLED AND FROZEN RAW FISH started in the 1940s. It is thus an important spoilage organism of gadoid fish such as cod. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms. Sh.

additional psychrotrophic species such as Shewanalla baltica. Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39). Shewanella oneidensis. unlike many other motile bacteria. with several new species being described. Shewanella frigidimarina. at the time of rejection. Shewanella affinis. 38). it was proposed that this organism. phosphoreum sufficient to cause spoilage was found. was capable of being in a significant numerical minority on the MAP fish but still able to yield the majority of the TMA thought to be responsible for the rejection (23. a population of Ph. putrefaciens growth has been collected (12.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). In 50% N2/50% CO2 MAP cod. phosphoreum has also been identified as responsible for histamine fish poisoning. Shewanella denitrificans. putrefaciens were identified as Shewanella algae. putrefaciens has been studied. Shewanella livingstonensis. owing to its cell size and activity. The ferric reductase activity of Sh. In fact. they are heterogeneous. phosphoreum were required for 50% of taste panellists to reject a sample. putrefaciens strains are characterised as microaerophiles or anaerobes. it is strictly respiratory. does not show carbon-source chemotaxis. and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. Shewanella olleyana. Sh. It is also able to produce hydrogen sulphide and a range of other off-odour compounds. Initially. Shewanella waksmanii. The prevalence of the bacterium in so many environments. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). mesophilic strains of Sh. This type of intoxication occurs when bacteria convert the histidine 14 . and recent taxonomic description using modern molecular methods bifurcated this species.how to reduce a molecule that they are not able to take into their cells efficiently. although. Another distinct feature of the physiology of Sh. Shewanella gelidimarina. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). Sh. Ph. 1. whereas >108 cfu/g Sh. putrefaciens lies in its chemotaxis: the bacterium. putrefaciens was not present in such numbers (23. putrefaciens were required. but Sh. like all members of the Pseudomonadaceae. In terms of its taxonomy. Research has shown that approximately 107 cfu/g of Ph. as the ferric iron is insoluble. Sh. the bacteria have a significant problem to overcome . This wide-ranging respiratory capability is thought to be unique (39). 41). Later. 41).3. Ph. and Shewanella profunda were described.5. baltica has been identified as the main H2S producer in cod during cold storing (37). and Shewanella aquimarina.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh.

CHILLED AND FROZEN RAW FISH present in fish into histamine. previous exposure and access to supportive treatment. Fewer still are capable of growing on fresh fish.6. E. thermosphacta did not produce off-flavours until 2 . 75% (with a 35% mortality rate) occurred in Japan.3 days after having reached 108 cfu/g (42).3. B. types B. human viruses. 5. Adult disease is caused by the production of the botulinum toxin.e. Acetate production has been reported as a good indicator of spoilage by this organism. (and very rarely F and G) have been reported to cause human disease. 6). MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). 1. age.5. but with proper cooking. they do not produce significant product spoilage). Recent studies have investigated the dominance of B. However. it is bacterial risks that increase after capture of the fish. Some Ph. C2. F and G) is well documented (44). thermosphacta is a well-characterised psychrophilic spoilage organism of meat. Growing evidence suggests a role for B. Types A. presents a potential risk. C1. botulinum. where a particular 15 . including antisera. Review of outbreak data suggests fresh fish to be safe. B. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. The heterogeneity of the types (A. E. a convenient but diverse species of bacterium with a number of different types.4 Brochothrix thermosphacta and lactic acid bacteria B. Huss (1. thermosphacta in the spoilage of some MAP fish. Of these. 1. lightly preserved fish products are associated with botulism. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. 5). bacteria and higher parasites all present some risk associated with fish consumption (1. When inoculated into VP corned beef and sliced ham. 1. B.3 °C (Group II) and these are non-proteolytic (i. 5) reports that fresh fish consumption has never been shown to cause human botulism. botulinum recorded up to 1963.6 Pathogens: Growth and Survival Endogenous chemicals. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. Of 404 intoxication outbreaks of type E C. these risks are substantially reduced (1. especially type E botulinum intoxication. E and F include bacteria with minimum growth temperatures of approximately 3. algal toxins. this is probably due to spoilage occurring before toxin elaboration.1 Clostridium botulinum C. D.

the majority are associated with products not cooked immediately before consumption. is small. In Japan. and proper cooking (boiling for 1 minute. which is believed to be required for virulence. Currently. due to the cultural preference for raw fish dishes. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. Generally. The first report of the organism being a foodborne agent was with shirasu. the majority of outbreaks are caused by consumption of raw fish products (e.2005). 1. parahaemolyticus is a marine organism.g. sushi) (5). Since 1996. vulnificus is associated with warm marine and estuarine waters. 11). I-sushi. The bacterium causes primary septicaemia.2 Vibrio parahaemolyticus and other vibrios V. whether stored aerobically or in modified-atmospheres. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). and North America. a Japanese boiled and semi-dried sardine dish.3 °C) will eliminate all risk. Russia. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. parahaemolyticus outbreaks. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. smoked. the risk posed to the consumer of fresh fish.6. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. Other lightly or semi-preserved (e. Southeast Asia. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45).FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. which was probably contaminated from an uncooked food source or an excreting foodhandler. France. 5). This strain has been responsible for 50 to 80% of all V. Proper chilling. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. and when in contact with contaminated water (no CDC reported cases from fish. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. salted or pickled) fish have also been associated with botulism. botulinum in the marine environment (isolated from around 90% of marine and environmental samples from northern European waters and its consequent presence in seafood (a surveyed incidence of as high as 65%) (2. parahaemolyticus cases have increased across the world. Only about 1% of marine isolates produce a thermostable haemolysin. Despite the ubiquitous nature of type E C. including epidemics in India. Proper temperature control (<3.g. It is generally undetectable in marine water below 19 °C but may grow in culture at temperatures as low as 5 °C and on food at 10 °C. storage and preparation (11). there is no specific guideline that describes a minimum level of V. Japan. again. accounted for the majority of cases (1. were associated with finfish between 1978 and 1998 (46). Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. 1978 . V. V. A unique clone of V. USA. parahaemolyticus O3:K6 is responsible for many of the recent V. which can cause human gastroenteritis.

000 17 . Although the psychrophiles are unable to grow at body temperature. 5.6. although lack of data concerning infectious doses and the possibility that foodborne virulence is linked to the immune state of the host mean that full risk assessment is impossible. listeriosis cases have been reportable to the CDC. monocytogenes has been well documented as a foodborne human pathogen. but proper cooking should substantially reduce risk of infection. V. with minimum growth temperatures around 0 °C and optima of 15 . and especially Aeromonas hydrophila. There is no risk associated with the consumption of properly chilled and cooked fresh fish.26 to 0. 1. are present on fish and can grow to significant numbers during storage (1. in its habitat. their potential to produce disease-causing exotoxin in food has not been fully examined (48). L.CHILLED AND FROZEN RAW FISH disorders). monocytogenes incidence has been between 0.55 cases per 100. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. Most cases have been sporadic rather than associated to large outbreaks. and proper cooking will substantially reduce risk. the causative agent of cholera. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. P.. 1. shigelloides is similar to Aeromonas spp. however. there is no risk from the consumption of properly handled and cooked fresh fish. coli on isolation media (50). The major virulence factor appears to be the production of toxins with enterotoxic.4 Listeria monocytogenes L. sodium channel blocking and haemolytic activity (48).20 °C. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. is historically associated with faecally contaminated water.6. and fish and shellfish are probably the major reservoirs for the organism (50). but the bacterium is known to survive and grow in the shallow marine. and especially estuarine environment. It is known that Aeromonas spp. They are aquatic organisms with an epidemiology that is yet to be fully understood. Chill storage may not eliminate growth. 17). proper chill storage will eliminate any risk associated with this organism. Similar problems as those encountered for Aeromonas spp. infection. Some isolates are true psychrophiles. shigelloides are encountered. cholerae O1. possibly due to under-reporting because of their likeness to E.3 Aeromonas Aeromonas spp. cytotoxic. are associated with human diarrhoeal illnesses. Again. Since 2000. which is often fatal (>50%). others are psychrotrophic mesophiles (49). In the US. Despite this. in assessing the risk of fishborne infection by P.

1. some Vibrio spp. it is not apparent to the consumer and it cannot be destroyed by cooking. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death.48 minutes in salmon have been reported (53). but more especially by Morganella morganii.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. Photobacterium spp. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. alcoholics. The organism poses a serious risk in chilled products not cooked before consumption. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). The most frequently reported parasites of human importance in fish are round worms of the genera Anisakis and Pseudoterranova and tapeworms of the genus Diphyllobothrium.. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. Like other toxins. D60 of 1.. but cooking will significantly reduce the risk of infection. It is regularly isolated from seafoods (51). Larvae of parasites such as Anisakis simplex are resistant to curing and marinating but can be easily destroyed by freezing at -17 to -20 °C for 2 hrs. neonates.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. Being a psychrotroph.6.pregnant women. there is a possibility for growth of the organism on chill-stored fish. Infections are mostly associated with the consumption of raw or 18 . Fortunately. are exceedingly difficult to produce within the legal limits of histamine as the temperatures used during production can lead to rapid growth of histidine decarboxylating bacteria. at risk groups .2002 (55)) and is also common worldwide. full risk analysis is not possible (52). Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . Lightly preserved products. 1. leads to the production of histamine. Klebsiella pneumoniae and Hafnia alvei.6. especially pickled fish.. foetuses. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). but proper cooking will reduce any risk.FISH AND SEAFOOD persons. If fish are not properly refrigerated. and Lactobacillus spp.98 minutes in cod and D60 of 4. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. which has a maximum permissible level (for fish products from fish species associated with high amount of histidine) of 200 mg/kg fish in the EC. Clostridium spp.

For fish from inshore or inland waters of doubtful microbiological quality. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. area or lot (CAC. Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease.IV Sampling plans and recommended microbiological limits for fresh and frozen fish (Adapted from ICMSF 1986) Limit per gram or cm2 Test Aerobic plate count E. FAO. Faecal coliform counts may be used instead of E.IV) (57). which in many cases will eliminate the risk. aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . Ababouch L.1997.. 2003. EC. long storage under refrigeration or one of the former prior to freezing. coli counts where this method is preferred. 1997) Huss H. No.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). it may be desirable to test for Salmonella and V. including parasites and/or quantity of their toxins / metabolites per unit of mass. parahaemolyticus Staph. Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers. parahaemolyticus (see Table 1.. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF). The most widely accepted microbiological criteria for chilled and frozen raw fish are those set for aerobic plate counts (APC) at 25 °C and E.H.7 Published Microbiological Criteria Microbiological criteria for food defines the acceptability of a product or a food lot based on the absence or presence or number of microorganisms. TABLE1. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. In summary. 444. especially in warm-water areas and where fish are to be consumed raw. Rome. volume. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it. Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. Gram L. 1. coli Case 1 4 Plan 3 3 n 5 5 c 3 3 m 5 x 105 11 M 107 500 Additional tests to be carried out when appropriate Salmonella V.

activities and safety. 121-37. 5 (1). Food Control.R. Codex Alimentarius Commission.H.J. Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms. 1996. 14.A. International Journal of Food Science and Technology. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres. 2. 334. Spoilage and shelf life extension of fresh fish and shell-fish. and natural toxins. Ed. 5.. Ashie I. 1992.. Davies A. Nychas G. Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef.. 2003. 37. Food and Agriculture Organisation. Armstrong D. 7. 243-60.FISH AND SEAFOOD 1. Microbiological spoilage of fish and fish products.8 1. Dalgaard P. Journal of Food Safety.. Rome. Gibbs P.. 20 . 2. Feldhusen F. Sivertsvik M. 6.. Food Control. Huss H.J..H. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth. 13. 2000.H. International Journal of Food Sciences and Techonology. 283-94.K. 1995. Gram L. References Huss H. 1651-60. Davies A. 12. Smith J. Control of indigenous pathogenic bacterial in seafood.A. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). Gram L.. 1996. Rome. 107-27. M. 24.. 1994.J. 8. Capell C. Italy. Rhodehamel E. Jeksrud W. 11. 348.N. 1994. 9. Kirby R. 8 (2). Huss H. Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review.H. Huss H. Food and Agriculture Organisation of the United Nations. Critical Reviews in Food Science and Nutrition. Codex Alimentarius Commission. International Journal of Food Microbiology. 19. Reddy N. 4.E. 3.T. 87121. Ed. Quality and quality changes in fresh fish. Fletcher R. Assurance of seafood quality. Jehanno D.A. 12. 1989. Ahmed F. Food and Agriculture Organisation of the United Nations. 67-71. 5-7. Simpson B. International Journal of Food Microbiology..P.R. 1992. 91-8. parasites. Kautter D... 87-118. Huss H. Davey K. 2002. 1993. 2001. Food Control.R.R. Incidence and growth of psychrotrophic Clostridium botulinum in foods. Microbes and Infection. 291-304.E. Rosnes J. 10. The role of seafood in bacterial foodborne diseases. in FAO Fisheries Technical Paper No.. International Journal of Food Microbiology.. 27. 1997. 36. Incidence of foodborne pathogens in European fish.S.H.. Food and Agriculture Organisation. 33. in FAO Fisheries Technical Paper No..K. 12.

Lipid oxidation in seafood. . 1992. Journal of Applied Bacteriology. Gibson D.M. 19-22..CHILLED AND FROZEN RAW FISH 15... 121-37. 22. Eds. 23. Bacteria active in the spoilage of certain seafoods..E.E. 27.. Huss H. 305-17.J. 1982.N. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products.A. Dalgaard P.J. Herbert R. Changes in purine nucleotide of red lateral muscle of rainbow trout. March 20. Børresen T.. Martin R. Quality and Quality Changes. Hebard C. 2007. 419-29.. 21. Hendrie M. Adopted. Microbiological spoilage of fish and fish products. Edinburgh. Westport. 1933. 34. Chemical composition. 1995. 20. http://seafood.H.J. Ed. Flick G. in Fresh Fish.M. Qualitative and quantitative characterisation of spoilage bacteria from packed fish. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout. Ed.. Martin R.C. Reginstein J.J.htm Anon. Washington D. Huis in't Veld J. 41-50. 245-67. The micro-determination of ammonia. 33. Shewan J..E.spoilage of fish. 22-36.J..E. 1. 1992. Khayat A. National Advisory Committee on Microbiological Criteria for Foods. 117-27. Davies A. Hebard C. 30.M. 26. 1995. Ratnayake W. Food and Agricultural Organisation of the United Nations.E. 1971. 19.H. Pedrosa-Menabrito A. Huss H. Conway E. Surette M. Gram L. Journal of Food Quality. Slade A. Arai K. Quality and Quality Changes. Byrne A.. Biochemical Journal.. Martin R. 36. in Advances in Seafood Biochemistry.M. 29.A. Microbial and biochemical spoilage of foods: an overview. Schwall D. Ackman R..E. Letters in Applied Microbiology.. in Chemistry and Biology of Marine Food Products. 18. 1983.edu/haccp/compendium/chapt08.J. Nature. International Journal of Food Microbiology.G.S. Lancaster. 17. International Journal of Food Microbiology. AVI Publishing. Leblanc P. 1996. Rome. International Journal of Food Microbiology. 1995. Journal of Agricultural and Food Chemistry. 1415. Gill T. Non-enzymatic oxidation of seafood lipids. 11. Huss H. 354-8. Yajima T. 1988. Flick G. An absorption apparatus for the micro-determination of certain volatile substances.H. Rome.J. 24.. 27. National Advisory Committee on Microbiological Criteria for Foods. in Fresh Fish. 25.a review.ucdavis. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere.. 33. Technomic. Part I . Saito T. 42-54. Food and Agricultural Organisation of the United Nations. 1-18.J. 1959. 26. Eds. Flick G. Gill T..R. 130-2. 184. 37. 21 16. 21.. Autolytic changes in fish post mortem.H. 28. Sea Fish Industry Authority. 1996. Shelf life extension of fresh fish . 1995. 1987. Food Technology. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage. 1985. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish.

.eu/LexUriServ/LexUriServ. Food and Agricultural Organisation of the United Nations. and regulation. Gram L. 262-6. 32. 40.R.R. Scanlan R. Quality and Quality Changes.M..M. 1995.M.. Brochothrix thermosphacta. 2002. FAO Fisheries Report. 36. Huss H. Archiv für Lebensmittelhygiene. International Journal of Food Microbiology. Myers C. 67. Occurrence and importance of proteolytic bacteria as spoilage indicators.H.. Nealson K.FISH AND SEAFOOD 31. Dalgaard P. Ed.E. Jørgensen B. Miller A.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J. Huss H. 41. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish. 334. Magno-Orejana F.. Myers J. Applied Microbiology. Annual Review of Microbiology.B. physiology. 323-9. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches. Role of proteolytic bacteria in fish spoilage. Saffarini D. 3429-38. Rome. Taussig M. Karnop G. 1984. Drosinos E.A. Cohen D. 1994. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens. 51-62.europa.H. 42. Gorczyca E. Rome. Iron and manganese in anaerobic respiration: environmental significance. Journal of Bacteriology. Fish spoilage bacteria – problems and solutions.J. 61-6. 6689-97. 1984. 1982. II. and an Achromobacter species.S.H. Journal of Applied Bacteriology. Kell D... 39. 311-43. Nychas G. Vogel B. Venkateswaran K. Processes in Pathology and Microbiology. Gram L. Blackwell Scientific Publications. 1989. 37. The inhibition by CO2 of the growth and metabolism of microorganisms. 328-9. 44.. 43. 317.. Sumner J. 1992. Italian Journal of Food Science.J.. 48.. 1973.. Pseudomonas fluorescens.. Oxford. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere. 18-21. 109-36. 33. 26. 34. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia. Applied and Environmental Microbiology.. 13. Dixon N. Satomi M. Dalgaard P. Environmental Biotechnology.F. Lee J.L. 22 . in Fresh Fish. 38.. 2005. 62-70. 36. 1996. Brady P. Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens. 9. 33. 1989. 35. Libbey L. 71. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. 174. 4. 72-80.. The effect of storage temperature on shelf life.H. Growth and activity of Shewanella putrefaciens isolated from spoiling fish. 1985.

. Microbial Ecology of Food Commodities. Embarek B.C..M. Lynch M. Woodruff R.M. 2000. International Journal of Food Microbiology. Microorganisms in Foods 2. 2005.... 51. Lindroth S.. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico.P. International Journal of Food Microbiology. Altekruse S. Springer. 181-96. Bej A. USA.. Majeed K. Ed. 172-249. 58. 51 (4). 49.CHILLED AND FROZEN RAW FISH 45. 47. E and F in salmon fillets stored under modified atmospheres at low and abused temperatures. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures.A. 181-8.J. 12. 100-3. 55 (SS10). detection and growth of Listeria monocytogenes in seafoods: a review. 57. Blackwell Scientific Publications. New York.K. Oxford... Myers M. 1993. Wadström T. Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B. 17-34. Lehane L.. Journal of Food Protection. Smith J.P.. International Journal of Food Microbiology. Thompson S. Vibrio parahaemolyticus Infections in the United States..W. Sampling for Microbiological Analysis: Principles and Specific Application. MacKinnon L. Garcia G. The International Commission on Microbiological Specifications for Foods. 1991. Loessner M. 1998-2002.. 181. Whiting R. 1661-6. 54. 1-34. 1973-1998.. Panicker G.K. International Journal of Food Microbiology. 52. 53.H. Histamine fish poisoning revisited. Presence. Miller A. Mac Rae I. Hammond R.K. 1994. Daniels N. Genigeorgis C. 2194200.J. 1986.. 50. Bishop R. Embarek B. 1987.. Surveillance for Foodborne-Disease Outbreaks . 2003. 20.L. Griffin P. Food Technology.N.L. 46.M. Jay J.. 2006. Morbidity and Mortality Weekly Report. Ljungh A.C. 2005. Olley J.. 1-37. 330-6. 55. Slutsker L.. The International Commission on Microbiological Specifications for Foods. 56. Journal of Infectious Diseases.. Applied and Environmental Microbiology. 1991. 23 . 12. Braden C. Huss H. Painter J. Kluwer Academic/Plenum Publishers. Modern Food Microbiology. Fish and Fish Products (Microorganisms in Fish Products).. The International Commission on Microbiological Specifications for Foods. 23. International Journal of Food Microbiology.. Wilson S.. Ray B. Aeromonas and Plesiomonas as food. 2000.A. 1997.H. 303-12. 48..United States. 69. 85-95. Golden D.and waterborne pathogens. 50. Bean N. in Microorganisms in Foods 6. Use of risk assessment to reduce Listeriosis incidence..

Cann D. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging.S. 1988. Rome. Florida. 13-32. Handling and processing of scad. Fisheries and Food. 1986. 65-72. Akiba M. International Journal of Food Microbiology. Studies on the microbiological ecology of mackerel stored by the method of partial freezing.E.2 Spoilage Shewan J. Torry Research Station of the Ministry of Agriculture.9... Aberdeen. Hobbs G.. McMeekin T. Cho Y. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. in Seafood Science and Technology. Estimation of the bacteriological quality of fish by automated conductance measurements.. 1992. 1997. Huss H. 1961. Aberdeen.M. Huss H. Gram L..G. 1. Olley J. 73-89.FISH AND SEAFOOD 1. Simard R.D. 317-21. Microbiological quality and shelf life prediction of chilled fish.M..H. Dalgaard P. in FAO Fisheries Report No. Gibson D. in Fish as Food. Smith G. Huss H. Technomic Publishers.M. 741-50.L. Ravn Jørgensen B. Ed. Trolle G. Kitts D. 51.H. Ogden I. 6.1 Further Reading Processing Anon. Sumner J. Jones Y. 1992. 271-85.. The Microbiology of Seawater Fish. International Journal of Food Microbiology. Gibson D. Ed. Food and Agriculture Organisation of the United Nations. 1. 317 (Suppl.. 295-307.9 1. Picard G. 24 . Application of predictive microbiology to assure the quality and safety of fish and fish products.. 1989. 1984.. Houston N..D.. Taoukis P. International Journal of Food Microbiology. Lancaster.H. Academic Press. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere.. 1. Changes in microflora and chemical compounds in mackerel stored by partial freezing. 1989.A.. 35. Eds.9.1983. International Journal of Food Microbiology. Shinano H.). 1985.. 15. Torry Research Station... 4. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. in Torry Advisory Note No 93. Bulletin of the Faculty of Fisheries (Hokkaido University). Food and Agriculture Organisation. 62-70. Journal of Food Science. Labuza T. 1984. Magno-Orejana F. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage..P. 55. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches.C. Torry Research Station of the Ministry of Agriculture. Fisheries and Food.. Shahidi F. Borgstrom G. Journal of Food Protection. Ross T.. Villemure G. 127-34.

Gram L.. Journal of Applied Bacteriology. A rapid method for detection of histamine producing bacteria. 20023.europa.M. Modified atmosphere packaging of fish and fish products. Sala F. 72. 4. in Quality Assurance in the Fish Industry.H. 1997.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. 1987. Blackie Academic & Professional.R. Jacobsen M.. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. International Journal of Food Microbiology. Huss H. 1992. Eds.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. 59.. 137-46. 322-6. Otero A.eu/LexUriServ/LexUriServ.. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. 2197-203. 1. Gram L. http://eur-lex.K. Amsterdam..L. Elsevier Science. Effect of culture age.. Garcia M. International Journal of Food Microbiology.. London. Ed. 5.J. Liston J. 1993. Hall G. 479-88. Klausen N. 65-72.3 Pathogens Condon S. Trolle G. 1. 1987. Applied and Environmental Microbiology. Huss H. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures.H..CHILLED AND FROZEN RAW FISH Einarsson H. in Fish processing technology.H. 25 .9.9. Huss H. 1992. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila.


colourings etc. The majority of seafood products produced require a cooking stage before consumption.). Sushimi tuna). due to the early cessation of spoilage. 27 . Even products where the seafood is only one ingredient within the recipe (composite food products) it should be noted that unlike other meat-based products. although recent developments in improved supply chains. The same used to be said for the difference between fresh and frozen products. and crustacea such as shrimp and crabs.g. smoked (cold) salmon. it does not necessarily apply to the toxins that may have been produced. but still acceptable and meeting all safety criteria. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e. Even though heating prior to consumption will eliminate pathogenic microbes.1 Introduction It was often thought that whilst the highest quality seafood products are processed and sold as whole fish and/ or fresh fillets (e. seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. emphasising the high quality of the seafood and the gourmet nature of the prepared products. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr.g. the exceptions being Sushimi (high quality raw fish).2. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2. clarity of eyes. products of lesser quality. This is no longer the case with many processed products now available in stores. smoked (hot) mackerel. colour of gills. smell etc. Few seafood products are sold ‘ready-to-eat’. and on-board freezing mean that products frozen at the point of capture are now perceived by the consumer as being of a better quality than a fresh product. This has the result that any microbes present in the product have the capability to grow (with subsequent spoilage) unless strict temperature controls are applied. or could be disguised by sauces.

including freshwater fish. the processing industry has developed a wide range of value added products. which can be farmed. where the preparation has already been carried out. Listeria spp. Prepared natural fillets are fillets of fish without any additional coating.. prawns. shellfish and molluscs. Seafood is the term used descriptively to include all categories of product. and are well known for their diet consisting of waste materials. as they tend to lose colour and sheen after defrosting. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. have shown the increasing importance of convenience foods. The expected shelf life attainable for this format is usually up to 8 days. in particular. and the consumer only has to heat (cook) the product. or in MAP depends on catch conditions and treatment. This includes all categories of seafood including trout and other freshwater species. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted. Microbiological hazards include Salmonella.FISH AND SEAFOOD Recent trends in the market for food and seafood products. they are usually of a better standard than species such as tilapia. Nevertheless. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. but these are now finding favour as products in their own right. and pathogenic Vibrio spp. 2. Shelf life on ice. rather than frozen. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP). At the same time. Frozen fillets may have a surface layer of 28 . often using the simplicity of a microwave oven.. and this has produced a greater availability and diversity of products in the marketplace. There is also a limited market for filleted material. but should be equivalent to cold-water products. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein. Doubts have been raised over the microbiological status and handling practices of reef-caught fish. etc. they are usually supplied filleted with head on. and it is worth noting the additional hazard to fish handlers from Vibrio vulnificus from injuries received during handling and filleting of spiny tropical fish. from both wild capture and aquaculture sources. They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. They are typically flown into export markets.2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. molluscs. and these are increasingly being presented with an ever-expanding range of accompaniments. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish.

tuna steaks) or band-sawn from a frozen block (e.2 days) longer than that of non-prepacked fish. The expected shelf life of these products is up to 8 days (or longer for some chilled. Fish steaks can be cut from a natural fillet or loin (e.g. resulting in a product that is ready-to-eat without any further cooking. which may.. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes. in addition. be flavoured with herb dips or peppercorns. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition.5%. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product. An alternative process using a liquid smoke in the brine is claimed to result in a similar flavour with reduced water loss in product and improved yields. cod). sawdust with other woods adjusted for flavour profile and burn duration). 29 .CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. etc. The cold smoking process does not cook the fish or destroy microorganisms so the expected shelf life is not significantly increased from that expected for wet fish fillets.g. salmon.g. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. and the product then shaped and heat-processed to cook the product before being chilled. or pre-packed in MAP or vacuum packaging (VP).5% in the aqueous phase. Some products may be salted for flavour only (e. mackerel. This process in effect cooks the product and reduces or eliminates microbial contaminants.g. Hot-smoked fillets are prepared by brining fish fillets in a 26% salt solution with added natural colours to provide a range of product types. the ingredients premixed. Fresh fillets are sold from wet fish counters. Added colours can be included with the brine solution to meet specific requirements for products e. Again. The result is fish with a typical residual salt content of 1. in VP or through mail order. and salmon. Pâtés are prepared with raw fish. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. swordfish steaks). a protective glaze may be added to the frozen steak. or pre-packed frozen with added glaze. mackerel. VP products) because of the high salt content acting as a barrier to spoilage. but may be increased (up to 1 month) by packaging methods like MAP or VP. annatto with smoked haddock.5 . The shelf life for MAP fillets is typically up to 5 days. natural fillets of fish such as herring. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. haddock. Cold smoking is essentially a flavouring process.2. with a minimum residual salt content of 3. They are sold from wet fish counters. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. which is approximately 30% (or 1 . Hot-smoked fish are prepared from brined.

They are pasteurised products and treated as high risk raw materials. fish nuggets/bites with dips and seafood-based snack products. and formed into round cake shapes. flavours. and seafood mixes. This ‘catch-all’ category includes seafood salads. Fish cakes are products produced by mixing fish with potato and herbs.or a five-stage process. rice. Band-saw dust and fish trimmings from fishblock preparation materials may be used. mixed whitefish. Prawns. and heat-fixed.g.g. with an expected shelf life of around 7 days. and in the shell. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process. There is an increasing market for high value fresh or thawed fish (e.g. Seafood ready meals (frozen or chilled) differ from seafood recipe meal centres in that they contain a source of carbohydrate such as a potato topping. haddock. With further processing. e. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. Other.FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate. The cakes are usually coated using a three. tuna. long-life seafood mixes using sorbate and benzoate as preservatives. coated in batter or breadcrumbs. rice or pasta. Natural prawns from cold or warm waters can include those peeled.) Raw material is typically fish fillet. Scampi are tails of Dublin Bay prawn or langoustine. ‘fingers’ before being coated in batter and/ or breadcrumbs. The protein source can be fish fillet or blocks. which may include molluscs. etc. Surimi is the insoluble protein fraction of fish after washing and processing. addition of ingredients such as colours. e. cephalopods. binders. Other ingredients may be natural or reconstituted. jellied eels. 30 .. salmon) coated and sold as chilled products. The majority are products made from various types of whitefish (cod. or cut from a preformed frozen fish blocks into shapes. It is often used as a protein supplement. pasta or potato. it can be used as a seafood analogue. Breaded fillets/steaks and battered fillets are either natural or shaped fillets. taramasalata. and may also include additional ingredients such as fresh herbs and spices.

Recent public opinion has favoured the use of dolphin.3 2. is not used for sushi or other high risk (and high value) products. storage.and turtlefriendly long lining to the less discriminatory netting. For a summary of background information on the hazards associated with fisheries products and ingredients. A primary role in fish spoilage has been assigned to this organism. 6). searing or bar marking). The edible tissue of some species of fish are more susceptible to gaping than others. 31 .I. and the potentially toxic by-products of histamine-producing bacteria (5. Only after death. or visible damage from gaff marks (4). as a result there is an increase in spoilage rate thereby reducing the product shelf life.3. and pathogenic flora associated with seafood. handling or preparation stage.e. Dalgaard et al. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. see Table 2.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. After capture and during any subsequent processing. will the saprophytic flora start to invade the surface flesh (1. it may be used for other products or markets depending on the product specification. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). Tuna that does not meet the highest of grades based on colour. product handlers. and ingredients added to the final product. i. the microbial flora of seafood is associated with the gills. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. which may become a focus of contamination deep within the muscle.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. including seasonal variation. on the whole. whilst gaping is also dependent on a range of other factors that effect the muscle composition. fat content. and intestines of the fish. species (Bluefin>Bigeye>Yellowfin>Albacore). Grading is based on size of fish (>25 kg). and processing conditions such as dehydration or poor filleting practices. sensory assessment of freshness etc. Contamination will. 2). The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). In summary. and have emphasised the origin and sustainability of the fish as key marketing points. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. skin.

I Spoilage and pathogenic microflora associated with common raw materials and ingredients used in prepared frozen and chilled products Raw material Vegetables (Untreated) including fresh herbs. Staph. staphylococcal toxin. cereus B. Aeromonas. toxin. for chilled products. histamine Cryptosporidium. cereus Staphylococcus aureus. shelf life determination. protozoa and viruses B. Fusarium. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter. Bacillus cereus.FISH AND SEAFOOD 2. and milk Cheese Water/Ice Bacillus. Vibrio cholerae.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. Pseudomonas. yeasts Lactobacilli.4 2. spore-formers. moulds Bacillus.I provides an overview of common raw materials. TABLE 2. and associated spoilage and pathogenic microorganisms. B. intermediate processing. product assembly. moulds. and so are the potential microbial contaminants associated with them. streptococci. In addition. anaerobic spore-formers. moulds Bacillus. Aspergillus.3. by the HACCP team or by a hazard and risk assessment programme. moulds Bacillus Bacillus. Moulds Enterobacteriaceae Salmonella. however. 32 . pseudomonads. Penicillium. Bacillus subtilis B. Clostridium perfringens. it is essential that during new product development a full hazard identification and risk analysis be conducted to ensure that the microbial risk for specific ingredients are identified. Shigella. yeasts Pathogenic microflora/toxins Salmonella. and. enteric viruses 2. be considered at an early stage during new product development. cream. mushrooms.4.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. Listeria. the effects of processing and combination or use of ingredients on identified microorganisms should also be considered with respect to both the safety and quality/shelf life of the product. Table 2. nuts and fruit Associated microflora Lactobacilli. coliforms. raw materials. cereus Bacillus licheniformis. Clostridium Lactobacilli. Clostridium botulinum. Enterobacteriaceae. Staphylococcus. Escherichia coli. coryneforms.

5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. Figures 2. up to -5 °C so as to improve the cutting and handling properties during processing. the core temperature. All have the same intention. It is recommended that the principles of HACCP are applied to novel processes.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. to evenly raise the temperature of the entire frozen product from -20 °C or lower. Where the raw materials are frozen. but it also poses a risk of cross-contaminating other products undergoing tempering. line extensions and process reviews. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. but it is assumed that any band sawing or dice process will introduce heat from blade friction. It is critical that the time and product temperature during tempering be controlled to avoid elevated temperatures (>5 °C) on the product surface allowing microbial growth or the onset of spoilage in raw material. Block-sawn dust that is required for use in fish cakes is swiftly transferred to a chiller to avoid water loss and spoilage from process contaminants. defined holding time at a chill temperature (0 – 3 °C). and contamination from equipment to clean fish muscle. 2. and potential opportunities for growth of foodpoisoning bacteria.1 Coated products Seafood products are coated with a cereal-based batter or crumb with up to 30 50 % of the final product weight accounted for by the batter or crumb.4.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. Examples of tempering techniques include the use of heated air. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. this may be an incidental effect. Band-sawn materials or fish dice are normally promptly further processed. Effects on the microflora are minimal during tempering (7).2. depending on the size of the fillet 33 . Rapid controlled cooling of heat-processed components will help to minimise spoilage.4. there are a number of distinct methods used.3 Preparation of fish component of chilled and frozen recipe dishes 2. but will also need to ensure that the optimum shelf life can be safely obtained.4.1 . or the use of microwave technology. For some components. Generic examples have been used to illustrate only the major points. 2. as the full complexity of the processes used for specific products cannot be covered in a general text. as a heat process is often required for functional or organoleptic reasons.3.

The processing of three examples types are described in Figure 2. quickfrozen.1C . 2.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). drained and then refrozen for storage until required for further processing. On arrival at the processing plant. frozen in a nitrogen blast freezer and weighed into sacks for later processing. with the coating heat-fixed. Fig.Shaped Fillets.3. after landing. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.1. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. This material is transferred to a processing facility with the ice replaced as necessary to ensure that the product is maintained at the temperature of melting ice. are placed into plastic boxes and covered in ice to preserve their condition. 2.Scampi.1A . by mincing and mixing with a binding agent (as described in section 2. chilled or sold chilled on defrost with an expected shelf life of 6 .1 (2.1B . The “blown” meat is graded.4. the scampi is washed in potable water to remove foreign bodies.Fish Cakes).FISH AND SEAFOOD processed. It is then size-graded.1.4. 2. Raw material preparation stages for chilled or frozen coated fish products 34 .2).3. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. may not rise above 0 ºC during processing.2.1. Frozen cores are usually batter-enrobed.7 days.

4. fish fingers). Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet.1.. additional spoilage organisms may be introduced. quickfrozen. as the physical structure of the material has been destroyed. 2. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. The mix is shaped and formed ready for packing or for enrobing. Where frozen blocks of fillets are used they will be “tempered” (temperature raised to around -7 °C) to facilitate efficient cutting with a band-saw to a specified thickness. from potato flake. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. This is especially the case with block-sawn dust.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. or those where the trimmings.4.3.2 Shaped products This type of product falls into two categories.3 Fish cakes Fish cake preparation is used as an example of the production of fish products with fresh or dried ingredients added during the process. The fish component of the cake will. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . pathogens such as Salmonella and Listeria. The remaining dried ingredients are mixed with the fish. that are to be reused for other foods. Waste material produced from this type of processing is often reused in the third category (fish cakes). but. are handled and stored as any other food product. and water is added to rehydrate the components.block-sawn dust. such as aerobic spore formers and Staphylococcus spp. with an expected shelf life of 6 . iii) Fresh/fillet/minced fish block. Therefore.1.g. unless it is poorly handled. chilled. those that use frozen blocks of whole fillets.off-cuts or belly flaps. 35 . material will need to be transferred swiftly to a chiller.3. Fish cake cores may be batter-enrobed. more seriously. with the coating heat-fixed. It is for this reason that all by-products from primary processing. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. spoil more rapidly than the dry ingredients. ii) Fish-filleting waste . because of its nature.7 days. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. or sold as a chilled product after thawing. or promptly processed to avoid further quality loss. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. once the dry components have been rehydrated.

as well as ensuring that there is minimal contamination from human processors. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. any excess is removed with an air knife. such as Listeria and Vibrio spp. raising agents. starch. Natural fillets d. The first stage. i. Batter contains water. and may include milk powder.4.2 illustrates the typical stages of enrobing or breading. quickfrozen. Products are preformed and presented to the process (either three. egg.7 days. e.FISH AND SEAFOOD In all of the above examples. IQF/pre-formed fillets c. flour. The batter in the enrober is continuously recirculated and fresh ingredients (normally a dry pre-mixed recipe) are constantly added to a pre-mix vessel with potable water. rusk or crumb. and if required. and to carry flavourings where required. perfringens could.g. or sold as a chilled product after thawing. Over time. A large variety of product formats and types can be batter-enrobed. because of the intensive handling required during processing. rapidly multiply if strict temperature control was not maintained. Tempered band-sawn materials b. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. with an expected shelf life of 6 . a. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process. with the coating heat-fixed. e. is the addition of a pre-dust. and Enterobacteriaceae from flour. and possibly Staphylococcus spp. some of this initial flora 36 . a characteristic flora will develop from the batter components. chilled. milk powder and spices. lactobacilli from the milk powder and flour.g. 2. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. Bacillus spp. which can occasionally be isolated from protein-handling processes. from flour. which typically consists of flour. absorb any excess moisture on the surface of the fish portion..or fivestage) frozen or chilled. spices. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. which is product-dependent and not always required. potentially. If a crumb coating is required. Spore-forming anaerobes such as C. fish cakes Portions or fillets may be batter-enrobed. Fish products. vibrios and other fishassociated spoilage organisms.e.4 Battering/enrobing process Figure 2. Initially. Breaded or battered fish fillets are fully heat-processed before consumption. The function of this is to improve the adhesion of subsequent coatings.

The origin of contamination for the two organisms is often unclear. but it is believed that poor hygiene regarding product and food handlers. This can be achieved by frequent clean-downs and adequate hygiene. in the case of B. cereus can proliferate under certain conditions. Flow diagram outline of a typical three. cereus. this is not of great significance to the microbiological standard of the finished product. aureus and B. dry batter mixes are all potential sources. and both of these can produce heat-stable toxins that can survive a batter heat-fix process. Fig. but because batter is heat-set. With inadequate temperature control of batter systems.CHILLED AND FROZEN PREPARED FISH PRODUCTS associated with the ingredients will be diluted out. Staph.2. replaced in part by process contaminants and bacterial flora from seafood.2. and. It is advisable that some control mechanism is employed to prevent toxinforming organisms from multiplying to dangerous levels. time-limitation or production run temperature control.and five-stage enrobing/breading process The length of production run and temperature of the batter are key factors in deciding the balance of the dominant flora within the enrober. The FDA has issued clear guidelines on acceptable 37 . contaminants such as Enterobacteriacae can proliferate and develop during the process.

but they are supplied without a source of carbohydrate e. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). rice.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. canned fish may be used in their preparation owing to it having wider availability and low wastage. For example.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe. A temperature no greater than 10 ºC for 12 hours cumulatively or 21.g. There are two main categories. Fig.3.2. The protein source can be fish fillet or block.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal.5.1 ºC for no more than 3 hours is recommended (1). to produce a specific product. pasta or potato. Flow diagram outline for recipe dish ingredients preparation 2. 2. The vegetables must withstand processing. 38 . and must not show evidence of disease. Figure 2. as this could introduce off-flavours and reduce shelf life.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes.4.4.

Although the processes are claimed to be superior to using unmodified hypochlorite for washing. However. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. 39 . acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. loss of quality. frozen block or canned fish.5. not just as accompaniments. colour and water will occur. Salmonella typhi. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components.lactobacilli. coryneforms. rice or pasta. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. i.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. if the process is not appropriately controlled. any processing stage. Once vegetables have been blanched. and mixed whitefish). A number of examples of direct faecal contamination of vegetables exist in the literature (8).2 Seafood ready meals (frozen or chilled) These differ from seafood recipe meal centres in that they contain a source of carbohydrate such as a potato topping. In an attempt to improve washing and reduce chlorine taint. with the accompanying release of additional nutrients by processing.3 log reduction in bacterial numbers on the surface of vegetables. 2. they should be stored under temperature-controlled conditions. for example. even those with pH-modified dressings. encourage more rapid decay and loss of structural integrity. and may include molluscs or prawns that may or may not have undergone a minimal heat process. and may include sauce accompaniments. chopping and mixing of material. The opportunity for growth of L. with cases of foodborne disease traced to Listeria monocytogenes. For example. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. 8) will. haddock. spore-formers. and protected from any post-processing contamination. (7. Blanching of vegetables can be an effective means of reducing or removing surface contamination. pastry. The presentation of vegetables has a significant effect on the product quality. coliforms and pseudomonads throughout the mix. For example. However. For premium products. treated as a highrisk ingredient. and E. Products must be washed thoroughly to remove pests and soil. Washing with a biocide will at best achieve a 2 . control is more onerous. or breadcrumbs.e. and reduce spoilage organisms and potential pathogens. will disperse the saprophytic flora of vegetables . but also as components in recipe meals. Any assumptions regarding the application of good agricultural practice to produce should be made with caution. The majority are products made from various types of whitefish (cod.4. coli O157. the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. all-year-round fresh vegetables are preferred. Raw material is typically fish fillet.

monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. cereus are well known and are still a common issue in the UK (9). Sauces can be multi-component preparations or more simple preparations. 11). rely on the use of microbiologically sound ingredients. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. can germinate and rapidly multiply. aureus toxin is well documented (10. 13). cereus).) to the product (12.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. Alternatively. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. a process equivalent to at least a 5D Listeria kill will be necessary.6. An incident involving Staph. Potato toppings made from flake or powder will normally have little associated spoilage flora. B.g. Placing such products in a standard chill store to cool down could result in condensation. Sauces or marinades are added to fish to broaden appeal or to accompany components. spore-formers can proliferate if adequate control of temperature is not maintained. If the sauce contains modified starches for freeze/chill stability. with the range always expanding as new tastes and cuisines are developed. For this reason all products. a heat process in excess of 73 ºC will be required to rehydrate this component fully. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. if subjected to temperatures between 20 and 40°C. and then cooled slowly allowing germination and re-growth. Appropriate handling facilities or equipment will be required to protect the product.4. such as butter garnish with black pepper. but temperature-abused pasta can also spoil rapidly by mould growth. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. where composite meals are heat-treated. cereus. This is a particular issue especially where products are heat treated to eliminate vegetative cells. Typically.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. especially in those products that have a vegetable component or rice accompaniments.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. Products or dried ingredients contaminated with either spores of aerobic bacteria (e.4. 2. 40 . should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. On hydration. and an increased risk of contamination from L.

herbs. Materials prepared with regard to good hygienic practice need only be assembled in a tray or pouch before chilling or freezing in readiness for distribution. Marinade-type products are becoming more popular. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. the sauce component will spoil rapidly and the possibility of vegetative pathogens from post-process contamination or raw components growing to significant numbers cannot be ruled out. suspended in an acidified sauce with an oil component. spices. for instance. respectively.4.5 illustrate the recipe dish ingredients preparation and assembly and packaging. It is expected that this product format would have a restricted shelf life to limit potential for growth of psychrotrophic bacilli or any survival of enteric pathogens (12.2. 41 .CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. Figures 2. flour products. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. etc. and handling of processed sauce can be less rigorous than. that of a dairy-based sauce.4 and 2. The format uses flavour compounds . Without barriers. 13).

so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. which may restrict either microbial growth or enzyme activity.FISH AND SEAFOOD Fig.4.4.2 Modified-atmosphere packaging Originally developed for the red meat industry. as it dissolves in the surface moisture to form a weak acid. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. 2.7. 42 . The oxygen-free conditions will however allow other anaerobic bacteria such as C. botulinum to grow. unless the temperature is strictly controlled to below 5 °C throughout the supply chain. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. generally carbon dioxide and nitrogen. Flow diagram outline assembly and packaging of recipe dish 2.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).7.2.4. and tuna loins are packed by placing the product in a plastic pouch and removing all the air.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. These include: 2. the method replaces air in the sealed packaging with a defined mixture of gases.5.

temperature control is important in preserving chilled integrity of ingredients. The case for growth and contamination of products with Aeromonas is stronger. but is not limited to.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. as this organism is a common contaminant of freshwater fish species (19). but they will also be readily adapted to the nutrient source and conditions. 43 . both organisms are capable of growth at chill temperatures. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’. by a loss of texture. It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage. The literature has good information on spoilage of individual components of recipe meals. The fish component of recipe dishes. This is also true of chill storage and the distribution chain. in numerical terms. 2. is typically uncooked or at best lightly processed to retain eating characteristics.10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15). However. At refrigeration temperatures (5 . except for some crustacea. Shewanella putrefaciens and Ph. (20). This is considered to be the one major difference between fish and meat products. and in influencing the final eating quality of the product. phosphoreum. followed by moisture loss. Nonetheless. the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. Beyond this. and finally flavour loss. Spoilage will be most readily detected in proteinaceous components of ready meals. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). and Yersinia spp.1 Chilled storage For chilled products or chilled components of frozen products.. Aeromonas spp. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18).5 Spoilage During processing. Not only will these organisms have a numerical advantage over other proposed spoilage organisms.5. Vibrionaceae will succeed (17 ). Spoilage of this material is not expected to differ greatly from the processes described elsewhere. at slightly higher temperatures (10 ºC). where the protein component is nearly always heat-processed. initially. but little on in-pack spoilage. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood.

Serratia spp. this should also take into account shelf life of product components used in the assembly of the final product.. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. As a note of interest. 44 . Still. Pseudomonas spp. 2. The conditions and freezing process most favourable for good-quality products will.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. for example. enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage. The type and nature of the enzymes and their activity will depend on the bacterial species. For example. and Aerobacter spp.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. producing characteristic off-flavours. eliminating nematodes in products that do not have a heat treatment step. Products that are developed with an extended shelf life beyond the 6 . which would warn the consumer of the potentially unsafe nature of the product. This reinforces the need for prompt processing and temperature control of ingredients and finished products. however. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. where the variety of components and ingredients used in manufacture is varied and complex. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e.. The ACMSF (22) issued clear advice on this topic and has recommended that the shelf life of products without barriers (23) is restricted to less than 10 days. botulinum. within dairy products are well known. Bacillus spp.g. auto-oxidation of fats/oils). freezing also has a protective role in improving the safety of raw materials by. However. These have been proved to impart off-flavours. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24).5. By retaining the intrinsic spoilage flora of seafood.5..FISH AND SEAFOOD 2. the shelf life of products is limited by the natural spoilage process described earlier. also be those that have the least effect on the viability of microorganisms (21).

and this issue may be of greater consequence for warmwater. These are C.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. or a heat shock followed by holding at a temperature suitable for growth. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage.. water. along with previous discussion. botulinum type E and Vibrio parahaemolyticus (25). Additional hazards from marine bio-toxins. any risk of C. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). with pathogenic microorganisms or viruses. in some cases. In addition. are dealt with elsewhere in this book. 27). air. and Campylobacter are recorded in filter feeding molluscs such as mussels. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. A number of other Bacillus spp. Counts of between 103 . cereus. Vibrio spp.e. Inappropriate holding times and temperatures will. 45 .2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. Hazards attributable to the non-seafood components are listed below and this information. Cases quoting contamination of seafood with organisms such as S.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. low oxygen or anaerobic conditions.6. agricultural run-off or sewage can contaminate in-shore species. have been linked with food poisoning and should also be considered. spices and herbs and vegetable shoots have been reported (12. especially molluscan filter feeders.3 Clostridium perfringens A study examining C. 2. perfringens contaminating seafood will depend on the source of raw material. etc. Therefore. farmed or free-living crustacea (30). and many raw or processed foods (26). may be of benefit within any risk assessment for chilled and frozen prepared fish products. botulinum when held in favourable conditions for growth (28) i.104/ g should be treated with some concern (26). potato. soil.6. Incidents in such diverse materials as cream. histamine intoxication. Germination of spores requires favourable conditions.1 Bacillus cereus This is a common contaminant of dust. 2. 13. 2. and some effects related to product type and time to toxin production have been noted.6. permit rapid growth of B. typhi.

6. and this is believed to be due to a tolerance to brine used in the smoking process (32). the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). It is unclear. will colonise factory environments in a similar fashion except for possibly Vibrio spp. Shigella spp. but under the appropriate conditions. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. aureus (35). which is not affected by any subsequent cooking process. indigenous or other. are considered primarily to be waterborne organisms but epidemiology indicates that contamination of vegetables and possibly in-shore caught seafood could act as vehicles for infection by Shigella (34). 2.5 Shigella spp.4 Listeria monocytogenes Interest in this organism has been characterised by a number of very serious incidents associated with a variety of product types (27). monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. 2. aureus enterotoxins in foods. Rates of contamination with smoked seafood are much higher than those with unsmoked products.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. An acceptance has now developed in the UK of the presence at low levels of L. and inadequate cooking or holding within the temperature growth range (27). These include inadequate refrigeration and preparing foods far in advance of planned service. infected food handlers.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. and will therefore cause “food poisoning” even in a product that is cooked before consumption. These conditions will allow the production of the toxin. L. However. Sources of contamination are many because of the widespread distribution of this organism in the environment. at the present time. 46 . the restrictions on growth rate seen with seafood will not apply. L. but. when introduced or inoculated into sterile components of chilled prepared recipe dishes. 2.6. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. if any other potentially pathogenic organisms. growth may occur in foods (26). Chilling may help Shigella to survive. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33).6. monocytogenes as a contaminant in foods (31).

III based on IFST Guidelines (39)) provide example guidelines for seafood products. 2. Microbial levels at the end of manufacturing (m) will be set much lower than those at the end of shelf life (M) which in turn should still be safe to consume. 2. but are levels that are determined as satisfactory by the customer (within the supply chain) as part of the specification of raw materials /final product. It should be remembered that 47 . such as batter mixes.II and 2. aureus enterotoxins and B. and not to final products. The following tables (Table 2. although due to long analysis times required.7 Viruses Concerns over contamination of seafoods with viruses (due to species-specific nature of viruses) are generally limited to molluscan shellfish from waters or crops contaminated with sewage.7 Published Microbiological Criteria Due to the complexity of many processed food products. as published by for example.6. and are a recognised means of providing data on manufacturing standards. (36). are not mandatory. and as such the monitoring is for verification purposes rather than control. although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts.CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). but these may be eliminated by a subsequent heat-treatment stage. with respect to the number and types of ingredients used. pastry. For instance. Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems. it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur. unless the criteria is for a pathogenic organisms where only non detection is acceptable. cereus emetic toxin are often confused owing to similar symptoms. . the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40). Microbial guidelines. Microbiological specifications do not necessarily relate to product safety. etc. and their association with similar materials. pasta. The criteria themselves will have a range of levels that are dependant on the point in the processing or supply chain at which they apply. but provide the industry with an indication of the microbial level at which action is required to ensure product safety. the fresh product is often consumed before the results are obtained.

fish products. coli GMP* <102 Maximum 104 TABLE 2. parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E. batter etc) should also be consulted when establishing the specifications for raw materials and final product. TABLE 2. fish fingers.g. pasta. coli GMP* <104 <102 <10 Maximum Product-dependent 104 103 GMP* Not detected in 25 g Not detected in 25 g <102 <20 Not detected in 25 g <50 ppm (<5 mg/100 g) Maximum Not detected in 25 g 103 104 103 102 <50 ppm (<5 mg/100 g) *GMP = Values found immediately after production under good manufacturing conditions.II IFST Guidelines for part-cooked foods – e. L. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp. aureus V.FISH AND SEAFOOD the microbial criteria for other products (rice. cereus Staph. coated fish. fish cakes Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Pathogens GMP* Maximum Criteria for absence not generally applicable Indicators and spoilage organisms Organism E. Maximum= Levels at end of stated shelf life 48 .III IFST Guidelines for processed foods . monocytogenes B.

373-8.R. 2.L. Letters in Applied Microbiology. Masuoka Y. Eds. . Gilbert R.J.C. Dalgaard P. Animals and Spoilage.R. 668 -70. Pvt. 8.. Fish and Fisheries Products: Hazards and Controls Guidance.. Kluwer Academic. Taylor and Francis. Matsuyama H. 167-93. Huss H. Hoshino T. 233-7. Ward D.R. 13. 24 (5). 38 (11). 1962. Microbiology of processed spices. London. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. 9. 67-73. Letters in Applied Microbiology. 11. Ohgiya S. 1997. Kumeta H. 12. Journal of Milk Food Technology. Recent Advances in Food Science.C. Hobbs G. 3. Journal of Hygiene. 1976.M. 663-87. Communicable Disease Report. 5 (3)... New York.. Journal of Food Science. 1993. in Fish Inspection Quality Control and HACCP: A Global Focus. Pathogenic microorganisms associated with fresh produce. 1991. Behling A.T. Latt T. 1986. 70-2. Sakamoto T. 2001.C.W... 1982..G.K. 7. Yumoto I. 1976. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. Mejholm O.. 1997. Powers E. Ishizaki K. 5. Communicable Disease Report. Food Science and Technology Today. 10. 47. London. Hackney C. Martin R. 49 4. 1997. Journal of Food Protection.... Lawyer R. Food and Drug Administration. 1.M.. 15.. Outbreaks of foodborne illness in humans. 1311-7. Aureli P. Brown T. The bacteriology of fresh and spoiling fish and some related chemical changes. International Commission on Microbiological Specifications for Foods. 96. 1997.R. References US Food and Drug Administration..CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). Collette R. Powers E. 2005. 14. 215. Toxin Related Diseases: Poisons Originating from Plants. Bacterial histamine production as a function of temperature and time of incubation.. Ltd. Shewan J. Cambridge. Microbiology of Marine Food Products.. Washington D. 166-73. England and Wales Quarterly Report.8 1. Murrel H. Oxford. Incidence and levels of Bacillus cereus in processed spices. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. 204-16.. Taylor S. Christiansen T. Slavin J.E.H..R.J.. IBH Publishing Co.L. Bartlett C.L.M. Journal of Milk Food Technology.A. Beuchat L. Woolaway M. 2. Van Nostrand Reinhold. Fish: microbiological spoilage and safety. 1991. Wienke A.. Importance of Photobacterium phosphoreum in relation to spoilage. 59 (2). Proper processing and handling of raw tuna. Bartram P. Tu A.. 6. 39 (10). 7 (24). 25 (1).

M.. New York. 1993. coli (VTEC) .W. 1986. 32. SAB Symposium Series No. 9. Jemmi T.J. 1979. Speck M. 27.J. U. Modern Food Microbiology.L. Anaerobes in relation to foods of plant origin. 1994. Blackwell Scientific Publications. What problems does the food industry have with spore forming pathogens B. Walker S. Ronner U.. Effects of freezing and storage on microorganisms in frozen foods: a review. Food Microbiology. 31. Microbiology in fishery science. Lund B. Food Control. Anon..A Literature Survey.J. Food Research International.H. 59-72. Brooks J. 274-86.FISH AND SEAFOOD 16. Clostridium perfringens in the environment. 155-9. 472-506. 28. Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants.. 1996. 2005. Halligan A. Gould G. Ed. 1990. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures. 65. Her Majesty’s Stationery Office. Barnes E. Aspen Publishers. 11. Liston J. Ray B. Eds. Food Taints and Off-flavours. Andersson A. 29. Matches J.. Leatherhead Food Publications. Springer.C. International Commission on Microbiological Specifications for Foods. 1995. 2000. 655-60. 19. Eds. 351-72. 25 (2).M.E. Blackie Academic Press. in The Microbiological Safety and Quality of Food. 50 17. Gaithersburg. 1992. Is Aeromonas spp. Lund B.A. 20. Huss H.. 309-16.C. International Journal of Food Microbiology. 333-6. Glasgow.J. 41-3. Trolle G. cereus and C. Fishing News Books Ltd. 30. Curran D.C. 34-40. 40 (5). 24. Gram L. Survey of the incidence of Aeromonas and Yersinia species in retail foods. in Advances in Fish Science and Technology.J. 22.. 1974. in Anaerobic Bacteria in Habitats Other than Man.R. 4. Keusch A... Leatherhead. London. 1996. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service. Liston J. Leistner L. Connel J. Advisory Committee on the Microbiological Safety of Foods. 18.H.. Oxford. Fresh and processed fish and shellfish . Loessner M. Jay M. 51-158 Saxby M. 1992. Golden D. The Emerging pathogens . perfringens? International Journal of Food Microbiology. 26. 1996. Food preservation by combined methods. 25. Blackie Academic and Professional. Mead G.K. 13 (1). Journal of Food Safety.R. Huss H... 1988.. PHLS Microbiology Digest. Baird-Parker T. 13. a foodborne pathogen? Review of the clinical data.V. Gram L.. . Granum P. Report on Vacuum Packing and Associated Processes. London. Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens.. 1987. 1977. Applied Microbiology.Yersinia. Aeromonas and Verotoxigenic E. Morgan D. Journal of Food Protection. 21. 4 (1). Wood L.. 23. 28 (4). 28.

Kraft A.A. Development and Use of Microbiological Criteria for Foods. http://www.pdf Shapton D. http://eur-lex. Varnam A. Shapton N. Chilled Food Association. 1992. Cambridge. 34.K. London. Institute of Food Science and Technology. Microbiological Criteria. 39.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. 35. Shigella as a foodborne pathogen. Journal of Food Protection.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology.. 51 .H. McCarthy S.uk/cday/posters2008/npd6. 41. 1991.A.K.F. Evans M. Foodborne Pathogens an Illustrated Text. 38. U.eu/LexUriServ/ LexUriServ. 372-6. 2006. 36. 40.co.chilledfood. Campden & Chorleywood Food Research Association.G. London. 60 (4). 1998.2. Chilled Food Association. 788-801. 37. Phil Voysey. Wolfe Publishing. Woodhead Publishing Ltd. Psychrotrophic Bacteria in Foods: Disease and Spoilage. 1999. British Retail Consortium. Smith J.L. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products..europa. 50 (9). http://www. Principles and Practices for the Safe Processing of Foods. CRC Press.campden. Journal of Food Protection. 1997. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs.A.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. 1987. U. Gloucestershire.


and 2.6 litres. the adductors. For example.7 litres of water per hour. Strong muscles.3. clams. Jr. Pelecypoda. The term shellfish in this context is limited to oysters. Pelecypods have little need for locomotion. S. snails. mussels. octopi. and Cephalopoda.S. slugs.E. they also trap bacteria. where it is eventually swallowed and digested. O. composed of two valves hinged together dorsally.1 Definitions General features of the mollusc The phylum Mollusca. abalones.1. MOLLUSCAN SHELLFISH Carlos Abeyta. cockles and scallops. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). scallops. oysters. mussels. clams. Plankton and algae form the bulk of their diet from seawater. Box 3012 Bothell WA 98041-4421 United States of America 3. The class having species of economic and public health importance is the Pelecypoda.1 3. Special tracts of cilia move this mucus toward the mouth. and shrimp. oysters and clams can process about 3. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. can hold the shells tightly shut against enemies (1). Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. respectively. while 53 . In the process of filtering water. Food and Drug Administration 22201 23rd Dr. the shell. In these forms. lobster. viruses. squid. In most species. Other species of shellfish such as crabs. The class Pelecypoda includes all of the bivalves such as clams. completely encloses the body. which includes oysters. cockles. P. The phylum consists of five classes: Amphineura. Many microorganisms ingested by shellfish survive the digestive process (3. mussels. chitons. cockles. and scallops. 4). chemical contaminants and other impurities on the mucus of the gills. Pacific Regional Laboratory Northwest U. whelks. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. lying well within the mantle cavity. Gastropoda. Scaphopoda.

South America Pacific – Japan Quahaug (also known as hard clam and hard shell clam) Coquina clam Hen clam Butter clam (also called the Washington clam) Mogal clam Little neck clam (also called rock Cockle Pacific littleneck) Atlantic – North American/Europe Atlantic – Europe/North America Japan Atlantic/PacificNorth America Donax spp.North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp. The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3. The habitat of oysters and mussels are sessile. Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 . but maintain contact with the water by means of a siphon tube (5). 3.1. Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific .I Partial list of bivalve molluscan shellfish products in commercial use internationally (5) Common name Clams Soft (shell) clam Pacific/Atlantic North America Pacific/Atlantic North America Pacific . are not as vulnerable to contamination through pollution of their beds as the bivalves. dwelling on the bottom or attached to structures in the water column.2 International commercial products Globally. Clams are benthic and live burrowed in the mud.I). many species of bivalves are edible (Table 3. Whatever contaminants are in the water will eventually get into the shellfish. and usually at higher concentrations than found in their habitat.FISH AND SEAFOOD commercially valuable.

Pacific – North America/ Australia/New Zealand Pacific – North America Crassostrea gigas Western oyster (also called Olympia oyster) Cockle Cockle Sydney rock oyster Rock oyster Dredged oyster Cockles Common cockle Ostrea lurida South America Japan Australia New Zealand New Zealand Ostrea chilensis Ostrea laperousei Crassostrea commercialis Crassostrea glomerta Ostrea lutaria North Atlantic – Europe/ North Africa Pacific – North America Cardium edule Cardium corbis Cardium aculeatum Cardium tuberculatum Spiny cockle Knotted cockle Mussels Blue mussel Atlantic/Mediterranean Atlantic/Mediterranean North Atlantic – Europe Pacific – New Zealand Pacific – North America Europe Atlantic/Mediterranean Mytilus edulis Common mussel Horse mussel Bearded horse mussel Mytilus californiaus Modiolus modiolus Modiolus barbatus 55 .MOLLUSCAN SHELLFISH Gulf clam Pismo clam Surf clam (also called bar clam) Freshwater clam Oysters Common oyster Portuguese oyster Blue point oyster (also called American oyster) Pacific oyster Europe Europe Atlantic – USA Ostrea edulis Crassostrea angulata Crassostrea virginica Gulf of Mexico Pacific – North America Atlantic – North America Titaria cordata Trivela stuttorum Spisula solidissima Japan Corbicula spp.

Most clams are only a few centimetres in their maximum dimensions. geoduck.2. extended above the surface to maintain water currents needed for respiration and feeding. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. hard-shell or soft-shell clam. laterally compressed. mostly marine bivalve molluscs. cherrystone. littleneck.1. The muscular foot is used to burrow in mud or sand. and other names. but 56 . Jacques Iceland scallop Bay scallop Weathervane scallop (also called Alaska scallop) Atlantic North East Atlantic North Atlantic West Atlantic Pacific – North America Pacific – North America Pacific – Japan Common scallop Sea scallop (also called giant smooth) Japan Atlantic – North America Pecten varius Pecten maximus Chlamys islandica Argopecten irradians Pecten caurinus Pecten aequisulcatus Pecten laquaetus Pecten yessoensis Penten magellanicus or Placopecten magellanicus Pecten meridionalis Pecten novaezealandiae Pecten jacobaeus Aequipecten gibbus Chlamys opercularis Chlamys varius Amusium balloti Mediterranean – South East Europe New Zealand Australia Mytilus galloprovincialis Mytilus canaliculus Mytilus planulatus Commercial scallop Australia New Zealand Great scallop Calico scallop Queen scallop Variegated scallop Scallop saucer Atlantic/Mediterranean Atlantic – North American Atlantic Atlantic/Mediterranean Australia 3. Different species are variously called quahog. Buried clams leave tubes.1 Clam Clam is the common name of a large group of often edible. or siphons. A mantle tissue encloses the body of the clam.

1. packed in weak brine. 57 . Canned: clams are steamed open and the meats removed from the shells.2 Oyster Oysters are marine bivalve molluscs. uncooked (shucked). either whole or minced. oyster soup. Canned: meats removed from the shell by steaming. Live: in shells. the meats are smoked and packed in oil in cans or jars. in cans with either brine or clam liquor. Soups: oyster stew. then heat-processed. uncooked. skewered with bamboo sticks. Oysters lack a foot or have a reduced foot. Frozen: in shell. 3. Canned soups include clam chowder. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. uncooked meats packed unprocessed.MOLLUSCAN SHELLFISH the giant clam of the Indian and Pacific oceans may be nearly 130 cm long and weigh 227 kg. Smoked: meats. or shelled meats. having rough and irregularly shaped shells that are closed by a single adductor muscle. Dried: Hoshigai (Japan) shelled meat.2. all in cans. and no siphon. after precooking and brining. clam madrilène and clam liquor. Live: in shell Fresh: in shell or shelled meats. tidal waters or in brackish water in the mouths of rivers. Fresh: in shell. oyster bisque. Frozen: shelled meats. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. sun-dried. or shelled meats. hermetically sealed. These molluscs feed on minute plants and animals carried to them by the current. Semi-preserved: meats cooked and packed in spiced vinegar. smoked meats are also canned. trimmed and packed. washed. Dried: shelled meats.

vinegar solution. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves.3 Mussel Mussels are marine bivalve molluscs that live either partially buried in the sea bottom or attached to rocky surfaces by means of byssus threads.1. Canned: cooked meats. smouldered and afterwards dried (Japan). in mussel liquor. or packed with jelly. The adductor muscle is the only edible part of the animal. Fresh: cooked meats. also with other ingredients. flattened ear or wing at the hinge area. cooked meats packed in dry salt for transport. are united. boiled. In some species. scallops possess a single large adductor muscle. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). scallops can swim or leap by a series of rapid closings and openings of the valves. and held apart.1. The familiar marine mussel is asymmetrical. Fresh: shelled meats (often only the adductor muscle and the roe are eaten. Live: whole with shells. The shells are fluted and round with a broad. 3. by interlocking tufts of hair-like cilia. or filaments. in which the individual branches. mussel in butter sauce. Internally. The posterior halves of the valves contain a large adductor muscle that holds the valves closed.2. They have filibranch gills. Semi-preserves: cooked meats with vinegar-acidified brine and spices.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. Salted: cooked meats bottled in brine.2. Smoked: meats may be canned in edible oil. sauces. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. mussel paste. gutted. and the anterior end smaller and pointed. with the posterior ends of the shells (valves) broad and rounded.) Dried: peeled. 58 . Unlike other sessile bivalve molluscs. Bait: used live extensively for baiting fish hooks for line fishing. The common scallop is about 5 cm long.FISH AND SEAFOOD 3. brine.

1. Live: in shell. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding. and butter. clams and mussels. depending on length of journey. in sauce. which affects the temperature.2. Whatever contaminants are in the water will eventually get into the shellfish (7). nutrient concentration and pollutants of the water column. For example sessile and benthic shellfish dwelling on the bottom or burrowed in marine sediment are especially vulnerable to pollutants (sewage and/or wastewater) introduced into the shellfish water beds. Salted: meats either lightly or heavily dry salted. pH. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms.MOLLUSCAN SHELLFISH Frozen: shelled meats. 3. Canned: in brine. also bottled in brine. and the environment. Raw and partially cooked shellfish.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. Vinegar cured: bottled in malt vinegar after brining. Cockles range from about 1 to 15 cm in length and have two valves of equal size.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves. Canned: in own juice. are vectors of disease transmission (8). The microflora are dependent upon a number of factors such as season. The following pathogenic microorganisms causing illness in humans are characteristically associated with molluscan shellfish: Vibrio parahaemolyticus Vibrio cholerae O1 Vibrio fluvialis Vibrio hollisae Vibrio vulnificus Vibrio cholerae non-O1 Vibrio mimicus Vibrio furnissii 59 . 3. they accumulate pathogenic microorganisms from polluted waters. mostly in oysters. Because molluscan shellfish are filter feeders. salinity. Fresh: meats removed from the shell by boiling.

Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. 12. Pseudomonas spp. Other microorganisms found in lesser numbers are the Achromobacter spp. rakes. responding to the environment in which they are harvested. dredges. that have never been cultured on laboratory media. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp..3 3. Temperature. Flow diagrams of the major processing steps of molluscs are presented in Figures 3.1.. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves.3. The largest groups found are the Pseudomonads. 13). Moraxella spp. and yeast cells of Rhodotorula rubra and Trichosporon spp. shellstock (live) which is shellfish in the shell. At harvest.3. Acinetobacter spp.1. 11. and shucked shellfish that is shellfish. There are microorganisms.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. clams.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. or by 60 . Micrococcus spp. Alcaligenes spp.2. mussels. 10. Flavobacterium spp. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. salinity. (9.1.000 /g. whole or in part. 3.. Bacillus spp. Water temperatures affect the concentrations of microflora in the water. Vibrios.1 Shellstock 3. and Aeromonads. mechanical harvesters. 15).1 Processing and its Effects on the Microflora Oysters..3. For example. nutrients. the initial flora composition and concentration of bivalves are highly variable. from which one or both shells have been removed. In summary. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100.. 3..1 and 3. They are termed viable non-culturable microorganisms.. Enterococci spp. Campylobacter coli Bacillus cereus Hepatitis non-A. but have never been cultured (14... and cockles The processing of bivalves takes two forms. observed microscopically. spirochetes of the genus Cristispira and Saprospira.

3. or in bay-approved shellfish-growing waters prior to sale or processing. Fig. Secondly. usually in open-mesh containers and/or burlap sacks. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. by truck or boat. There are three primary reasons for wet storage: first. or from other sources approved by the State Shellfish Control Agency (SSCA). Shellstock may be wet-stored at the processing facility in tanks of clean seawater. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock.1). Thirdly. If by boat. for transportation to the point of sale. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. The shellstock is transported dry. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. Generic flow diagram of live shellfish from harvester to point of sale 61 . wet storage also may be used to de-sand those shellfish species that tend to accumulate sand in their mantles and gills thus making them more palatable.MOLLUSCAN SHELLFISH hand (Figure 3. after harvesting. or immediately placed in refrigerated trucks. Water used for shellstock washing is from approved growing areas. in containers from which they can be retrieved easily.1. Shellstock are placed in drycooler storage containers in the facility.

without adequate refrigeration.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock.1.1. The washing of sediment and detritus soon after harvesting will prevent further contamination. bivalves tightly close their shells thus trapping water and microflora. synthetic seawater for the purpose of de-sanding and salt uptake. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. Furthermore. and the sanitary practices applied during harvesting and shipping. and thus contaminating the shucked meat at the point of sale. with the exception of A. 24. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. Harvest conditions are another factor that influences the growth of bacteria in shellstock. If long delays occur before reaching the processing facility and/or refrigeration. At harvest. with prolonged storage the microbiological quality of the bivalve decreases. Generally. some pathogenic organisms will survive and multiply in shellfish (20. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. Generally. handling. and Citrobacter species) and Vibrios. to avoid possible contamination of the shellstock. however.2 ºC (45 ºF) or below (19). The most important factor in controlling bacterial growth in shellstock is temperature. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. hydrophila (18). An outbreak of infectious hepatitis in Sweden in 1956. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). for a considerable length of time. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. shellstock are placed in refrigerated trucks for transport or placed in wet storage. vulnificus. 23. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. coli. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18.3. 25). 16. including V. Enterobacter. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7.FISH AND SEAFOOD 3. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . Over a period. when it reaches the processing facility. 17). At this temperature the multiplication of faecal coliforms (E. are prevented. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. the bacterial counts in the meats will increase. shellstock are not refrigerated on the harvest boat. Klebsiella. there is evidence that in the absence of adequate refrigeration. 21. and temperature storage of shellstock to point of sale. Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. 26). Water used for shellstock washing should be of good sanitary quality. 22).

Due to the nature of the shucking operation.3. for the effects of microflora during the processing of shellstock.2.3. If shellfish are not reasonably clean at the time of processing. whereas cotton become wet and 63 .3. perforated tray in which shucked shellfish are spray washed and/or drained) or in a blower-washer (a tank-like device for immersion washing of shucked shellfish). Thus. In addition. the finished product may become contaminated. washing. thus contributing to high bacterial counts in the final product (28). and fish. or subjected to splash from foot traffic. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). After washing of meats. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation.2.1. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank.2 Shucked shellfish 3. Containers are placed in packing boxes for shipping at refrigerated temperatures. Under these conditions if the shuckers enter the packing room. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. In the blower-washer.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. washing and culling shellfish prior to storage is essential. clothing becomes very soiled. 3.1. shellstock is subjected to contamination when stored on the floor in standing water. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. the tank is opened through a drain gate and chute.2). removal of shellfish (shucking). Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported. through which the washed meats are discharged for rinsing and then packed in containers.1. To remove the shellfish attached to the remaining valve. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station.1 Processing description The processing of shucked shellfish is limited to washing of shellstock. and packing shellfish into containers (Figure 3. lobsters. The process of shucking involves forcing a knife between the valves and cutting the adductor muscle of one valve freeing the shellfish from one valve. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab.2 Effects of microflora during processing The same considerations as previously discussed. Studies have shown the use of rubber gloves reduces the contamination level. are applied to the shucking operation of shellfish. 3. Shuckers routinely use either cotton or rubber gloves during the shucking operation.

and sanitised before each filling.2 ºC or less. Shucking containers are rinsed clean with running water. within one hour. Fig 3. Shucked meats are thoroughly drained.2. 2. Separate rooms or lockers for storage of clothing. 3. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. and gloves help prevent the contamination of shellfish.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. which may lead to contamination of product. There is a tendency to store such articles on the shucking benches or in packing rooms. 4. 64 . Generic flow diagram of shucked shellfish from harvester to point of sale Shellfish are subject to contamination during the shucking and packing process when the following are not adhered to (19): 1. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. aprons.

shucking pails. Temperature abuse will result in high bacterial counts. skimmers. Flavobacterium. thus producing acid and gas (38). streptococci. showed that organisms such as Achromobacter. a carbohydrate. 65 . 36). or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. 3. Pseudomonas. spoilage will occur. and packing the shellfish in a timely manner. Much of the work has been done with oysters and clams. whereas bacterium such as Pseudomonas. As described previously. The spoilage process in shellfish may be divided into three stages: increase of acidity. where there is a breakdown of temperature control or prolonged storage of shellfish. For example. Shucked shellfish are an excellent medium for the growth of bacteria. Shellfish. blowers and other utensils that have cracked. All equipment coming in contact with the shellfish such as colanders. The initial flora found in shellfish is the major factor of spoilage in bivalves. in oysters. is stored in the bivalve as a source of energy. at the turn of the century. 34. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. In summary. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended.47 to 6. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. anywhere during the process of shellfish from harvesting to the point of sale. Achromobacter and Flavobacterium levels decrease during storage. The environment has an effect on the bacterial population found in shellfish. Micrococcus. Alcaligenes. Controls that should be applied include using clean packing equipment. particularly oysters stored at 7 ºC.2 ºC or less. Lactobacilli seem to be the predominant microflora in shellfish. 35. Glycogen. Glycogen is hydrolysed by the initial flora found in shellfish. rough.2 ºC. and proteolysis (10).MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. are an excellent medium for growth of bacteria. Proteus. The high incidence of proteolytic bacteria and other types capable of fermenting glucose in the natural flora may be responsible in postmortem spoilage of shellfish.8% (37). This process changes the bacterial profile to microorganisms favouring low pH. Shucked shellfish are held and transported at temperatures of 7. glycogen levels range from 0. These organisms may play a significant role in the spoilage processes. 31.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. abundant gas production. particularly shucked meats. clean gloves. and yeasts. these include lactobacilli. 32). 33. Earlier studies.

The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. V. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. however.5. This bacterium is present in all the major processing steps of a shellfish plant. parahaemolyticus possibly producing illnesses (40). V. and to the point of sale. ranging from epidemic cholera to sporadic cases of diarrhoea. although non-hemolytic strains have been associated with illness (43. 44). diarrhoea. shellstock and packing of shellfish in jars) (29). from harvesting to finished product (i. lot and retail outlet. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. only a small percentage of strains in the marine environment are potentially pathogenic (45). Major outbreaks are associated with the warmer months. nausea. and estimates based from a hospital study show less than 0. Products involved were from the same processor. prevention of cross-contamination. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. of V. The incidence of V. Between May and September 1997. 3.5.1 Vibrio parahaemolyticus V.e. primarily in brackish or saltwater. during processing.000 population/year (46). 3.48 hours with abdominal pain. It is a common marine bacterium. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). where outbreaks occur regularly. 66 . parahaemolyticus causes gastroenteritis lasting 24 .5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish.5 cases/100. These organisms can be present in the shellfish from the point of harvesting.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. headache and fever. and very common in Japan. Fortunately. 41). Sporadic cases are frequent along the coast of the United States. parahaemolyticus. They can be introduced to the shellfish from the environment or added during processing. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). there is a recent report of low levels of V. approximately 23 cfu/g.FISH AND SEAFOOD 3. One of 250 cases reported involved shucked products with low levels. Diseases caused by Vibrios are enteric in nature.1. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. and harvesting of shellfish in the cooler months. parahaemolyticus from shellfish is low. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. coastal areas). widespread in both polluted and unpolluted waters.

chills. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). are particularly susceptible to primary septicaemia.MOLLUSCAN SHELLFISH Control of V. parahaemolyticus growth in shellfish meats is temperature dependent. vulnificus is by rapid refrigeration during warm weather months. High-risk individuals. cholerae. cholerae non-01 has been associated with septicaemia. V. Illnesses are usually associated with the consumption of raw oysters as described in Table 3.000 population/year (8. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. Oysters are the most hazardous food associated with V. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). 55. Inadequate refrigeration will accelerate the growth of V. leukaemia. 54. like V.7 days. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. It is one of the most severe foodborne infectious diseases. Again. cholerae non-01 is ubiquitous in the estuarine environment. Onset of symptoms occurs in 48 hours and can be severe. or immunosuppression. which have been implicated in shellfish-related disease outbreaks (31). 57. The organism attaches to the intestine and release of a toxin is suspected. 56). vulnificus.II (8. In one study. parahaemolyticus.5.8 case/100. and nausea within 24 .1. 53. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. V. these individuals are advised not to eat raw shellfish. Hazards from this bacterium can be controlled by thorough cooking of shellfish.1. 3.3 Vibrio cholerae non-01 Illness caused by V. and rapidly become inactive at low storage temperatures. abdominal cramps. 58). with a fatality rate of 50% for individuals with septicaemia. diabetes. Healthy individuals are susceptible to gastroenteritis. and preventing cross contamination once the product is cooked. Control of oyster-associated 67 . the organism has been found in the Gulf of Mexico. 46. The occurrence of V. parahaemolyticus the estimated annual incidence rate is very low. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. those who have liver disease. 23 of 24 major USA-West Coast estuaries were positive for V. 46. fever. Death has been reported to occur within 36 hours from onset. lasting 6 . All isolates are considered pathogenic. or primary septicaemia. V. in the range of 0.5. gastroenteritis. They are sensitive to cold. Control of V. vulnificus are fever. however. cirrhosis. cholerae non-01 (51).0. and vomiting.2 Vibrio vulnificus The symptoms of V. nausea. 49). The bacterium is an estuarine species that causes wound infections. 48). The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. Primarily. 3. It causes diarrhoea.4 .48 hours of onset.

FISH AND SEAFOOD V. They are common marine bacteria.5. and nausea. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. including V. associated with shellfish-borne illness outbreaks. e. and illness lasts 7 . There are several other Vibrio spp. V. cholerae 01 is transmitted by faecal contamination of food or water. drying. hollisae. Gulf coast environment. Illness can include abdominal cramps. V. and the transmission of the disease by consumption of raw. imported product from a country experiencing an epidemic.2 Campylobacter jejuni C. 3. parahaemolyticus and V. nausea. In developing countries. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. dehydration. The frequency of infection of these organisms is no different from other Vibrios. jejuni requires reduced levels of oxygen (3 . cholerae non-01 illnesses is by cooking seafood thoroughly. 60). cholerae 01 establishing in the U.10% carbon dioxide for optimal growth conditions.5. 68 . and shock.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. 63). low pH. The presence of these strains appears to be in low numbers and non-virulent (59). 61. (8. or cross-contaminated shellfish (57. Illnesses are associated with the warmer months. processors should know the product source. and V. these organisms cause gastroenteritis lasting 24 . however.5%) and 2 . particularly in oysters.g. There is evidence of strains of V. Poor sanitation and contaminated water supplies will spread the disease. widespread in both polluted and unpolluted waters. undercooked. preventing crosscontamination of cooked seafood.1. The control of V.48 hours with abdominal pain. In general.5 Other Vibrios spp. cholerae 01 in molluscan shellfish. It causes diarrhoea. acidity and heat. is the same as previously discussed for the non-01 strains. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. fluvialis.1. 62. 3. Control of infection from these organisms is no different from the other Vibrios. respectively.S. cholerae 01. vulnificus. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). headache and fever. nausea. vomiting. V. fever. 3.10 days.5. Death may occur after severe fluid and electrolyte loss. mimicus. salinity. In addition. Onset of disease is 6 hours to 5 days. Onset of illness is from 3 to 5 days after ingestion of contaminated food. the pathogenic severity is lower than V. and southern Asia including India and Bangladesh. abdominal pain. diarrhoea.

Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. fluviallis.II Molluscan shellfish associated outbreaks caused by Vibrio spp. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. 84. 72. 71). In one case.3 Other organisms of concern 3. however. hydrophila group (75. 80) are ubiquitous in the marine environment.3. jejuni does not multiply in shellstock at refrigerated temperatures. 81. 78) and P. infection occurred among participants of a firemen's banquet.5. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. Although. V. ranging from 0. C. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. control of C. cholerae 01 and non-01. Human feeding studies suggest that about 400 . and their immediate proper refrigeration.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No.5. Campylobacter appear to survive well in oysters during storage. of Incidents 4 279 0 1 0 No.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. C.500 cells/gram of Campylobacter may cause illness (64. jejuni were recovered. V. 77. 82. 70. 66). vulnificus. of Cases 4 362 0 1 0 a Includes V. 68. however. This finding is significant in that C. jejuni begins in the harvest of shellstock from approved waters. 76. 85). 83. 65. 73). To reduce the risk of shellfish-borne campylobacteriosis. they survive well at refrigerated temperatures. whereas at 10 ºC Campylobacter survive only for 5 days (71). Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC.MOLLUSCAN SHELLFISH TABLE 3. The seasonality of 69 . 3. Recently. and V. have been isolated from shellfish beds throughout the world (67. mimicus. parahaemolyticus. They have been implicated in limited shellfishassociated gastroenteritis cases (8. Campylobacter spp. hollisae Infective dose varies with host susceptibility and virulence of the strain. 69.4 to 114 cells. from Pacific shellstock oysters purchased from a retail grocery store (74). shigelloides (79. V. V. although their role as a pathogen is highly controversial and circumstantial.

negative for heat-labile toxin (LT) or heat-stable enterotoxin (ST) 1 Negative for the presence1 Negative for the presence of toxin producing 01 or non-01 organisms 1 <10. coli Salmonella V. They are easily destroyed with mild cooking temperatures. parahaemolyticus Staph. dried milk/whey drink powders. FDA/Administrative Guidelines 3 Source (91). prepared sandwiches.000 to 3. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. In one study. and where indicated are intended to be used for official control purposes. hydrophila group stored at refrigeration temperatures. coli Enterotoxigenic E. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. vulnificus in molluscan shellfish.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. Similar results were seen by other investigators (86. 2 Source (90). cholerae V. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. Caution should be exercised by individuals with haematological malignancies (88). Note: Although molluscan shellfish are not included in the selected high risk food. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. jejuni V. 70 . Control of these organisms in shellfish products is limited. non-dairy frozen desserts.0001 negative for the presence3 Under review4 Source (89). FDA/ Administrative Guidelines. aureus C.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. soy products. 87) of the outgrowth of A. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. Test Standard Plate Counts E.000 cfu/g1 <230/100 g2 <1000 per gram.000/gram (29). 3.600. being isolated more often during the warmer month.

. 64. Nauman R. Effects of harvesting waters and storage conditions on yeast population in shellfish. Smith F. 46.C. 13.C.S. Vanderzant C. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. Van Nostrand Reinhold. Colwell R. Eds. Hackney C. Ray S.N.S. water and sediment from Galveston Bay. 58. Pfeifer D. 245-7.K. U. 6. 4. 36. A typhoid fever epidemic caused by carrier bootlegging oysters. 755-76.. 419-25. in General Zoology.R. Cook D. 1970.E.B. 105-8. Saunders Company. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). 1994.R. 1926. Galtsoff P. 7.E.W. References The Phylum Mollusca. 7. 1991.. Galtsoff P. 1984.D. 1995. Journal of Milk and Food Technology. 14. 1963. 1981. 29. Organisation for Economic Co-operation and Development. Veliger.S. Hood M. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. 11. Wiley-Blackwell.R. 231-51. 16. 1960. Rowse A. Microbiology of bivalve molluscan shellfish.R. Thompson C. Viability and release Salmonella charity and Escherichia coli from oyster faeces..K. 12. Journal of Food Protection. Walker W.M. 633-40. in Microbiology of Marine Food Products. Cook D. 82-7. 1968. 104-9. Fleet G.. 6. Lee J. 13. Tubiash H. 19-34. Eds. 10. 33643. Gill S. Fish and Wildlife Service. 71-6.. Eliot C.F. in Fishery Bulletin 1964 No. 1964.H. Schmidt C.A. Ward D. Government Printing Office.W. 17. 42. USA. 1975.. 1973. 72-8.. Biology of the oyster in relation to sanitation. 15.A. Philadelphia. Fate of enteric bacteria in estuarine sediments and oyster faeces.R. The American Oyster. Proceedings of the National Shellfisheries Association... Infectious diseases associated with molluscan shellfish consumption. 30. Microbiological characteristics of Dungeness crab (Cancer magister). Tall B. Bacterial flora of the market oyster.. in Multilingual Dictionary of Fish and Fish Products. 33-6.R. Applied Microbiology. American Journal of Public Health.J. Washington D. New York. Colwell R. Ed. 3. 1940.7 1. Villee C. 5. 2. 30. Liston J. 9. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. American Journal of Hygiene.L. 1982.S. Applied and Environmental Microbiology. Lovelace T. Applied Microbiology.A. Jr. 71 . W. 44. American Journal of Public Health. Rippey S. Bernard F. 1983. Clinical Microbiology Review. 447-52.MOLLUSCAN SHELLFISH 3. 8. 8.. Jr. Applied and Environmental Microbiology.. 26. Journal of the Mississippi Academy of Sciences. Old H. 544-8. 1936. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas).

J. in National Shellfish Sanitation Program Manual of Operations.S..G. Applied and Environmental Microbiology. 29. 123-8.. 30.H. Jr.A. Sadaguchi G.B.D. Ruple A.E. Handling and Shipping Shellfish.C. Blake N. LaRock P. Eds. 5..W. Public Health Service Technical Report. Survival of enteric organisms in shellfish. 167-71. 72 .. 31. U. Washington D. 26.E. Stott R. 994-1002. Public Health Report. 989-1003. 1992.. crabs and mullet.F. Hepatitis epidemic conveyed by oysters. 1205-10.A. Applied and Environmental Microbiology. Hood M. 27. during depuration.W. 24. and Manillal clams (Venerupis japonica). Svenska Lakartodningen.A..A.M.H. Ltd. 25.W... 1961. Koburger J. Food and Drug Administration.. Cook D. 1983. Sakazaki R. Williams L. 1221-8. Influence of U. Tokyo. Roos B. 32.. 1954.FISH AND SEAFOOD 18. 50. Crassostrea commercialis. Saikon Publishing Co. Liston J. Maryland. Vasconcelos G. Journal of Food Protection. Food and Drug Administration. Colburn K. Timetemperature effects on the bacteriological quality of stored shellfish. U.W. Takeda Y.. 1956. 1490-5... Presnell M. 19. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters.. Bacteriological changes in live shellfish: Pacific oysters (Crassostrea gigas). Fleet G. Food and Drug Administration. Bacteriological studies of commercial shellfish operations on the Gulf Coast. 53. 1989..B. seafood-handling procedures on Vibrio parahaemolyticus. Wekell M. 22. Arcisz W. Incidence of Samonellae in clams. 52..A. Effects of storage on microbial loads of two commercially important shellfish species.J. Son N. 20. Olympia oysters (Ostrea lurida).W. 1995. Trost P. Journal of Food Protection. P-1. Ness G. Kelly C.. Temporal occurrence of Vibrio species and Aeromonas hydrophila in estuarine sediments. 1964. Establishment of critical control points in west coast oyster processing plants. 343-5. 28. Vaughn M.B. Special Report U.S. Public Health Service. and storage. in International Symposium Vibrio parahaemolyticus. Abeyta C. Ericksen T. Applied and Environmental Microbiology. oysters.S. The influence of time and temperature on the bacteriological quality of shell oysters during processing and shipping. Ed. Journal of Food Science. American Society for Microbiology. 1985. Beck W. re-laying.. Part II Harvesting. Rodrick G.. Behaviour of pathogenic bacteria in the oyster. 1214. Bacteriological survey of an oyster bed in Tangier Sound. Hoff J.A. 47 (5). Presnell M. Fujino T. 1980. 69. 1984. 1973. Chesapeake Science. U. Native Littleneck clams (Protothaca staminea). Kaysner C. 1971.. 1967. 23. F61-9.. Jones W.C. FDA Gulf Coast Technical Services Unit. 45. Crassostrea virginica and Mercenaria campechiensis. 40. 343-9.T.S.J. I. 324.S. 4. Kelly C. Fraiser M. 21..

Crassostrea virginica. 1989. U. Journal of Clinical Microbiology. Geographic and monthly variation in composition of oysters. 27. 38. 1. 1969.M. 47...A. 45. Hlady W. Stroh E. 33. 1989. Hoge C. Grodner R. 53. Journal of Infectious Disease. Kaysner C.L. 1997. Volume 1. Watsky D. In vitro haemolytic characteristics of Vibrio parahaemolyticus: Its close correlation with human pathogenicity.. Journal of Clinical Microbiology.. Academic Press. Microbiology of shellfish decomposition.. Akiyama S. Berry F. 42. Applied Environmental Microbiology.A..P. Food and Drug Administration. 41. Serological and hemolytic characteristics of Vibrio parahaemolyticus from marine sources. 1979. Tanikawa E. 2. Bacteriological control of oysters during processing and marketing. Borgstrom G. Sample T.. 204-5. 38. Sidwell V. Thompson C. 27.B.A.N. Shellfish Specialist. 34. Jr.W.. 2891-3. Joseph M.. Leitch J. Peeler R... Kato T. 1937.. 36.E. Bacteriological findings in Baltimore oysters. Libonati J.. Fieger E.M. Abbott S. Bothell. The bacterial content of market oysters. Bulletin of the Johns Hopkins Hospital. Loomis A.D.. Epidemiology and spectrum of Vibrio infections in a Chesapeake Bay community. 353-7. 1954. The bacterial flora of market oysters. DePaola A.G. Journal of Food Protection.. Bacteriological examination of oysters stored at low temperatures. Takikawa I. 1958. 160. 44. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States west coast.M. Joseph S.S. Miyamoto Y. Vibrio infections associated with raw oyster consumption in Florida. Kelly C. Vanderzant C. Obara Y.. in Fish as Food. 1976.. 41. Journal of Bacteriology. 37. Yamai S. Abeyta C.T.. 985-93. 69.. Ed. Powers C. 100. Yokohama Medical Bulletin. 25. Zentralblatt für Bakteriologie. Wekell M. 273-80. 1916. Kelly M. 97.W.. Ishibashi M.F. 1987. Takizawa K.. 35. 107-8. Ward W. Parasitenkunde.C.. 1926..MOLLUSCAN SHELLFISH 32. 39. Stott R. U. New York. Emergence of a restricted bioserovar of Vibrio parahaemolyticus as the predominant cause of vibrio-associated gastroenteritis on the West Coast of the United States and Mexico.L. 128-31. 1349-51. 60. Israel E.G. Novak A.S.. 133-47. Public Health Reports. Takeda Y.D.M.. Jr. 1914.M... (2). Vibrio parahaemolyticus illnesses. 43. 313-22. Marine Fisheries Review. Journal of Bacteriology. Kaysner C. 40.. 2820-2. 1147-9. 41. West Coast States and British Columbia. Geiger J. Morris J. 73 . Studies on pathogenic halophilic bacteria. Jacobson M. 1997... 46. Janda J. kanagawa-negative Vibrio parahaemolyticus from patients and the environment in the Pacific Northwest. 1989. 13-7.A. Urease-positive. Washington Report October 14. Journal of Infectious Disease. Arcisz W. 1961. Infectionskrankheiten und Hygien. Journal of Food Science.

C. Annals of Internal Medicine.T. 60. Hug M.M. Conte J. Siebeling.C. Hunt J. Lee J.M...E...J. McCay C.V. 1988. Stott R. Non-01 Vibrio cholerae bacteraemia: Case report and review.G. Blake P.. 299-312.. Kaper J.P.... Abeyta C.M.. 5972. 1981. 51.G. Incidence of Vibrio cholerae from estuaries of the United States West coast... Wazenski T. sediment and waters of the U. Bradford H. Hollis D. 59. Peeler J. Review of Infectious Disease. 46. Roberts N. Presnell M. 802-6. Leitch J.. 1979... Colwell R. 91-103. epidemiologic. 41. and laboratory characteristics of sporadic cases. Applied and Environmental Microbiology.. John Wiley & Sons Inc.... and enterotoxin production of Vibrio cholerae in Chesapeake Bay. Jr.. Brenner F...I. 1475-8.. Johnston J.. Jacobs R.E. Roderick G. 74 . DePaola A.A. Spite G. Feldman R. R. Vibrios in the Louisiana Gulf Coast environment. West P. Colwell R. 1344-8.F.W. Applied and Environmental Microbiology.. Bradford H.. Ecological relationship between Vibrio cholerae and planktonic crustacean copepods. 1985. Davis B.A. Colwell R.O.. Microbial Ecology.J. 558-61. 1983. 49.O. 55.D.. Gulf Coast. Wilson T. Vibrio vulnificus: Man and the sea.S.M.. Riddle C. Journal of Infectious Disease. Jr. 56.W.FISH AND SEAFOOD 48.. Kaysner C.D. Black R..R. Feeley J. Rahman R.G. Non-01 Vibrio cholerae in shellfish. 10. 1985.A.G.M. Morris J.. 149.M.... 1980.. 37. New England Journal of Medicine..L.. Jr. McFarland L..T.. 305-9. cholerae in Louisiana. 253. 1984. 1982.G. 53. Maden J.. Applied and Environmental Microbiology. Plaard R. Applied and Environmental Microbiology.. Adams M. Jr.. Tacket C. Lancet. 53. An epidemiological study of V. 58.. serology..E. Duran W. 305.A. Wathen H.A possible endemic focus in the United States. 1012-7. Cholera and other vibrioses in the United States. Characterisation of Vibrios cholerae isolated from oysters.. 1983.B.. Blake P.M. Illness caused by Vibrio damsela and Vibrio hollisae. DePaola A. Allegra D. Morris J.K.N. Francis D. Hug A..A. Ed..R.. Craig J.T. New York.G. Hickman F..W. 52.B..A. Barrett T. Hebert S.E. Journal of Food Protection. Small P. Joseph S. Wilson R. Morris J. 1. 1987.G. Puhr N.. 61. 343-50. Pons V. 1294-7. 1981. Jr. Cholera . Weaver R.R.. Journal of the American Medical Association. 312.F. Snyder J. in Vibrios in the Environment. Blake P. Twedt R.. 8. 45. Non-O group 1 Vibrio cholerae gastroenteritis in the United States: Clinical. Safrin S.. Clinical features and an epidemiological study of Vibrio vulnificus infections. Jr.M. 50. 1984. Morris J. 57. Miller H.W.. New England Journal of Medicine. McPhearson R.B.F. Jr.. Kaper J..A. Wekell M. 54. Blake P. Twedt R...J.. Caraway C..O.M. 2050-3.T.. 275-83. 1982. Ecology. 656-8. Motes M.T.R. 94. Lockman H. Becker S..A. Tacket C. Zywno S.G. McFarland L. Becker R.A. Wachsmuth I..

Eds. 4.J. 1983. Matsuda M.F.. 99. Newell D. Moore J..G.. Blaser M.. 1995. Plenum Press. 1987. 71.M. 157. Fitzpatrick M. Helicobacters and Related Organisms. Moore J.G. British Medical Journal.A. Journal of Rapid Methods and Automation in Microbiology. Journal of Infectious Disease.. Public Health Laboratory Service.G. 1996. EF-6. a newly recognised Vibrio species.J.R. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N. International Journal of Food Microbiology. Jr.M.R.P. Davis B.A..E.R..B. 72.R.X. Wilson I..W..M. 11.. Hunt J. 68.L. Feldman R. Isolation of vibrio-like group. Annals of Internal Medicine. Shandera W.A.. Kaysner. Murphy P.D. Campylobacter gastroenteritis associated with raw clams. coli and lari) from shellfish and the marine environment.E. Proudford R.. 4. 75 .S.. in Campylobacters. 73. 64.. Eds. 63. Deeter F. Blaser M..I. Griffin M. 4.J. 257. Journal of Food Protection. Jr. in Campylobacters.. Huq M..A. 1981. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness.. Infective dose of Campylobacter jejuni in milk.M. 67. Abeyta C.. Arumugaswamy R. Levine M.. infections in man.D.M.E. Stott R. Abeyta C. Dalley E. Kaysner C.R. Wilson I...H. Wareing D. Kaysner C. 69. Levine M. Plenum Press. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp. Wekell M. Abeyta C.. 80.. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation. 101-4. Skirrows M. Rowe B. Wilson T.. Johnston J. 621-4.. Wilson T. 1988. 65. (jejuni. Disease from infection with Vibrio mimicus. Wekell M. Ireland.G. New York. in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections.A.J. Murphy P. Newell D.. Austin S.K.. Davis J. 1991. Alam A. Huges T. Journal of Clinical Microbiology.K. 13. Ketley J.A. Jr.G.P..M.M. Ketley J. 66. Journal of the Medical Society of New Jersey. Morris G.MOLLUSCAN SHELLFISH 62.. 1993. Blake P.A.E. Pearson A. 1584.G.. 607-9.. 282.M. Eds.. Feldman R.M. C.G.. Robinson D. 256. 1996. Hughes T. Clements M. Black R.A..S. 472-9. Studies of Campylobacter jejuni infection in volunteers. Brenner D. 6-9 September 1983. 169-71... Wareing D. Black R. 1983... Clements M. 1980. Helicobacters and Related Organisms.. Microbial Ecology in Health and Disease.L. 1980. S41.. 56. 70. New York. from patients with diarrhoea. 323-5. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster. A method for isolation of Campylobacter spp.K. 51-64.. Brussels.A. London. Experimental Campylobacter jejuni infection in humans. Jones D.

. Wekell M. Plesiomonas shigelloides: An opportunistic food and waterborne pathogen. 690-4. Hood M. Journal of Applied Bacteriology. The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish. 55. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness. 78. Robins-Browne R.C.W. 849-55.N. 47. Seidler R.J. 1984.A. 105. epidemiology. Jr. 48. 85.R..M. Fliermans C... Partridge K.T. 1989. 1988... Lockman H.M.L.. 979-85. Tall B. Stott R.W.. Kaper J. Levine M..M. Journal of Rapid Methods and Automation in Microbiology...R.M. Department of Health and Human Services/Public Health Service/Food and Drug Administration. Journal of Food Protection. 52. 343-9. 53. 359-77.D. 87. Kaysner C.L. 50. 449-57. Infection and Immunity. Wachsmuth I. Wekell M.H.. 39... Fla.M. Kaysner C. Sullivan J.A. Esch G. Miller M. 76. Hirsch R. Kaysner C.. 223-47.... Colwell R.. 77. 1981.. Effects of processing and storing oyster meats on concentrations of indicator bacteria. Ruple A. Report (1/13/84).FISH AND SEAFOOD 74. 83. Journal of Food Protection. Journal of Food Protection. Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species.. Journal of Clinical Microbiology. 1989.. Applied and Environmental Microbiology. 1987.. Kain K.. 84. 76 . 998-1001. Hunt J.A. Van Damme L. Vandepitte J.A..W. Gracey M. Aeromonas hydrophila: ecology and toxigenicity of isolates from an estuary.J. 598-601.A. 1985. Koburger J. Abeyta C. 1980. Herrington D. Abeyta C. Stelma G.F.A. Jr. 75. Prevalence and distribution of Aeromonas hydrophila in the United States. Frequent isolation of Edwardsiella tarda and Plesiomonas shigelloides from healthy Zairese freshwater fish: A possible source of sporadic diarrhoea in the tropics. Plesiomonas enteric infections in the United States.D. 1997.. 1978.. 5. Wekell M.D. Farmer J.A. Singelton F. 7-12. Cooper M.. Abeyta C. Clinical features..B. III. 52.A.M.J.. 36. Incidence of motile aeromonads from United States west coast shellfish growing estuaries. Blake P. Herrington T. Trost P. Beaman J. Applied and Environmental Microbiology.J. Burke V. Hickman-Brenner F.K. 1984. Bark D.. Applied and Environmental Microbiology. Robinson J. Kelly M. 1986. Petersburg. 79. Jr.. Baker R. Peterson D. 86.... Atlanta. 1990.. Oyster borne disease outbreaks in St.. Annals of Internal Medicine.. Journal of Food Protection. 47.C. 82. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters... 475-9. 1146-9. Holmerg S. and treatment of Plesiomonas shigelloides diarrhoea. 80. Tenge B.M. 81. 731-8. GA. Tzipori S. Hazan T.L. Journal of Food Protection.P. Cook D. vibrios and Aeromonas hydrophila. 27. In vitro and in vivo pathogenicity of Plesiomonas shigelloides.

L. Smith J. U. 63-8. Fish Inspection. Ed.C. Washington D. Baird-Parker T. Chapter 5: Foods. Aspen Publishers Inc. Food and Drug Administration.S.MOLLUSCAN SHELLFISH 88. Food and Drug Administration. Food and Drug Administration Compliance Program Guidance Manual. Program 7303. Inc. The Microbiological Safety and Quality of Food. Ed.H.600 (CPG 7119. U. Food and Drug Administration.S. Food and Drug Administration Compliance Policy Guides. Martin R. Collette R. in National Shellfish Sanitation Program Manual of Operations. U. U. Food and Drug Administration. 1995. Pittsurgh.S.R. 89.S. National Academy Press.C.E. Washington D... Gaithersburg.S. Food and Drug Administration. and HACCP: A Global Focus. 7. fresh or frozenadulteration by bacteriological contamination Sec. Quality Control.M.. 560. Seafood Safety..A... 1987.C. oysters. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. 1991.. 90.S.S. 77 . Part I Sanitation of Shellfish Growing Areas. Sherlock C.8 Further Reading Ahmed F. 2000. Government Printing Office . Washington D. 1995. Slavin J. U. Ed.030.Pathogen monitoring of selected high risk foods (FY 88/89). Food and Drug Administration. in U.E. 3.C.W.1987. 91. Lancaster.12). in U. Food and Drug Administration.. 1997. Lund B. mussels.. Technomic Publishing Co. Food and Drug Administration. U.S. Clams. Burdge D.W. Gould G.


and available in 0. crabs. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. M.Ph. Such products are usually only partially cooked before freezing.g. the front pair ends in claws. packed and frozen in 0. Shrimps.5 kilo pack (boxes or bags). and frozen in 1 and 2. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption.0 kg polythene bags. with additional water to form a glaze. Species eaten from tropical waters include Penaeus spp. peeled product. Pandalus montagui and Crangon crangon (inshore shrimp). Crayfish. CRUSTACEAN SHELLFISH Linda Nicolaides.0 kg polythene bags. Peeled ready-to-cook shrimp are raw. They possess a segmented body. 79 . (deep-water shrimp). Pandalus borealis (deep-water shrimp). and are usually aquatic. lobsters.4.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda.5 and 1. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. lobsters and crabs have ten legs.. jointed limbs and a chitinous exoskeleton.. and Trachypenaeus spp.5 and 1. Parapeneopsis spp. Peeled and cooked shrimp are ready-to-eat after controlled thawing. crayfish and shrimp (1).. with shells and heads on or off are usually packed. e. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi).

Shewanella putrefaciens and 80 . particularly after it has been cooked.g. 4. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. e. particularly for members of the genus Vibrio (2). or frozen crab. Following capture or harvest. tropical waters. If these products are subjected to temperature abuse. lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption. hence. the geography of the area they are captured from or are grown in. shrimp die immediately after capture. lobster tails and cooked crabmeat presented decoratively in their shells. during the peeling of cooked shrimp. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. their feeding and living habits. As with finfish. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria. and the temperature and quality of the waters in which they exist. so that spoilage may occur during landing and transportation to the processing plant. storage above the recommended storage temperature. stored and transported as frozen product. Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. the potential exists for crosscontamination from both the processor and from the environment. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed. e.. However. packed into polythene bags. Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function. Pasteurised crabmeat is edible meat removed from the exoskeleton by hand.2 Initial Microflora As with finfish. only live carcasses are processed. with the increasing demand for ready-to-eat/cook crustacean products. Cooked and chilled. In comparison. traditionally. it is important that shrimp are handled and iced correctly in order to keep them in as fresh and uncontaminated a state as possible.FISH AND SEAFOOD Crabs. Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance. boiled to kill the crustaceans and cook their meat prior to consumption.g. which include the environment from which they have been captured or harvested. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors. spoilage bacteria are of little importance as. which means that the haemolymph can be a reservoir of bacteria. For example. the season. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp. The structure of the circulatory system of crabs is not closed. for crabs and lobsters. Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP).

The rate of spoilage will be a function of time and temperature. lobsters and crayfish are harvested from cages or pots containing bait. Pseudomonas spp. When the shrimp arrive at the processing plant they are washed and then sorted into size. Micrococcus spp. chilled 81 . corynebacteria. from the time of capture and processing.10 days for temperate water species (3). with “head on” or “head off” whereby the edible tail is removed from the head. Shrimp might be further processed with the shell on. Hence. up to 16 days for tropical species compared with 8 . A large proportion of shrimps are produced by aquaculture. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected. As with finfish.for example. the rate of deterioration of the crustacean shellfish by these organisms is reduced. especially if they are stored in ice or iced water..1. They are then taken to market alive in either holding tanks.. or cooked. chilled sea water.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. 4. glazed and frozen (see Figure 4.1). graded according to size. so they are either sold on the local market or are processed in local processing plants. the genera associated with tropical species include Enterobacteriaceae. After capture and transport to the processing plant.1 Prawns and shrimps An example of the process used for preparing frozen. or are found in brackish water around the shores. uncooked prawns and shrimp is presented in Figure 4. Vibrio spp.3. however. The claws are usually bound together to prevent the animals fighting and causing damage to each other. they are washed. whether GHP are in place. or transported directly to processing plants. peeled and cooked. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. bacterial contamination and spoilage are inevitable.. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . The psychrotrophic bacteria responsible for causing spoilage are not present on freshly captured crustaceans from tropical waters in such high levels as are found in crustaceans from cooler waters and. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. or with the shell and central vein removed. gills and thorax. and Achromobacter spp.3 Processing and its Effects on the Microflora Crabs. Shrimp will then be glazed and frozen in a raw state. thus. 4. Alcaligenes. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. These animals can survive out of water if the surrounding environment is kept cool and damp. Shrimps can be processed whole. In comparison shrimps die quickly after capture. In contrast.

Flow diagram . After cooking. The cooking stage will eliminate the intrinsic microbial flora of the shrimp as well as any contaminants that have been added during handling and transportation to the processing plant. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour. should be followed to ensure that the product is not re-contaminated with microorganisms present in the processing environment or from the workers.frozen uncooked prawns 82 .1. spices and cooking batter. and packed to meet the customer’s specification. Fig. Breaded products introduce an added risk of spore forming bacteria. including GHP.FISH AND SEAFOOD frozen. Good Manufacturing Practice (GMP). 4.

If the tail drops. If boiled.3. If hygienic procedures are not followed. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. with the growing demand for ready-to-eat products from crab and lobster meat. An offensive odour is also produced. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. bright and dry appearance. whilst boiled crustaceans have a pleasant slightly sweet smell. 4. 4. there is no indication of life. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. there is no associated spoilage problem. contaminating microflora may include enteric bacteria. Once removed and cleaned. putrefaciens and Pseudomonas spp. the meat is either repacked and sold in the shells.4. 4. or in hermatically sealed cans that have undergone an appropriate heat treatment.1 Prawns and shrimps Freshness indicators for prawns and shrimps are for the shells to be crisp and dry and feel sharp and cool to the touch. since crabs and lobsters are traditionally consumed immediately after cooking. wet and ‘soapy’ or ‘jammy’ to the touch. they include Sh. dull and sticky. immediately prior to boiling.2 Cooked crabs and lobsters As indicated earlier. Heat is produced if the shrimps decompose in a confined space. for crabs. and a strong ammoniacal odour is produced. However. and have a sweetish.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. bacteria of public health significance. In comparison. the joint between the tail and carapace is open and the limbs hang loosely.4 Spoilage The main genera of Gram-negative bacteria responsible for the spoilage of crustacean shellfish are the same as those that cause spoilage in finfish. The shells have a crisp. The shell will be soft.CRUSTACEAN SHELLFISH 4. spoiling prawns and shrimps have shells that are soft. viruses or parasites. slightly iodine smell. 83 . or packaged and processed.4. With spoiled crabs and lobsters. putrefaciens.

Moreover. Following this. nearly 30 years elapsed before the potential hazard of L. as well as for Salmonella.cooked prior to consumption. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. Salmonella species are more commonly found in crustaceans reared by aquaculture or growing in faecally contaminated brackish water compared with species of marine origin (4. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. An import alert was put in place in June 1987. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. 7). animal manure. However. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . fish fingers. monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. Although foodborne illness had not been associated with the product. in domestic/imported shrimps (15). They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. Hence. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. and usually for both. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). although efficient cooking will quickly eliminate it (6). in 24. 16).5% of frozen raw shrimps from different countries (13). later surveys in 1990 recovered the pathogen from retail raw. 6. 4.5.FISH AND SEAFOOD 4. The presence of L. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments.5 4. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). Salmonella cannot be completely removed from raw shrimp during processing. monocytogenes in cooked ready-to-eat foods. shrimp and shellfish (11).1 Pathogens: Growth and Survival Salmonella spp.2 Listeria spp. even if GMP is followed. 5. monocytogenes was absent from 40 samples of cooked prawns. cooked and smoked fish. The above results prompted the FDA to develop a compliance programme for the pathogen. shrimps and cockles sold (17). a common pond fertiliser used in aquaculture. Further surveys identified Listeria spp.5.

4. However. as part of the natural bacterial flora. pasteurisation and cooking.5. 25). which can become colonised by V.5. Once the competing microflora has been removed by a heat treatment. cholerae is divided into two groups. aureus is not a member of the intrinsic flora associated with crustacean shellfish. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. particularly crabmeat (18. no incidence of foodborne illness caused by L.5 Vibrio spp. 22) as well as from undercooked crustacea when harvested from contaminated waters (23. Crabs are covered in a chitin exoskeleton. the causative agent of cholera. it is important that the cooking stage is controlled to eliminate the pathogen from the product and that procedures are in place to prevent any cross-contamination of the cooked product. Control should be achieved by promoting GHP and staff training programmes. combined with the degree of handling of these types of product.g. is widely distributed in aquatic environments. pregnant women and the immuno-compromised. especially in products that have been vacuum packaged. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. The pathogen has resistant spores. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. 85 .3 Staphylococcus aureus Staph. although contamination by types A and B may occur from the environment. An epidemic that began in South America in 1991. about 50 . monocytogenes has been attributed to crustaceans. so is able to survive heat treatments that eliminate vegetative cells. 4. aureus because of the ubiquity of this organism in the processing environment. botulinum type E at 10 °C but not at 4 °C (20).70% of healthy individuals carry toxin-producing strains of Staph. cholerae non-O1. was associated with seafood consumption. the pathogen can grow at temperatures as low as 3 °C. e.4 Clostridium botulinum C. therefore. whilst boils and cuts are also common sources of the pathogen. Researchers have demonstrated that inoculated shrimp supported toxin production by C. V. 24.CRUSTACEAN SHELLFISH old. The main source of contamination is the hands of processing personnel. 4. Vibrio cholerae. cholerae O1 and V.5. cholerae. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. botulinum type E is commonly associated with fish and fishery products. aureus on their hands. Crustaceans have been linked with food intoxications attributed to Staph. 19). The former. V. To date.

South America.5. Japan. but has the potential to cause fatal foodborne illness (26). vulnificus is an invasive and lethal pathogen associated with wound infections. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). jejuni has not. to date. 4. Hepatitis A virus presents the most serious hazard. hydrophila and A. However. Contamination by this organism can be controlled by rapid chilling of crustaceans after harvesting and avoidance of time/temperature abuse during distribution and processing chains. V. 4.6 Campylobacter jejuni and Campylobacter coli C. hydrophila has been isolated from prawns (29). occurs in crustaceans from warm environments. whilst 5.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. However. Caliciviruses.5. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates.2 °C) so that GHP coupled with controlled low temperature storage is essential to prevent the pathogen from reaching levels that may present a hazard.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. the potential hazard exists as C.8% of samples contained C. Astroviruses and non-A. however. Parasitic infections are controlled by either cooking infected material prior to consumption. hydrophila is able to grow well at refrigeration temperatures (0 .5.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. It is more commonly implicated in countries where crustacean shellfish are eaten raw. particularly trematodes (6).5. a marine halophile. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. e. coli. Norovirus. in a survey carried out in the United States between 1977 and 86 . Of these.7 Aeromonas hydrophila and Aeromonas sobria A.g. Quantitative levels were below limits of detection in all cases (<0. 4. parahaemolyticus.30 MPN/g) (27). 4. V. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. non-B Hepatitis viruses (31). 32). A. or freezing at a temperature of -23 °C for 7 days (30. been implicated in a case of foodborne infection associated with crustaceans. 31. although A.

A summary of microbiological criteria for crustacean shellfish is listed below. No foodborne incidents have been attributed to this source to date.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. 4.5. 2073/2005. They can survive low pH levels. 4. produced or imported into the European Union (EU).6. or chlorine concentrations similar to those found in drinking water. only 0. are included in the Commission Regulation (EC) No. consistent with that in the stomach. although it is one of the European Union (EU) legislated allergens. Selected parts from this regulation is presented below. The toxin causes amnesic shellfish poisoning (ASP) when consumed. Norwalk viruses can also withstand heating to 60 °C.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). The risk of such hazards is far higher with fin fish and molluscan shellfish.11 Chemical pollutants and biotoxins Researchers have demonstrated that crabs feeding in waters contaminated with algae do have potentially high levels of algal toxin domoic acid (DA) in their viscera.CRUSTACEAN SHELLFISH 1981. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). 87 .6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. 4. from farm to fork (37.5. which could be transferred to the meat during unskilled removal from the shell (35). 38. 4. risk-based system based upon the seven principles of Hazard Analysis and Critical Control Point (HACCP) supported by pre-requisite or good practices programmes at all stages of the food chain.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. Provisions laid down concerning the use of additives would also apply to these types of products. which results in a persistent and apparently permanent loss of short-term memory (36). 39).

FISH AND SEAFOOD Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) Chapter 1 - Food safety criteria

L. monocytogenes Prepared crustacean products fall into the category described in the regulation as “Ready-to-eat foods available to support the growth of L. monocytogenes other than those intended for infants and for special medical purposes”. During capture and preparation there is a zero tolerance for the presence of L. monocytogenes on the product. However, to take into account the issues of natural habitats being contaminated with low levels of this pathogen there is a slight loosening of the regulation at the point of sale and for the duration of the products shelf life (refer to Table 9.I, Food Category 1.2). If the level of L. monocytogenes should be above this level then the consignment is considered to be unsatisfactory. Salmonella All cooked crustaceans and molluscan shellfish should be free from Salmonella at the point of sale and for the duration of their shelf life (refer to Table 9.I, Food Category 1.16). Should any sample test positive for Salmonella then the product is declared to be unsatisfactory. Chapter 2 - Process hygiene criteria

Process hygiene criteria for E. coli and coagulase-positive staphylococci in shellfish and shucked products of cooked crustaceans and molluscan shellfish are laid down (refer to Table 9.II, Food Category 2.4.1). 4.6.2 International requirements

International requirements are based upon having a risk-based, preventive management system in place at all stages of the supply chain. The requirements for this preventive approach is described as follows (37): • Fish and fishery products should be prepared in plants certified by the local competent authority. All certified plants should comply with the GHP (39); • The fisheries industry should take responsibility, implement and maintain safety management systems based upon HACCP; • The national competent authority is responsible for the certification of fish processing and manufacturing plants, verification of effective systems, and



issuing of certificates of compliance for export products. This includes auditing and inspection programmes; • National surveillance and monitoring programmes should be in place to demonstrate that all identified hazards are under control, e.g. biotoxins, and to identify potentially emerging hazards; Within this requirement is the flexibility for countries and trading blocks to set specific standards required to deliver an equivalent requirement with those at national level. Such standards need to be science-based, especially if they are more stringent that those recommended by the Codex Alimentarius Commission. Verification that GHP have been followed during the handling and preparation of crustacean shellfish for consumption is usually done by assessing the level of E. coli and/or coagulase-positive staphylococci in the products. Legal requirements generally require that these groups of indicator organisms are present in levels <100 cells/gram of product. Pathogenic bacteria such as Salmonella spp. should be absence is 25 grams multiplied by the number of subsamples tested. United States The FDA brought into force a Federal Mandate Seafood Rule in 1995 which forms the foundation for good practices required for the handling and processing of imported fish and fishery products into the U.S. The best practices include GHP and HACCP (37). All fish and fishery products produced in and imported into the U.S. after 17 December 1997 are required to demonstrate that HACCP principles were applied to their handling and processing. The USFDA has a zero tolerance for Salmonella in fish and fishery products; this includes farmed shrimp. The US FDA considers that the presence of pathogenic microorganisms in seafood is an indicator of adulteration (32). 4.7
1. 2.

Encarta 1997. Encarta 97 Encyclopedia World English Edition. Microsoft. The International Commission for Microbiological Specifications for Food. Fish and Fish Products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. The International Commission for Microbiological Specifications for Food. New York, Kluwer Academic/Plenum Publishers. 2005, 174-249. Cann D.C. Bacteriology of shellfish with reference to international trade, in Handling, Processing and Marketing of Tropical Fish. Ed. Tropical Products Institute. London, Ministry of Overseas Development. 1976, 377-94. D’Aoust J.Y., Gelinas R., Maishment C. Presence of indicator organisms and the recovery of Salmonella in fish and shellfish. Journal of Food Protection, 1980, 43, 679-82.




FISH AND SEAFOOD 5. 6. 7. 8. 9. 10. 11. Fraiser M.B., Koburger J.A. Incidence of salmonellae in clams, oysters, crabs and mullet. Journal of Food Protection, 47, 343-5. Reilly P.J.A., Twiddy D.R., Fuchs R.S. Review of the occurrence of Salmonella in tropical shrimp. FAO Fisheries Circular No. 851, 1992, (FIIU/C851). Reilly P.J.A. Salmonella and Vibrio cholerae in brackishwater cultured tropical prawns. International Journal of Food Microbiology, 1992, 16, 293-301. Twiddy D.R. Antibiotic-resistant human pathogens in integrated fish farms. ASEAN Food Journal, 1995, 10, 22-9. Olsufjew N.G., Petrow V.G. Detection of Erysipelothrix and Listeria in stream water. Zhurnal Mikrobiologii Epidemiologii i Immunobiologii, 1959, 30, 89-94. Anon. First Listeria finding in crabmeat confirmed by FDA. Food Chemistry News, 1987, 29 (14), 38. Ryser E.T., Marth E.H. Incidence and behaviour of Listeria monocytogenes in fish and seafood, in Listeria, Listeriosis and Food Safety. Eds. Ryser E.T., Marth E.H. New York, CRC Press. 2007, 617-54. Anon. Mexican crabmeat, Greek pasta automatically detained. Food Chemistry News, 29 (19), 43. Anon. FDA checking imported domestic shrimp, crabmeat for Listeria. Food Chemistry News, 1987, 29 (24) 15-7. Archer D.L. Review of the latest FDA information on the presence of Listeria in foods. WHO Working Group on Foodborne Listeriosis, Geneva, Switzerland. 1988, 15-9. Anon. FDA to sample shrimp for Salmonella, Listeria. Food Chemistry News, 1988, 30 (36), 9-10. Anon. Monitoring of changes to listeriosis problem urged. Food Chemistry News, 1989, 31 (20), 13-7. Richmond M. Report of the Committee on the Microbiological Safety of Food. London, HMSO. 1990, 133-7. Swartzentruber A., Schwab A.H., Duran A.P., Wentz B.A., Read R.B. Jr. Microbiological quality of frozen shrimp and lobster tail in the retail market. Journal of Food Protection, 1980, 40, 765-9. Wentz B.A., Duran A.P., Swaartzentruber A., Schwab A.H., McClure F.D., Archer D., Read R.B. Jr. Microbiological quality of crabmeat during processing. Journal of Food Protection, 1985, 48, 44-9. Garren D.M., Harrison M.A., Huang Y-W. Clostridium botulinum type E outgrowth and toxin production in vacuum-skin packaged shrimp. Food Microbiology, 1994, 11, 467-72. Molitoris E., Joseph S.E., Krichevsky M.I., Sindhuhardja W., Colwell R.R. Characterisation and distribution of Vibrio alginolyticus and Vibrio parahaemolyticus isolated from Indonesia. Applied and Environmental Microbiology, 1985, 50, 1388-94.

12. 13. 14.

15. 16. 17. 18.





CRUSTACEAN SHELLFISH 22. Varma P.R.G., Iver T.S.G., Joseph M.A., Zacharia S. Studies on the incidence of Vibrio cholera in fishery products. Journal of Food Science and Technology, 1989, 26, 341-2. Madden J.M., McCardell B.A., Morris J.G. Jr. Vibrio cholerae, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 525-42. Twedt R.M. Vibrio parahaemolyticus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 543-68. Oliver J.D. Vibrio vulnificus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 569-600. Pearson A.M., Dutson T.R. Quality Attributes and Their Measurement in Meat, Poultry and Fish Products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994. Reinhard R.G., McAdam T.J., Flick G.J., Croonenberghs R.E., Wittman R.F., Diallo A.A., Fernandes C. Analysis of Campylobacter jejuni, Campylobacter coli, Salmonella, Klebsiella pneumoniae and Escherichia coli 0157:H7 in fresh handpicked blue crab (Callinectes sapidus) meat. Journal of Food Protection, 1996, 59 (8), 803-7. Buchanan R.L., Palumbo S.A. Aeromonas hydrophila and Aeromonas sobria as potential food poisoning species: a review. Journal of Food Safety, 1985, 7, 15-29. Palumbo S.A., Maxino F. Williams A.C., Buchanan R.L., Thayer D.W. Starch ampicillin agar for the quantitive detection of Aeromonas hydrophila in foods. Applied and Environmental Microbiology, 1985, 50, 1027-30. Wekell M.M., Manger R., Colburn K., Adams A., Hill W. Microbiological quality of seafoods: viruses, bacteria and parasites, in Seafoods: Chemistry, Processing Technology and Quality. Eds. Shahidi F., Botta J.R. Glasgow, Blackie Academic and Professional. 1994, 196-219. Schantz J., McKerrow J.H. The dangers of eating raw fish. New England Journal of Medicine, 1989, 320, 1143-5. Ahmed F.E. Seafood Safety. Seafood Safety. Committee on Evaluation of the Safety of Fisheries Products, Food and Nutrition Board, Institute of Medicine. Washington D.C., National Academy Press. 1991. Blackwell J.H., Cliver D.O., Callis J.J., Heidelbaugh N.D., Larkin E.P., McKercher P.D., Tayer D.W. Foodborne viruses: their importance and need for research. Journal of Food Protection, 1985, 48, 717-23. Kapikian A.Z., Chanock R.M. Norwalk group of viruses, in Virology. Eds. Fields D.M., Knipe D.M. New York, Raven Press. 1990, 359-77. Horner R.A., Postel J.R. Toxic diatoms in western Eashington waters (US west coast). Hydrobiologia, 1993, 269/270, 197-205. Perl T.M., Bedard M.S.M., Kosatsky T., Hockin J.C., Todd E.C.D., Remis R.S. An outbreak of toxic encephalopathy caused by eating mussels contaminated with domoic acid. The New England Medical Journal, 1990, 322, 1775-80.

23. 24. 25. 26.


28. 29.


31. 32.


34. 35. 36.


FISH AND SEAFOOD 37. Ababouch L., Gandini G., Ryder J. Causes in Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. Australian Government. Australian Quarantine and Inspection Service Hazard Analysis and Critical Control Point (HACCP). A Guide to Compliance with Export Control (Fish and Fish Products) Orders 2005. FAO/WHO. General Hygiene Requirements. Rome, Food and Agriculture Organisation of the United Nations. 2003.



4.8 4.8.1

Further Reading Spoilage and pathogens

Gram L., Huss H.H. Fresh and Processed Fish and Shellfish, in Microbiological Safety and Quality of Food Volume 1. Eds. Lund B.M., Baird-Parker A.C., Gould G.W. Maryland, Aspen Publishers Inc. 2000, 472-506. Ababouch L., Gandini G., Ryder J. Causes of Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. International Commission on Microbiological Specifications for Foods. Fish and fish products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1998, 130-89. Sutcliffe P., Disney J. Handling, Processing and Marketing of Tropical Fish. London, Tropical Products Institute. 1977. Connell J.J. Control of Fish Quality. Oxford, Fishing News Books. 1995 Ward D.R., Hackney C. Microbiology of Marine Food Products. New York, Van Nostrand Reinhold AVI. 1991 Pearson A.M., Dutson T.R. Quality Attributes and their Measurement in Meat, Poultry and Fish products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994.

4.8.2 Pathogens - general
International Commission on Microbiological Specifications for Foods. Microorganisms in Foods 5. Characteristics of Microbial Pathogens. Eds. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1996. Varnam A.H, Evans M.G. Foodborne Pathogens - An Illustrated Text. London, Wolfe Publishing Ltd. 1991. Doyle M.P. Foodborne Bacterial Pathogens. New York, Marcel Dekker. 1989.


Fish curing is practised worldwide. drying and smoking processes. split and opened out flat. and a preliminary operation to some smoking. their combinations and/or variations. Fish can be salted in three different ways or their combination: Dry salting.5. and marinating processes. drying. and salting is done by stacking the fish in layers. where the salt replaces moisture in the fish flesh. 93 . SMOKED AND DRIED FISH Dr.1 Definitions Curing is the preservation of fish by removing or displacing available water in the fish flesh to a level that discourages microbial growth. and before significant spoilage takes place. and is sometimes known as ‘pickling’. Usually. rather than for preservation itself. dependent on the country or the type of product. fish is immersed for a few hours in a relatively low concentration brine. It includes the salting. both to preserve and to create a variety of new products. alternating fish layers with a layer of salt. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. Wet salting or brining. The fish is headed. and the extracted moisture is allowed to drain away. This can be called ‘kench’ curing and is mainly used for non-fatty white fish. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. CURED. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. The processes take anywhere from a few hours to a few days or even months. Salting is both a method of preserving fish.

Usually. the ice sublimes and the vapour is removed from the fish by vacuum pumps. usually vinegar. After the freezedrying process. or boiling it with dry salt and water. depending on the type of product. it can also be named ‘pickling’ or ‘blood pickling’. Salt. Food products can be dried in three different ways: Air or contact drying. are added to the marinade. Traditionally. is used as a preservative instead of salt. thus making the product unsuitable for microbial growth. Salted-boiled fish are the products obtained on either boiling fish in a brine. The fish is mixed with dry salt but in a watertight container. Freeze drying. evaporated water is removed by vacuum pump. Vacuum drying. prior to or during the boiling process. under normal pressure. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. the preservative effect is due to the low pH used in the process. thus reducing the water activity of the product. Salt-boiling. When the process is running in a closed system. such as ‘gravad’ fish. Fish in contact with refrigerated plates freeze. Smoking is an ancient process for preserving and flavouring food. Heat is transferred to fish from heated air or a heated surface. The original principle of preservation was due to a combination of lowered water activity of the product and the uptake by the product of bactericidal and antioxidant components 94 . this was done naturally by exposure of the fish to the air. where an organic acid. as concentrated brines are formed. thus delaying spoilage for a few days.FISH AND SEAFOOD Dry / wet salting. spices). Marinating or pickling. As the water is drawn out of the flesh by the salt a brine is formed. without other preservation methods applied after boiling. as well as other flavourings (sugar. The products are usually consumed raw. This is the common method of drying fish. The product is dried by contact with heated surfaces or by radiation. Thus. and the fish can be immersed in such brines for long periods. This is a mixture of a dry and a wet salting process. the product rehydrates quickly but does not retain reabsorbed moisture on cooking. and the air movement removes the vapour. Marinades are acidified brines. Included in this group are some lightly preserved products where no acid is added to the marinade. Longer shelf life is achieved in the products when salt is added. It is not commonly used for fish products. Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms. this ‘brine’ can be collected and placed again over the fish several times. It has found limited application with fish.

Usually. The different spices used in the immersion brines are also a potential source of microorganisms. including pathogens. or added to the brine with other flavourings. and the preservative effect is only slight. which impart the flavour of smoke to fish. SMOKED AND DRIED FISH of smoke. fish can be smoked by means other than the direct action of smoke. Cold-smoking. Smoked-flavoured fish. smoking is now used mainly to give the fish a characteristic flavour. process temperatures are over 60 °C. 5.CURED. leaving only elevated levels of carbon monoxide. and filtered to remove tars and particulate matter. or by means of smoke-flavouring processes with liquid smoke. Clear smoke. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour. The smoke itself may contain spores of moulds. temperatures below 30 °C are used. However. In this process. It can be smoked either traditionally. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. With the exception of some Asian countries. Smoking can be either traditional. by exposure to the wood or sawdust smoke. the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. Usually. Is an illegal method (in the European Union (EU)) employed to treat tuna to ensure a bright red colour is maintained during storage and transport. Smoked fish is prepared by subjecting the fish to the direct action of smoke from wood or wood sawdust burning. the fish is subjected to heat for a period that does not coagulate protein.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. such as injection or dipping in a solution of liquid smoke. Also. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke. or liquid smoke. After the salting step. The secondary objective is to impart sensory characteristics such as smoke colour and smoke flavour to the product. The process was designed so that the smoke is filtered to remove all smoke particles. or by means of smoke flavouring processes. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. 95 . sporeforming bacteria or osmophilic moulds. Hot-smoking. the salt used in the processes may contain halophilic bacteria. It is commonly used in place of traditional smoke. In this process. It can be injected into the product. by exposure to the wood or sawdust smoke.

sometimes the salt crystallises onto the surface of the fish.0 158.FISH AND SEAFOOD 5.2 105.3 Processing and its Effects on the Microflora Owing to the wide variety of products. Different brine strengths are required according to the curing processes and/or the type of product. or salt in the water phase. Therefore. about 50º (132.4 184. only the most common processes can be described here. and salt content.I Brine strength at 18 °C (after Bannerman (2)) Brineometer degrees 10 20 30 40 50 60 70 80 90 100 96 Weight of salt (g/litre of brine) 26.0 g of salt per litre of brine). Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.6 264.4% salt (Table 5. Salt in the water phase links the two conditions of moisture and saltiness.8 211. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. a 0º brine saturation means pure water.80º brine (184. is employed. processes and producers’ private recipes.0 . The strength of the solution is described as degrees of saturation (brineometer degrees). A 100º brine solution means that the solution is saturated and no more salt can be dissolved.6 g of salt or more per litre of brine) may cause salt burn in this way. At 18 °C a 100º brine is produced when the solution contains 26.I).8 211. it causes swelling because the fish absorbs water from the salt solution (1).8 79. When a 100º brine is used. etc. For smoking salmon. % Salt in the water phase = % salt content % salt content + % moisture x 100 Another important point to remember when wet salting is used is the concentration or strength of a brine. When working with salting.2 g of salt per litre of brine). it is important to understand the concept of salt concentration. smoking or marinating processes. it is impossible to establish standardised processing stages in the curing of fish. all using salting steps. When a milder brine. the fish is usually salted with a 70 . TABLE 5. Salt concentration means the percentage of salt in the water contained in the fish. This should be distinguished from the salt content. The dissolution of salt in water is dependent on the temperature. it is important to state the temperature.6 132.2 237. a 90º or above brine (237.4 52. Thus.

The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. and essentially free from iron and copper (3).1 ppm and 10 ppm. but the concentration of copper and iron must be kept under 0. respectively (1). respectively. See Figure 5. the fish is restacked with fresh salt after about a week.1. However. are recommended to help make the product whiter. addition of levels of 0. outgrowth of halophilic bacteria may occur. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. if very pure sodium chloride is used for curing purposes. mainly in tropical conditions.3. The calcium and magnesium impurities. caught in North Atlantic waters. and for a heavy salting a ratio of 1:3 salt:fish is used (1). lean white fish is used for this purpose. the fish is mixed with one part of salt per eight parts fish.1 Salting Salting of fish for preservation purposes has been used since pre-history.1 Dry salting / Kench curing Usually. calcium and magnesium ions of impure salts can form a barrier to the passage of sodium ions from the surface to the bulk of the fish. which include calcium and magnesium. which consequently inhibits the diffusion of the water from the bulk of the product to the surface.15 . when the air humidity is high. The salt used should be of food-grade quality. ‘Salt burn’ might occur when the surface of the fish dries out too quickly as a result of the small grain size of the salt. producing characteristic pink patches. When pure salt is used. having a short section at the end of the tail. it tends to lead to a slight yellowing and softening of the product.1. On the other hand. 5.3. and such delays. This is because fatty fish turn rancid quickly. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). ‘Green cured’ split cod with about 60% of the backbone removed. however. SMOKED AND DRIED FISH 5.CURED. Usually it is followed by other preservation techniques such as drying and/or smoking. This condition is known as ‘putty fish’ (4). For a light salting. are likely to cause hydroscopicity of the product and. 97 . Salting can be done in a number of different ways. and salt can speed this oxidation process.15% calcium and magnesium. and the salt has penetrated throughout the fish (3). It takes about 3 weeks until proper salt penetration and moisture loss are achieved. and a ‘putty’ fish product is also obtained (1).0. The most common fish used in the northern hemisphere is cod (Gadus morhua). can be dried further to produce ‘klipfish’ or ‘bacalão’.35% and 0. for which there is a high demand in Mediterranean countries. low in calcium and magnesium. Also.

FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat.1.5. such as 98 . Where fish are salted in their whole form. so that the risk of spoilage is high.1. eviscerated and/or split). Fig. the process is mainly for flavouring purposes prior to the use of other preservation processes.90 are rarely achieved. Food-poisoning organisms. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. Processing of dried salted cod 5. The greatest barriers to salt penetration are the skin and scales.3. For large fish it is advisable to split it before salting. it appears that water activities much less than 0. although toxin production would be unlikely (4). such as Staphylococcus aureus could survive and grow at such water activities in the product.2 Wet salting or brining As referred to above. when a light brine is used. Only small fish can be salted whole.

a saturated brine is used.3 days.64 4.70 6.49 4. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.61 3.51 5. headed and gutted.5 4.23 2. Fresh sardines.03 2.II.76 4.26 2.66 5.1.76 4.0 2.33 5.51 3. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 . The fish are kept immersed in their own brine near saturation point for several days. See Table 5.73 1.96 1.41 5. Anchovies are the main fish used for these purposes.76 4.41 3.03 4. 5.11 9. the liquid that comes out of the fish forms a brine.23 8. can be cured in a saturated brine solution for 6 days and.17 4.80 5.30 2.04 5. the fish enzymes play an important role by maturing or ripening the product. Histamineforming bacteria do not generally grow at water activities below 0.54 5. There are several variations of this process.86 2.09 8. but mackerel.41 4. and a red burgundy colour will develop in the product.06 4.0 4.70 3.45 3.0 3. but as it is placed in containers.26 1.3 Dry / wet salting In this process.05 7.5 3.10 1.66 5.78 6.78) (1). SMOKED AND DRIED FISH smoking.11 3.91 2.98 5.88 5. the brine is then replaced over the fish.65 2.22 6.23 3. limiting the amount of rancidity that can develop.17 5. with a shelf life of about 4 weeks (water activity (aw) about 0. they can be stored at 28 °C. herrings and other fatty fish can be used.23 2. This is why it is the preferred method for fatty fish.35 4.60 2.85 3. the fish is dry-salted.11 4. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.90 (1).47 6.76 4.56 6.41 6.26 7. the product obtained is called gaspé-cured (5).16 7. sardines. Here. The main objective of the brining method of salting fish is to limit air penetration during processing. and can be collected and replaced again over the fish.61 3.25 5.00 3.85 3.95 3.5 10.98 6. Histamine formation is of importance when using scombroid and some clupeidean species.09 8.98 6. TABLE 5.19 3.3. with pressure being applied to the top of the container to help the brine to drain away.44 2. For preservation purposes.78 2. For example. thus. if split fish is dry-salted.25 5.76 99 .46 4.84 1.II Salt in the water phase (after Ian Doré (5)) Moisture (%) 40 45 50 55 60 65 70 75 80 85 90 Percentage salt in the flesh (by weight) 2.74 9.44 2.26 3.26 4.CURED. after being drained and pressed at 465 kg/m2 for 15 hours.5 3.76 4.99 2.

up to 1 . In a second step. 5. All these products are ‘semi-preserves’. which is dipped into a boiling brine for 10 . They will have a salt content >6% (w/w) in the water phase.4% of salt) for distribution and sale (1). giving a cooked appearance to the product.5% 120grain distilled vinegar. only the nonspore-forming organisms are eliminated in the process and. because of the salt content of the product. and even death. a Peruvian product.6 days without refrigeration (1). Once cooled. Herring can also be cured for several days in an 80 to 90º brine with 2. outgrowth of bacterial spores after processing may occur (4). ‘ikan kaju’. even if the product is hermetically sealed. Other preservation techniques can be used for extending the shelf life of these products. containing 1 kg of salt for every 25 kg of fish. until the flesh is soft. The fish can be soaked in a high concentration of vinegar (about 4 .2% acetic acid and 2 . Traditional jellied eels are a product where pieces of fish are cooked in boiling water.3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. However.3. The lime juice plays the role of the vinegar.FISH AND SEAFOOD 5. and spices) is added to the brine.7%) for as long as 3 weeks. then packed in barrels with a 70º brine. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. Another variation uses sour cream instead of the second brine step.30 minutes. After this. or can be boiled by being placed in a basket. usually flavoured with onions and hot chillies. but it has further smoking. herring or other fatty species are used. usually requiring refrigeration below 10 °C for longer-term storage. The first process can give a longer shelf life. As referred to above. and lasts for 3 . and 6% salt. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. is made of different varieties of fish. Another salt-boiled Indonesian product. salt. they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. placed in a marinade of lime juice. it can be repacked in milder brine (1 . they are packed for consumption within 2 weeks at chill temperature (6). Usually. Afterwards. whereas the latter method preserves the fish for only a few days. and packed (3).2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. There is epidemiological evidence that these products have been the cause of some 100 . Ceviche. or a pH <5 and they are stabilised for a limited period. A much firmer product is obtained when the fish is salted prior to acidification. drying and salt-boiling steps until a hard translucent block is obtained (4). underlines the need for careful control (4). flavouring (sugar. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten.3.2 months. has a shelf life of several years. the occasional implication of pindang in cases of foodborne illness.

There are several products named marinades (matjes. Provided the product is kept in dry and fairly cool conditions. In Scandinavian countries. Italy and Spain (5). which are placed in a mixture of salt. At pH levels below 4. mixed with salt. marinated products will keep for several months at a temperature of 4 °C.CURED. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. etc. These can be considered as ‘lightly preserved’ products with a salt content <6% sodium chloride (w/w) in the water phase and low acid content (pH >5. Matjes are made of fat young herring. some fish species such as moray eel and varieties of dog fish. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). Stockfish products have a golden brown colour and are very hard. are likely to survive in these mild-processed products. depending on the thickness. This bacterial and enzymic action can produce a typical flavour desirable in some semi-preserves in which salt or a mixture of salt and sugar is used to preserve the fish (8). the fish is dried after salting or brining. Usually. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. gutted. Shark pectoral fins. 101 . Stockfish (dressed. the temperature and the amount of pressure applied (5). sugar and spices. and the flesh will break down. and the stacks are placed in the refrigerator.) where no acid is added to the process. it will last for many years (5). by using either solar heaters or mechanical dryers. a very fine delicacy for Oriental food lovers. Gravad fish is a Scandinavian product made of salmon fillets. such as squid. after frying. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). Foodborne pathogens.0) (7). sea cucumbers and seaweeds. 5. jellyfish. It is important to assure the quality of the raw material.5. It takes about 2 days to marinate completely. In the first case.3. bacterial toxins and parasites (7). Dried octopus is also a very common dish in the interior of the country. The most common method used for fish products is the air-drying process. Some bacteria and enzymes are still active in the presence of acid and salt. They are an important dried product. gravad fish.4 Air drying This is probably the oldest method of fish preservation. Dried shrimps are widely used in Asia. split and dried gadoid species) are traditionally made in cold climates. still made without prior salting on a large scale. are generally air-dried at sea. and a fine product is obtained. but sometimes it is dried without prior salting. Stockfish is extensively used in Portugal. Other products. are commonly dried in Asian countries. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. South America and the Caribbean. Fresh dill is placed between the layers of fish. the fish is dried naturally in a relatively cold environment. In some coastal areas of Portugal. are sundried and. air flows naturally. used as an appetiser. under a light pressure. such as Listeria monocytogenes and Clostridium botulinum.

However. tarpaulin. In conclusion. skin up. Spain. It is important to balance the drying rates and quality/safety issues.g. some of which are listed in Table 5.III. and the water must be changed several times before cooking. To prevent this. A highly demanded dried fish product in Mediterranean countries is dried salted cod. They must be soaked in water. Iceland and Canada. and it is exported to Portugal. nor subsequent cooking operations will affect the presence of toxins (e. the skin will form a hard case (case-hardening). C.83 (9)). neither it. 102 .g. The product is dried to a final water content of about 25 to 38% (3). but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). When no more water can be removed from the fish. more commonly nowadays. the protein must be set or denatured with low-temperature drying before applying higher temperatures. and increasing amounts of such products are making their way into the market place. and has the primary objective of creating a product with specific taste and textural characteristics. if the rate of water removal is too high in comparison with the diffusion rate. Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources. Whilst the production of dried fish products is tightly controlled in industrialised countries. France. typically in stews and soups. In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. Italy and South America. that may have contaminated the product during the drying process. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain. Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s. making a water barrier and consequently.FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. aureus) or spores (e. by warm air circulating in a drying chamber. botulinum). the fish must be dried slowly enough to avoid case-hardening. toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0. the quality is more difficult to control. The amount of moisture transfer from the fish to the air is dependent on the relative humidity . the product can be held for months at room temperature. Dried salted products must have a water activity below 0. botulinum growth might occur in the bulk of the product.75 (1). there are many regions of the world where air drying is the only means of ensuring that a day’s catch can be preserved for later use. the better the moisture transfer. there is an increasing demand from people that have emigrated to industrialised nations for traditional products. Whilst the low aw of the final product will inhibit microbial growth. Staph. After dry salting. the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or. The fish must be stored in reasonably dry conditions to avoid moisture absorption. the diffusion of the bulk water to the surface of the fish is blocked. which leads to mould growth (aflatoxins. spoilage and possibly C. or wooden drying rack to dry in the sun. Thus. dried salted cod (bacalão) is now mainly produced in Norway.the lower the humidity.

softwood colours the product quickly. herring. Maintaining the temperatures of pyrolysis between 425 and 200 °C. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10). white mangrove and abura has an inhibitory effect on some microorganisms. botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. trout. oysters.III Hazards and sources of contamination associated with air drying Hazard Enterobacteria Salmonella Source Sewage Reptile Birds Humans Humans Environment Airborne Applied to combat insect infestation Environment Human Vector Human Insect Staph. It can be used for cold or hot smoking. haddock. In the mechanical kiln. cod roes. as referred to above. The quality of the smoke is related to the type of wood used. SMOKED AND DRIED FISH TABLE 5. tuna. Blends of various woods can be formulated to produce sawdust with specific flavours. It has been reported that the smoke from mahogany. using electrostatic filtration of the smoke. Top-quality fish must always be used. mussels and many others from tropical countries. Aroma and flavour are a blend of smoke components (3). aureus (toxin) C.5 Smoking Smoking today. 5. cod. There are many fish species that are used for smoking purposes . Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. eel. Hardwoods are considered better because of the milder flavour they impart to the food.salmon. clams. and the use of smoke generated by superheated steam or liquid smoke. swordfish. aureus and Saccharomyces cerevisiae (10).3. However. sprats. such as Escherichia coli. Staph. making full description impossible here.CURED. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns. The smoking procedures vary widely. is mainly a process for flavouring fish rather than a preservation technique. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish. Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . mackerel.

covering the fish surface completely (10). If frozen fish is used. Whether the product is prepared headed. smoked cod fillets and finnan haddock are usually cooked before consumption. Drying should be minimal at the start. Thus. it will reduce the salty flavour. Brining at 10 15 °C is recommended (10).and hot-smoking is the temperature of the process.e.3.or hot-smoked. However. as it retains more moisture. pasteurisation) temperature is not achieved in any stage of the process. as well as many other products prepared in the same way. Cod roes.18 hours at 4 °C. This difference is crucial for safety issues because the flesh is not cooked. Also. are not cooked prior to consumption. tuna and swordfish. are boiled before smoking (12). Depending on the type of product. wooden boxes or master cartons (10). When frozen/thawed fish is used. it will take up salt twice as fast as fresh fish (10). temperatures must be kept under 30 °C throughout the process. whether cold. The salting period is dependent on the size of the fish and its presentation (whole. the following step is salting. 104 . All the smoked products.5. or packed in retortable foil pouches. canned or bottled. an attractive glossy pellicle on the fish fillet surface is achieved after draining. Some herring products. In cold-smoking. but a ‘safety’ (i. The maintenance of temperatures below 30 °C avoids protein coagulation. If wet salting is chosen. low temperatures will slow the uptake of salt. but this concentration will again depend on the type of product and the length of the brining stage.10% of the weight of the fish. they can be vacuum packaged. The fish can be dry-salted using a weight of salt 5 . 70 – 80 º brine is commonly used.FISH AND SEAFOOD water flow system with chlorinated water (25 .1 Cold-smoking The major difference between cold. spices. usually dry-salted and cold-smoked. which is important for the appearance of these products (4).50 ppm) should always be done just before process initiation (3). shrink packaged. all cold-smoked products should be cooked before consumption. Sugar. the excess salt is removed with water. such as smoked salmon-trout. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. Sugar can be added together with the salt. skin off or on. The length of the smoking process for such products is much greater than for the hotsmoked fish. the highly desirable cold-smoked salmon. and the cold smoke is denser. giving the product the desired appearance and flavour (4). or quick-frozen and stored at -18 °C (2). 5. thus allowing the deposit of smoke on the fish surface. skin on or off). it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11). permitted colourings or smoke flavours can also be added to the product in the brining step. however. but it will increase towards the end of the process (10). after being smoked need to be chilled immediately to 4 °C before being packed. filleted. Wet salting or brining is preferable to dry-salting when smoking fillets because. Thus. gutted or filleted. owing to the elution of soluble proteins. thawing must be done slowly for 12 .

a drying period at 30 °C. can grow with salt levels of 10% in the water phase. The exception is vacuum packed fish. Smoked products can be also packed under controlled-atmosphere packaging. secondly. if present. but they should be cooled to a temperature of 10 °C within 3 hours. Length of the smoking process at each stage will depend on the species and the type of product. which underlines the danger of storage at abuse temperatures of >10 °C (13). gravad salmon (also called gravad lax or gravlax). Although other human pathogens that can grow at refrigeration temperature. further cooled to 3 °C within 12 hours and then packed.CURED. a partial cooking period at 50 °C. the products are cooled. a final cooking period at 80 °C (12). if they are taken from a chill room at 3 °C and packed. as. and their sensitivity to salt (13). Furthermore. Hot-smoked products require a longer cooling stage. condensation will occur inside the pack. If the refrigeration chain is maintained. Pathogenic vibrios. Neither the smoking temperature nor the salt content is enough to kill the organism. 5. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). Smoked salmon fillets are usually vacuum packed whole or after slicing. See Figure 5. refrigerated storage and vacuum packaging still allow its growth (12). SMOKED AND DRIED FISH During cold-smoking. thus allowing any psychrotrophic C. in order to produce toxin. can be isolated from seafood.2. During smoking.2 Hot-smoking Hot-smoking must use temperatures between 70 and 80 °C at any stage of the process to ensure protein coagulation throughout the product. are of concern when smoking fish from tropical waters (13). The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. there is no point in the process that can fully assure the absence of L. These products should be vacuum packed at room temperature. and hotsmoked salmon (kippered) (3). Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. botulinum type E present an even longer time than before to grow and produce toxin (14). and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). risk from the presence of Bacillus cereus and Clostridium perfringens can also be excluded as potential hazards because of the relatively high temperatures they require to grow to. To avoid growth of non-proteolytic C. botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. Vacuum packing is recommended only for coldsmoked salmon.3. such as Yersinia enterocolitica. thirdly.5. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). stored at 4 °C and packed. if a 105 . typically mesophilic. This temperature should be maintained during storage and distribution (3). but the proteolytic types A and B. monocytogenes. A typical temperature set is as follows: firstly. On the other hand. Afterwards.

Fig. 5. it will have a reduced shelf life.2. and moulds will grow on the product (12). Process of cold-smoked sliced salmon 106 .FISH AND SEAFOOD product is packed too warm without being cooled to room temperature.

then hot smoking and finally sun drying it. They will keep well frozen. Growth of these moulds on the tuna results in the extraction of moisture. white fish about 8 days. The product is removed from the drying cycle. Aspergillus oryzae. Hot-smoked products made from white fish will usually keep better than those from fatty fish. rewetted and remoulded to give the desired shape and consistency. They are generally not cooked prior to consumption.3½ hours using the typical temperature set for hot-smoking referred to above (12). 5. are used. In terms of safety.CURED. Aspergillus tamarii) and several Penicillium spp. cooking in brine. 23). but seems. apart from the improved appearance. a very expensive and fashionable smoked product in the Netherlands.6 hours and smoked for 3 . The composition of commercial liquid smoke can vary widely depending on the method of manufacture. Natural smoke extracts. Aspergillus ostianus. but mechanical drying is used instead of sun drying. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). and substances unconnected with smoke but with a smoky flavour or smell. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). and many closely related Aspergilli (Aspergillus ochraceus. synthetic smoke flavours. Katsuobushi. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. oysters and mussels are usually wet-salted in 50º brine for 5 minutes and hot-smoked at 80 °C for less than 1 hour (12). Mackerel. 17. The production method is similar to masmeen. including ‘umami’ (16. moulding it into long thick sections. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). are usually 80º brined for 5 . 21. When refrigerated at 3 °C. to have no further advantage over other packing methods for other fish (12). including Eurotium repens and Eurotium rubrum. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. resulting in a characteristic flavour (20. chilled or freeze-thawed. The classical liquid smoke is 107 . Eels. similar to masmeen. break down of fats and the moderation of smoke flavour. Vacuum packaging retards the onset of rancidity when fatty fish is used. 18).3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. Boiled or steamed clams. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). 22. (Penicillum cyclopium and Penicillium puterellii) (19). fatty fish will remain in good condition for 6 days.5. which means that any bacterial contamination and growth after the heat treatment is a serious hazard that must be controlled by Good Manufacturing Practices (GMP) and factory hygiene/sanitation after the heat treatment (7). such as yeast derivatives.3.

applying it as a dip after brining. In experiments comparing traditionally smoked fish with liquid smoke. Fresh fish spoilage may be measured by 108 . According to some recent research. and less than 0.3.6 to 2.4 Spoilage Brining. injecting it into the fish flesh. It has been shown that liquid smoke can reduce the formation of PAHs (3). 5.5. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. The process relies on the generation of the smoke to produce elevated levels of carbon monoxide (CO) which interact with haemoglobin in the muscle to form the very stable bright pink/red carboxymyoglobin complex. nutritional neutrality and antioxidant activity (27). Liquid smoke in combination with sodium chloride in a hot-process smoke flavoured fish was found to be effective in preventing growth and toxin production associated with the growth of C. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). Liquid smoke was considered to provide advantages of safety. smoking. the result of this process is seen as a manipulation of the natural degradation of tuna colour that hides the true nature of the product quality and therefore has been made illegal in the EU.8% and still provide protection for 7 days (25). or applying it as an atomised spray within a modern automatic smoke kiln.4 Clear smoke treatments The “Clear” or “Tasteless” smoking process is a processing technique whereby smoke is generated as in a normal food smoking process but is filtered to remove the characteristic smoke flavours before application on to otherwise untreated (uncured) tuna. the salt level could effectively be reduced from 4. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25).FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). 5. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. When liquid smoke was used. or smoke extracts in organic solvents. The synergistic effect of liquid smoke with other preservatives could reduce the amount of salt in the product when light diets are necessary whilst still meeting the necessary safety criteria (26).06 μg/kg of benzoanthracene in the final foodstuff (28). pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. consistency of flavour and colour. The European Scientific Commitee for Food. so as to impart an unnatural red colouration to the flesh.03 μg/kg of benzopyrene. drying. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. both of which are considered potential carcinogens.

. Most marinated and smoked fish products are first brined or salt-pickled. and coryneform bacteria. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). For some products. rock and manufactured salts did not usually contain halophilic bacteria (29). contaminant bacteria from other materials used to produce the product. the bulk of the microflora comprises Bacillus spp. both of these will affect the spoilage patterns. Some brines are re-used over a period of days and a load of 10. which are found in the salt. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure). the shelf life may be estimated and investigation made into the causes of the loss of sensory quality. but. The use of drying and/or salt to lower the water activity. When heavy brining or salting is important for product shelf life. therefore. a very dry product is required to produce 'shelf-stability'. Stronger and longer brining has an effect like that of pickling. Once these changes are defined. chemical oxidation. spoilage is due to endogenous microorganisms and enzymes of the fish. and high concentrations are usually required to inhibit their growth fully. which may also be produced to describe the quality attributes and their loss over time for processed fish. also. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). SMOKED AND DRIED FISH grading schemes or quality indices. a product may become undesirable for consumption as a result of changes in its sensory characteristics. Spoilage needs to be carefully defined for each product. Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2. the fish are cut and splayed to allow greater surface contact with the salt. limits the types of microorganism capable of growing. Often. pickling and dry-salting all introduce bacteria to the fish. Upon salt packing. for some. and of smoke to add chemical preservatives. or their products require refrigerated storage. these bacteria tend to continue to dominate the flora in the liquid. these halophilic organisms may grow and cause spoilage.g. however. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime.. Within the limits of safety. When acid marinating or smoking. as the fish are added. marination or drying). In salts. otherwise. Even saturated brines allow some halophilic organisms to grow. or extended refrigerated storage. Once placed into brines. the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. contact with the bacteria 109 . Micrococcus spp. for others. the production process will modify the microbial flora present on the product. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. The microflora attached to the fish is relatively unaffected by light brines used before smoking.3%). Brining. the consequent reduction in the pH will help to inhibit spoilage organisms. a process similar in microbiological effect to that of saturated brining.000 cfu/ml of brine may build up. and. Usually. the fish flora are transferred to the brine mix.CURED.

and Aspergillus penicilloides. repens and E. Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration. yeasts. The bacteria form coloured colonies on the surface of salted fish prior to drying. including A. gaspé). and Halobacterium spp. 34) have shown that P. Basipetospora halophila. it was found in about 50% of samples examined. powdery growth. most notably Penicillium citrinum and Penicillium thomii. The metabolism of the organisms will produce off-odours. ‘dun’ is not characterised by the production of offensive odours. pisce was the main fungus isolated.g. including hydrogen sulphide (26). Studies conducted on dried and salted fish from Indonesia (31.. like the ‘pink’ organisms. Aspergillus wentii. (syn. or after drying if the fish is stored in excessive humidity. Spoilage of dry-salted and pickled fish is mainly due to mesophilic bacteria (those that grow with optimum temperatures around human body temperature and tend to grow poorly or not at all at refrigeration temperatures). as little if any growth will occur below 5 °C on salted fish (29). Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. 33). Aspergillus sydowii. A number of spoilage organisms can grow and produce conditions such as ‘sliming’. protein breakdown. it is possible to wash off the infected parts. as the water activity of saturated brine solutions is low enough to inhibit most bacteria. halotolerant or halophilic micrococci. A. The microbiology of heavily salted products is dominated by Gram-positive. and still produce a reasonable dried product. which. together with a range of Aspergilli. spore-forming bacteria. rubrum are also common isolates from dried fish. flavus. or fat breakdown) caused by enzymes rather than by microbial growth. B. rancidity due to lipolysis. Several Penicillium spp. were also isolated. halophilica was isolated 110 . spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. may also be found in the salt used in production. ‘pink’ and ‘dun’. Unlike ‘pink’. The bacteria are osmosensitive and will lyse in water. though only the latter two species were considered likely to have grown on the fish (32). 32. the best way of controlling the organisms is to refrigerate the fish. Therefore. At the pre-drying stage. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. W. (syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. although it may occur in other salted fish. E. This condition is often associated with case-hardening. sometimes covering the whole of the fish with a whitish. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage. ‘Pink’ describes a condition associated with surface growth of the pigmented halophilic bacteria Halococcus spp. The condition describes the growth of the halophilic moulds (Wallemia sebi. Sporendonema sebi)). sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). Eurotium amstelodami. lactic acid bacteria (LAB) and moulds. yeasts and fungi.FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. niger. if not too contaminated. Only halophilic microbes may grow.

but this will not prevent the growth of spoilage bacteria. Fish processed by cold-smoking. which does not create the salty conditions required for spoilage by those halophilic microorganisms.IV. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. 36). not the growth of spoilage organisms.CURED. Smoked fish products tend to have a light brine treatment.. where a high internal temperature is reached. Marine vibrios were the dominant flora in smoked salmon that was drysalted. Cold-smoked fish will contain microbial flora representative of that of the raw fish. including A. The brining to 3 . LAB. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. SMOKED AND DRIED FISH infrequently at low levels. The preservative effect of the smoke will inhibit some spoilage microorganisms. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations. able to grow on dried salted fish. has been isolated from dried fish (35). Aspergillus spp. so the bacteria spoiling these products will be psychrotrophic or psychrophilic.5% sodium chloride in the water phase will inhibit some bacteria. around the water activity of saturated sodium chloride. Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. but. and microbial spoilage is more likely. Aspergillus clavatus and Aspergillus niger. Vacuum packing limits chemical oxidation. however. as already stated. The salt in the water phase must be high enough to inhibit the growth of C. 33.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). In Vietnam. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life.747. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. will have a different residual post-smoking flora from 'hot-smoked' fish. botulinum type E. with a lower aw limit for growth of 0. The heat during hot-smoking will kill vegetative microorganisms but not all spores. Pseudomonadaceae. Table 5. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. it was. flavus. It has been reported that the type of salting (dry. Refrigeration is necessary. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. For more information on spoilage see Table 5. 111 .

.W. sebi and B. marinated and smoked fish Spoilage controlled by pH Heat Ant1 Ref2 Process Product Aw Probable spoilage organisms None Fresh fish spoilage organisms Halophilic spore formers and PPC4 Halophilic spore formers and PPC Halophilic organisms: Pink-Halococcus spp. halophila Halophilic spoilage organisms Halophilic spoilage organisms. boiled. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. smoked and dried Citrus marinade Dried Brined or salted then liquid or cold smoked Brined or salted then liquid or hot smoked 1 2 +++ Jellied eels + Ikan kaju Ceviche Stockfish Coldsmoked fish Hotsmoked fish +++ +++ ++ +/++ + + ++ ++ - ++ + + + ++ ++ ++ ++ + ++ + ++ Antimicrobial compounds added Refrigeration applied 3 v.IV Spoilage of salted.variable. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted.FISH AND SEAFOOD TABLE 5. LAB. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. Dun. + to +++ strength of preservative effect 4 Post-process contaminants 112 .not used/unchanged. and Halobacterium spp.

5-10 4. In the United States.5 5. The concentration required to prevent growth at room temperature can vary from as low as 3. sodium 113 . the products were heavily salted. smoked and dried.5-10. It has been shown that. SMOKED AND DRIED FISH TABLE 5. botulinum type E Staph. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13).CURED. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. botulinum type E.97 0. aureus Yeasts Moulds Lactic acid bacteria pH 5.0-9. and C. although the combination of the two factors.8 1. could not grow and produce toxin. when present in the raw material. types A and B are not affected by 3 . but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. pressure on the traditional methods of curing.97 0. botulinum type E and production of toxin.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. respectively.8 0. with this change.75 0. popular demand for healthy foods has led to a change in the tastes preferred by the public and.5-8. the fermented products being the most dangerous group (7).5 5. Since neither the smoking nor the drying parts of this curing process are particularly severe. under optimal conditions.5. However.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. 9 and 8 of the outbreaks.5 4. the importance of cold-chain integrity must be emphasised.5-8. the non-proteolytic type E strains can grow in up to 5% sodium chloride. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C.5 0-11 3. Before.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). botulinum. salted and pickled products accounted for 10.9 0. but if the fish products are stored at temperatures <10 °C. botulinum Staph. The presence of salt in cured products has a great effect on the growth of the bacteria.61 Organisms Most Pseudomonadaceae C. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14).86 0. and as botulism is a serious hazard in foods.5% to 5%. Examination of 165 outbreaks of botulism caused by fish products showed that smoked.

5. if storage temperatures are over 10 °C.5% salt in the water phase.FISH AND SEAFOOD nitrite can be added to the brine but this is not permitted in EU countries because nitrites might lead to formation of the carcinogenic nitrosamines. the organism may contaminate the product. However. after processing. they do not grow well in the raw material.0 . As with other organisms.5. Sliced smoked fish products. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . aureus growth.3 Enterobacteriaceae The pathogens Salmonella. they will grow rapidly (7). Usually.30% of the population positive for enterotoxin-producing strains (41). but requires higher temperatures for toxin production (>10 °C). related to the consumption of a raw smoked fish meal. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. if it contains sodium nitrite ≥100 ppm.94 (7). these parameters change as a result of the interference of food components and other limiting factors that can act in an additive or synergistic way to prevent Staph. as they require more handling. coli all occur on fish products as a result of contamination from animal/human reservoirs. Shigella and E. are particularly susceptible to contamination and. 5. 5.5. more importantly. Once the product is contaminated. An outbreak of Shigella in an Italian cruise ship. aureus. it should contain not less than 3.0% salt in the water phase (30). and the minimum pH for growth is 4. the risk of infection can be eliminated by proper cooking.5. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. but. The organism is mesophilic with a minimum growth temperature of 7 °C. when the bacterial load of the product is lower. if recontamination occurs and storage conditions are favourable. Thus.or cold-smoked fish should have at least 3. has been reported (42). Infection by Shigella sonnei has increased in the UK. but. It is a halotolerant bacterium able to grow at water activities as low as 0. In foodstuffs. growth and toxin formation may occur. As they are poor competitors.4. if the person handling the product is a carrier of Staph. throat and skin. the human nose.4 Listeria monocytogenes The psychrotrophic L. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). the organism can survive most of the light curing processes.5. The FDA established that hot.86 and 10% sodium chloride. gills and guts as well as in several lightly preserved fish 114 .

cold-smoked fish. salt levels of 8 . This includes matjes herring. widely used as starter cultures in food processes. or nematodes. which means that the process must conform to a level of ‘no detectable L. marinades (ceviche) (43) and gravad fish (44). The use of LAB in fish preservation is a topic that has been studied in projects (UP. SMOKED AND DRIED FISH products such as cold-smoked fish. Pseudoterranova dicipiens. such as ‘lightly preserved’ fish products. in combination with curing agents and refrigerated storage. and the tape worm Diphyllobothrium spp.the round worm Anisakis spp. are considered unsafe.2. Some LAB. do not survive 1 min at 60 °C. They have complicated life cycles. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). from freshwater fish. monocytogenes pathogenic for only specific segments at risk. favours growth of psychrotrophic LAB.45 °C. the CT 3162-FAIR program. the USA established ‘zero tolerance’ for the organism. and in CT95-1207FAIR). both being detected in cold-smoked salmon. a short period of freezing either of the raw material or of 115 .92 . has also been detected in cod (7). in the EU Fisheries R&D-FLAIR program. which considered L. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. gravad fish. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. monocytogenes in levels at or above 102 per gram (46). The Health Protection Agency (HPA) in the UK considered that it was unacceptable for readyto-eat foods (including all the products that are consumed without prior cooking) to contain any serogroup of L. pH 4. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). Unlike some European countries. lightly salted caviar. a number of fish products.514. As they are heat-sensitive. Thus. there were no reported outbreaks related to cured fish.12%. 5. the only concern to public health is when eating raw or uncooked fish products. from seawater fish. Its limiting conditions for growth are 0 . a round worm. which include intermediate hosts. and other local traditional products (7).9. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. have been shown to have an anti-listerial activity in many food products. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. Two outbreaks in the United States have been reported to be linked to salmon (10). Of microbiological concern in the northern hemisphere are two parasites . monocytogenes’ in 25 g of the finished product.0.95 (45). which. most of them are of little concern to public health.5. The round worms.CURED.6 and water activities below 0.5 Parasites Despite the presence of parasites in fish being very common.8 . Until 1997. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes.

48h ± 2h) Indicator organisms‡ Enterobacteriaceae E. for example.<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . E.<104 104 . cereus and Bacillus subtilis# <20 <20** <20 <10 20 . smell. cholerae Vibrio parahaemolyticus L. aureus C.100 >100 N/A Pathogens Salmonella spp.<104 20 .<100 200 .<100 10 . and unpasteurised yoghurt.6 Published Microbiological Criteria TABLE 5. coli (total) Listeria spp. and the levels or absence of indicator organisms or pathogens.VI Guidelines for the microbiological quality of ready-to-eat smoked fish. the organism should be identified # N/A denotes not applicable 116 .<104 100 . soft cheese. coli O157 & other VTEC Vibrio. monocytogenes Staph. monocytogenes) (cfu/g unless stated) <106 106 . (not L. If the Bacillus counts exceed 104/g. Acceptability is based on appearance.<100 20 . not detected in 25 g Campylobacter spp.<100 >104 >100 N/A N/A <20 20 .<107 ≥107 N/A <100 <20 100 .<103 N/A 100 . ‡ On occasions some strains may be pathogenic.<100 20 . taramasalata and cooked shellfish sampled at the point of sale (adapted from (11)) Criterion Satisfactory Borderline limit of acceptability (cfu/g unless stated) Unsatisfactory Unacceptable/ potential hazard* (cfu/g unless stated) (cfu/g unless stated) Aerobic plate count† (30 °C. These foods fall into category 5. 5.<105 ≥105 * Prosecution based solely on high colony counts and/or indicator organisms in the absence of other criteria of unacceptability is unlikely to be successful. † Guidelines for aerobic colony counts may not apply to certain fermented foods. salami.FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). texture. perfringens B.

Flick G.7 1. 9. Fernandes R. * Staph. Handling and Processing Eels.H. 381-406. 1997.F. 1984. 334. Hall G. 37(revised). Horne J. Torry Advisory Note No.M. USA. Birnie K. 1979. 8. 254-82 5. Bannerman A. 1991. Food spoilage fungi. Doré I.A.. Chapman & Hall. Regenstein C.E. aureus * Vibrio cholerae V. botulinum * Limit of tolerance 1x 103 ETEC/g. 117 . Flick G.. Wareing P. Marinades. 120-38. 2007.W.CURED. Leatherhead Publishing. Torry Advisory Note No. 56. Leatherhead. Introduction to Fish Technology.VII US Food and Drug Administration (FDA) guidelines related to ready-to-eat fishery products (45) Product Ready-to-eat fishery products (minimal cooking by consumer) Organisms and/or toxins Enterotoxigenic E. 82 (revised).. parahaemolyticus Vibrio vulnificus C. Ed.. 1993. Van Nostrand Reinhold. Eds. or presence of toxin 0. Torry Advisory Note No. Catching. Wareing P. SMOKED AND DRIED FISH TABLE 5. 7. References Regenstein J. 4. in The Seafood Industry. coli (ETEC) L. Martin R. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. The Smoked and Cured Seafood Guide. Preservation of fish by curing (drying. monocytogenes Salmonella spp.. in Micro-Facts: The Working Companion for Food Microbiologists. London. 3. 6. 2. 1990. except in the viscera of dungeness crab. Hot Smoking of Fish.M. 1994.8 ppm (saxitoxin equivalent) 20 ppm domoic acid.F. Smoked. Fernandes R. salting and smoking). McLay R. New York. LT or ST positive Presence of the organism Presence of the organism Positive for enterotoxin or ≥104 (Most Probable Number) Presence of toxigenic 01 or non-01 ≥ 1 x104 (Kanagawa positive or negative) Presence of the organism Presence of viable spores or vegetative cells in products that will support their growth. in Fish Processing Technology. Eds. Van Nostrand Reinhold. 3273.. New York. Huss H. cured and dried fish. FAO Fisheries Technical Paper. Urner Barry Publications Inc. Moody M. Assurance of Seafood Quality. Horner W. 1982.

Flavour constituents of dried bonito (Katsuobushi). 5 (revised). Food Chemistry. Biotechnology and Biochemistry. 13. 553-72. Food Control. 1998.. Ogiwara Y. The discovery of umami. Food Reviews International.C. 2002. 1982. 1980. 1995.. Microbiological control for fish smoking operations. 840-6. Botulism and Fishery Products. Yajima I. Embarek P. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors. 19. Technomic Publishers.M. Lindemann B. Perusing Asian soup options.. Biotechnology and Biochemistry.FISH AND SEAFOOD 10. 958-60. 51-81. 14. Teeny F. 1992. Doi M. Doe P.. Chapman & Hall.... 11. Agricultural and Biological Chemistry. Pelroy G. 1997. Druschitz A..E. Yamauchi H. 22 (revised).M.. Food Product Design. Bioscience. and Biochemistry. Torry Advisory Note No. 15. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour. 1981. 45 (10). 118 .. 843. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor. Doi M. Doi M. Matsui M. 27 (9). 1051-5. 17 (9). What’s in a name? Are MSG and umami the same? Chemical Senses. 59 (12). Doi M. Torry Advisory Note No.. 25.C. 2007.F. Hosokawa M.. Cann D.. Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish.M. Biotechnology. 6 (4). 17.. Journal of Food Protection. Hayashi K. 1386-7. Kanayama T. Shuto Y. 18. Eklund M. Recommendations for the Preparation of Smoked Salmon. 2324-5. Kinoshita Y. 53 (4). London.. Torry Advisory Note No. in Fisheries Processing. Biotechnological Applications. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour. Mitou M. Huss H. Lancaster. 21. 1994.. Processing Cod Roes. Matsui M.. 2008.H. Torry Advisory Note No.. Halpern B. Bannerman A.. 22. Shuto Y. Kinoshita Y. Bioscience. 23. Martin A. 1977.P. Paranjpye R. Shiemori Y. Ninomiya Y.. Dillon R. 1989. Fish Drying and Smoking: Production and Quality. Bioscience. 16. 935-41.R. Aoshima H. Jeppesen V. Shuto Y. 1990. Control of biological hazards in cold smoked salmon production. 88. 61 (8). Martin A.K. Packing Fish in a Modified Atmosphere.W.. Ed. 18. 1984. Horne J. Cann D. 335-40. Bannerman A. 108 (3).. 20..A. O-methylation of 2... 6 (6). 24. Patel T. Ninomiya M. 12. Peterson M. Sakakibara H... 1995.B. 56 (6). 508.. Yokayama S. Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species.E.

. Luxembourg. Ed.. 1996. Seafood Safety. Wheeler K. 1961.I. Modern Food Microbiology. 34.R. V. 1986..V. 1993. 29. 1991. Jay J.. Pszczola D. Fungi and Food Spoilage. Borgstrøm G. 27686. 33. Smoke flavourings. Hansen L. Maga J. Commission of the European Communities. Nahrung. Hocking A.O. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. 1994.D. Pitt J. 181-8. 36. New York.I. 1995. 74. Food and Drug Administration.T.. Taylor L. ceviche. 1985. Scientific Committee for Food. 31.. 123-9. Letters in Applied Microbiology. Washington DC.A.F. Food Technology. Golden D. 38 (3). Cann D. 32. 70-2.. 12.B. 164-9.A. 1995. 74 (2). Fungal flora of South Vietnamese fish and rice.. 35. Hocking A. Hocking A. 1979. 42. 1991. Pitt J. Food and Drug Administration. Sprouse R.M. 1941. Boca Raton.. Shewan J. Fungi associated with Indonesian dried fish.K. Cox J. Pitt J. 197-208. Diyaolu S. 88-90. Institute of Medicine. Røntved S. Food Research International. National Academic Press.. Importance of autolysis and microbiological activity on quality of cold-smoked salmon. Blackie.. Gill T. Incidence of Listeria monocytogenes in an acidified product. US Food and Drug Administration. Wheeler K. Ahmed F. 119 . Lucas F. 22. Ed. 1971. Hocking A.. Simini B. Loessner M. 30.. Interactions among xerophilic fungi associated with dried salted fish.S. Ed. 43. Press Inc. Academic Press. in Reports of the Scientific Committee for Food (34th Series). Huss H. Antimicrobial properties of smoke. Journal of the Fisheries Research Board of Canada. 742. An outbreak of Shigella. in Fish and Fishery Products Hazards and Controls Guide.H. 107-11.E. 1997. CRC.F.I. Studies on Sporendonema epizoum from ‘dun’ salt fish. Committee on Evaluation of the Safety of Fishery Products.C. 3 (4).A.1988. Sirvas S. The control of botulism in hot-smoked trout and mackerel.D. Ed. SMOKED AND DRIED FISH 26. Anggawati A.M. Tour highlights production and uses of smoke-based flavours. Lancet. in Fish as Food. Studies on salt fish. Journal of Food Technology. 98100. 49 (1).Y. Oxford. 2005. Fuchs R. London. 1994. Adebajo L. Frank M. 5. 40.. Hess E. 14. 41.J.M.E. 28. Journal of Applied Bacteriology. 38. New species of fungi from Indonesian dried fish. 348. Food Microbiology. 39. Mycotaxon. 218-25. 74. CEC.D. 1996. Journal of Tropical Medicine and Hygiene. Springer Science. 37.CURED. 27. Smoked and smoke-flavoured fishery products.. 311-7. Rockville.D. 29 (2).. The microbiology of sea-water fish.A. in Smoke in Food Processing.. 351-7... Townsend J.

Hurst A. 1991. 1996.C. Bolton F.W. Danielsson-Tham M-L. Davies A.L. Fletcher R.. 37 (1). 58 (8).. Richards J. 833-41.S. 5-7. Hughes A. Donovan T.. Garren D. in Quality Assurance in the Fish Industry. 2001. 2000. Round Worms in Fish.. 1982. Amsterdam. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions. 1992..R. Roberts D. Appendix 5: FDA and EPA Guidance Levels.FISH AND SEAFOOD 44. 55 (4).fda.J. http://www. 1989... Association of Food and Drug Officials.. Wooten R. in Fish and Fishery Products Hazards and Controls Guide. Ribeiro C.N.W. in smoked and "gravad" fish... Eklow A. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus. 26 (4). Ed. Unnerstad H. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon. 45 (9). Jacobsen M..M. Nye K. Pelaez C. Journal of the Association of Food and Drug Officials. 13-8. 1980. 49-59... Eds. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale. Clostridium botulinum and its importance in fishery products.cfsan. 197-209. Acta Veterinaria Scandinavia. 5 (1)..A. Canadian Journal of Microbiology. 46. 135-85. de Louvois J. Elsevier Science Publishers B. Cann D.. Beltran A. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects. US FDA Centre for Food Safety & Applied Nutrition. Journal of Clinical Microbiology. US FDA Centre for Food Safety & Applied Nutrition.. Torry Advisory Note No.C. 1994. 863-6. 1976.V.D. 1980. Pathogenic microorganisms of importance in seafood. Peterson M.J. Pelroy G.. Eklund M. 163-7. Zeitschrift für Lebensmittel-Untersuchung und –Forschung. Harrison M. Paranjpye R... 232-6.. Cured. 1995.A.gov/~comm/haccp4x5. Tham W. 1997. Hobbs G. Washington D.. An outbreak of listeriosis suspected to have been caused by rainbow trout. Advances in Food Research.. Gibbs P. 120 . 47. 48. salted. 5. Journal of Food Protection. 3. 188 (3). Loncarevic S.A. 35 (11) 2904-7. 507-10. Moral A.M. Incidence and growth of psychrotrophic Clostridium botulinum in food. Loncarevic S.html Gilbert R. Tham W.. Danielsson-Tham M. Food and Drug Administration. Huang Y. Communicable Disease and Public Health.8 Further Reading Gibson D. 80.. Journal of Food Protection.. Mentzing L-O. 45. Persson I.. Liston J. Huss H. Little C. Susuki E.H.. 22. Food Control. and smoked fish establishments good manufacturing practices... Ericsson H.E. Prevalence of Listeria monocytogenes and other Listeria spp..

. Ben Embarek P. 1992. 42. 502-8. Beuchat L. Food Microbiology. 1987. van Spreckens K. Patel T. Pelroy G. Fuchs R. Poysky F. Competition of food bacteria with Listeria monocytogenes during enrichment culture. Poindexter C. Elsevier. 293301.R.A. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. Taylor S. detection and growth of Listeria monocytogenes in seafoods: a review. Presence. 866-70. International Journal of Food Microbiology. International Journal of Food Microbiology. Journal of Food Protection. Journal of Food Protection. 58 (5). Food Technology. 23 (1). Davies A.J. 309-16. 14.P.. 11. and temperature on the growth of Vibrio parahaemolyticus.. Journal of Food Safety.A. Rørvik L.. Keusch A. Hudson J.N.. Applied Microbiology. Beuchat L. 1994.E. Marine toxins of microbial origin. Combined effects of water activity. Histamine production by psychrophilic flora..178-80. Eds. 19. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. International Journal of Food Microbiology. Ratnam S. 27 (6). Bayliss A. Letters in Applied Microbiology. in Seafood Quality Determination. solute. SMOKED AND DRIED FISH Sutherland J. 19-27. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. 1973. 61 (12). temperature. and salt concentration on growth and survival of Vibrio parahaemolyticus. Paranjpye R. Peterson M.. Caugant D. 1734. 1974.. 21 (3).. 1075-80. Tran T.. 1995. Applied Microbiology. Jemmi T. Prevalence of Listeria in smoked fish.W. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants.. 121 . Slade A. Lashbrook L. in a salmon slaughterhouse and smoked salmon processing plant. 4389-95. 1994. 25.R. 1995. 217-36.. Roberts T.. 844-6. Dallas H.. Nicolaides L..K. Letters in Applied Microbiology. pH and sodium chloride. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants.C.S. Incidence of Listeria in hot and cold-smoked fish. 11. 1991. 55 (11). 394-6. 1995. 1994... 309-18.CURED.A. Interacting effects of pH. Yndestad M.T. Letters in Applied Microbiology.R. 25 (5). 94-8. 1992. 1988. Dillon R. Kramer D. Liston J. Eklund M.. New York. Hitchins A. 1994. Contamination pattern of Listeria monocytogenes and other Listeria spp.


added during preparation. Carbohydrates are added to some fish sauces during their preparation. FERMENTED FISH Prof. typically in the range 15 . The fish sauces are produced on the largest scale. 123 . North American and other markets and are. though some are produced elsewhere.1 Definitions As applied to fish products.25% and are used mainly as a condiment. producing the fish sauces and pastes. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. Essentially. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. For example. they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. Fermented fish products are largely confined to east and south-east Asia. The relative importance of these two activities depends on the formulation of the product. koji. there are numerous variants of some common themes and a host of local names used to describe them.1.6.I. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. lactic fermentation can play a significant role. they are exported to European. the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6. Those to which only salt is added tend to be mainly autolytic. 6. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. As with most traditional products that are still produced principally on a cottage industry or domestic scale.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. whereas in those where a carbohydrate source such as rice or sugar is also added. the most important of all fermented fish products. in economic terms. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu.

III.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. to products analogous to other lactic fermented foods such as salami. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. who had adopted them from the Greeks. may be direct descendants. made from fish livers in Greece. where the bacterial production of lactic acid is a major feature. jeotkals (shrimp) nam-pla (pa) Budu Patis nam-pla nuoc-mam padec Belachan (shrimps) bagoong kapi mam-ca man-tom (shrimps) Laos Malaysia Philippines Thailand Vietnam Even within the limitations of fish and salt as the sole ingredients. and the more widely known Worcestershire sauce a very distant relative.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. In Europe.1. 6. although garos. These have long since fallen from use. salty taste. A selection of Asian products is presented in Table 6.FISH AND SEAFOOD TABLE 6. cheese-like aroma Burma Indonesia Ngapi ketjap-ikan Paste Red/brown salty paste nga-ngapi trassi-ikan trassi-udang (shrimps) prahoc mam-ruoc (shrimps) Kampuchea nuoc-mam nuoc-mam-gau-ca (livers only) Korea myulchljeot jeots. Fermented fish products are of very minor importance outside Asia. and pissala. 124 . a variety of products can be produced within a single country.II. made from whole fish in France. This is well exemplified by some of the fish/salt products of Thailand described in Table 6.

crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 . pla-som. burong-ayungi.FERMENTED FISH TABLE 6. mam-pa-kor. som-pa-keng pekasam. mam-chao. burong-dalag. som-fak/ som-fug 125 Malaysia Philippines Thailand . crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 . pla-chao.12 months 10% raw sugar added. funa-suchi. shrimps. som-pa.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. salt/fish ratio Production period Type of use Hoi-dong Molluscs Whole or without shells Whole - Main dish Kapi (fish or shrimp paste) Nam-pla (fish sauce) Nam-budu Small fish. mam-seeing sikhae som-kay-pa-eun. tai-suchi phaak. burong-bangus pla-ra. ayu-sushi.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 .g.g.8 months Main dish TABLE 6. pa-khem. e. pa-chao. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps. e.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. cencalok burong-isda.3 months Main dish Tai-pla Bowels 1:3 6 .

Penaeus indicua Many of the fish used in fermentation are uneviscerated.IV Fish species used in lactic fermented fish products of the Philippines Product names Burong ayungin Burong bangus Burong dala Burong gurami Burong hito Burong kanduli Burong tilapya Balao balao Burong hipon Local Ayungin Bangus Dalag Gurami Hito Kanduli Tilapya Tagunton Suwahe English Silver perch Milkfish Mudfish Goramy Catfish Sea catfish Tilapia Shrimp Shrimp Scientific name Therapon plumbeus Chanos chanos Ophicephalus striatus Osphronemus goramy Clarias batrachus Arias manillensis Tilapia nilotica Macrobrachium spp. For example. or buried salmon. 126 .FISH AND SEAFOOD In Northern Europe. freshwater fish are the usual raw material.the flooded rice fields. in Sweden. it was found that several different sources were used for the fish . which employ lower levels of salt than Asian products. which has been increasing in popularity over the last 50 years. especially in fish/salt products. some fermented fish products remain popular. Lactic fermented fish products are often associated with inland areas such as the Central Luzon region of the Philippines and the north-east of Thailand. in a survey of fermented fish products in north-east Thailand. for example.2 Initial Microflora Composition of the initial microflora of fish is described in some detail elsewhere in this Handbook and will not be reiterated here. fermented trout. states that sugar is often included with the salt. malt or flour (1). and old descriptions of their production often refer to the addition of whey. made from herring. in Norway and surstrømming. 6. The diversity of fish used in fermented products (see. These are often described as fish/salt products. relatively mild-tasting product.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. is a traditional. and counts ranging from below 104 /g to in excess of 107 have been reported. Table 6. and their microflora tends to reflect their local environment more than that of marine species. and the initial microflora is likely to be considerably larger in these cases. paddy ponds beside rice fields used for collecting fish when the field has dried up. TABLE 6. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. A modern account of their production. however. There are also more heavily fermented products. Norwegian gravlaks. Here. rakefisk or surfisk. and a large local freshwater reservoir (2). the most popular varieties of which are rakørret.

When this is a simple carbohydrate source such as sugar.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. Hansenula. In some cases. or boiled or roasted rice. principally species of the genera Rhizopus. These preparations contain a mixed microflora of moulds and yeasts. either directly or as the partially saccharified rice they produce. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). they can contribute to the initial microflora. and the anaerobic conditions that prevail in the bulk of the material.FERMENTED FISH Where other ingredients are used. the Philippine fish sauce. The amount of salt used is often only vaguely defined but is generally sufficient to reduce the aw below levels at which most bacteria associated with the raw material will grow and. This. 6.74 or below.3. It also demonstrated their significant proteolytic activity during the first month of fermentation.4. traditional starter culture preparations such as koji (Japan). the fish substrate is mixed with dry salt and the product is packed into containers and left to ferment for an extended period of weeks to months. were found to be the predominant isolates. Other halophilic species have been identified from fish/salt products. More recently. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. however. angkak (Philippines and Thailand) and look-pang (Thailand) are used. Endomycopsis and Saccharomyces . after one month. Mucor. however. in many cases.3 6.1 – 6. Generally. Similar results were seen with patis. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. sufficient to saturate the water phase present and produce an aw of 0. and studies 127 . work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. mean that microorganisms have been generally held to play little or no role in the process that follows. 4). Bacillus spp. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. In another microbiological study of four fish sauces.1 Processing and its Effects on the Microflora Fish sauces and pastes Detailed flow charts for several fish/salt fermented products are presented as Figures 6. which included nam-pla and patis. (3.

12).2. Fig.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 . Patis (Condiment) 128 . 6. 6. Belacan (Condiment) Fig.1.

6. To produce semi-solid fish pastes as opposed to fish sauces.FERMENTED FISH Fig.3. however. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues.6. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are. 14). Ika-Shiokara (side dish) Fig. In fish sauce production.4. thought to be largely the result of autolytic breakdown of the fish tissues. the 129 . and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. eviscerated fish are sometimes used to slow the autolytic process.

2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. Fig. for example.5. filtered and bottled. In contrast to free-swimming fish.6 – 6. Hoi-Malangpu-Dong (side dish. snack) 130 . 6.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids.8. Some examples of products and their formulation are presented in Table 6. 6. volatile fatty acids and nucleotides. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. shellfish such as mussels often have appreciable levels of carbohydrate in their tissues and in these cases addition of carbohydrate is not necessary for a lactic fermentation to occur (see. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. Figure 6.3.5). which is eventually separated from the residual fish tissues. particularly glutamate.V and Figures 6.

3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 .6. NE.FERMENTED FISH TABLE 6.6.12 months Shelf life Use 1 . N.12 days 3 . the prefix 'pla' is replaced by 'kung'. Kung-Chom (staple food.7 days 5 .03 Preparation time 6 .12 months Main dish 2 Pla-jao C. 3:1:0. C NE. Fig. NE 10 . S.2-4 Fish (pieces): salt:khaomark (fermented rice).20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. north-east. north.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 . In these cases. N. N S 5 . C NE.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. C. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. N.5-1. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. central. NE Formulation Fish (whole or pieces): salt: roasted rice.4 are also produced using shrimps. south of Thailand. side dish) 131 .

2.3. typically in the range 2 . This is readily explained by the susceptibility of LAB to elevated salt concentrations. Phillipines Journal of Food Science and Technology. If acidity is a desired sensory property of the product. (from Arroyo et al.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. but there is considerable variation in this. of the two factors contributing to shelf life . It also indicates that. This can be clearly seen from the data on reported shelf lives of a number of fish salt carbohydrate products and their respective pH ranges and salt contents (Table 6.7. the salt is of far greater importance in determining the product’s long-term stability.. then this will take longer to develop than if lower salt levels are used. giving a reduced rate of acid generation. while they are able to grow at high salt levels. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. Red burong isda 132 .10%. The LAB isolated from these products are those with more marked salt tolerance but. 2.1 The use of higher salt levels results in a longer production phase but a better keeping quality product. these products are salted at lower levels than the fish sauces and pastes. Generally.salt and lactic acid.VI). their growth and metabolism are nonetheless slowed.6. 106) Fig.

6. 2.5.8 .9.8 .3 5.FERMENTED FISH (from Arroyo et al.2 4. White balao-balao TABLE 6.5 .5.6 3.0 Shelf life 3 weeks 6 . Philippines Journal of Food Science and Technology.0 .4.8 pH 4.3 years 1 .6. 106) Fig.1 .5.9 .5 3.4. 133 ..2 .2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).5 .1 4.8 2.2 4.3 .6.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.0 .4 .0 .9 4.8.9 4.12 months 1 .5.

0 . a common ingredient in these products. alternatively. Although the ability to ferment starch is not widespread in the LAB.5) despite other factors. Rice is the most common carbohydrate source used. the rice may be added in a partially saccharified form such as kao-mark (Thailand).1 . they increase the amount of soluble sugars produced and thus the range of LAB that can grow.9) compared with the white product (pH 4. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity.2 Inclusion of more carbohydrate gives a faster fermentation and a stronger acid taste Increasing the proportion of carbohydrate in the formulation decreases the product’s buffering capacity and provides the LAB with more substrate on which to act. TABLE 6. such as salt content. also lower the unit cost of the product and this is probably a significant factor in many cases. 19).VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. 21). Addition of carbohydrate will. nare sushi pla-chao pla-paeng-daeng pa-chao Country Philippines Japan Thailand Thailand Laos Saccharified rice kao-mark Kha-namak Starter culture ang-kak koji ang-kak - 134 . a number of amylolytic species have been described and an amylolytic strain of Lactobacillus plantarum has been isolated from burong bangus in the Philippines (17). of course.2. inulin. This results in a more active lactic fermentation and a lower pH.FISH AND SEAFOOD 6. Where they are used. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6.3. When added as boiled or roasted rice alone. In som-fak 9% of LAB isolated were able to ferment rice whereas 19% could ferment garlic and omission of garlic from the formulation resulted in lower acid production during fermentation (20. For example. being similar (18.VII). Garlic. also serves as a valuable source of the more readily fermented carbohydrate.4.3. although the final population of LAB may be similar. In several products.

but they are likely to behave similarly to lactic fermented products produced from other commodities. and there is a marked regional variation in the preferred type. in turn.8 . and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24).4 Spoilage The ASCA handbook on fermented foods in the region ascribes shelf lives to various fermented fish products ranging from 2 weeks in the case of some of the low-salt lactic fermented products to 5 years for some of the high salt sauces and pastes (16). Some authors have claimed that the function of the roasted rice added during pla-ra production is to improve texture and slow the fermentation rather than to serve as a substrate for lactic fermentation (22). Generally. Results of a product survey were consistent with this. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. allowing other spoilage organisms to grow. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak.5. markedly extending the shelf life (25).FERMENTED FISH Sikhae. The growth of yeasts and moulds could. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. There is little published information on the factors that limit the shelf life of the low-salt products. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23).5 in the first 2 . the level of salt employed is on the high side for fish/salt/carbohydrate products. which ranged from 7. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. is something of an exception since millet is used along with garlic as the carbohydrate source.17.9%. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet.3 weeks at around 20 °C.2 at the end of fermentation.3 days. Traditional packing techniques used with lactic fermented fish products. such as wrapping in banana leaves. The same product can range from a brown turbid liquid where the fish has been subject to extensive autolysis to brown partly dried pieces of fish. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. will not reliably exclude air. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products.7 up to 6.2. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. 6. Excessive growth and acid production by the LAB could render products too sour for acceptability. the Korean fermented fish product. thereafter declining to about 4. the pH drops from 6. lead to an increase in pH. Clearly. presumably correlating with the salt content. 135 . around 12%. In a successful fermentation. The mixture is allowed to ferment for 2 . This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products.7 to below 4. reporting the product’s pH to range from 4.5:9:3.

fish products are the most common source of botulism. The bacterial pathogens most commonly associated with the living environment of fish. There is little in the way of documented information associating fermented fish products with foodborne illness. and consumed without cooking. such as soaking the fish at temperatures below 5 °C before fermentation and the use of starter cultures to ensure a rapid and reliable decrease in pH.FISH AND SEAFOOD Products where a substantial population of moulds and yeasts is deliberately added in the form of saccharified rice or a starter culture would spoil as a result of their continuing activity breaking down the rice and producing unacceptable levels of alcohol in the product.8 and at salt levels as high 10%. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. although these limits apply when all other factors are optimal (26). but we are unaware of any reports of this happening. however. viable numbers will probably decline quite rapidly. typically 30 °C. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. outbreaks of 136 . Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. parahaemolyticus has been reported growing down to a pH of 4. In Japan. under conditions inimical to growth and high ambient storage temperatures. As in Norway. Because of the severity of the illness it causes.87. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. the high salt concentrations preclude the growth of all food-poisoning bacteria and those initially present would not survive well the prolonged exposure to high salt during production. 6. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. During fermentation. After capture. V. Several measures have been suggested for reducing the risk of botulism.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. which accounted for 88% of outbreaks in the period 1952 . The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). In the fish sauces and pastes. are Vibrio parahaemolyticus and Clostridium botulinum.

FERMENTED FISH botulism have become less frequent in recent years. Liston J. while increasing the acetic acid content of the formulation had little or no effect (32). though 27% survival was still recorded (33). Fermented Fish Products Study (Thailand) Phase 1. With this particular organism. and salmon heads and eggs. which will persist into the product. 105-10. particularly in south-east Asia and the western Pacific regions (30). Greenland and Alaska. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. Oslo. 2. Journal of Food Science. Raw materials subject to this process include seal flipper. Novus Press.6 1. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). walrus nose. 1966.. References Riddervold A.. Puwastien P. and warm weather can result in rapid growth of C. Thailand. Foodborne trematode infections are emerging as a major public health problem. Dollar A. botulinum. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. 1990.O. giving an alkaline fermentation. Microbiology and chemistry of fermented fish. Mahidol University. Dhanamitta S. 6.M. which is generally consumed raw (29).. Lutefisk. Baseline Study (1987-1988). One study with the trematode Haplorchis taichui. beaver tail and paw. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31). 3. affecting an estimated 40 million people. Institute of Nutrition. Kasemsarn B. botulinum and the production of toxin. whale meat. Many of these products are described as fermented. Yhoung-Aree J. 1989. they could act as an important vehicle of transmission.. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). Saisithi P. Salting levels are often insufficiently low to prevent growth of C. Where these products are consumed without final cooking. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. 137 . Studies to determine the effect of traditional processing techniques on the infectivity of metacercariae was identified as a priority research area by a WHO study group (30). although the absence of any source of fermentable carbohydrate means that the process is putrefactive. and these are often used in the production of fermented fish products.. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada. salt content was found to have the greatest impact on the viability of larvae. 31. Rakefisk and Herring in the Norwegian Tradition.

Nov. SK33 isolated from fish sauce fermentation. 16. 10. New York. Hiraga K.V. 6338. Im W. isolated from jeotgal. Kaneko Y..A..FISH AND SEAFOOD 4. 7... Hull R. Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme. Protein hydrolysate from fish. Nov.. 1986. 57. 12.R. Lim J.. Phil.. 138 . Rodriguez-Valera F. Plenum Press.. Suntinanalert P. Rattagool P. Dymsza H. Tsunemi M.R. Orejana F. Liston J.J. Kudo T.. 16A. 1976.W. Rodtong S. 2007. Some chemical changes in fish silage.. 11. Effects of gamma radiation on sensory qualities. Lee S.M.H. 14. 1975. 29. 63.. 43. 2007. 124-30.. Takada K.. Reddi P. 934-9. Production and characterisation of NaCl-activated proteinases from Virgibacillus spp. Oda K. 381-8. Journal of Food Science. 3. 80. International Journal of Systematic and Evolutionary Microbiology. 353-63.T.S. a traditional Korean fermented seafood.... Olympia M. Pakdeeto A. 1991.M. Journal of Fermentation and Bioengineering. Catheptic activity of fish muscle. Halophiles in Thai fish sauce (Nam pla).. Moonmangmee S. Sands A.K. Dong-Ho K. Saono S. Adams M. Tanasupawat S. Crisan E..K. Sinsuwan S. Fermented fish products of south east Asia. Tropical Science. Fukuda H. Cooke R. SR5-3 isolated from fish sauce: purification and characterisation.. Process Biochemistry. 2008. Tanasupawat S. Jae-Hun K. 37. Constantinides S... Bioscience. 15. A halophilic serine proteinase from Halobacillus spp.... 25. 141-2. 17-30. 106-8.. Yongsawatdigul J. Yook H. International Journal of Systematic and Evolutionary Microbiology.. 6. 1985.. Aslam Z. 11. Canberra... 5. Biotechnology and Biochemistry... Namwong S. Ahn H..R.H. Itoh T. 9. Backhoff P. in General and Applied Aspects of Halophilic Microorganisms. Salinococcus jeotgali spp. from fermented shrimp paste in Thailand. Applied Microbiology. 1395-401.D. Takano M. Microflora of four fermented fish sauces. 61-73. 2000. Ono H. Journal of Food Science and Technology.. 1979. 57. 8. 2006. Byun M. Journal of Food Protection. 1972. Agents of proteolysis and its inhibition in patis (fish sauce). Chung Y. Kyong-Haeng L. Thawai C.. 3649. Thongthai C.T. Velankar N. 185-92. 17. Ed.. 13. 1995. Yasir M. Government of Australia. Lentibacillus kapialis spp. Dhamcharee B. Journal of Scientific and Industrial Research (India). Journal of Food Technology. 1957. microbiological and chemical properties of salted and fermented squid. 643-7.. 70.. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries.

. Benjakul S. 19. a Thai low-salt fermented fish product. Food Chemistry. Journal of Agricultural Science (Tokyo).... Avurhi J. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation.H.A. 1-4. Tanaka M. 26. in the Philippines. 305-22. WHO Study Group. 28. Hauschild A. 92. 30.W. ASEAN Food Journal. 23. Control of foodborne trematode infections. Dodds K. Eds.B. 1983. Ed.H.L. Isaac-Renton J.D.. Journal of Agricultural Science (Tokyo). Dodds K.. 20. Eds. Owens J. 1996. 105-12. 103. 1992.H. The fermented fish food. 28 (6). Caldo G. Visessanguan W. Yamprayoon J.W. 1992.D.. burong-isda. Fung C.. Sakai H.. Paludan-Müller C.H. Wedell-Neergaard C. Huss H. in Clostridium botulinum: Ecology and Control in Foods.W.H. Valyasevi R. Saisithi P.H. Marcel Dekker. 138-44. Erikson U. Moody L. Characterisation of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation. Kozaki M. Paludan-Müller C.. Rustad T... 274-86. 69-104. 28.. in Microorganisms in Foods. Volume 5: Microbiological Specifications of Food Pathogens. 139 . Marcel Dekker. 2007. 1998.. Hauschild A.A. 307-14. Riebroy S. Hauschild A.. Traditional fermented fish products with special reference to Thai products. Blackie. in Clostridium botulinum: Ecology and Control in Foods. World Health Organisation Technical Report Series.K. 21. 38. 25.FERMENTED FISH 18... New York. 27. Gram L... Vibrio parahaemolyticus. 28. Ben Embarek P. International Commission on Microbiological Specifications for Foods. Fermentation and spoilage of som fak. Yeast-flora in red burong-isda a fermented fish food from the Philippines.L.. Huss H. Wainwright R.B. 219-29. Journal of Applied Microbiology. Kozaki M. Two outbreaks of botulism associated with fermented salmon roe – British Columbia. Hazards from northern native foods. 104-6. 22. 1815. International Journal of Food Microbiology... International Commission on Microbiological Specifications for Foods. 3-10. 3. 1987. Huss H. Tropical Science. Gram L.. New York.M. 46. 24. Caldo G... 2002. Patrick D. 27. 1995. Martin J. Gram L. 29. Ostergaard A. Journal of Food Science and Technology. Canada Communicable Disease Report. Sakai H. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper.. Epidemiology of human foodborne botulism. London. 1983. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak. a Thai low-salt fish product. 2002. 426-35. 1990.. Dawar M. 1999.

International Journal of Cancer. 2001.R.FISH AND SEAFOOD 31...R.. Todoroki T. Horst K. 117. Srivatanakul P. Progress in Industrial Microbiology. northeast Thailand. 32. Hanai S. Honjo S. Survival of Anisakis larvae in marinated herring fillets. 1995. Huss H. 1986. Gaithersburg. Survival of Haplorchis taichui metacercariae in lab-pla. Uchida K. Fermented flesh foods.. Nout M. 854-60. 110-1. 15998. 29. Nithikathkul C. Fermentation and Food Safety... Jedpiyawongse A. 6. 36. Sripa B. Ruamsuk A. 2005. Wongsawad C. Kittiwatanachot P. 2005. Thai traditional food preparation... Kikukawa H.. 33. 661-70. Bloemsma B. Sriplung H.. International Journal of Food Science & Technology..J.H... Southeast Asian Journal of Tropical Medicine and Public Health. Roepstorff A. Chuboon S..7 Further Reading Adams M. Aspen Publishers Inc. 140 . Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom. Miwa M. Deerasamee S..R. Adams M.. 23...

and a second updated edition dealing with the pharmacology. Increasingly sophisticated instrumentation (e. 5). and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. Although most of these antibody-based test kits have yet to receive international regulatory approval. including the well known marine toxins 141 . more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins.C.3) and some practical applications for detecting shellfish toxins have been reported (4. The manual on harmful marine algae and their products.D.. Another book dealing with the clinical effects of a variety of poisons and toxins. most notably the azaspiracids and the cyclic imines as well as some additional polyethers. Scientific and Cultural Organisation) has been updated (7). Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins.M.7. Since the original chapter in this series was published (6). produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. physiology and detection of marine and freshwater toxins has also appeared (8). Wright. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues.. An emerging technology is the use of immunosensors (1 . Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. It remains to be seen if this approach gains momentum in the future. FISH AND SHELLFISH TOXINS Jeffrey L. known and unknown. Nevertheless. Ph. C.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. While some of these toxins only occur in seafood from a restricted geographical region. several scientific reports have described their efficacy and potential.g. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7. a number of new toxins and toxin families have been identified.

with intoxications occurring in many countries even today (15). some are the result of biotransformation processes by the filter-feeding host. There is a recommendation from a joint FAO (Food and Agriculture Organisation of the United Nations)/IOC/WHO (World Health Organisation) report that marine biotoxins be classified according to their structural chemistry group rather than the symptoms they induce (e.2. All PSPs degrade rapidly in alkaline conditions. In addition.FISH AND SEAFOOD has been produced (9). Many of these species are extremely common and are found throughout the world in a variety of cold. even crustaceans such as lobsters. Gymnodinium catenatum. and a review on the risk assessment of marine toxins has also been produced (14). An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). can become contaminated with these toxins. Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. Around twenty PSP toxins are known.g.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. Saxitoxin Group instead of the Paralytic Shellfish Poisons). The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). and another. 142 . neoSTX and Gonyautoxins (GTX) 1-4. and a tropical species Pyrodinium bahamense (16). Consequently. temperate. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). Not all the toxins found in shellfish extracts are microalgal products.1). The Ncarbamoyl sulfates are readily hydrolysed to STX. all of which contain a tetrahydropurine skeleton (Figure 7. and sub-tropical waters and habitats (15). a great variety of shellfish from all latitudes. 7.2 Paralytic Shellfish Poisoning (PSP) Toxins The PSP toxins are arguably the most recognised group of marine toxins and continue to be regarded among the most dangerous. a number of useful and comprehensive reviews and book chapters have appeared dealing with various topics related to individual toxin groups and these are referred to in each appropriate section. 7.

Neurophysiological studies have shown that PSP toxins all possess the same mode of action though some derivatives such as STX and neoSTX are considerably more potent than the C-toxins (15). Human symptoms begin with tingling and numbness of the lips. 143 .2 Toxicology PSP toxins affect the central nervous system of mammals. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis. PSPs bind to site 1 of sodium channels reversibly blocking sodium conductance in nerve and muscle membranes by binding to a specific receptor site located on the outside surface of the membrane close to the ion channel opening. and humans seem to be particularly susceptible. tongue. However. 7.2.FISH AND SHELLFISH TOXINS STX GTX2 GTX3 GTX5 (B1) C1 C2 NEO GTX1 GTX4 GTX6 (B2) C3 C4 R1 H H H H H H OH OH OH OH OH OH R2 H H OSO3 H H OSO3 H H OSO3 H H OSO3 - R3 H OSO3 H H OSO3 H H OSO3 H H OSO3 H R4 H H H SO3 SO3 SO3 H H H SO3 SO3 SO3 - Fig.30 minutes of consumption. and fingers within 5 .1. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. this is never sufficient to render the seafood safe to consume.

3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. The mouse bioassay has been in place for decades and the method has been standardised. China. the detection limit (4 µg/g) of the method is close to the safe level and does not provide any information on the profile of PSP toxins present in the extract. including Japan. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. and while simple and relatively inexpensive to operate.2).8 µg/g) of shellfish or seafood tissue. Thailand. specifically 144 . 7. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. While the chemical detection methods are most promising for the future. a criticism has been that such methods are time consuming. After some development and various trials. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis.3. 15. a pre-column method has received Association of Analytical Communities (AOAC) approval (18). Two approaches have been taken here. 17). 7. and islands of the South Pacific. and has also been evaluated by a number of EU laboratories.FISH AND SEAFOOD 7.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. 23. Most intoxication events have occurred in South East Asia. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). However it remains as the monitoring method of choice in many countries. TTX most commonly occurs in many species of puffer fish. as well as other parts. Since its discovery.2. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). 24) (Figure 7. Taiwan. and over ten analogs are known (22. The latter test is accepted by the United States Food and Drug Administration (US FDA) for use in the United States National Shellfish Sanitation Program. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). other derivatives have been reported. Alternative HPLC methods have subsequently reduced this to a single HPLC run.

25. horseshoe crabs. In Japan. Since then. respiratory failure and coma. hypotension. 26). Fig. and blocks diffusion of sodium through the sodium channel.2 Toxicology Like the PSP toxins. 23.2 mg of purified toxin can be lethal.3. and in mild cases results in distal muscle weakness. molluscs. The onset of symptoms usually occurs rapidly.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. the blue-ringed octopus. where puffer fish of the genus Fugu are a delicacy. nausea and vomiting. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. it is served in speciality restaurants with qualified chefs to properly prepare the fish. Typically this has been accomplished using HPLC followed by post 145 .2. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. 7. and even newts. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. 27). Structure of tetrodotoxin 7. raising speculation that TTX is produced by a bacterial source. and once again the mouse bioassay has been used to detect the presence of the toxin. and more recently by an actinomycete Norcardiopsis dassonvillei (26). gonads and roe (24). 7. A review of TTX detection methods has been published (12.FISH AND SHELLFISH TOXINS in the liver. An oral dose of 1 . Chemical methods for TTX detection have also been investigated and are mainly based on its smooth conversion by heating in an alkaline solution to a detectable fluorescent (C9) base.3. preventing depolarisation and propagation of action potentials in nerve cells. Severe poisoning causes paralysis and respiratory failure. All of the observed toxicity is secondary to the action potential blockade.

FISH AND SEAFOOD column oxidation to the fluorescent derivatives. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA.4. but 146 .3. Immunoassay methods have also been investigated. Structure of domoic acid and three naturally occurring photoisomers 7. need further development (24). These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). 7. a polar glutamate agonist. However this method has been criticised for being complex.3 for examples). and more rapid LC-MS methods [(M+H)+ m/z 320] have been reported for the detection and identification of TTX and its various derivatives (27). and while promising.4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). but it is known that such congeners are less toxic than DA itself (30). but all of them are considerably less toxic than DA itself (29). both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. 7. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28). Fig. Currently 12 isomers of DA have been reported (see Figure 7.

Recently. 33). nausea.FISH AND SHELLFISH TOXINS only three strains (P. disorientation and coma.39). another diatom species Nitzschia navisvaringica. and in locations around New Zealand (32. and sea animals such as sea lions and even whales (35). the Mediterranean. the wide distribution of toxin-producing strains of the diatom have resulted in the frequent occurrence of DA in many filter feeding organisms such as mussels. The oral absorption of DA is around 5 . and this has led to ASP toxicity in birds. the Philippines. as does impaired renal function which increases serum concentration and residence time of the toxin. To a great extent. Indeed. The original 1987 event was followed some years later by a more widespread episode along the western coast of the US and today toxin-producing strains of Pseudonitzschia have been found in the Gulf coast of the US and along the western shores of the US and Canada. Penetration of the blood brain barrier is poor in healthy subjects. seizures. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. which has been well documented in studies along the west coast of the US. Pseudonitzschia and DA have been detected in many other maritime locations in the north Atlantic and the North Sea where it appears around the coast of the UK. southern and western portions of Ireland. but any impairment of the blood brain barrier results in additional risk. Pseudonitzschia australis. including persistent short term memory deficits (36 . Levels of DA in the other strains are generally <1 pg/cell. Another factor is the movement of the toxin through the food chain.. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). found in the Pacific around Vietnam. 7. Victims display a variety of symptoms including headache. and review of the toxicologic pathology of DA has been published (36). 147 . cormorants and pelicans (34). and clams. has been reported as a medium-level producer of DA (33). and is mainly distributed in the blood. When DA was identified as a new marine toxin it was quickly determined to be a glutamate agonist that binds to a sub-class of neuronal receptors called kainite receptors. scallops. it has been reported that cooking or steaming contaminated mussels merely has the effect of spreading the toxin to other tissues of the shellfish as a result of disruption of the digestive gland. For example.10% of the administered dose.4. Okinawa and more northerly parts of Japan. multiseries.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming.

australis.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue).3 Detection methods DA is water soluble and can be extracted from shellfish tissue by the acidic PSP extraction method although this may result in some decomposition. although in the latter study the probes for Pseudonitzschia pungens. and this is typically the method of choice. DTX-2 was discovered in contaminated Irish shellfish (46). The benthic Prorocentrum spp. and these methods have been reviewed (40. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. dinophysistoxin-1 (DTX-1). California. are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. Fortunately DA is conveniently detected by a variety of LC-linked analytical methods and this is the recommended approach for DA monitoring in EU countries. Dinophysis spp. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. in Monterey Bay. Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). and consequently were often overlooked as the real source of the toxins.4. DA can be detected by the mouse bioassay but the regulatory limit for shellfish is set below this level at 20 µg/g (20 mg/kg) shellfish tissue. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. though this method is most commonly used to analyse plankton and seawater (42). The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. 48. the heterotrophic dinoflagellates Dinophysis spp. as can LC/MS. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present.FISH AND SEAFOOD 7. 7. produce a variety of other 148 .4). and an immunosensor method has also been reported (4). are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. 41).. A rapid immunodiagnostic kit is now available to detect DA in phytoplankton and is reported to work satisfactorily in this application in the field (43).. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. To further complicate matters. Alternatively. However.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. but just over 15 years ago a third toxin. 49). Since then. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . and dinophysistoxin-2 (DTX-2) (Figure 7. In the context of this chapter. Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective.

The parent acids are the only DSP congeners to be found in shellfish tissue.4. Dinophysis fortii. although occasionally fatty acyl esters linked through the 7-OH group (referred to as DTX-3 compounds) are also found and are likely a result of bioconversion in shellfish tissue. Prorocentrum maculosum. In this case Dinophysis was suspected as the culprit organism. Dinophysis caudata. and Dinophysis norvegica among others (47. 49). Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp.1 Origins and distribution The first member of the group was identified in contaminated Japanese shellfish in 1985 and DSP toxins continue to be a threat in those areas. have been referred to as lipophilic shellfish poisoning toxins (LSPs or LSTs) (49) because they are found in the same lipophilic fractions as the DSP toxins and are described in more detail later in this chapter. In comparison. are also recognised as a threat to human health. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. DSP toxins produced by benthic Prorocentrum spp.. Lately these compounds. Dinophysis acuta. which include the pectenotoxins and the yessotoxins. While these esters are not toxic in vitro. 149 . such as Prorocentrum lima. Today over ten Dinophysis spp. and include Dinophysis acuminata. 7.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. are recognised as DSP toxin producers. Structure of okadaic acid and the two related DSP toxins 7. Fig.5. This was probably not the first DSP episode. DTX-5. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). and Prorocentrum belizeanum.

2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. Human symptoms include diarrhoea. although a comparative study of in vitro phosphatase inhibition with mouse bioassay data has shown that OA is about twice as potent as DTX-2 (51).5. it does not provide information on the DSP toxin profile. The 7-O acyl derivatives (DTX-3) of OA that can form in shellfish also show poor cross reactivity and this further exacerbates the problem. most progress in DSP toxin detection has been using new or improved LC/MS methods and a detection limit of 10 µg/kg has been reported.3 days. DSP episodes have been reported from Japan. or indeed on the presence of other associated LSTs that can occur with DSP toxins and which result in a positive mouse bioassay result (vide infra). and while this provides critical seafood safety information. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. vomiting. DSP-Check is a commercially 150 . but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. 7. Due to the lipophilic nature of the toxins. even with the minimal of clean up steps (48). and DTX-2.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . A regulatory level of 80 . Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. New Zealand and Chile. various European countries. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). Since the late 1970s.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. nausea. and a detection limit of 100 µg/kg has been reported (12). 7. DSP episodes have been reported worldwide and are included in many monitoring programs (47). and abdominal pain.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins.5. it was agreed that the established mouse bioassay (52) with an observation time of 24 hours is currently the preferred method for the detection of the acute toxicity of acetone soluble DSP toxins. In recent years. both critical enzymes involved in many key metabolic pathways in mammalian cells (50) and consequently affect a host of other biological processes in a cell including tumor promotion. which can lead to an underestimation of the total DSP content in a sample.

with a claimed detection limit of 20 µg/kg.6 Lipophilic Shellfish Toxins (LST) Lipophilic extraction of toxic shellfish for DSP toxins often results in the simultaneous extraction of other lipophilic polyethers such as the pectenotoxins (PTXs) and yessotoxins (YTXs). 7.6. In yet another approach. Some representative examples of both groups of toxins are shown in Figures 7. 58). with good agreement with the mouse bioassay and LC–based methods (54. but with the increased availability of pure toxins permitting detailed toxicology studies it is now recognised that both these groups of compounds possess a different toxic profile from the DSP toxins. However. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. and the latter group are usually found to be responsible for any reported shellfish toxicity (59).5. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. and another rapid antibodybased test kit for DSP toxins has also been reported (53). 55). If a European collaborative study of the fluorometric protein phosphatase inhibition method is successful it may eventually be approved for regulatory purposes in the EU. Some doubt remains as to the significance of these compounds as marine or shellfish toxins with respect to human health since the PTXs for example are co-extracted with the DSP toxins. Fig.5 and 7. Within this group. 7. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). Indeed. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 .FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. for many years these compounds were included in the DSP toxin complex.

which may co-occur in blooms with other organisms. eastern Canada. 58).6.1 Origins and distribution It is now well recognised that PTXs are co-produced with DSP toxins by various strains of the dinoflagellate Dinophysis and hence the distribution of both families of toxins in shellfish is similar (49).FISH AND SEAFOOD Fig. Structure of the two parent yessotoxins. YTX and homo-YTX 7.6.6. 7. As is typical for many algal species. Japan. Oxidation of the methyl group at C-43 to a carboxyl function as in PTX-6 significantly reduces toxicity (500 µg/kg body weight) as does hydrolysis to PTX-2 seco acid (>5000 µg/kg body weight) a common metabolite found in shellfish extracts. many parts of Europe and the Mediterranean.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). New Zealand and South America (49. While 152 . the toxin content and profile may vary among different strains. PTX-1 and PTX-2 are frequently the most common derivatives to occur and PTX-2 is the most potent following intraperitoneal (IP) injection (230 µg/kg body weight). 57. 7. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum.

a regulatory level of 1 mg/kg shellfish tissue has been set. 58). 7. some of which are found as glycosides and sulfates. chemical analysis is best carried out by LC/MS and several approaches have been described (57. The crude extract may be used directly in the mouse bioassay. which aids detection by LC/UV. Although there is no evidence that yessotoxins are responsible for human illness. Estimates of acute toxicity of the parent compound following IP injection vary considerably. it may be compromised by other biological materials in the matrix. Their toxicology and mechanism of action remain uncertain. Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds. The YTX group has not been reported to cause human illness.6. Because of the number and complexity of the YTX group. The regulatory level of 160 µg/kg shellfish tissue has been established and the mouse bioassay is often the primary detection method although LC/MS methods are preferred and this requires some simple clean-up of the crude extract prior to chemical analysis.FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. There are no clear cases of human illness resulting from ingestion of PTXs (59). While LC/UV detection is possible for PTXs. Some of the PTXs are potent cytotoxins in vitro (60).3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue. Many YTX toxins contain a conjugated diene system. which is further compounded by differences in the experimental design. but a fluorescent derivative has been prepared for others. 64). but less so following oral dosing (61). The structures and chemical data including the molecular weight of each member have been collated (56). The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. the PTXs show considerably less toxicity when administered orally. the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present. However. 7. For many years it was believed that 153 . but various congeners are acutely toxic in the mouse bioassay following IP injection.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7.7). though its cell effects may be linked to modulation of calcium channels (62).

Interestingly. PbTx-1 and PbTx-2. 154 . These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). whelks. 7. and one of the brevetoxin metabolites BTX-B2 7. A complicating factor both in terms of establishing the profile and toxicity of brevetoxin-contaminated shellfish has been the discovery of the so-called “brevetoxin metabolites”. In the Gulf of Mexico.FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida. verruculosa off the mid Atlantic coast of the U. oysters. Structure of the two parent brevetoxins. Many of these species are associated with massive fish kills. 66). was chemically characterised as PbTx-2 (67).S. but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65. Fig. and Heretosigma akashiwo. which are found in Japanese waters (66). and off the coast of New Zealand (65).1 Origins and distribution The brevetoxins are produced by the dinoflagellate Karenia brevis (formerly known as Gymondinium breve and Pytochodiscus brevis) a dinoflagellate found in the Gulf of Mexico. 66). and cockles (65. the same toxins have been reported to be produced by three genera of Raphidophytes. and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf.7. namely Chatonella marina.7. brevetoxins are found in mussels. They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. Fibrocapsa japonica.

The mouse bioassay is used as the primary detection method. and mammalian body fluids without pretreatment or dilution (72). this family of toxins has proven to be a challenge both to identify and monitor. the competitive ELISA method correlated most closely with the mouse bioassay results. 7. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. with hot spots in the coral reef areas of the Caribbean and French Polynesia. 69). with a mean duration of symptoms of around 1 day. The mean time to onset of symptoms is around 3 hours. While the mouse bioassay continues to be used. 7.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). and in severe cases seizure. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. and an action level has been set at 800 µg/kg (or 80 µg PbTx-2 equivalents/100 g or 20 MU/100 g or 4 µg/mouse) of shellfish tissue to trigger regulatory action. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. which can result in a cascade of associated events in a cell (66). Ingestion of brevetoxins results in a variety of symptoms including parathesia. vertigo.7. and diarrhoea. The human health implications following exposure to NSPs have been reviewed (69). 7.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). nausea. In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. shellfish homogenates. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . In this application. malaise.7. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). 66. The brevetoxin metabolites are reported to be less toxic than the parent compound (70).2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater.

Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red. 7. and only a few structures have been identified. a group of closely related polyethers that are considered the source of ciguatoxins. Fig. A detailed comprehensive review of ciguatoxins has recently been published (74). The diversity of benthic phytoplankton that exist on reefs further complicates the picture. CTXs are large.1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins.8). Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin.8. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. Distribution through the food chain begins with 156 . These include species belonging to the genera Ostreopsis and Coolia. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. lipid soluble polyethers.8.FISH AND SEAFOOD transferred through the food chain to the larger carnivores. complex. but more are suspected to exist. green and brown seaweeds and also occur in sediments and coral debris.

Another symptom that can develop over time is extreme fatigue. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . 75). and although the gastronomical problems may dissipate within a day or so. Like the brevetoxins.3 Detection methods As a consequence of their potency.76). there are neither standards nor an official testing program. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. 75. barracudas.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. which can be confused with chronic fatigue syndrome. The onset of symptoms can occur rapidly within 24 hours. and moray eels. and there are suggestions that such incidents are increasing and spreading worldwide (74). and it is estimated that ingestion of as little as 0.8. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. 78). the neurological symptoms can persist for days. and the sensation of loose teeth (77). only trace amounts of CTXs can cause illness following consumption of contaminated fish.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. although not all CTXs are equipotent. Local island populations attempt to avoid poisoning by catching only smaller fish. arthralgia. which can return following the consumption of alcohol and certain foods such as oily fish (74.1 µg of ciguatoxin can result in human illness (74. where cold objects feel hot and vice versa. groupers. The recommended method to extract fish tissue has been described in detail (74) and the initial warm acetone extract is partitioned against hexane and diethyl ether where the CTXs are concentrated. 7. myalgia. Patients have reported symptoms lasting weeks and even months. which in turn are consumed by carnivores such as snappers. 7. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. weeks or even months (74 . all of which can be linked to the effect of CTX on excitable membranes. sharks. Nevertheless CTX poisoning continues to be a threat to local and visiting populations.8. and in fact CTX poisoning is often only identified after consumption of contaminated fish. pruritus. These include paresthesias (numbness and tingling of the extremities). though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. Even when suspect samples are available. Other symptoms include a metallic taste. and temperature sensation reversal. Globally.

and following synthesis of the molecule some time later.80%).30%) and AZA3 (5 . specific for CTX activity. 7.9). and another dealing with the chemistry and detection methods has been published (85). and only a few structures (AZA 1 . and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. Since the original discovery and identification of the toxin. which features characteristic fragment ions corresponding to sequential losses of three molecules of water. another in vitro sodium channel-based assay. 84). The majority of these have been proposed based on LC/MS analytical data. minor revisions of a few stereocenters were made. following consumption of contaminated mussels Mytelius edulis (82). and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. Chemical detection methods are severely compromised by the lack of standards. The original structure proposed revealed an entirely new class of shellfish toxin.85). Immunoassays may be an alternative detection method (79). at C-3 and C-23). which are based on an LC-MS/MS approach. Recent reviews of this new group of toxins have appeared (83. Consequently.g.9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). followed by clean-up on silica and aminopropyl cartridges. although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). only a few laboratories are able to undertake such analyses. but this only has application in a clinical setting (81). Acetone extracts of fish tissue are subjected to solvent partitioning.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis.9).20%) present in lesser amounts (Figure 7. Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. 158 . Recently. with AZA2 (10 . has been developed. high-throughput screen (74).FISH AND SEAFOOD Lethal Dose (MLD) 0. This is then subjected to LCMS/MS analysis.25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. and the corrected structure was published in 2004 (86) (Figure 7. Usually the major AZA derivatives found in shellfish are AZA1 (50 . almost 20 additional derivatives have been reported (83 . A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine.

Structure of the three principal azaspiracids found in shellfish 7. scallops (Pecten maximus). There has been considerable debate over the source of azaspiracids. and most recently in a crustacean (Cancer pagurus). and to date this has been the only source from which these toxins have been found at toxic levels. it has been impossible to establish that P. In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. and some time after the original episode in Ireland. Mytilus galloprovincialis). 84. cassipes produces AZAs in culture. vomiting.9. diarrhoea and stomach cramps. which are ingested by P.9. which can persist for 2 . Donax spp. these molluscs were never considered to be the biogenetic source.FISH AND SHELLFISH TOXINS Fig. and to date no long-term effects have been recorded (83. but current data indicates that unlike the DSP compounds. Portugal. AZA toxins do 159 . 7.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. 90). it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). Following the original episode. Ensis siliqua. oysters (Crassostrea gigas. Scandinavia. These symptoms are not unlike those experienced following the consumption of DSP toxins.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. While this result is not necessarily in debate. AZA occurrence has been reported by several European countries including France. 88.3 days. However. as in other cases of shellfish toxins. clams (Tapes philipinarum. 7. Ostrea edulis). The bivalves affected include mussels (Mytilus edulis. cassipes (83). and off the coast of Morocco (83. cockles (Cerastoderma edule).9.). 89). and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp.

Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. the lack of sufficient material and other logistical problems have prevented complete studies of any chronic effects arising from long term exposure to low levels of the AZA toxins. Using the same approach. 91. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic.10 – 7.12). Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. though the possibility exists that they may inhibit other phosphatases. The mouse bioassay has been used to monitor shellfish extracts. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. and necrosis of epithelial cells on the microvilli (91). abnormalities in the thymus and spleen. which are particularly amenable to detection by positive ion electrospray mass spectrometry. interferences can arise when analysing a matrix as complex as a crude shellfish extract and it is recommended that a simple solid phase extraction (SPE) step be introduced prior to LC/MS analysis (93). Importantly these pathological outcomes were reported as being different from those caused by DSP. 92). the pinnatoxins.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. it is recommended that the entire shellfish be used for AZA analysis since it has been shown that these toxins can migrate to other tissues. 7. Oral administration of AZAs at substantially higher doses than those used in IP injection studies did not result in any mouse mortalities after 24 hours. 85). but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. Based on IP minimum lethal dose data. PSP. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . However.9.10 Cyclic imines These toxins are represented by several distinct structural types. 7. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. the spirolides. AZAs also display cytotoxicity towards various mammalian cell lines (83. and erosion and bleeding in the stomach. but all contain a spirocyclic or cyclic imine function within their structure. and closely related ptierotoxins (Figures 7. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. parenchymal cell pyknosis in the pancreas.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. and ASP. Once again. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg).

gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7.10. cockles..10. At the time of the initial observation of gymnodimine. Structure of the cyclic imine gymnodimine 7. it was proposed that a dinoflagellate identified as Gymnodinium spp. surf clam and New Zealand abalone. In only two cases (gymnodimines and spirolides) has a planktonic source been identified. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. 95).520 Da (94. Pacific oyster. 7. they are sometimes referred to as emerging toxins of uncertain human health concern.10). The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . 7.1. but they are included here for completeness. Some time later. Fig. selliformis (97). Cyclic imines are often referred to as fast acting toxins due to their “all or nothing” effect in the mouse bioassay over a very narrow dose range. two additional congeners. 95). 161 .1 Gymnodimines The observation of neurotoxic symptoms in the mouse bioassay of lipophilic extracts of dredge oysters (Tiostrea chilensis) collected from around the South Island of New Zealand.FISH AND SHELLFISH TOXINS health. Subsequent taxonomic studies revised this again to a new dinoflagellate K.10. scallops.1 Origins and distribution The original observation of toxicity in oysters led to a wider survey of other shellfish species in New Zealand when it was discovered that the toxin was present in a variety of bivalve molluscs including mussels. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin.

together with a lack of evidence of human intoxication.to10-fold reduction of toxicity (LD50 600 . Since their original discovery which resulted in the characterisation of spirolides A-F (99.10.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). 7.100). Oral administration of the toxin to mice by gavage resulted in about an 8. and this is the detection method of choice.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. finishing with 100% methanol. the first step in the process is typically an initial clean-up step using a short C18 cartridge and elution with water containing increasing amounts of methanol. almost a dozen derivatives in total have been identified to date (94. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. However.10.FISH AND SEAFOOD 7. 50%) of 96 µg/kg. Whether isolated from dinoflagellate cells directly or from shellfish tissue.11). The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. They were originally discovered in extracts of mussels (M.3 Detection methods The characteristic “fast-acting. In particular. The time lapse between ingestion and death is between 5 and 15 minutes. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94.1. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. A later study using purified toxin established an LD50 dose (Lethal Dose. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction.1. abdominal breathing. paralysis of the hind quarters and eventually complete immobility. 95). it has been stated that regulation against gymnodimine is not required (98). and so spectroscopic detection by UV methods is not favoured.900 µg/kg).10. and lack of responsiveness to stimuli. Based on this diminished oral toxicity. There is a rapid onset of symptoms including respiratory distress. 7. Most cyclic imines lack an extended chromophore. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. Reduction of the imine group eradicated the toxicity. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. 95) (Figure 7.

Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. 7. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 .1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. 95). All data reported here are for desmethyl spirolide C. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 .11.11.2 Toxicology The spirolides are the most studied group of the cyclic imines. the gulf of Maine.2. fasted mice). the western coast of Scotland.166 µg/kg.10.707 µg/kg. While oral toxicity is considerably less than by IP injection.10. Fig.2. 7. Structural differences among the various spirolides result from variations in the number of methyl groups and the sequence of spiroketal rings 6-5-5 in one group and 6-6-5 in the other. by gavage (87 .8 µg/kg). fasted mice). their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. those spirolides possessing the vicinal dimethyl grouping (see Figure 7. Interestingly. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101).FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). Structure of four spirolide toxins A-D 7. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). southern Ireland. or by oral ingestion (381 . in fjords in Denmark and Norway. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98).

Structure of the pinnatoxin group and the closely related pteriatoxins 164 . Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. 102). more useful information on the amount and distribution of toxins present in a shellfish extract is obtained by LC-MS analysis usually following a simple clean-up step. A number of methods following this procedure have been described (94.10.FISH AND SEAFOOD (100).12). Three members of this group have been reported. 7. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. However.B contain a spiro imine moiety similar to that found in the spirolides. To date.12. the mouse bioassay can be used to detect these fast acting toxins. four members of the group have been identified – two of them being the C-34 diastereomers. 95. While the mechanism of action remains to be clearly defined. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C. The compounds are zwitterionic and therefore somewhat more polar than the spirolides.3 Pinnatoxins and pteriatoxins Pinnatoxins A .3 Detection methods As discussed for gymnodimine. and in fact these compounds share a number of other structural features.10. 7.2. consistent with modulation of acetyl cholinergic receptors (103). which includes two diastereomers. Fig. 7.

7.1 Origins and distribution Shellfish of the genus Pinna. which occupy tropical waters around Okinawa and the south east coast of China. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures.100 µg/kg). new toxin groups such as the azaspiracids and cyclic imines have also been reported. 7. 7. With increasing knowledge of the chemistry of toxins.3.3. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105).10. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other.10. like the gymnodimines and spirolides. However. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). Despite this progress. and more advanced means of detection.3 Detection methods No formal detection methods for these compounds have been reported. 99%) values for pinnatoxins A-C are in the range (22 . which in many cases are now found to be more widespread than previously believed.3. there is no compelling evidence to suggest that they have resulted in human toxicity (98). while pinnatoxin D was essentially inactive. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins.11 Conclusion There has been considerable progress in the detection of known marine toxins.FISH AND SHELLFISH TOXINS 7. This is generally a result of increased monitoring and continually improving detection technologies and methods. have long been suspected of causing human illness after ingestion. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. While this is recognised as a real possibility for a wide range of toxins. to date action or 165 . but this remains to be conclusively established (106). arise from an algal source .2 Toxicology There is little information describing the toxicology of these compounds (98).180 µg/kg).most likely a dinoflagellate.10. research and certified standards of the key toxins are often unavailable or in short supply and in the future considerable logistical effort will be required to address this issue. Coupled with this. The LD99 (Lethal Dose. The LD99 dose for the pteriatoxins is reported to be in the range (8 . There is strong circumstantial evidence to suspect that these toxins. Only the acute toxicity in mice following IP injection has been reported.

Seafood and Freshwater Toxins.M. Hallegraeff G.. 5. Recent advancements in surface plasmon resonance immunosensors for detection of small molecules of biomedical. In most cases this is due to the paucity of material that is usually available for any single group of toxins.P. Even if they do not completely replace other methods such as the mouse bioassay or chemical analysis that require sophisticated chemical analytical instrumentation.. While the mouse bioassay continues to be the mainstay of most monitoring programs. 7. Furlong C. Enzymatic recycling-based amperometric immunosensors for the ultrasensitive detection of okadaic acid in shellfish. O’Sullivan C. 166-74. thus significantly reducing the number of mouse or chemical analyses that have to be performed. Toxicon. Guilbault G. CRC Press. Anderson D.C. 2007. Pravda M. Miura N.. Marty J-L. Wekell J.C. 6. 2008. 2003.M.. 10.D.. Biosensors and Bioelectronics. 2004. Shankaran D. Marty J-L. Stevens R. Trainer V.. 2007. Harmful Algae. Novel electrochemical immunosensors for seafood toxin analysis. 2008. de la Igelsia P.L. Talanta. Sensors and Actuators B: Chemical.. Biosensors to detect marine toxins: Assessing seafood safety. Le Berre M.. Gobi J.L. 716-22. 72. Botana L.K. Fish and shellfish toxins. and in particular the ruggedness of such methods.C. Manual on Harmful Marine Microalgae. Campas M. Campas M. Leatherhead.. Lawley R.. 7. 166 4. Wright J. 1735-45. 179-99.. Diogene J..M. Rome. Prieto-Simon B.M. Paris. FAO Food and Nutrition Paper 80. in order to fully study the synergistic effects of multiple toxin types. Eds.. Cembella C.. 3. 2006. in Microbiology Handbook: Fish and Seafood. 40. Soelberg S. and environmental interest. Food and Agriculture Organisation. Gibbs P. ... they can provide a valuable “yes/no” filter for shellfish and finfish extracts.12 1. never mind the amounts required of several groups.A. The challenge facing such approaches is the variety of matrices that must be examined. Boca Raton. an encouraging development is the use of antibody-based methods to detect the presence of marine toxins. 1267-74. Toyofuku H.. 121.L.FISH AND SEAFOOD regulatory levels for toxins continue to be assessed only for individual purified toxins. Chinowsky T.. 24 (4). 8. but these are gradually being overcome. 6. Joint FAO/WHO/IOC activities to provide scientific advice on marine biotoxins (research report). Kane M. Detection of toxin domoic acid from clam extracts using a portable surface plasmon resonance biosensor. food. 9.V.. 884-95.M. 2007. Marine Pollution Bulletin. 2. Marine Biotoxins. Leatherhead Food RA. Eberhart B-T. Spencer S. 2002. UNESCO. References Kreuzer M. Food and Agriculture Organisation of the United Nations. 52...R. 1998.E. 15871.G..

Ed. Christian B. 505-8.. Cembella C.H. Anderson D. Naoki H. Food and Agriculture Organisation of the United Nations. Ben-Gigirey B. 117-34. 2008. Van Amerongen A. 13. 177-96.M.. Gaithersburg... Boca Raton.L.. 10-7.R. 2008. Aune T. 290. Jellet J.. Barrett R. Shoji Y...M. and liquid chromatography/tandem mass spectrometry.. Paralytic Shellfish Poisoning. Yotsu-Yamashita M. Detection of paralytic shellfish poisoning (PSP) toxins in shellfish tissue using MIST Alert. in Rapid Methods for Biological and Chemical Contaminants in Food and Feed. Electrospray ionisation mass spectrometry of tetrodotoxin and its analogs: Liquid chromatography/mass spectrometry.. in Methods of Analysis of the Association of Official Analytical Chemists. Botana L. Krys S. Food and Agriculture Organisation of the United Nations. 391 (1). in FAO Food and Nutrition Paper 80. 18.M.M. Luckas B.. 16. 24. UNESCO. Eds. Barug D.A. Lauwaars A. 2008. Mackintosh F. Tetrodotoxin – distribution and accumulation in aquatic organisms and cases of human intoxication. Endo A. Determination of marine biotoxins for regulations: from the mouse bioassay to coupled LC-MS methods. 21. 19. 349-71.. Rapid detection methods for marine toxins: needs and limits. 22. Association of Official Analytical Chemists. 2008.J. Pure and Applied Chemistry. prechromatographic oxidation and liquid chromatography with fluorescence detection. Turrell E. Marine Drugs. 283-317.E. Wageningen. Risk assessment of marine toxins.. 51. 40. 1995.V.L. Quilliam M. Marine Biotoxins.. Toxicon. 6. 12. Toxicon. Neurotoxins from marine dinoflagellates: A brief review. Yasumoto T. Lacaze J-P. Yasumoto T. Roberts R. 17... Botana L. Analytical and Bioanalytical Chemistry.. in Seafood and Freshwater Toxins. 2008. 2002.M.. in Manual on Harmful Marine Microalgae. 2008. CRC Press. Ed. 21. 3-20. Stobo L.A. Arakawa O.06. 23. Chemical analysis. 61... Wang D-Z. tandem mass spectrometry. Boca Raton. Evaluation of MIST Alert rapid test kits for the detection of paralytic shellfish poisoning toxins in shellfish. Rome. Ed. Marine Drugs. 5-52. Interspecies distribution and biogenetic origin of tetrodotoxin and its derivatives.. 230-42. 15. method 2005. 2006. 14. 219-27. Eds.C.. Villar-Gonzalez A. 1407-25. Larsen J.A. Paris. Fukuyo Y. Surveillance of algal toxins in shellfish from Scottish waters. Taylor F. CRC Press. Petrie J. Wageningen Academic Publishers. 1989.. Laycock M.A. Hallegraeff G.. Miyazawa T. Murata M. Analytical Biochemistry. 2005. 2002. 6. Noguchi T.. 2001. Scott A. Yotsu M. 20. Paralytic shellfish poisoning toxins in shellfish. 635-48. in Seafood and Freshwater Toxins. Taxonomy of harmful dinoflagellates. Journal of Shellfish Research. Food and Agriculture Organisation. 2004. 455-60. AOAC International.FISH AND SHELLFISH TOXINS 11.F. in parallel with the regulatory AOAC mousse bioassay. 167 . Smith E.C. a new rapid test.

Gulland F. Doucette G. 2005. 2000..J.D. de Freitas A.A. Journal of Toxicology – Toxin Reviews.G. Walter J... isolated from the ovaries of puffer fish Fugu rubripes..L. Hanic L.E. 1992. Kotaki Y. CRC Press.. Atergatis floridus. Walter J. 168 . Xie L. Haulena M. Fritz L... Ed.P. 46. Canadian Journal of Fisheries and Aquatic Sciences. 1203-15. Foxall R. 99. Odense P. Ed.. Canada. Silver M. Jeon J. 481-90. Botana L. Moeller P.. Miller P. Trainer V. Wang S.. McInnes A.A. Smith J. Canadian Journal of Chemistry. 1-8.A. Barr B.A. Boca Raton... McCulloch A...A.. Boca Raton..C. Wright J. Amnesic Shellfish Poisoning (ASP). Wright J. de Freitas A..C.. McLellan W. Mortality of sea lions along the central California coast linked to a toxic diatom bloom. Beale A. Hashimoto K.H...A. Norcardiopsis dassonvillei.A.. Loscutoff S. Tasker A. Sugita H... Epidemiology of domoic acid poisoning in brown pelicans (Pelicanus occidentalis) and Brandt's cormorants (Phalacrocorax penicillatus) in California. 35... Xia G. 31. Identification of domoic acid... 218.. 2004. 1993.. Rowles T. Odense P. Rome. Foxall R.A.V. Ecobiology of amnesic shellfish toxin producing diatoms. McCulloch A.. Busman M. Laycock M. in FAO Food and Nutrition Paper 80. Nie Y.. Sim P. Food and Agriculture Organisation.. Dewar D. Lipscomb T.. Todd E... Walter J.. 80-4.A. Ed. in toxic mussels from eastern Prince Edward Island. Gilgan M.S.. Noguchi T. Gilgan M. 2001. Botana L. Wright J.FISH AND SEAFOOD 25.. 33.J... 29.. 397-429. Huang X. 2008.A. 1986. Cordaro J.. 1989.M.C.... 30. 28. Interaction of domoic acid and several derivatives with kainic acid binding sites and AMPA binding sites in rat brain...A. a toxin in shellfish from eastern Prince Edward Island. Chavez F.A. Marine Biotoxins. Hampson D. Powell C. 851-9. 24. Ragan M. pharmacology.M... Falk M. Boyd R.C..P.. Bird C... Nature. Lowenstine L.D.. Van Dolah F.. 27. Silvagni P.. 311-4. 54-62. Shida Y.K. Domoic acid: Detection methods. DeLong R..H.C..L. Quilliam M. Current methodologies for detection of tetrodotoxin.D. 35-50.. Scholin C.L. Noguchi T. Wells J. 20...V. CRC Press.S..C.G.A.W..W. and toxicology. in Seafood and Freshwater Toxins.. in Seafood and Freshwater Toxins. Sim P.W.K. Doucette T. Occurrence of tetrodotoxin and anhydrotetrodotoxin in Vibrio spp.L. Mahmud Y.S.L. Benson S.. Marin R. Pennate diatom Nitzschia pungens as the primary source of domoic acid. Deguchi Y.A. De Vogelaere A. 67. Richard D. 2008.A. Work T.. 97-135...M.A.. 32. Hu J. isolated from the intestines of a xanthid crab.J. III.W.. Toxicon.M.. Zhang R. Thibault P. Food and Agriculture Organisation.G. Pathak V. Subba Rao D... Pocklington R.. 403. 383-96.J.. 1989... European Journal of Pharmacology. a neuroexcitatory amino acid. 26. Quilliam M. Journal of Biochemistry (Tokyo). 34. Journal of Zoology and Wildlife Medicine.R. Wright J.. A new tetrodotoxin-producing actinomycete. Arakawa O. Spraker T. Lefebvre K.. Harvey J. Bates S.W.. Johnson G. Wu Z.. Zhang J. 45. Jamieson W... Food and Agriculture Organisation of the United Nations. Quilliam M.

2008.D.A..S. 2008. Remis M.. Murata M. in Seafood and Freshwater Toxins. Trends in Analytical Chemistry. Stobo L.R. 1990.. Cembella C.C. 38. Quilliam M.. Clardy J..L.E. 180-219.M. Wright J. Kennedy G.J.J. Milley J. Harmful Algae...M.. Walter J. LC-UV and LC-MS methods for the determination of domoic acid. Marine Drugs. 45.. New England Journal of Medicine. 47. 48... Jeffery B. Boca Raton.A. Ed. Boyle C.... Pseudonitzschia in New Zealand and the role of DNA probes and immunoassays in refining marine biotoxin monitoring programs. 53-96.... Rhodes L.D.A..S. 1019-25. Yasumoto T. Anderson D. 37.K. Tetrahedron. Eds. Hallegraeff G.. 443-58. in FAO Food and Nutrition Paper 80. McGovern E. Perl T... Trace determination of domoic acid in seawater and phytoplankton by high performance liquid chromatography of the fluorenylmethoxycarbonyl (FMOC) derivative. Turrell E. 6. 2004. Neurologic sequelae of domoic acid intoxication due to the ingestion of contaminated mussels. 1998.C. Doyle J. Hu T. 42. 169 .L. 322..M.. 44. 41. Pulido O. 41. Morris S. Chemistry. deFreitas A. 1775-80. 24.M.W. McMahon T. 2005. New England Journal of Medicine. Sano M.. Journal of the Chemical Society Chemical Communications. Diarrhetic shellfish toxins.M. Matsumoto G..FISH AND SHELLFISH TOXINS 36. CRC Press.A. 113-33.M. 351-68... 1985. Methods for domoic acid the amnesic shellfish poison.. 39.. Ed. 209-28. Pocklington R. Brown J. Oshima Y... Hess P. Carpenter S. McNabb P...A.. International Journal of Environmental and Analytical Chemistry. Food and Agriculture Organisation. Gjedde A. 1992. Detection of Pseudonitzschia (Bacillariophyceae) species and amnesic shellfish toxins in Scottish coastal waters using oligonucleotide probes and the Jellet Rapid Test. Gallacher S..C. Jackson D. Young P. Food and Agriculture Organisation Diarrhoeic Shellfish Poisoning (DSP).C.. Todd E... Cashman N. Bird C. 1781-7. Bedard L. Food and Chemical Technology. Natural Toxins. Quilliam M. 1990. Isolation of a new diarrhetic shellfish poison from Irish mussels.. 7.A. Zatorre R. Gendron D. Amnesic shellfish poison. Collins L. 40.. Pleasance S. Kotsatsky T.... Nixon E.B. in Manual on Harmful Marine Microalgae. metabolism and chemical analysis of okadaic acid group toxins. UNESCO..G. Wright J. Botana L.C.. 1995. 2004. Bates S. Marine Biotoxins. Moizer K. 46.S.. Slattery D. 105-11.C. 2008. Quilliam M.. Food and Agriculture Organisation of the United Nations. Brown N. Garthwaite I.. Graham N. Teitelbaum J.. 322. Marr J.. 1990. Hockin J. Bresnan C. McEvoy D. Evan A. Paris. Rome. 43..G. 38. Scholin C. 42.. 6. 545-57. Barlow T. Domoic acid toxicologic pathology: A review. 358-67. 39-41. Paul S. An outbreak of toxic encephalopathy caused by eating mussels.

6. Botana L.. 2008. Fleming L. 73-102. Hammond R. Alfonso A. metabolism and chemical analysis. 170 .. Botana L.M.E. 2008. 55. 141-75. 5. 315-27.V. Ed.M. Jellet J. Ed. Toxicology of the pectenotoxins. Oshima Y. 44. 2006. Boca Raton.A. 1997. Munday R.. a group of marine polyether toxins: an overview. Bulletin of the Japanese Society of Science and Fisheries. 361-9. Marine toxins. Vieytes M.. Botana L. Toxicon. 371-80.. 343-9. Harmful Algae.. Fontal O. in Seafood and Freshwater Toxins. Boca Raton. Part II.. 909-22. 59. 2008. Boca Raton. Pharmacology of pectenotoxins.M.F. 258-64.M. Bean J. 2008. Fattorusso E. Occurrence of a new type of shellfish poisoning in the Tohoku District. Natural Toxins and Drugs.J. 58. 287-314. CRC Press.FISH AND SEAFOOD 49.. De Wolfe F.R. First report of a new rapid assay for diarrhetic shellfish poisoning toxins.. 74-8. Bornancin A. 56. 2008. Espina B. Amsterdam.. Boca Raton. in Seafood and Freshwater Toxins. Fernandez J. Laresen S. 52. in Seafood and Freshwater Toxins. 51..A.. 1-7. Easy D. Franco J.. 57.. Analytical Biochemistry. Chemistry. 50. in Seafood and Freshwater Toxins. in Seafood and Freshwater Toxins. Baptista de Souza J.H.. Yasumoto T.. Donovan M. Ed. Vilarnino N.M. Botana L. Boca Raton.. Norte M. Pizzaro G. CRC Press. 2001. CRC Press. Aune T. Ed. CRC Press. 2008. Boca Raton. Rossini G.M. 2008. Yessotoxins. Biological detection. Ed.M.L. Reich A. CRC Press.J. Relative toxicity of dinophysistoxin-2 (DTX-2) compared with okadaic acid based on acute intraperitoneal toxicity in mice.. Riobo P. Boca Raton. Toxicology of the yessotoxins. Aune T. metabolism and chemical detection methods of pectenotoxins.. in Seafood and Freshwater Toxins.O. 63. Hungerford J... Neurotoxic shellfish poisoning. Munday R.P. Baden D. Daranas A. in Seafood and Freshwater Toxins. Ciminiello P. Rehman N.. 248. Sosa S. Planktonic dinoflagelates that contain polyether toxins of the old “DSP complex”. 2008. Watkins S. 431-55. Laycock M. CRC Press. Alfonso C.B. Hess P. Mountfort D.A. in Handbook of Critical Neurology: Intoxications of the Nervous System.. in Seafood and Freshwater Toxins.. Truman P. Protein phosphatase inhibition assay adapted for determination of okadaic acid in mussels. CRC Press. 2008. Ed. Botana L. Reguera B. Elsevier. 2008. 54. Botana L. 49.. Suzuki T. Ed. 53. Tubaro A. Ed.. Botana L. 39. Botana L. Botana L. 64.M. 2007. A fluorescent microplate assay for diarrheic shellfish toxins. 1249-55.. 257-84. 329-39. Yessotoxins. Paz B. 62.M. Yessotoxins.. CRC Press.A. Aasen J. Ed. Boca Raton. Pharmacology and toxicology of diarrhetic shellfish toxins. Leira F. 61.. 6. Yamaguchi M.. Fleming L. Suzuki T.M. 1995. 60.G... Marine Drugs. Satake M. 1978. Toxicon..M..M. Marine Drugs.E. Chemistry. Pharmacology and mechanism of action.. 229-53.

Marine Drugs...S. shellfish and mammalian body fluid.... Baden D. Harmful Algae. Naar J. Lenzi A. The molecular and integrative basis to brevetoxin toxicity. Zaias J.. 77. in Seafood and Freshwater Toxins. 67. Cheng Y.M.H. in Seafood and Freshwater Toxins.J. Backer L.. Ed.. 23.. Wanner A. Poon-King T. 707-23. Henry M. Dechraoui M-Y. Jacocks H. Granholm A. Ramsdell J. Naar J.FISH AND SHELLFISH TOXINS 65. 2002. 187-94. Botana L. Abraham W. Steidinger K. 71.. 99-115. and detection. Buonocore M. Brevetoxins.. 110. Backer L.R. in FAO Food and Nutrition Paper 80.M. 171 . Ciguatera toxins: Chemistry. Kubanek J...E. Watkins S. 76. 6.. 50. Abbott J. 2000. 73. Wang Z. Literature review of Florida red tide: implications for human health. Boca Raton. Ed. Dickey R. Rome.S. Gannon D.G.S.P. 68. Weisman R. Tomas C. 220-6.E. Pierce R. Wolny J.. Bourdelais A. 72. 110.. Muscle Nerve. Naar J.G.. toxicology. Environmental Health Perspectives.D.. 70.P. Mild ciguatera poisoning: case reports with neurophysiological evaluations. Kirkpatrick B. 1598-603.G. 2004. 2007. Clark R. Locatelli C. Botana L. 69. in Seafood and Freshwater Toxins. 2004. Schrank K.M... Fleming L..... 479-516..G. Gandini C. Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2. 117-71. 551-8.. 519-50.B.. Fleming L. Benson J... Poli M. 50. New fish-killing alga in coastal Delaware produces neurotoxins. Boca Raton.. Bienfang P. 66. Aerosolised red tide toxins (Brevetoxins) and asthma. Ed... 2008. like ciguatoxins. Dickey R. Pierce R.. 131.G.... Ciguatera fish poisoning: treatment prevention and management..P... CRC Press.. 2002. Freidman M.. Backer L. Bean J.. 2008. 456-79.. Landsberg J.. Food and Agriculture Organisation of the United Nations... Ayyar R.. Granade R. Chest.. Toxicon. Fernandez M. 2007. Bourdelais A. Chen A.. Baden D. Reich A. Boca Raton.A. Kubanek J.. Zaias J. Johnson D. 2004. West Indian Medical Journal. Bottein M-Y. Baden D.S..A. 75.M.. Tomas C. Kirkpatrick B. Fleming L.J. Flewelling L. Abraham W.D. are potent ichthyotoxic neurotoxins that accumulate in fish.E. 2007. 465-70.C.W... 74.A. Ramsdell J.N. Toxicon. Environmental Health Perspectives. Marine Biotoxins. 3. 179-85. CRC Press. Lancaster J. Botana L. Manzo L. 53. Prockop L. Neurologic Shellfish Poisoning (NSP)...C.M. Squicciarini D. Naar J. Food and Agriculture Organisation.. Cheng Y.A.. Baden D. Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater... Pierce R... Baden D. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. Reich A. Detection of brevetoxins in the twenty-first century.E.E. Flewelling L.. 825-34.. 2008. CRC Press. Butera R. Whitney P.. Ed.. 2008. Bossart G. Food and Agriculture Organisation. Poon-King C..

FISH AND SEAFOOD 78. 79. Galziou P., Marti P.M.V. Study of factors that influence the clinical response to ciguatera fish poisoning. Toxicon, 1993, 31, 1151-4. Hokama Y., Ebesu J.S.M., Nishimura K., Oishi S., Mizuo B., Stiles M., Sakamoto B., Takenaka W., Nagai H. Human intoxication from Hawaiian reef fishes associated with diverse marine toxins. Journal of Natural Toxins, 1996, 5, 235-47. Park D.L. Detection of ciguatera and diarrheic shellfish toxins in finfish and shellfish with Ciguatect kit. Journal of AOAC International, 1995, 78, 533-7. Bottein-Dechraoui M.Y., Wang Z., Turquet J., Chinain M., Darius T., Cruchet P., Radwan F.F., Dickey R.W., Ramsdell J.S. Biomonitoring of ciguatera exposure in mice using blood collection cards. Toxicon, 2005, 46, 243-51. Satake M., Ofuji K., Naoki H., James K.J., Furey A., McMahon T., Silke J., Yasumoto T. Azaspiracids, a new marine toxin having unique spiro ring assemblies, isolated from Irish mussels Mytilus edulis. Journal of the American Chemical Society, 1998, 120, 9967-8. Twiner M.J., Rehmann N., Hess P., Doucette G.J. Azaspiracid Shellfish Poisoning: A review of the chemistry, ecology and toxicology with an emphasis on human health impacts. Marine Drugs, 2008, 6, 39-72. Food and Agriculture Organisation of the United Nations. Azaspiracid Shellfish Poisoning (AZP), in FAO Food and Nutrition Paper 80. Marine Biotoxins. Ed. Food and Agriculture Organisation of the United Nations. Rome, Food and Agriculture Organisation. 2004, 173-84. James K.J., O’Driscoll D., Fernandez J.G., Furey A. Azaspiracids: chemistry, bioconversion and determination, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 763-73. Nicolao K.C., Koftis T.V., Vyskocil S., Petrovic G., Ling T., Yamada T.M.A., Tang W., Frederick M.O. Structural revision and total synthesis of azaspiracid-1, Part 2: Definition of the ABCD domain and total synthesis. Angewandte Chemie International Edition, 2004, 43, 4318-24. James K.J., Moroney C., Roden C., Satake M., Yasumoto T., Lehane M., Furey A. Ubiquitous ‘benign’ alga emerges as the cause of shellfish contamination responsible for the human toxic syndrome azaspiracid poisoning. Toxicon, 2003, 41, 145-51. Magdalena A.B., Lehane M., Krys S., Fernandez M.L., Furey A., James K.J. The first identification of azaspiracids in shellfish from France and Spain. Toxicon, 2003, 42, 105-8. Vale P., Bire R., Hess P. Confirmation by LC-MS/MS of azaspiracids in shellfish from the Portuguese north-western coast. Toxicon, 2008, 51, 1449-56. Ryan G., Cunningham K., Ryan M.P. Pharmacology and Epidemiological impact of azaspiracids, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 755-61. Ito E. Toxicology of azaspiracid-1: Acute and chronic poisoning, tumorigenicity, and chemical structural relationship to toxicity in a mouse model, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 775-84.

80. 81.








89. 90.



FISH AND SHELLFISH TOXINS 92. Twiner M.J., Hess P., Bottein-Dechraoui M-Y., McMahon T., Samons M.S., Satake M., Yasumoto T., Ramsdell J.S., Doucette G.J. Cytotoxic and cytoskeletal effects of azaspiracids-1 on mammalian cell lines. Toxicon, 2005, 45, 891-900. Moroney C., Lehane M., Brana-Magdalena A., Furey A., James K.J. Comparison of solid phase extraction methods for the determination of azaspiracids in shellfish using electrospray liquid chromatography-mass spectrometry. Journal of Chromatography, 2002, 963, 353-61. Molgo J., Girard E., Benoit E. Cyclic imines: An insight into this emerging group of bioactive marine metabolites. Phycotoxins, 2007, 319-35. Cembella A., Krock B. Cyclic imine toxins: Chemistry, biogeography, biosynthesis and pharmacology, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 561-80. Seki T., Satake M., MacKenzie L., Kaspar H., Yasumoto T. Gymnodimine, a new marine toxin of unprecedented structure isolated from New Zealand oysters and the dinoflagellate Gymnodinium sp. Tetrahedron Letters, 1995, 36, 295-9. Haywood A.J., Steidinger K.A., Truby E.W., Bergquist P.I., Adamson J., Mackenzie L. Comparative morphology and molecular phylogenetic analysis of three new species of the genus Karenia (Dinophyceae) from New Zealand. Journal of Phycology, 2008, 40, 165-79. Munday R. Toxicology of cyclic imines: Gymnodimine, spirolides, pinnatoxins, pteriatoxins, prorocentrolides, spiro-centrimine, and symbioimines, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 581-94. Hu T., Curtis J.M., Oshima Y., Quilliam M.A., Walter J.A., Watson-Wright W., Wright J.L.C. Spirolides B and D, two novel macrocycles isolated from the digestive glands of shellfish. Journal of the Chemical Society, Chemical Communications, 1995, 2159-61.


94. 95.





100. Hu T., Burton I.W., Cembella A.D., Curtis J.M., Quilliam M.A., Walter J.A., Wright J.L.C. Characterisation of spirolides A, C, and 13-desmethyl C, new marine toxins isolated from toxic plankton and contaminated shellfish. Journal of Natural Products, 2001, 64, 308-12. 101. Cembella A.D., Lewis N.J., Quilliam M.A. The marine dinoflagellate Alexandrium ostenfeldii (Dinophyceae) as the causative organism of spirolide shellfish toxins. Phycologia, 2000, 39, 67-74. 102. Ciminiello P., Dell’Aversano C., Fattorusso E., Magno S., Tartaglione L., Cangini M., Pompei M., Guerrini F., Boni L., Pistocchi R. Toxin profile of Alexandrium ostenfeldii (Dinophyceae) from the northern Adriatic Sea as determined by liquid chromatography-mass spectrometry. Toxicon, 2006, 47, 597-604. 103. Gill S., Murphy M., Clausen J., Richard D., Quilliam M., MacKinnon S., LeBLanc P., Mueller R., Pulido O. Neural injury biomarkers of novel shellfish toxins, spirolides: a pilot study using immunochemical and transcriptional analysis. Neurotoxicology, 2003, 24, 593-604. 104. Uemura D., Chou T., Haino T., Nagatsu A., Fukuzawa S., Zheng S.Z., Chen H. Pinnatoxin A: a toxic amphoteric macrocycle from the Okinawan bivalve Pinna muricata. Journal of the American Chemical Society, 1995, 117, 1155-6. 173

FISH AND SEAFOOD 105. Takeda N., Uemura N., Suenaga K., Uemura D. Structural determination of pteriatoxins A, B, C, extremely potent toxins from the bivalve Pteria penguin. Tetrahedron Letters, 2001, 42, 3495-7. 106. Kuramoto M., Arimoto H., Uemura D. Bioactive alkaloids from the sea. Marine Drugs, 2004, 1, 39-54.


Rhea Fernandes and Dr. Peter Wareing Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 8.1 Introduction

The Hazard Analysis Critical Control Point (HACCP) system is a structured, preventative approach to ensuring food safety. HACCP provides a means to identify and assess potential hazards in food production and establish preventive control procedures for those hazards. A critical control point (CCP) is identified for each significant hazard, where effective control measures can be defined, applied and monitored. The emphasis on prevention of hazards reduces reliance on traditional inspection and quality control procedures and end-product testing. A properly applied HACCP system is now internationally recognised as an effective means of ensuring food safety. The HACCP concept can be applied to new or existing products and processes, and throughout the food chain from primary production to consumption. It is compatible with existing standards for quality management systems such as the ISO 9000-2000 series, and HACCP procedures can be fully integrated into such systems. The new ISO 22000 food safety standard formally integrates HACCP within the structure of a quality management system. HACCP is fully integrated into the British Retail Consortium (BRC) Global Standard for Food Safety, and is one of the ‘fundamental’ requirements of that system. The application of HACCP at all stages of the food supply chain is actively encouraged, and increasingly required, worldwide. For example, the Codex Alimentarius advises that "the application of HACCP systems can aid inspection by regulatory authorities and promote international trade by increasing confidence in food safety". In many countries, there is a legal requirement for all food business operators to have some form of hazard analysis based on HACCP as a means of ensuring food safety. For example, within the European Union, Regulations 852/2004 and


853/2004 require a fully operational, and maintained HACCP system, according to Codex, to be in place. 8.2 Definitions

Control (verb) - To take all necessary actions to ensure and maintain compliance with criteria established in the HACCP plan. Control (noun) - The state wherein correct procedures are followed and criteria are met. Control measure - An action and activity that can be used to prevent or eliminate a food safety hazard or reduce it to an acceptable level. Corrective action - An action to be taken when the results of monitoring at the CCP indicate a loss of control. Critical Control Point (CCP) - A step at which control can be applied and is essential to prevent or eliminate a food safety hazard, or reduce it to an acceptable level. Critical limit - A criterion which separates acceptability from unacceptability. Deviation - Failure to meet a critical limit. Flow diagram – A systematic representation of the sequence of steps or operations used in the production or manufacture of a particular food item. HACCP - A system which identifies, evaluates and controls hazards which are significant for food safety. HACCP Plan – A document prepared in accordance with the principles of HACCP to ensure control of hazards which are significant for safety in the segment of the food chain under consideration. Hazard - A biological, chemical or physical agent in, or condition of, food with the potential to cause an adverse health effect. Hazard analysis - The process of collecting and evaluating information on hazards and the conditions leading to their presence to decide which are significant for food safety and therefore should be addressed by the HACCP plan. Monitoring – The act of conducting a planned sequence of observations or measurements of control parameters to assess whether a CCP is under control.


The application of methods. Figure 8. tests and other evaluations. operation or stage in the food chain including raw materials.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. hygiene managers. Establish procedures for verification to confirm that the HACCP system is working effectively. 5. Determine the CCPs.1 is a flow diagram illustrating this 12-stage logic sequence. ingredient and packaging buyers. The team should include members familiar with all aspects of the production process as well as specialists with expertise in particular areas such as production. Verification .Obtaining evidence that the elements of the HACCP plan are effective. 8.1 Assemble HACCP team HACCP requires management commitment of resources to the process. 6. from primary production to final consumption. The team should also include 177 .A point. It is recommended by the Codex Alimentarius that the practical application of the HACCP principles be approached by breaking the seven principles down into a 12-stage logic sequence. Conduct a hazard analysis. procedures. 4. procedure.3. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. in addition to monitoring to determine compliance with the HACCP plan.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. Establish a system to monitor control of the CCP. food chemistry or engineering. food microbiology. 8. Validation . Establish documentation concerning all procedures and records appropriate to these principles and their application. 2. 3. quality assurance or control. Each stage is discussed below in detail. 7. Establish the critical limit(s).

biological.g. chemical and/or physical). it may be obtained from external sources.1.FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations. 8. The scope of the plan should be determined by defining the extent of the production process to be considered and the categories of hazard to be addressed (e. Logic sequence for application of HACCP 178 . If expert advice is not available on-site. Fig.

pH. is a cooking process required?). composition.1. there may be species-specific hazards related to certain fish and shellfish. heat treatment. These factors are likely to have a significant effect on some potential hazards. 8. etc.g. naturally present on fish from warm waters (> 15 °C).3. The description should include information such as the product name.4 Construct a flow diagram The flow diagram should be constructed by the HACCP team and should contain sufficient technical data for the study to progress.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. such as enteric viruses in shellfish. packaging. marine toxins associated with particular fish and shellfish species.). or in aquaculture operations. which should be described in detail.g. If the processing does not include a kill step. such as children or the elderly.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.2 Describe the product It is important to have a complete understanding of the product. storage and distribution conditions and instructions for use. However.3. In addition. freezing. physical and chemical structure (including water activity (aw). 8. contaminated fish and shellfish should not be used. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . It is also important to identify the consumer target groups.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments. 8. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation. aquaculture drug residues and potentially pathogenic Vibrio spp.). etc. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. Expertise on such inherent hazards. 8.3. shelf life.3.3. processing conditions (e. Vulnerable groups of the population. including practices onboard fishing vessels. may need to be considered specifically. fermentation.3.

features of equipment design. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. It is essential that the diagram is accurate.5 On-site confirmation of the flow diagram The HACCP team should confirm that the flow diagram matches the process that is actually being carried out. and therefore a detailed description of the procedure applied must be available. 180 ● ● . The team should then conduct a hazard analysis to identify which hazards are of such a nature that their elimination or reduction to an acceptable level is essential to the production of safe food. it is essential that the details of each step are fully appreciated and recorded. and identify any measures to control identified hazards The HACCP team should list all hazards that may reasonably be expected to occur at each step in the production process.6 List all potential hazards associated with each step. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. it should also include details of CCP steps. If HACCP studies are applied to proposed new process lines or products. 8. or circumstances of any reclaim or rework activity. Ideally.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly.3. time/temperature data. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. Many fish-processing operations have relatively few steps and the flow diagrams appear simple.3. cleaning and hygiene procedures and storage conditions.4. survival or multiplication of pathogenic microorganisms.3. It may include details of the factory and equipment layout. For example. once determined. However. 8. ingredient specifications. conduct a hazard analysis. the qualitative and/or quantitative evaluation of the presence of hazards. The operation should be observed at all stages. because the hazard analysis and decisions regarding CCPs are based on these data. 8.FISH AND SEAFOOD product.

Hazards introduced during processing of fish products depend very much on the characteristics of the process. The HACCP team should then determine what control measures exist that can be applied for each hazard. Some hazards may require more than one control measure for adequate control and a single control measure may act to control more than one hazard. the hurdle effect.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products. For example.g. since this may interfere with the analysis.). Therefore it is not possible. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin. There may also be inherent hazards from parasites such as the roundworm Anisakis simplex and from marine toxins such as ciguatera in reef fish (derived from microalgae) and scombrotoxin (biogenic amines) development in fish containing high levels of histidine. or desirable. The incidence of psychrotrophic. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. and viruses). the number of consumers potentially exposed and their vulnerability. 181 . and the potential growth of Listeria monocytogenes on smoked fish. Salmonella spp. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards.g. where a hazard is repeated. For example.3. Pathogens may be part of the resident microflora of the living animal (e. Vibrio spp. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. and there are an equally varied range of potential hazards associated with them. any food safety objectives or manufacturer’s food safety requirements. botulinum. 8.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. or may originate from polluted water or from post-capture contamination (e. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage. One control measure may be relevant to several process steps. Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials.6. Note: it is important at this stage that no attempt is made to identify CCPs.

biological or sensory parameters.8 Careful selection of sources for raw materials Adequate temperature control Effective sanitation Food handler hygiene Prevention of cross-contamination Establish critical limits for each CCP Critical limits separate acceptable from unacceptable products.8. then the process must be modified to include a control measure.FISH AND SEAFOOD 8. application of the decision tree should be flexible. However. Criteria used to set critical limits must be measurable and may include physical. However. If a significant hazard has been identified at a step where control is required for safety. given the enormous variety of fish products and processes in use.3. More than one critical limit may be defined for a single step. 8. Where possible. it can be said that effective control measures are likely to include the following: • • • • • 8. and its use may not always be appropriate. chemical. and the reader is referred to the appropriate product chapter. For example.3. Some examples relevant to fish products are: • • • Time and temperature limits for pasteurised and cooked products Temperature limits for chilled fish Brine concentration in cured fish 182 . critical limits should be specified and validated for each CCP. it is not possible to generalise on likely CCPs.1 Fish and seafood Again.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits.7. but for which no control exists at that step or any other.3. structured approach to decision making.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. 8.3. it is necessary to specify both time and temperature for a thermal process. It is prudent to set stricter limits (often-called target or process limits/levels) to ensure that any trend towards a loss of control is noted before the critical limit is exceeded.2) to provide a logical. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively.


• • • 8.3.9

Water activity (aw) values for dried fish pH values in fermented fish Microbiological quality of water in shellfish production areas Establish a monitoring system for each CCP

Monitoring involves planned measurement or observation of a CCP relative to its critical limits. Monitoring procedures for target levels must be able to detect loss of control of the CCP, and should provide this information with sufficient speed to allow adjustments to be made to the control of the process before the critical limits are violated. Monitoring at critical limits should be able to rapidly detect when the critical limit has been exceeded. Monitoring should therefore either be continuous, or carried out sufficiently frequently to ensure control at the CCP. Therefore, physical and chemical on-line measurements are usually preferred to lengthy microbiological testing. However, certain rapid methods, such as ATP assay by bioluminescence, may be useful for assessment of adequate cleaning, which could be a critical limit for some CCPs, for example pre-start-up hygiene. Persons engaged in monitoring activities must have sufficient knowledge, training and authority to act effectively on the basis of the data collected. These data should also be properly recorded. 8.3.10 Establish corrective actions

For each CCP in the HACCP plan, there must be specified corrective actions to be applied if the CCP is not under control. If monitoring indicates a deviation from the critical limit for a CCP, action must be taken that will bring it back under control. Actions taken should also include proper isolation of the affected product, repair of defective equipment, and an investigation into why the deviation occurred. A further set of corrective actions should relate to the target levels, if process drift is occurring. In this case, only repair of the process defect and investigation of the fault are required. All corrective actions should be properly recorded. 8.3.11 Establish verification procedures

Verification usually involves auditing and testing procedures. Auditing methods, procedures and tests should be used frequently enough to determine whether the HACCP system is being followed, and is effective at controlling the hazards. These may include random sampling and analysis, including microbiological testing. Although microbiological analysis is generally too slow for monitoring purposes, it can be of great value in verification, since many of the identified hazards are likely to be microbiological. In addition, reviews of HACCP records are important for verification purposes. These should confirm that CCPs are under control and should indicate the nature


of any deviations and the actions that were taken in each case. It is also useful to review customer returns and complaints regularly. 8.3.12 Establish documentation and record keeping

Efficient and accurate record keeping is an essential element of a HACCP system. The procedures in the HACCP system should be documented. ● ● Determination of critical limits The completed HACCP plan

Examples of documented procedures include: ● ● The hazard analysis Determination of CCPs

Examples of recorded data include: ● ● ● Results of monitoring procedures Deviations from critical limits and corrective actions Records of certain verification activities, e.g. observations of monitoring activities, and calibration of equipment.

The degree of documentation required will depend partly on the size and complexity of the operation, but it is unlikely to be possible to demonstrate that an effective HACCP system is present without adequate documentation and records. The length of time that records are kept will be as per company policy, but should not be less than one year beyond the shelf life of the product. Three to five years is typical for many food companies. 8.4 Implementation and review of the HACCP plan

The completed plan can only be implemented successfully with the full support and co-operation of management and the workforce. Adequate training is essential and the responsibilities and tasks of the operating personnel at each CCP must be clearly defined.



Fig. 8.2. CCP Decision Tree (Adapted from Codex Alimentarius Commission, 2003) Finally, it is essential that the HACCP plan be reviewed following any changes to the process, including changes to raw materials, processing conditions or equipment, packaging, cleaning procedures and any other factor that may have an effect on product safety. Even a small alteration to the product or process may invalidate the HACCP plan and introduce potential hazards. Therefore, the implications of any changes to the overall HACCP system must be fully



considered and documented, and adjustments made to the procedures as necessary. Triggered reviews/audits should occur as a result of changes, whereas a scheduled review/audit should be annual, as a minimum. 8.5 References

Wareing P.W., Carnell A.C. HACCP – A Toolkit for Implementation. Leatherhead, Leatherhead Food International. 2007. Bernard D., Scott V. Hazard Analysis and Critical Control Point System: use in controlling microbiological hazards, in Food Microbiology: Fundamentals and Frontiers. Eds. Doyle M.P., Beuchat L.R. Washington D.C., ASM Press. 2007, 971-86. Su Y-C., Daeschel M.A. Sanitation and HACCP, in Surimi and Surimi Seafood. Ed. Park J.W. Boca Raton, CRC Press. 2005, 141-62. Huss H.H. Considerations in the application of the HACCP principles to seafood production, in Assessment and Management of Seafood Safety and Quality. Eds. Food and Agricultural Organisation, Huss H.H., Ababouch L., Gram L. Rome, FAO. 2004, 153-77. Chen H-C., Chang P.C-M. Frozen seafood safety and HACCP, in Handbook of Frozen Foods. Ed. Hui Y.H., Cornillon P., Legaretta I.G., Lim M.H., Murrell K.D., Nip WK. New York, Marcel Dekker. 2004, 339-63. Mortimore S., Mayes T. The effective implementation of HACCP systems in food processing, in Foodborne Pathogens: Hazards, Risk Analysis and Control. Eds. Blackburn C.deW., McClure P.J. Cambridge, Woodhead Publishing Ltd. 2002, 229-56. Mayes T., Mortimore C.A. Making the Most of HACCP: Learning from Other’s Experience. Cambridge, Woodhead Publishing. 2001. Mortimore S.E., Wallace C., Cassianos C. HACCP (Executive Briefing). London, Blackwell Science Ltd. 2001. Chartered Institute of Environmental Health. HACCP in Practice. London, Chadwick House Group Ltd. 2000. Jouve J.L. Good manufacturing practice, HACCP, and quality systems, in The Microbiological Safety and Quality of Food, Volume 2. Eds. Lund B.M., BairdParker T.C., Gould G.W. Gaithersburg, Aspen Publishers. 2000, 1627-55. Stevenson K.E., Bernard D.T. HACCP: A Systematic Approach to Food Safety. Washington D.C., Food Processors Institute. 1999. National Seafood HACCP Alliance. IQF cooked shrimp processing HACCP model, in HACCP User's Manual. Ed. Corlett D.A. Gaithersburg, Aspen Publishers. 1998, 454-73. Mortimore S., Wallace C. HACCP: A Practical Approach. Gaithersburg, Aspen Publishers. 1998.


HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Khandke S.S., Mayes T. HACCP implementation: a practical guide to the implementation of the HACCP plan. Food Control, 1998, 9 (2-3), 103-9. Forsythe S.J., Hayes P.R. Food Hygiene, Microbiology and HACCP. Gaithersburg. Aspen Publishers. 1998. Food and Agriculture Organisation. Food Quality and Safety Systems: A Training Manual on Food Hygiene and the Hazard Analysis and Critical Control Point (HACCP) System. Rome, FAO. 1998. US Food and Drug Administration. Fish and Fishery Products Hazards and Controls Guide. Rockville, Food Drug Administration. 1998. National Advisory Committee on Microbiological Criteria for Foods. Hazard Analysis and Critical Control Point Principles and Application Guidelines. 1997. Gardner L.A. Testing to fulfil HACCP (Hazard Analysis Critical Control Points) requirements: principles and examples. Journal of Dairy Science, 1997, 80 (12), 3453-7. Martin R.E., Collette R.L, Slavin J.W. Fish Inspection, Quality Control and HACCP: A Global Focus. Proceedings of a Conference, Arlington, May 1996. Lancaster P.A., Technomic Publishers. 1997. Codex Alimentarius Commission. Hazard Analysis Critical Control Point (HACCP) System and guidelines for its application, in Food Hygiene: Basic Texts. Ed. Codex Alimentarius Commission. Rome, FAO. 2003, 21-31. Garrett E.S., Jahncke M.l., Tennyson J.M. Microbiological hazards and emerging food safety issues associated with seafoods. Journal of Food Protection, 1997, 60 (11), 1409-15. National Seafood HACCP Alliance. HACCP: Hazard Analysis and Critical Control Point Training Curriculum. Raleigh, North Carolina State University. 1997. Dillon M., McEachern V. HACCP and quality assurance of seafood, in Fish Processing Technology. Ed. Hall G.M. London, Blackie. 1997, 224-48. Savage R.A. Hazard Analysis Critical Control Point: a review. Food Reviews International, 1995, 11 (4), 575-95. Pearson A.M., Dutson T.R. HACCP in Meat, Poultry and Fish Processing. Glasgow, Blackie. 1995. Huss H.H., Food and Agriculture Organisation. Assurance of Seafood Quality. FAO Fisheries Technical Paper No. 334. Rome, FAO. 1994. Bonnell A.D. Process control, in Quality Assurance in Seafood Processing: A Practical Guide. Ed. Bonnell A.D. London, Chapman & Hall. 1994, 39-53. Pierson M.D., Corlett D.A., Institute of Food Technologists. HACCP: Principles and Applications. New York, Van Nostrand Reinhold. 1992. Hudak-Roos M., Garrett E.S. Monitoring, corrective actions and record keeping in HACCP, in Quality Assurance in the Fish Industry: Proceedings of an International Conference, Copenhagen, August 1991. Eds. Huss H.H., Jakobsen M., Liston J. Amsterdam, Elsevier. 1992, 521-31.


1992. 1988. World Health Organisation. 1992. Blackwell Scientific Publications. Food Control. Microorganisms in Foods. International Commission on Microbiological Specifications for Foods. WHO. 14-9. International Journal of Food Microbiology.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H.L. Development and use of the HACCP concept in fish processing. Oxford. 33-44. 1992. 3 (1). 15 (1/2). Mayes T. 188 . Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety. Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality.H. Bryan F. Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. Geneva..

with the intention of introducing consistency and clarity throughout the food production chain from primary production. Hygiene is a general concept that covers a wide subject area. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials.9. from structural conditions in the factory or process facility. The new legislation was designed to establish conditions. to sale or supply to the final consumer. It is generally considered that the principles of Good Manufacturing Practice (GMP) and application of Hazard Analysis Critical Control Point (HACCP) systems are of greater importance. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . under which food is produced to optimise public health and to prevent. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. the existence of microbiological standards alone cannot protect consumer health. including microbiological criteria. The general food hygiene Directive 93/43/EEC and other Directives on the hygiene of foodstuffs and the health conditions for the production and placing on the market of certain products of animal origin intended for human consumption.1 Introduction Hygiene is an important aspect of ensuring food safety. and one that plays an important role in most countries’ food legislation. transport and delivery vehicles requirements. final product specifications. A number of standards are provided in food legislation. to personnel requirements. however. EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr. have been replaced by several linked measures on food safety rules and associated animal health controls. eliminate or acceptably control pathogen contamination of food. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. and conditions of raw materials.

existing hygiene Directives including those below were repealed: • Council Directive 91/492/EEC of 15 July 1991 laying down the health conditions for the production and the placing on the market of live bivalve molluscs (Off. handling and offering for sale or supply to the consumer. manufacturing. 7. transportation.2 Legislative Structure From 1 January 2006.1).B. p. p. and sets out what those enforcing the provisions have to do. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. Council Directive 91/493/EEC of 22 July 1991 laying down the health conditions for the production and the placing on the market of fishery products (OJ L 268.1991. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption Regulation (EC) No. 24. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. Council Directive 92/48/EEC of 16 June 1992 laying down the minimum hygiene rules applicable to fishery products caught on board certain vessels in accordance with Article 3(1)(a)(i) of Directive 91/493/EEC (OJ L 187. storing. A number of more detailed implementing and transitional measures have been adopted at EC level. as last amended by Regulation (EC) No 806/2003. distribution. J.15). the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. p. packaging.7.9. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. processing.1991. as last amended by Regulation (EC) No 806/2003.9. 41). Subsequently. 9. European Communities L 268. 190 • • . 24. N.1992.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation.

19. with primary production.I.1993. As regulations. The regulation lays down general hygiene provisions for which food business operators carrying out primary production must comply with as laid down in Part A of Annex 1. 9. Primary products are defined as products of primary production including products of the soil. which will be covered later in this chapter. Wales and Northern Ireland). 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175.11) The EU hygiene regulations apply to all stages of food production including primary production. 191 . processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. national legislation in the form of a Statutory Instrument (S. and of hunting and fishing. for example. right through to the implementation of procedures based on HACCP principles. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. e. The Food Hygiene (England) Regulations 2006 (S. 852/2004. and equivalent legislation in Scotland. the legislation is directly applicable law. Additionally the requirements of EC Regulation 853/2004 must be complied with. domestic preparation or handling. and binding in its entirety on all member states from the date of entry into force. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. 2006 No. food storage that is for private or domestic consumption. Although EU food hygiene regulations are directly applicable in the individual Member States there are some aspects where Member States are required or allowed to adopt certain provisions into their national laws.14. There are some exemptions.I. Although the regulations have the force of law. Likewise Regulation 852/2004 will not apply to collection centres and tanneries meeting the definition of food business because they handle raw material for the production of gelatine or collagen.3 Regulation (EC) No. of stock farming. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland.7.g. setting offences. penalties and powers of entry. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. etc. 2073/2005.) in England. 1 . revocation of existing implementing legislation. p. is required to give effect to the EU regulations. Wales and Northern Ireland.



Annex I – Primary production

Annex I (Part A) relates to general hygiene provisions for primary production and associated operations covering: (a) the transport, storage and handling of primary products at the place of production, (b) the transport of live animals, (c) for fishery products - transport operations to deliver primary products (which have not been substantially altered) from the place of production to an establishment Food business operators have the responsibility to ensure primary products are protected against contamination. Any community and national legislation relating to the control of hazards in primary production such as measures to control contamination resulting from surroundings e.g. air, soil, water etc., and measures relating to animal health and welfare, and plant health that may impact on human health should be complied with. Requirements for record keeping are also laid down. This relates to animal feed (nature and origin), veterinary medicines administered to animals (date given and withdrawal periods), any diseases, analysis of samples from other animals that might impact on human health as well as reports on animal checks performed. Part B of Annex I contains recommendations for guides to good hygiene practice. 9.3.2 Annex II – Stages other than primary production

Annex II of the regulation lays down additional general hygiene requirements that must be met by food business operators carrying out production, processing and distribution of food at all stages following primary production. A summary of Chapters I to IV of Annex II is provided as follows: Chapter I Chapter I applies to all food premises, except premises to which Chapter III applies. Food premises must be kept clean and maintained in good repair and condition. Their layout should allow for this. The environment should allow good hygiene practices and provide temperature-controlled handling and storage conditions where necessary ,and enable foods to be kept at correct temperatures and be monitored.




Additionally, there are requirements for provision of adequate lavatories, basins, ventilation, lighting and drainage. Chapter II Chapter II applies to all rooms where food is prepared, treated or processed, except dining areas and premises to which Chapter III applies. The design and layout of rooms should allow for good hygiene practices between and during operations. Therefore floor and wall surfaces, ceilings and windows should be constructed to prevent dirt accumulating. Surfaces where food is handled must be well-maintained and allow easy cleaning and disinfection, preferably using smooth, washable corrosionresistant and non-toxic materials. There should be facilities for cleaning or disinfecting, and for storing working utensils or equipment. Clean potable water and adequate provision for washing food is needed.


- Chapter III Chapter III applies to temporary premises (e.g. marquees, market stalls, and mobile sales vehicles), premises used primarily as a private dwelling-house but where foods are regularly prepared for placing on the market, and vending machines. Premises and vending machines should be practically sited, designed, constructed and kept clean and maintained in good repair and condition so as to avoid the risk of contamination, in particular by animals and pests. Facilities should allow adequate personal hygiene, and surfaces in contact with food should be easy to clean. Enough potable water and storage arrangements for hazardous or inedible substances are required as well as adherence to food safety requirements.

- Chapter IV Chapter IV applies to all transportation. Conveyances and/or containers used for transporting foodstuffs are to be kept clean and maintained in good repair and condition to protect foodstuffs from contamination and are, where necessary, to be designed and constructed to permit adequate cleaning and/or disinfection. Food should be maintained at appropriate temperatures.



FISH AND SEAFOOD Chapter V Chapter V refers to equipment requirements. Adequate cleaning and disinfection is to be carried out frequently for articles, fittings and equipment in contact with food where contamination needs to be avoided. Equipment should be installed to allow adequate cleaning, and be fitted with the required control device.

- Chapter VI Chapter VI refers to food waste. Food waste, non-edible by-products and other refuse is to be removed from rooms where food is present as quickly as possible to avoid accumulation. Such waste is to be deposited in closable containers, to allow easy cleaning. Refuse stores should allow easy cleaning and be free of pests. Waste must be eliminated hygienically in accordance with European Community legislation.

- Chapter VII Chapter VII refers to the water supply. There must be an adequate supply of potable water, which should be used whenever necessary to ensure foodstuffs are not contaminated. There are also requirements for recycled water, ice in contact with food, use of steam and for the water used in the cooling process for heat-treated foods in hermetically sealed containers. Specific requirements are that clean water may be used with whole fishery products. Clean seawater, may be used for live bivalve molluscs, echinoderms, tunicates and marine gastropods; and for external washing. If it is used, there must be adequate facilities to ensure the supply does not create a source of contamination for the foodstuff. Recycled water used in processing or as an ingredient must not present a contamination risk. It must be of the same standard as potable water, unless the competent authority is satisfied that the quality of water cannot affect the wholesomeness of food in the finished form.



Additionally, ice that comes into contact with food or which could possibly contaminate it needs to be made from potable water or, in the case of chilling whole fishery products using clean water. Ice must be made, handled and stored under conditions that protect it from contamination.


LEGISLATION Chapter VIII Chapter VIII sets out personal hygiene requirements for those working in a food handling area including clean protective clothing and the requirement that those carrying or suffering from a disease are not permitted to handle food. Chapter IX Chapter IX covers provisions applicable to foodstuffs. A food business operator should not accept raw materials or ingredients, other than live animals, or any other material used in processing products, if they are known to be contaminated with parasites, pathogenic microorganisms or foreign substances, to be toxic or decomposed to such an extent that, even after the business operator applied normal hygienic processing , the product would be inedible. Raw materials must be kept under appropriate conditions throughout production, processing and distribution. In particular, temperature control (i.e. cold chain and food thawing) requirements are laid down.

- Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. Chapter XI Chapter XI lays down heat treatment requirements for food that is placed on the market in hermetically sealed containers. The main relevant process parameters (particularly temperature, pressure, sealing and microbiology) must be checked by the food business operator to ensure that the required heat treatment has been achieved. Also, the process used should comply with an internationally recognised standard (e.g. pasteurisation, ultra high temperature or sterilisation). Chapter XII Chapter XII states training requirements for food business operators to ensure that food handlers are trained in food hygiene matters and in the application of HACCP principles.





The Regulation requires that food business operators must notify the relevant competent authority of each establishment under their control that carries out any of the stages of production, processing and distribution of food, with a view to the registration of each establishment. Food business operators must also ensure that the competent authority always has up-to-date information on establishments, including the notification of significant changes in activity, and closure of an existing establishment. Food business operators must ensure that establishments are approved by the competent authority, following at least one on-site visit, when approval is required by the national law of the Member State in which the establishment is located, or under Regulation (EC) No. 853/2004, or by a separate decision adopted. Separate rules apply for businesses producing products of animal origin. 9.3.4 HACCP

Food business operators, other than at the level of primary production, and associated operations must put in place, implement and maintain a permanent procedure or procedures based on principles of the system of hazard analysis and critical control points (HACCP). Emphasis is placed on risk-related control, with responsibility placed on the proprietor of the food business to ensure that potential hazards are identified and systems are developed to control them. Under HACCP, food business operators must amongst other considerations, identify hazards to be prevented or eliminated or reduced to acceptable levels, identify critical control points (CCP) at which control is essential to prevent, eliminate or reduce hazards and establish critical limits at these points, implement effective monitoring procedures, and establish corrective actions in the case where a CCP is out of control. Procedures must be followed to confirm the above is in place and up-todate, as well as providing the competent authority with documents and records as evidence, when required. 9.4 Regulation (EC) No. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin

Regulation (EC) No. 853/2004 lays down hygiene rules for products of animal origin that apply in addition to the general hygiene rules of Regulation (EC) No. 852/2004. 9.4.1 Definitions

The following definitions apply for fishery products and live bivalve molluscs:


filleting. Mechanically separated fishery product means any product obtained by removing flesh from fishery products using mechanical means resulting in the loss or modification of the flesh structure. and chopping. heading. mincing or processing.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. shelling. whether whole or prepared. slime or sand. gutting and removal of fins and. purification centres or dispatch centres in tanks or any other installation containing clean seawater. slicing. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing. live tunicates and live marine gastropods. Conditioning means the storage of live bivalve molluscs coming from class A production areas. after preparatory work such as bleeding. Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. skinning. Fresh fishery products means unprocessed fishery products. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness. such as gutting. heading. parts and products of such animals. and all mammals. or in natural sites. followed by wrapping or packaging. including products packaged under vacuum or in a modifiedatmosphere that have not undergone any treatment to ensure preservation other than chilling. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. slicing. in particular as a result of feeding on plankton containing toxins. where appropriate. 197 . live echinoderms. reptiles and frogs) whether wild or farmed and including all edible forms. Purification centre means an establishment with tanks fed by clean seawater in which live bivalve molluscs are placed for the time necessary to reduce contamination to make them fit for human consumption. if necessary: chilling or freezing: filleting. to remove mud. where necessary. shucking.

1.FISH AND SEAFOOD 9.4. (iii) Food business operators must have in place systems and procedures to identify food business operators from whom they have received and to whom they have delivered products of animal origin. (ii) The mark must indicate the name of the country in which the establishment is located and written out in full or shown as a two-letter code in accordance with the relevant ISO standard. the mark must show the approval number of the establishment.2.4. (iv) If the identification mark is applied in an establishment within the Community. An identification mark can only be applied provided the relevant requirements as stated above in relation to Regulation (EC) 852/2004 have been met in particular for the registration and approval of establishments.2 Fishery product requirements 9. Section I of Regulation (EC) 853/2004 are given as follows: 9.4. and the characters easily decipherable and be clearly displayed. please refer to Regulation (EC) 853/2004.1. the food business operator may apply the same identification mark to both types of food. EF. ES. EK. EB or WE. Extracts of the requirements of Annex II. the mark must be oval shaped and include the abbreviation CE. EY. Section I of Regulation (EC) 853/2004.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. (ii) However. (iii) Additionally. the new mark must show the approval number of the establishment where this occurred. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. the code would be UK. EK.2.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. For the UK.2 Form of the identification mark (i) The mark must be legible and indelible. EO. EG. EÜ. 198 .4. EC. when a product's packaging and/or wrapping is removed or is further processed in another establishment. In this case. a new mark must be applied to the product.2. 9. for other codes.

the approval number of the establishment and the abbreviation for the community where appropriate. (v) When the mark is applied directly to products of animal origin.1.LEGISLATION 9. echinoderms. i.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. specifically relating to fishery products are given below. Extracts of the requirements of Annex III.3 Method of marking (i) The mark is to be applied directly to the product. Parts A. This section does not apply to bivalve molluscs. (iii) In the case of liquid. Part A and Chapter V apply to retail. as amended. reference should be made to the actual Regulation. a) requirements for establishments. it is sufficient to apply the mark to the exterior of that package only. and fishery products carried in bulk. tunicates and marine gastropods when placed on the market live. (ii) For products of animal origin that are placed in transport containers or large packages.2. they would need to be obtained following Section VII on live bivalve molluscs. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. wrapping or packaging in another establishment. granulate and powdered products of animal origin carried in bulk. it applies to those animals when not placed on the market live. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. the mark may be applied to the outer surface of the container or packaging. Chapter III. 9.4. the colourings used must be EC authorised colours. Section VIII of Regulation 853/2004.4.e. including vessels. for full requirements.2. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . or be printed on a label affixed to the product. Chapter IV. With the exception of Chapters I and II. Therefore. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. the wrapping or the packaging. C and D. processing. its wrapping or the packaging. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. and are intended for further handling. The requirements of this section accompany those in Regulation (EC) 852/2004.

Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 . refrigeration and wrapping. The definition of fishery products covers primary production as well as its associated operations. in that record keeping requirements stated by point 7 in Part A of Annex I of Regulation 852/2004 do not apply to operators involved in small-scale coastal fishing (as defined through Regulation (EC) 1198/2006) if the activity is under 24 hours. gutting. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products. i. from the place of production to the first establishment of destination.e. 2. it must be placed somewhere that will not contaminate the water supply. There is a derogation from point a. bleeding. (a) primary production covers the farming. A. heading. if a vessel has water coming in that is used with fishery products. removing fins. or to handle or process them after harvesting.1 Chapter I .) 9. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. Structural and equipment requirements refer to the following points 1.2.2. clean seawater may be used for the handling and washing of fishery products.. fishing and collection of live fishery products with a view to their being placed on the market. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material.FISH AND SEAFOOD c) for the water supply. Chapter VII of Regulation 852/2004. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking.4. this supplements the requirements of Annex II.Requirements for vessels Food business operators are required to ensure that they meet the structural and equipment requirements as well as hygiene requirements required for vessels used to harvest fishery products from their natural environment.

recording of temperatures. 3. any waste processing unit needs to have a separate hold to store the waste. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. tanks must incorporate devices that allow a uniform temperature throughout the tanks. (iii) In vessels equipped for chilling fishery products in cooled clean seawater.LEGISLATION (i) Such vessels need to have holds. 201 . where necessary. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C. 4. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas. Requirements for factory vessels a) Factory vessels (i) These must have at a receiving area reserved for taking fishery products on board so each catch can be separated (ii) There must be a hygienic system for conveying fishery products from the receiving area to the work area (iii) Large work areas are needed to allow fishery products to be prepared and processed hygienically. and not more than 0 °C after 16 hours. Such devices must achieve a chilling rate that ensures that the mix of fish and clean seawater does not exceed 3 °C six hours after loading. to ensure that melt water does not stay in contact with the products. (ii) Holds must be separated from the engine compartments and from the crew quarters by partitioning that prevent any contamination of the stored fishery products. where necessary. Also. A temperature recording device must be present and placed where the temperature in the hold is the highest. Storage areas for the finished products also need to be sufficiently large and cleanable. and allow the monitoring and.

d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. landed as soon as possible. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. where fishery products are displayed for sale. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. other than those kept alive. c) Fishery products.Requirements during and after landing 9. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. (ii) In the case of food business operators responsible for auction and wholesale markets etc.2.FISH AND SEAFOOD (vi) A water tank needs to be placed where there is no contamination of the water supply and hand washing taps should be designed to prevent contamination. there need to be lockable facilities to refrigerate detained fishery products and separate lockable facilities to store fishery products that are unfit for human 202 . if this is not possible. provided no damage occurs to the flesh. The brine must not be a source of contamination. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. and products washed thoroughly. e) In cases where whole fish intended for canning is frozen in brine. B. chilled and wrapped. the product must be kept at no more than -9 °C.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. Handlers may use spiked instruments to move large fish or fish that might injure them. must be chilled or. The unloading and landing process is to be done quickly to avoid contamination (but at the same time not causing bruising to the fish) and to ensure fishery products are placed in a protected environment at the temperatures stated in Chapter VII. Chapter II . b) Factory vessels on board which crustaceans and molluscs are cooked. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption.2. Hygiene requirements Hygiene requirements refer to the following: a) Parts of vessels used for storage of fishery products must be cleaned and maintained in a good state with no contamination.4.

Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. Fillets and slices must then be removed from work tables as soon as possible after preparation.2. Requirements for frozen products An establishment on land that freezes fishery products needs to have equipment that meets that for freezer vessels as stated in Section VIII. Additionally. packaged and chilled as quickly as possible after preparation. they must be stored under ice in appropriate facilities. (v) Containers for dispatching and storing unpackaged prepared fresh fishery products kept under ice should ensure that melt water does not remain in contact with the products.2. C. Chapter I. including vessels. dispatched. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels.4. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . 854/2004. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. Food business operators need to co-operate with the relevant competent authorities so as to allow them to carry out official controls as laid down in Regulation (EC) No. B. C. They may also continue to be transported in cooled water after landing and be transported from aquaculture establishments until they reach the first establishment on land that carries out an activity other than transport or sorting.Requirements for establishments. part I. 9.LEGISLATION consumption. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. points 1 and 2. prepared or processed immediately after arrival at an establishment on land. and where necessary. (ii) Heading and gutting must be carried out hygienically.3 Chapter III . They should be wrapped. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed. handling fishery products A.

(iii) A document from the manufacturer. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. they must be cooled rapidly. must accompany fishery products referred to above when placed on the market. stating the type of process they have undergone. and (c) marinated and/or salted fishery products. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. if the processing is insufficient to destroy nematode larvae.2.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. if they are to undergo a cold smoking process in which the internal temperature of the fishery product is not more than 60 °C: (i) herring. (iii) sprat. and whole fish must be gutted and washed beforehand. If no other method of preservation is used.2. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. or a stabilising treatment.4 Chapter IV .20 °C in all parts of the product for at least 24 hours (this treatment must be applied to the raw product or the finished product): (a) fishery products to be consumed raw or almost raw.Requirements for certain processed fishery products A. Once produced. except when supplied to the final consumer. raw materials must have no guts. (b) fishery products from the following species. D. cooling must continue until a temperature 204 . (iv) (wild) Atlantic and Pacific salmon. (ii) mackerel. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . 9.4. (ii) Compliance with the above is not necessary if epidemiological data indicates that the fishing grounds of origin do not present a health hazard in relation to the presence of parasites and this has been authorised by the competent authority.

provided that the freshness criteria are met and the total volatile basic nitrogen (TVB-N) value of the unprocessed fishery products does not exceed the limits set out in point 1 of Chapter I of Section II of Annex II to Regulation (EC) No. 2073/2005. raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. 2074/2005). pressing. C.5 Chapter V . B. 9. (ii) The production process for fish oil must ensure that all raw material intended for the production of crude fish oil is subject to a treatment including (where relevant) heating. centrifugation. Organoleptic properties of fishery products An organoleptic examination of fishery products must be carried out to ensure that fishery products comply with any freshness criteria (such criteria have yet to be set). processing. 2073/2005). separation. However. B. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. Shelling/shucking is to be done hygienically.4. and the raw material is processed within 36 hours after loading. 205 . 2074/2005.LEGISLATION approaching that of melting ice is reached. the following conditions need to be met: A. there is an exception: food business operator may refrain from chilling the fishery products when whole fishery products are used directly in the preparation of fish oil for human consumption. Total volatile nitrogen Unprocessed fishery products cannot be placed on the market if chemical tests reveal that TVB-N or TMA-N limits are exceeded (limits are set in Regulation (EC) No.2. refining and purification steps before being placed on the market for the final consumer. and after this. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No.2.

thawed unprocessed fishery products. prepared and processed fishery products belonging to the family Gempylidae. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice.FISH AND SEAFOOD D. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. The scientific name must accompany the common name on the label. 9.6 Chapter VI . Molidae.2. Diodontidae and Canthigasteridae. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum.Wrapping and packaging of fishery products (i) Containers in which fresh fishery products are kept under ice must be waterresistant and ensure that melt water does not remain in contact with the products. it should be kept at no more than -9 °C. Fishery products contaminated with parasites may not be placed on the market for human consumption.2. 206 . Fresh.7 Chapter VII . (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. and when fishery products are wrapped on board fishing vessels. may only be placed on the market in wrapped/packaged form and must be appropriately labeled to provide information to the consumer on preparation/cooking methods and on the risk related to the presence of substances with adverse gastrointestinal effects. 9. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product.4. food business operators must ensure that wrapping material is not contaminated and is stored in such a way that it is not exposed to contamination and is easy to clean where necessary.Storage of fishery products (i) Fresh fishery products. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption. In the case of whole frozen fish in brine intended for the manufacture of canned food.4.2. E.2.

The temperature of frozen fishery products must not fluctuate above -15°C. Sebastichthys capensis. 2074/2005. (ii) If fishery products are kept under ice.2.4.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar.). The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. provided that the journey is short and the competent authority allows this.8 Chapter VIII . 9. species belonging to the Gadidae family. melt water must not remain in contact with the products. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. The following in relation to TVB-N limits is extracted from the regulation: Unprocessed fishery products shall be regarded as unfit for human consumption where organoleptic assessment has raised doubts as to their freshness and chemical checks reveal that the following TVB-N limits are exceeded: (a) 25 mg of nitrogen/100 g of flesh for the species: Sebastes spp. Note: (i) Frozen fishery products do not need to be kept at a temperature of -18 °C if they are being transported from a cold store to an approved establishment to be thawed on arrival for the purposes of preparation and/or processing.4.2. species belonging to the Merlucciidae family. 9. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them.3 Other information: Rules relating to visual inspections for the detection of parasites in fishery products are stated in Annex II (obligations on the competent authorities) of Regulation (EC) No.2.Transport of fishery products The above points for Chapter VII apply in relation to transport. (d) 60 mg of nitrogen/100 g of whole fishery products used directly for the preparation of fish oil for human consumption as referred to in the second sub-paragraph of Part B(1) of Chapter IV of Section VIII of Annex III to 207 . Helicolenus dactylopterus.

they supplement the requirements of Annex II to that Regulation. reference should be made to the actual regulation. Chapter IX applies to pectinidae harvested outside those areas. 9. Apart from the provisions on purification. this Section also applies to live echinoderms. VIII and IX. The requirements of this Section accompany those laid down in Regulation (EC) No 852/2004: (a) In the case of operations that take place before live bivalve molluscs arrive at a dispatch or purification centre. where the raw material complies with points (a). as amended. (b) In the case of other operations. tunicates and marine gastropods.3. specifically relating to bivalve molluscs are given below. and point 3 of Chapter VII applies to retail.4. for full requirements. Extracts of the requirements of Annex III. Chapters V. Section VII of Regulation 853/2004. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. (b) and (c) of Part B(1) of that Chapter. taken from at least three different points and mixed together by grinding. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. they supplement the requirements of Annex I to that Regulation. please refer directly to the regulation. The reference method to be used for checking the TVB-N limits involves distilling an extract deproteinised by perchloric acid as set out in Chapter III For distillation apparatus and routine methods which may be used to check the TVB-N limits. Live bivalve mollusc requirements - 9. The copy of 208 .1 Chapter I . shellfish species etc.3 The regulation details specific hygiene requirements for bivalve molluscs. The sample must consist of about 100 g of flesh. however. The section is split into 9 chapters for which a summary is provided as follows: Chapters I to VIII apply to animals harvested from production areas that the competent authority has classified in accordance with Regulation (EC) No 854/2004.4.FISH AND SEAFOOD Regulation (EC) No 853/2004. The information required in the registration document is given in point 4 and includes the health status of the production area.General requirements for the placing on the market of live bivalve molluscs Live bivalve molluscs can only be put on the market for retail sale if done through a dispatch centre. VI.

3. processing or relaying. (ii) Provided the requirements of chapter V are met.4. Requirements for harvesting and handling following harvesting This includes requirements that any further handling should not cause contamination or excess damage to shells/tissues of the live bivalve molluscs that might affect their suitability for purification. collection from: .Class A means they can be placed directly on the market . Requirements for production areas (i) The competent authority classes (classed A. 9.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified.2 Chapter II . poles or other fixed means are to identify the boundaries of the sites. B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with. C. Transport needs to allow sufficient drainage for best survival conditions. B.Class C means relaying over a long time before it can be placed on the market For bivalve molluscs not subject to Class B or C treatments. 209 . they need to be sent to a processing establishment and undergo specified treatments such as sterilisation in hermetically sealed containers or specific heat treatments such as immersion in boiled water so the internal temperature of the mollusc flesh reaches at least 90 ºC and this temperature is maintained for at least 90 seconds.Hygiene requirements for the production and harvesting of live bivalve molluscs A.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market . Requirements for relaying live bivalve molluscs Only areas approved by the competent authority can be used for relaying live bivalve molluscs with a requirement that buoys.

5 Chapter V . (ii) amnesic shellfish poison (ASP). Before dispatch.4 Chapter IV . B. 160 micrograms of okadaic acid equivalents per kilogram.3. 210 . the purification system to allow them to resume /maintain filter-feeding activity. and there are requirements for tanks and water storage containers.4. (iv) yessotoxins. containers and a label to identify purification.4. Requirements for purification centres There are requirements for live bivalve molluscs to be washed. requirements if there are many batches of live bivalve molluscs. the levels of marine biotoxins should not exceed the following limits. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. 1 milligram of yessotoxin equivalent per kilogram.Hygiene requirements for purification and dispatch centres A. 9.3.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. dinophysistoxins and pectenotoxins together. Requirements for dispatch centres Handling should not result in contamination to any degree.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements.3 Chapter III . 800 micrograms per kilogram.4. shells of live bivalve molluscs are to be washed throughout with clean water and the molluscs are to come from one of the following: (i) Class A production area (ii) Relaying area (iii) Purification centre (iv) Another dispatch centre 9.3. for: (i) paralytic shellfish poison (PSP).FISH AND SEAFOOD 9.

the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’. Finally.7 Chapter VII .2. the species of bivalve mollusc and date of packaging (at least the date and month) are required.3.4.Other requirements (i) The temperature at which live bivalve molluscs are stored/ transported in should be at one that does not adversely affect food safety or their viability.3. 9. Requirements of Chapters III and IV are to be complied with.9 Chapter IX .Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.3. 211 .4.8 Chapter VIII .Identification marking and labelling In addition to general identification marking (contained in 9. must be waterproof. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre.4. dispatch centre or processing establishment.4.4. 9. they cannot be re-immersed in or sprayed with water. by way of derogation from directive 2000/13/EC.3. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction. the label.LEGISLATION (v) azaspiracids. including the identification mark. Also. 160 micrograms of azaspiracid equivalents per kilogram. Registration document requirements also apply.1). 9. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.6 Chapter VI . (ii) Individual consumer-size packages of live bivalve molluscs must be closed and remain closed after leaving the dispatch centre and until presented for sale to the final consumer.Specific requirements for pectinidae harvested outside classified production areas Requirements for harvesting and handling following harvesting and health standards for live bivalve molluscs must be met.

they derive from: (i) poisonous fish. This refers to official controls or production and placing on the market of fishery products. Section VIII. echinoderms. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. Chapter V of Regulation 853/2004 apply.5. Fishery products in particular must comply with the requirements of Annex III of the regulation. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation.FISH AND SEAFOOD 9. 9. Therefore the health standards for fishery products as previously stated in Annex III.5 Regulation (EC) No. In this regulation. they contain in their edible parts contaminants or residues in excess of the limits laid down in Community legislation or at levels where the calculated dietary intake would exceed the acceptable daily or weekly intake for humans. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption Regulation (EC) 854/2004 gives requirements for official controls on products of animal origin and states requirements for those enforcing the provisions. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. or - - 212 .1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given. chemical. It is a requirement that food business operators give assistance to ensure that official controls carried out by the competent authority can be done properly. or (iii) bivalve molluscs. (ii) fishery products not complying with the requirement concerning biotoxins. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004.

LEGISLATION - the competent authority considers that they may constitute a risk to public or animal health or are for any other reason not suitable for human consumption.000 E. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. that microbiological criteria and chemical contaminants amongst other requirements are met. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. Also. coli per 100 g of flesh and intravalvular liquid. The reference method for this analysis is the five-tube. 4. 000 E. in 90% of the samples. as previously stated. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. three dilutions MPN test specified in ISO 16649-3. live echinoderms. Regulation (EC) 854/2004. live bivalve molluscs must not exceed 46. live tunicates and live marine gastropods. In the remaining 10 % of samples. - - - - B. 213 . three dilutions Most Probable Number (MPN) test specified in ISO 16649-3.600 E. coli per 100 g of flesh and intravalvular liquid. Sampling plans and frequency are required.2.Official controls concerning live bivalve molluscs from classified production areas A. Live bivalve molluscs from Class C areas must not exceed 46. chapter V is reinforced.5. Annex II 9. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. Classification of production and relaying areas: This section reinforces that specified in Annex III.5. A summary is given as follows: 9. section VII.2 This states official controls for the production and placing on the market of live bivalve molluscs. The reference method for this analysis is the five-tube. coli per 100 g of flesh and intravalvular liquid. Live bivalve molluscs from Class B areas must not exceed.1 Chapter II .

5.6 Regulation (EC) No. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. Decisions after monitoring There are specific requirements whereby if sampling results indicate that health standards for molluscs are exceeded or there is a risk to human health. which has applied since 1 January 2006. 9.2 Chapter III . E. the competent authority is to close the production area. Recording and exchange of information Requirements are stated for competent authorities in relation to establishing and maintaining a current list of approved production and relaying areas and their role to inform interested parties of any changes of location.2.FISH AND SEAFOOD C. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. 2073/2005. Additional monitoring requirements This concerns the competent authority in relation to monitoring of classified production areas where bivalve mollusc harvesting is forbidden and of relaying and production zones to ensure there is no malpractice etc. establishes microbiological criteria for a range of foods. the Member States may permit the import of bivalve molluscs and fishery products from the countries listed respectively in Annex I and Annex II to regulation (EC) 2076/2005. F. 9. Procedures for re-opening a closed production area are stated D. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. 214 .5. 9.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. boundaries or class of a production area or its closure. opening or closing a production area.

I. The relevant criteria are outlined in Table 9. area or batch. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. 9.4 referring to fishery products. and/or on the quantity of their toxins or metabolites. a batch of foodstuffs or a process. The criteria are outlined in Table 9.LEGISLATION The aim of this legislation is to complement food hygiene requirements. with chapter 2.6. Two kinds of criteria have been established: food safety criteria. ensuring that foods being placed on the market do not pose a risk to human health. presence or number of microorganisms. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. 9.1 Food safety criteria Chapter 1 of the regulation focuses on food safety criteria that cover foods such as ready to eat foods and fishery products and live bivalve molluscs.6.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. volume. and the legislation applies to all businesses involved in food production and handling. per unit(s) of mass. based on the absence. 215 .II.

I Food safety criteria Sampling plan(1) n 10 5 Absence in 25 g EN/ISO 11290-1 c m M Products placed on the market during their shelf life Limit(2) Analytical reference method(3) Stage where the criterion applies Food category Microorganisms 1. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1. monocytogenes L. other than those intended for infants and for special medical purposes EN/ISO 11290-1 Products placed on the market during their shelf life Before the food has left the immediate control of the food business operator.2 Ready-to-eat foods able to support the growth of L.TABLE 9. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1. monocytogenes. other than those intended for infants and for special medical purposes(4) (8) L.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life .3 Ready-to-eat foods unable to support the growth of L. monocytogenes 1. monocytogenes.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L.

coli (15) 1.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1.25 Live bivalve molluscs and live echinoderms. manufactured from fish species associated with a high amount of histidine (17) Histamine . tunicates and gastropods 1(16) 0 230 MPN/100 g of flesh and intravalvular liquid 100 mg/kg 200 mg/kg HPLC (19) ISO TS 16649-3 Salmonella 1. tunicates and gastropods 9 (18) 2 E.17 Live bivalve molluscs and live echinoderms.27 Fishery products which have undergone enzyme maturation treatment in brine.Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.

when he is not able to demonstrate. c = number of sample units giving values between m and M For point 1. monocytogenes.92. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . Malle P. Scombresosidae. 79. soft drinks. honey and confectionery. Pomatomidae. according to which the whole batch is to be deemed unsafe.. wine. spirits and similar products.1 . 5 This criterion shall apply if the manufacturer is able to demonstrate. when recontamination is not possible after this treatment (for example. products heat treated in their final package).4 or aw ≤ 0.1. Clupeidae. Other categories of products can also belong to this category. excluding sprouted seeds. Assay of biogenic amines involved in fish decomposition.. products with a shelf-life of less than five days shall be automatically considered to belong to this category. Bouquelet S.25. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. shall not apply. Malle P. to the satisfaction of the competent authority. Single samples may be taken at retail level.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. including cocoa and chocolate products. This criterion shall apply to products before they have left the immediate control of the producing food business operator. The operator may fix intermediate limits during the process that must be low enough to guarantee that the limit of 100 cfu/g is not exceeded at the end of shelf life. Bouquelet S. Regular testing against the criterion is not required in normal circumstances for the following ready-to-eat foods: — those which have received heat treatment or other processing effective to eliminate L. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. 1996. subject to scientific justification.. The most recent edition of the standard shall be used.. Duflos G. Coryfenidae. 43-9. References: 1. 2. — bread. Valle M. — — bottled or packed waters. cider. that the product will not exceed the limit 100 cfu/g throughout the shelf life. biscuits and similar products. E. m = M. uncut and unprocessed vegetables and fruits. J.0 and aw ≤ 0. Dervin C.94. Products with pH ≤ 4. sugar. A pooled sample comprising a minimum of 10 individual animals.. AOAC Internat. Engraulidae. beer. Particularly fish species of the families: Scombridae. coli is used here as an indicator of faecal contamination. products with pH ≤ 5. — fresh. to the satisfaction of the competent authority. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. live bivalve molluscs.

— unsatisfactory. if any of the values are > the limit. if the following requirements are fulfilled: 1. tunicates and gastropods in relation to testing E. no values observed exceed the limit of M.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. 1999. 82. monocytogenes in ready-to-eat foods able to support the growth of L. if the presence of the bacterium is detected in any of the sample units. if all the values observed are ≤ the limit. Histamine in fishery products from fish species associated with a high amount of histidine: — satisfactory. where the limit refers to a pooled sample. if the presence of the bacterium is detected in any of the sample units. L. a maximum of c/n values observed are between m and M 3. The test results demonstrate the microbiological quality of the batch tested(1). J. (1) 219 . the mean value observed is ≤ m 2. — unsatisfactory. if all the values observed indicate the absence of the bacterium. if all the values observed indicate the absence of the bacterium. monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory. — unsatisfactory. — unsatisfactory. if the presence of the bacterium is detected in any of the sample units. excluding live bivalve molluscs and live echinoderms. The test results may be used also for demonstrating the effectiveness of the hazard analysis and critical control point principles or good hygiene procedure of the process. L. monocytogenes before the food has left the immediate control of the producing food business operator when he is not able to demonstrate that the product will not exceed the limit of 100 cfu/g throughout the shelf life: — satisfactory. coli. Interpretation of the test results relating to Table 9. Salmonella in different food categories: — satisfactory. — unsatisfactory. AOAC Internat. monocytogenes in other ready-to-eat foods: — satisfactory.I The limits given refer to each sample unit tested. if all the values observed indicate the absence of the bacterium. L. 1097-101. if the mean value observed exceeds m or more than c/n values are between m and M or one or more of the values observed are > M.

1 Shelled and E.TABLE 9. coli 5 shucked products of cooked crustaceans and molluscan shellfish 2 100 cfu/g 1000 cfu/g EN/ISO 6888-1 or 2 FISH AND SEAFOOD Coagulasepositive staphylococci 5 Improvements in production hygiene 220 .II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2.4.

7. 2006 No. and the rest of the values observed are ≤ m. The test results demonstrate the microbiological quality of the process tested. For example.7 Food Hygiene (England) Regulations 2006. had not been 221 . The regulations prescribe a chilled food holding temperature of 8 °C or less. 2 Interpretation of the test results relating to Table 9.1 In the U. which overrides the 8 °C requirement. 14 (Hygiene requirements specific to the U.K) Temperature control requirements 9. the defendant will need to prove that the food was on service or display. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health.I. — unsatisfactory. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. — acceptable. The defendant may be required to produce well-founded scientific proof to support his claims. 14 details temperature control requirements for foods in general.II The limits given refer to each sample unit tested. — acceptable. if one or more of the values observed are > M or more than c/n values are between m and M. and the rest of the values observed are ≤ m. The regulations provide for defences in relation to upward variations of the 8 °C temperature. c = number of sample units giving values between m and M The most recent edition of the standard shall be used. 9. with chill holding tolerance periods. tolerance periods for chill-holding of foods and hot-holding variations. E. S.LEGISLATION 1 n = number of units comprising the sample. if one or more of the values observed are > M or more than c/n values are between m and M. — unsatisfactory. S. if a maximum of c/n values are between m and M. Schedule 4 of the Food Hygiene (England) Regulations 2006.I. Hot-held foods (food having been cooked or reheated and is for service or on display for sale) must not be kept below 63 °C. if all the values observed are ≤ m. if a maximum of c/n values are between m and M. 2006 No.K. if all the values observed are ≤ m. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature.

9. it would need to be proved that the food was being transferred to or from a vehicle used for the activities of a food business.8 Guidance In the U. S. the Food Standards Agency has published guidance notes on the requirements of the EU hygiene and microbiological criteria regulations which can be accessed at the following link: http://www.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment.K.I. and was kept at above 8 °C or the recommended temperature. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S. including: Shelf life: where the minimum durability or 'use by' indication is required according to Regulation 20 or 21 of the Food Labelling Regulations 1996 (form of indication of minimum durability and form of indication of ‘use-by date’) the period up to and including that date.S. For other food. 2006 No. or. 2006 No. 14) do not apply to any food covered by EU Regulation 853/2004 on hygiene of products of animal origin or any food business operation carried out on a ship or aircraft. Alternatively. or temporary breakdown of equipment For Scotland there are separate provisions to include requirements to hold food under refrigeration or in a cool ventilated place. such as that below. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature.gov.I. 3). preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. for a limited period consistent with food safety.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. or at a temperature above 63 °C and to reheat food to a temperature of at least 82 °C (The Food Hygiene (Scotland) Regulations 2006. was kept at above 8 °C or the recommended temperature for an unavoidable reason.food. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions.

2076/2005. 852/2004 is now set out in a Corrigendum (OJ No. 14. 1. 1662/2006. 25.2004. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. L226. 2076/2005. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption (OJ No.10 Further Reading Regulation (EC) No. 25. 2074/2005. 25. 1441/2007. as read with the corrigenda at OJ No.6. The revised text of Regulation (EC) No. 55). 2076/2005. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. 30.12. 9. 30. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. The revised text of Regulation (EC) No. Regulation (EC) No.4. The revised text of Regulation (EC) No. 223 . L139.2004. 1). L226. L139.2004. L283. 2074/2005. 2074/2005 and Regulation (EC) No. 2075/2005 and Regulation (EC) No. 62) as amended by Regulation (EC) No. these are not be discussed here. 853/2004 is now set out in a Corrigendum (OJ No.LEGISLATION 9. Regulation (EC) No.2006. 1688/2005. 2073/2005 on microbiological criteria for foodstuffs (OJ No.6.9 Other Relevant Legislation The following regulations about fish designations need to be considered: • Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products Regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products • Note: Requirements for preserved fish are laid down elsewhere. L226. namely.6.4. 30.2004. 2076/2005.2004. Regulation (EC) No. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. L139. Regulation(EC) No.2005. 83) as amended by Regulation Nos 882/2004. 3) as amended by Regulation (EC) No.4. 22. laying down common marketing standards for preserved sardines. 206).10. Regulation (EEC) 2136/89. 1019/2008 and as read with Regulation 2073/2005. as amended. 22) as amended by Regulation (EC) Nos. Regulation (EC) No. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. L338. 1663/2006.2004. Commission Regulation (EC) No. 854/2004 is now set out in a Corrigendum (OJ No. 2074/2005. Regulation (EC) No.

22. 22. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.1.12. (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004 ( OJ L 338.2001.2000. 22) Commission regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products as amended by Commission Regulation (EC) No 1792/2006 of 23 October 2006 (OJ L 278.FISH AND SEAFOOD Commission Regulation (EC) No 2074/2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC)No 882/2004 of the European Parliament and of the Council.2005. 23. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004.10.2005. 6) 224 .12. 21. derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004.

5 minutes.1. 225 .1 10.1.2 Oxygen requirements Facultative anaerobe.3. Some death will occur at this pH if the temperature is 4 °C.0 x 1. PATHOGEN PROFILES 10.3 . 10. a 10 to 1.3 minutes.5. and D-values at 45 and 51 °C have been reported as 29. respectively (1). At pH 4. 10. 0.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.5. Morphology Gram-negative rods. 10.1. At 5 °C.5 pH Under otherwise optimal conditions.3 Temperature Aeromonas grows at temperatures of 2 .1 Aeromonas spp. The optimum growth temperature is 28 °C (2).1. no growth is observed (1).10.1.45 °C (1).5 µm 10.1. under optimal incubation temperatures. Aeromonas has the potential to grow over a pH range of ca 4 . and 2.000-fold increase in number is observed after one week in food systems (1).0 . The minimum pH value for growth is thought to be pH 5.10 (3).

6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C.5% NaCl (4).2.1 Morphology Gram-positive. or at 90 ºC for a few seconds. Thus an aerobically packed product may not support the growth of the organism on the surface. F are the types that are responsible for most cases of human botulism (5. 10.5 . being capable of slow growth and toxin production at low temperatures .2.0 µm. but the interior of the food may do so. but Group II strains are psychrotrophic. B.2. 10.2 Clostridium botulinum Seven different types of C. but the spores of this organism are more so. All C. 10. 226 . botulinum are known. 10. Group I will not grow at temperatures of 10 ºC or less. botulinum is prevented.4 Heat resistance The vegetative cells of C. A to G. botulinum are not particularly heat resistant. botulinum types produce heat labile toxins. types B and F can be either. botulinum grow reasonably well in the temperature range of 20 45 ºC. 6).3 Temperature All strains of C.2 Oxygen requirements Although C. 10. forming at least seven different toxins. All type A strains are proteolytic and type E strains are usually non-proteolytic.even as low as 3 ºC (7).7 . Types A.2% sodium chloride (NaCl) (1). spore-forming rods. There are four main groupings of the organism.22.2.2. E and.FISH AND SEAFOOD 10. which may be inactivated by heating at 80 ºC for 20 . but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different.1. at 85 ºC for 5 minutes. to a lesser extent.4 x 1. and groups I and II are those responsible for the botulism cases. 0. and is sensitive to > 4.30 minutes. botulinum is an obligate anaerobe.

0 .5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4.3.97 for Group II (5). In commercial situations.3 10. 10.2. where oxygen levels are reduced. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. therefore. 10. safety margins must be introduced to allow for process variability.0 µm. botulinum spores The most heat resistant spores of Group I C. It will not. The organism will grow well in the centre of meat or poultry dishes.0 and aw > 0. 10.21 minutes) (9).is an obligate anaerobe. 0. These values have been established under carefully controlled laboratory conditions.94 for Group I and 0. pH and temperature.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. grow on the surface of foods unless they are vacuum or gas-packed. perfringens .1 Clostridium perfringens Morphology Gram-positive spore-forming rods. 10. These concentrations correspond to limiting aw of 0. Note also that if humectants other than salt are used.7 Characteristics of C. respectively (8. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. 227 .g.0. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I.10. 9).PATHOGEN PROFILES 10. However. particularly by cooking. the minimum aw for growth may be lower. they may survive mild heat treatments (70 .2. parcooked meals with pH value > 5.9 x 2. vacuum packed.97) (10.2.1. 11)). botulinum depends on solute.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e.like other clostridia .3 . botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0.2 Oxygen requirements C.3.6 and pH 5.

perfringens in roast beef at 60 ºC is 14. 10. Recorded heat 228 .FISH AND SEAFOOD 10. 0 . 15. (It is unusual for spores to be formed in foods after the growth of this organism (14)). Some studies indicate that the presence of 3% NaCl delays growth of C. 13). the limits for aw are affected by temperature. In addition.3. perfringens in vacuum packed beef (18). although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. The D-value for C. vegetative cells die rapidly (12).10 ºC. and their heat resistance is also affected by the heating substrate.3.3. The lowest aw recorded to support the growth of C. Its optimum temperature for growth is 43 .8% inhibit growth of most C.7 Characteristics of C. etc. 10. 16).50 ºC. lower concentrations may be effective in combination with other factors. perfringens can vary quite considerably in their heat resistance. type of solute. At cold temperatures.3.3 Temperature The most significant characteristic of C. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures.6 Aw/Sodium chloride C.it is destroyed by heating at 60 ºC for 10 minutes (14. 10. 18). perfringens strains. While some growth can occur at 50 ºC.3. Under otherwise ideal conditions. perfringens has the potential ability to grow within the temperature range 15 . very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. the enterotoxin is not heat resistant . 10.45 ºC. Salt concentrations of 6 . perfringens appears to be 0.5 pH C. however.93 and 0. will survive greater extremes of pH (and aw) (12.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C.5 minutes (17).3. pH.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. perfringens spores The spores of C. It grows best at pH values between 6 and 7 (the same pH as most meats). Spores. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. perfringens is not a tolerant organism with respect to pH. although C. 13). As in the case of other factors limiting the growth/survival of this organism. depending on strain and other conditions. perfringens is not tolerant of low water activities. perfringens.

the organism is able to grow at pH values well below pH 5 (pH 4.6 . or even below 0 ºC.19 minutes. Under ideal conditions. using hydrochloric acid as an acidulant).4 Heat resistance L.0. the lowest limit for growth is likely to be considerably higher .especially at refrigeration temperatures. The upper temperature limit for the growth of L.albeit slowly . monocytogenes in crawfish tail meat at 55.2.3 is the lowest value where growth has been recorded. D-values for L. In foods. it is not a sporeformer.3 . 10. -0. being potentially capable of growing .at refrigeration temperatures down to.5 x 0. 229 .2 has been suggested as the lowest working limit (22). Listeria spp. However. however. 60 and 65 ºC were reported as 10. Morphology Gram-positive short rods. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.23.4. 10.64.4 ºC is probably a more likely minimum for growth in foods (19).5 .98 and 0. is between 30 and 37 ºC. respectively (21).2.4 . monocytogenes is not a particularly heat resistant organism. 10. requiring days or weeks to reach maximum numbers. and temperature. so can be destroyed by pasteurisation. however.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic.2 Oxygen requirements Aerobe or microaerophilic. Its optimum growth temperature. 1. growth at low temperatures can be very slow. monocytogenes is reported to be 45 ºC (20). 0.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17.4. pH < 5. 10.0 µm.8 minutes for heat sensitive spores (12).4.5 pH The ability of Listeria to grow at different pH values (as with other bacteria) is markedly affected by the type of acid used. and where acetic acid is used as an acidulant.4 10.0 minutes for heat resistant spores to 1.

3.0 µm.6 10. It is likely to survive.5. at salt levels found in foods (10 .5. 0. but has been shown to be able to grow at an aw level of 0.2 Oxygen requirements Facultative anaerobe.7 x 1.FISH AND SEAFOOD 10. monocytogenes is quite tolerant of high NaCl/low water activities.5 10. 10.3 µm.5.5 (24. or even grow.6 Aw/Sodium chloride L.8.6.1 Plesiomonas Morphology Gram-negative.5.90. 10.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24. 10.3 Temperature Plesiomonas can grow in the temperature range of 8 . 230 .1 Salmonella spp. 10. 25). It grows best at aw of ≥ 0.8 .1.5 . 0. 10.97. 25).5 pH Plesiomonas has a pH range for growth of pH 4.5. The bacterium may survive for long periods at aw values as low as 0.83 (20). Morphology Gram-negative short rods.0. peritrichous flagella.45 ºC. 10.4.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24). 10.0 x 1 .5.5 . the optimum being 30 ºC (23).12% NaCl or more).0 .

10. a heat resistant organism. pasteurisation and equivalent treatments will destroy the organism under normal circumstances.93 (as reported in dried meat and dehydrated soup). The optimal pH for Salmonella growth is between 6. some strains are able to grow at temperatures as low as 4 °C (26). The growth of Salmonella is generally inhibited by the presence of 3 .5. Salmonella senftenberg 775W is about 10 . under otherwise ideal conditions. or possibly 0.3 Temperature Salmonellae can grow in the temperature range of 7 .6 Aw/Sodium chloride Where all other conditions are favourable. and with an appropriate acid. Furthermore.2 Oxygen requirements Facultative anaerobe.8 . substrate.48 ºC. Salmonellae are quite resistant to drying. depending on the type of acid and temperature. with a z-value of 4 . depending on serotype. It is not. Some death will occur at pH values of less than about 4.945.5 ºC.6.6. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments.6.7. D60 ºC values normally range from about 1 to 10 minutes. However. 10. However.PATHOGEN PROFILES 10.37 ºC. temperature and pH.4 Heat resistance Salmonella is not a spore-forming organism. Salmonellae are quite resistant to freezing. strains vary in their ability to withstand heating. 10. the optimum being 35 . although salt tolerance increases with increasing temperature (28). Growth is slow at temperatures below about 10 ºC. and appropriate heat treatments must be determined experimentally for low-aw foods.9.6. therefore. Salmonella has the potential to grow at aw levels as low as NaCl.5 . 10. 231 .5 pH Salmonella has a pH range for growth of pH 3.20 times more heat resistant than the average strain of Salmonella at high aw (27).

As is the case with other bacteria.2 to 9.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH.5 pH The pH at which a staphylococcal strain will grow is dependent upon the type of acid (acetic acid is more effective at destroying Staph.7. etc.37 ºC (29).0 . under otherwise ideal conditions (29.7. fat content.7 .20 minutes.2 Oxygen requirements Facultative anaerobe. The limits for toxin production are also narrower than for growth.7. NB: The growth of Staph.5). 10. aureus is more limited under anaerobic than aerobic conditions. aw. aureus viability. 10. the organism is heat sensitive and will be destroyed by pasteurisation.7.3 Temperature Under otherwise ideal conditions. Under most circumstances.3 (optimum 7. stressed cells can also be less tolerant of heating. Freezing and thawing have little effect on Staph.0. aw and temperature (sensitivity to acid increases with temperature).7. 10. aureus than citric acid).7 10. 0.).9 µm diameter.FISH AND SEAFOOD 10. It can survive well at low temperatures.1 Staphylococcus aureus Morphology Gram-positive cocci. The following relate to limits for growth only. 31). Staph. with an optimum of 35 .7. however. but may cause some cell damage (30). 232 . Most strains of staphylococci can grow within the pH range 4. the D60 ºC value is 2 . depending on aw. In meat macerate.5 ºC. 10. aureus can grow within the temperature range 7 – 48.

In general.99 aerobically under otherwise optimal conditions.5 .8. 10.0 . However.2 Oxygen requirements Facultative anaerobe. 10.>0. * dependent on temperature.7.2. an aw of 0. Staphylococci are more resistant to salt present in foods than other organisms. In general.0.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.6 Aw/Sodium chloride Staph. 0. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31.3 Temperature The optimum temperature for growth has been reported as 30 ºC.99 little or no toxin production in anaerobically packed foods.87 and > 0. 31) between 0. but certain strains can grow in 20% NaCl. It can grow over the aw range 0. and solute. aureus can grow in 7 .8 x 1.6 µm 10.7.7. strain. with a temperature range for growth of 10 .2 .83 .5) (29. Staph.9.1 Vibrio cholerae Morphology Gram-negative curved rods.10% NaCl. 10. 233 .7 Limits permitting toxin production 10 . especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.8 10.86 is the generally recognised minimum in foods (32). atmosphere.43 ºC (35).8. An effect of increasing salt concentration is to raise the minimum pH of growth.8.PATHOGEN PROFILES 10. 34).0 (optimum 7. pH.4 .

10. It can grow in the presence of 0. 234 . under otherwise favourable conditions.97 .0% NaCl (35).9 10.FISH AND SEAFOOD Also.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media. declining in number during storage. 10. cholerae in crab meat homogenate at 60 ºC is 2. it has a pH range for growth of pH 5 .8. and it is sensitive to refrigeration temperatures (0 .4 Heat resistance V. with an optimum of 0.6. 10.8. parahaemolyticus is easily destroyed by drying.0.2 Oxygen requirements Facultative anaerobe. 10.984 (35). cholerae may persist on frozen fish for >180 days (36).9.6 (35. cholerae can grow over the aw range 0. V.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods.4. 10.998. but not acid conditions.6 Aw/Sodium chloride V.9. are 5 and 43 ºC. cholerae is sensitive to heat at temperatures of >45 ºC.9.65 minutes (35).5 pH This organism is tolerant of high pH values.5 ºC). there is evidence that V.8. 37.1 . with optimum at 7.9. 38). respectively. The D-value for V. but only moderately sensitive to freezing (40). with an optimum of 37 ºC (39). 10.

10.922.5 minutes.4 Heat resistance V.02 . 41).8. respectively.6 (41).10.29 minutes. with a temperature range for growth of 8 . The organism can survive for long periods at freezer temperatures (42). vulnificus is heat sensitive.5 pH The pH range for growth is 4. but the optimum concentration is 2 .9. parahaemolyticus will grow at aw levels as low as 0. The D-values at 55 ºC for clam and crab homogenate are 0. 10. 10. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.43 ºC (42). with an optimum pH of 7. depending on solute.9. parahaemolyticus is unable to grow in the absence of salt.6 Aw/Sodium chloride V.9.11. 41).3 Temperature The optimum temperature for growth has been reported at 37 ºC. 10.4 Heat resistance V.5% NaCl.10 Vibrio vulnificus 10.PATHOGEN PROFILES 10. V. 10. and 2.6 .1 Morphology Gram-negative curved rods.98 (35.8 .2 Oxygen requirements Facultative anaerobe.0. and are readily destroyed by cooking. 10. The optimum concentration of NaCl equates to aw of approximately 0. 10. parahaemolyticus are not heat resistant. 235 .4% (39.10. respectively (39).10. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).

Betts G.. 9. ASM Press. Ed. Blackie. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.. 6 (1). Dodds K.. Eds. 1994. Novak J.FISH AND SEAFOOD 10. 357-67.N. 1-19.W. Eds. Gorman J. Clostridium botulinum. 10.11 1.R.. Beuchat L.W.10. in International Handbook of Foodborne Pathogens. Hui Y.D. 236 . New York.A. Abeyta C. 111-89. Lund B.N. vulnificus grows over the range 0.L. Cliver D. Marcel Dekker. Volume 1.D. 4..5.R. Clostridium botulinum. Marcel Dekker. Great Britain.H.. Diseases Caused by Bacteria. 1-27. Juneja V. Eds. 5-19.L.. Doyle M.. Ed. 3. Hauschild A.H.H. 5.K. in Foodborne Pathogens.H.D.J. 2. New York.L. 279-303. 1993. Gorham J.. New York. 121-76. in Foodborne Disease Handbook.E.. 1995.5 . 97-131..0% NaCl. Caister Academic Press. 383-408. London. 1994. Eds.M. Murrell K. Stelma G. Clostridium botulinum. Doyle M. International Commission on Microbiological Specifications for Foods. Aeromonas hydrophila group. with an optimum pH of 7.. with an optimum of 2. in Foodborne Bacterial Pathogens. Doyle M. Cliver D. Palumbo S. Faris A..H. 6. 10.W. 1989. Eds.6 Aw/Sodium chloride V. Volume 1: Diseases Caused by Bacteria.P.. Hui Y. Aeromonas hydrophila. Principles of control. Hauschild A.. 329-49.D. Gaze J. Aeromonas. 2001. Marcel Dekker. Notermans S.O. Bhunia A.W. Washington D. 1993. New York. Dodds K.H. Microbiology and Molecular Biology... New York. Foegeding P. in Clostridium botulinum: Ecology and Control in Foods. 57-63. Marcel Dekker Inc. Marcel Dekker. 1996. Johnson E. Eds.L.M.R.O. Kim J. in Food Microbiology: Fundamentals and Frontiers. Ed. Clostridium botulinum and Clostridium perfringens. Montville T. Stelma G.10.. Smith J. 2005.5% (42).5 pH The pH range for growth is 5 . Hauschild A. Kovacek K. 11. Marcel Dekker. in Clostridium botulinum: Ecology and Control in Foods. Miliotis M. 7.. New York. 1989. Dodds K..C. Aeromonas species. in Foodborne Disease Handbook.M. Fratamico P. Volume 5: Microbiological Specifications of Food Pathogens..10. Potential hazards associated with REPFEDS. 10. Eds.W. Austin J. in Foodborne Bacterial Pathogens. New York.P. 2003. Food Control. Murrell K. in Microorganisms in Foods. Bier J. 8.P... References International Commission on Microbiological Specifications for Foods. Marcel Dekker.8 (42). Peck M.

201-8. Cliver D. Journal of Applied Bacteriology. Ed. Doyle M.. 1990. Eds. New York. Clostridium perfringens. 18.. Marcel Dekker.PATHOGEN PROFILES 12.P.A.J. in Foodborne Diseases. Volume 5: Microbiological Specifications of Food Pathogens.. Doyle M.. 735-42. 237 . Beuchat L. Ed.A.J. Hui Y. Lancet. Ed. 17.P. Diseases Caused by Bacteria. 23. Cliver D. Emery D.. in Foodborne Bacterial Pathogens... Koburger J. International Commission on Microbiological Specifications for Foods.O. 112-25.H. Growth of Listeria monocytogenes at refrigeration temperatures. 1990. Clostridium perfringens. 191-243.H. Eds.M... Labbe R. Kim C.C. in Microorganisms in Foods. Blackie.. 19. in Microorganisms in Foods. 16. in Foodborne Bacterial Pathogens. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs.. Wolfe Publishing Ltd. Cliver D.G.A. Murrell K. Lund B. Listeriosis and Food Safety. Halvorson D. 26. in Food Microbiology . 1996. Archer P. 13.R. International Commission on Microbiological Specifications for Foods. International Commission on Microbiological Specifications for Foods. Volume 1.T. Academic Press.H. Clostridium perfringens. Doyle M. in Foodborne Disease Handbook.P. Juneja V. Washington D. Walker S. Plesiomonas. Washington D.. 2001. London. Plesiomonas shigelloides. Eds. 1996. Marth E. Gorham J. Listeria monocytogenes.O.D. Listeria.G.. Clostridium perfringens gastroenteritis. 15. 141-82. Marcel Dekker. Eds. 2005. Wrigley D. New York.R. New York.J. 1991. 68 (2). Evans M. McClane B.V. 2006.A. Ed. Listeria monocytogenes. 351-82. Blackie. 20. Plesiomonas shigelloides.. Blackie. 1989. Montville T. in Foodborne Infection and Intoxication. Montville T. Foodborne disease due to Bacillus and Clostridium species. 33. Nagaraja K. Avian Disease. 137-64. Evans M.A.O.K. 208-13. 22. 311-25.H. 14. Banks J. Volume 5: Microbiological Specifications of Food Pathogens. International Commission on Microbiological Specifications for Foods. Ed.C. Varnam A. 1989. ASM Press. Volume 5: Microbiological Specifications of Food Pathogens. in Food Microbiology: Fundamentals and Frontiers. Clostridium perfringens. Varnam A. 1989. London. 336 (8721). in Foodborne Pathogens: An Illustrated Text. Marcel Dekker.M.G. Ed. 21. 24. 2007. 129-38. London.J. in Microorganisms in Foods.An Introduction. 1996. International Commission on Microbiological Specifications for Foods. Ryser E. 1994. New York. Riemann H. London. Elsevier. 133-67.. London.. 25. 982-6. Labbe R. Marcel Dekker. Clostridium perfringens food poisoning. 1990. Matthews K. International Commission on Microbiological Specifications for Foods.. London. 229-40.R. 157-62.P. Eds. Rinke H.. ASM Press. Johnson E.

Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. Cholera: current epidemiology. 543-68. 1996.C. Academic Press. 642. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems.. Ed. 1983. Food and Environmental Sanitation. New York. 426-35. Ed. Vibrio parahaemolyticus. Cleland D. Smith J.S. 85-106.K. CDR Review. 29. 35. London.FISH AND SEAFOOD 27. Crowcroft N. Ed. 39. New York. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures. Wymondham. Jay J. 414-25... Ed.A. Doyle M. Griffiths M. in Microorganisms in Foods. 31.62. International Commission on Microbiological Specifications for Foods. Microbiological Specifications of Food Pathogens. Cliver D.. Bergdoll M. Staphylococcus aureus. 30. Volume 5. Blackie.. Salmonellae..O. Academic Press.O. 1930.S. 327-445. 28. 33. Staphylococcal gastroenteritis. Blackie. Springer Science. in Foodborne Bacterial Pathogens.. Bergdoll M. Ed.M. Volume 5: Microbiological Specifications of Food Pathogens. in Modern Food Microbiology.. Applied and Environmental Microbiology. 2005. 32. 1996. 37.M.P. Volume 5.L. Vibrio cholerae. Fratamico P. London. Jay J. 2005. Woodhead Publishing Ltd. 523. D’Aoust J-Y. Marcel Dekker. Golden D. 45 (3). Reed G. in Foodborne Infections and Intoxications.R.H.P. London. 331-57.A.S. 545-66. 1996. 1993. Wilkinson B. Warner R. Tohyama Y. Ed. 273-84. Eds. in Foodborne Disease. Twedt R. 1989. 1994. London. Microbiological Specifications of Food Pathogens. Marcel Dekker. Staphylococcal Food Poisoning. Lee-Wong A. Golden D. Japanese Journal of Experimental Medicine. International Commission on Microbiological Specifications for Foods. 40. 2005. 327-52. Salmonella. 1989. 985-98.M. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. 2005. Stewart G. 13 (11). in Microorganisms in Foods. Gustafson J. in Foodborne Pathogens. 38. Ed. Eds. International Commission on Microbiological Specifications for Foods. London.. Eds. Loessner M. 1990. International Commission on Microbiological Specifications for Foods. Cliver D.J.M. International Commission on Microbiological Specifications for Foods. Staphylococcal intoxications. 36. Loessner M. Stress Response and Resistance. 238 . Oliver J. in Foodborne Pathogens: Microbiology and Molecular Biology. Bhunia A. Riemann H. R15764. Dairy. Cambridge. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). 34. Caister Academic Press.P. Doyle M. 4 (13). in Understanding Pathogen Behaviour Virulence.J. International Commission on Microbiological Specifications for Foods.D. 8.A. New York. Vibrio parahaemolyticus. in Microorganisms in Foods.. Blackie. 217-64.C.

55 (12). Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters. New York. 436-9.D.. Springer Science.W.A. 1996. Foodborne gastroenteritis caused by Vibrio. International Commission on Microbiological Specifications for Foods. Journal of Food Protection. 657-78. Jay J. Loessner M. 1992..M. Ed. 2005.. 985-9. Volume 5. 239 . Golden D.. Yersinia.. Microbiological Specifications of Food Pathogens. in Microorganisms in Foods. Cook D.M. Jay J. International Commission on Microbiological Specifications for Foods.PATHOGEN PROFILES 41. Blackie.A. and Campylobacter species.J.J. Golden D. Ruple A. Loessner M. Vibrio vulnificus. London. 43. in Modern Food Microbiology. 42.


gov.uk or v.holt@ifm.org.gsi.uk Institute of Fisheries Management (IFM) 22 Rushworth Avenue West Bridgford Nottingham NG2 7LF United Kingdom Tel: + 44 (0)115 9822317 Fax: + 44 (0)115 9826150 Email: info@ifm.gov.defra.uk Web site: www.ifm.uk Web site: www.uk/Topics/Fisheries Seafish Industry Authority 18 Logie Mill Logie Green Road Edinburgh EH7 4HS United Kingdom Tel: + 44 (0)131 5583331 Fax: + 44 (0)131 5581442 Web site: www.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: cawoods@denholmseafoods.gov.org.org.scotland.org/ .seafish.co.uk/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: helpline@defra.gov.uk Prepared Fish Products Association (PFPA) Federation House 6 Catherine Street London WC2B 5JJ United Kingdom Tel: + 44 (0) 208 78362460 The Scottish Government Fisheries Group Environment and Rural Affairs Department Pentland House 47 Robb's Loan Edinburgh EH14 1TY United Kingdom Tel: + 44 (0) 8457 741741 or + 44 (0)131 5568400 Fax: + 44 (0)1397 795001 Email: ceu@scotland.gsi.uk Web site: www.

org/fishery/ European Association of Fish Producers’ Organisations H.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: post@fiskeriforskning. Andersens Boulevard 37.is/ 242 .dk Web site: www.is Web site: www.suite.FISH AND SEAFOOD Department of Agriculture and Rural Development.uk Web site: www.dk Danmarks Fiskehandlere H.fao.gov.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.fiskeriforskning.ie/templates/homepage.matis.org Web site: www.eapo.gov.dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 . Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www. Northern Ireland Fisheries and Rural Policy Division 4th Floor North Dundonald House Stormont Estate Belfast BT4 3SB United Kingdom Tel: + 44 (0) 28 90524999/ 4991 Fax: + 44 (0) 28 90378323 Email: dardhelpline@dardni.uk/index/fisheriesfarming-and-food.fiskehandlerne.difta.dardni. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo.no Web site: www.bim.ifpea@eircom.dk Web site: www.asp Food and Agriculture Organization of the United Nations Fisheries and Aquaculture Department Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57053152 Email: FAO-HQ@fao.C.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: matis@matis.com/ Danish Institute for Fisheries Technology and Aquaculture (DIFTA) illemoesvej The North Sea Centre DK-9850 Hirtshals Denmark Tel: + 45 98 944300 Fax: + 45 98 942226 Email: difta@difta.com Web site: www.htm Irish Fish Processors and Exporters Association 25 Kincora Avenue Clontarf Dublin 3 Eire Ireland Tel: + 353 1 833 7882/ 02770249 Fax: + 353 1 833 7882 Email: tormg.

co.uk Institute of Food Science and Technology (IFST) 5 Cambridge Court 210 Shepherds Bush Road London W6 7NL United Kingdom TeI: + 44 (0) 207 6036316 Fax: + 44 (0) 207 6029936 Email: info@ifst.uk Web site: www.gov.org Other Sources of Information British Standards Institution 389 Chiswick High Road London W4 4AL United Kingdom Tel: + 44 (0) 208 9969001 Fax: + 44 (0) 208 9967001 Email: cservices@bsigroup.com Web site: www.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.csiro.au Web site: www.org.gsi.ac.nz Web site: www.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: reception-cmar-hobart@csiro.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: generalenquiries@fdf.org Web site: www.fdf.au New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: info@seafood.co.uk Web site: www.uk/en/index.CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: contact@nfi.ifr.org.hpa.dh.cmar.bsi-global.gov.org.com/ 243 .nz/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.ifst.uk Institute of Food Research (IFR) Norwich Research Park Colney lane Norwich NR4 7UA United Kingdom Tel: + 44 (0) 160 3255000 Fax: + 44 (0) 160 3507723 Web site: www.seafood.

uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: hpsenquiries@hps.The Swedish Institute for Food and Biotechnology Box 5401 SE-402 29 Göteborg Sweden Tel: + 46 31 3355600 Fax: + 46 31 833782 Email: info@sik.euro.hps.asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.int/foodsafety World Health Organization (WHO) Headquarters CH .codexalimentarius.int.co.scot.php Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: admin@sofht.euro.uk Web site: www. Web site: www.de 244 .FISH AND SEAFOOD Society for Applied Microbiology Bedford Heights Brickhill Drive Bedford MK41 7PH United Kingdom Tel: + 44 (0) 1234 326661 Fax: + 44 (0) 1234 326678 Web site: www.se/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: fao-who-cc@bgvv.nhs.scot.who.uk/scieh.sik.who.int/ SIK .se Web site: www.uk Web site: www.sfam.nhs.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.uk/index.who.sofht. jsp Food Safety 10 .int/en/ WHO Regional Office for Europe Scherfigsvej 8 DK-2100 Copenhagen Ø Denmark Tel: + 45 39 171717 Fax: + 45 39 171818 Email: postmaster@euro.int Web site: http://www.co.org.who.org Web site: www.who.net/web/indexen.int Web site: www.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: foodsafety@euro.

cfsan.cdc.org/ The Association of Food.ie/ Food Science Australia http://www.org Web site: http://www.eu/pol/food/index_en.FoodSafety.foodprotection.htm Centre for Disease Control and Prevention (CDC) (US) http://www.org/ Royal Institute of Public Health (UK) http://www.org.org/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.fsai.Center for Food Safety and Applied Nutrition 5100 Paint Branch Parkway College Park MD 20740-3835 United States of America Tel: + 1 888 SAFEFOOD (+ 1 888 723 3366) Web site: www.ift.au/ International Association for Food Protection http://www.ift.co.sofht.foodscience.csiro.uk/ Society of Food Hygiene and Technology (UK) http://www.int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa.gov/foodsafety/ Food Safety Authority of Ireland http://www.fpa-food.uk/ 245 . Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: info@ift.gov/ Institute of Food Technologists 525 W.gov/ World Health Organization (WHO): Food safety programmes and projects http://www.CONTACTS US FDA .org/ Institute of Food Technologists http://www.fda.riph. Beverage and Consumer Products Companies http://www.who.


breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. 69-70 Aeromonas hydrophila.INDEX Achromobacter spp. to test microbiological criteria 19 Aeromonadaceae. as autochthonous bacterial flora of fish 3 as spoilage organisms in oysters 65 as spoilage organisms in temperate crustacean shellfish 80 as spoilage organisms in tropical crustacean shellfish 81 in molluscan shellfish 60 Aerobic plate counts. pathogenic microorganisms in molluscan shellfish 59-60 as contaminant in enrobing process 37 as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45 growth in vegetable and rice 247 . planktonic source of spirolides in shellfish 163 Allergens. in crustacean shellfish 87 Alteromonas spp. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. preserved by dry/wet salting 99 Anisakis spp. 111 Astroviruses. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. in crustacean shellfish 86-7 ATP. growth and survival in crustacean shellfish 86 Aeromonas sobria. source of PSP toxins 142 Alexandrium ostenfeldii. 102 Bacillus spp. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. causing spoilage of dried fish 110. as autochthonous bacterial flora of fish 3 Aeromonas spp. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp.

cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. smoked and dried fish 113-6 hazard in mild-processed marinated products 101 hazard in sun-dried products 102 non-proteolytic – risk of growth in fish recipe meals 44 248 . 98-9 Brochothrix thermosphacta. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. 113. 137. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. of fish – effects on microflora 5-6 Carbohydrate. caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. 131 Carbon dioxide. of recipe fish meals 43 Cholera. preserved by marinating 100 Chemical detection methods. for PSP toxins 144 Chemical indicators. associated with consumption of fish/fish products 16. in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage. as cause of spoilage of MAP fish 15 Calciviruses.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. of fresh fish spoilage 11 Chemical pollutants. as autochthonous bacterial flora of fish 3 Clostridium botulinum. 226 Breading process 36-8 Brevetoxins. in crustacean shellfish 86-7 Campylobacter coli. 136. fish sauce 123 Basipetospora halophila. importance in lactic acid fermentation in fish products 130. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang. 68-9 Capture and handling. concern with respect to use of MAP 7 growth and survival in chilled and frozen prepared fish products 45 growth and survival in chilled and frozen raw fish 15-6 growth and survival in crustacean shellfish 84 growth and survival in cured.

INDEX pathogen profile 226-7 Clostridium perfringens. for detection of DA 148 ELISA kit (DSP-Check). description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. 152 Diphyllobothrium. 149-50. as contaminants in scampi 36. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). processing 334 Cockles. to determine fresh fish spoilage 11 Corynebacteria. smoked and dried fish 114 Enzymic spoilage. hot-smoking 107 ELISA assay. as autochthonous bacterial flora of fish 3 Crabs. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. 106 Contacts 241-5 Conway method. 37 as spoilage organisms in chilled products 43 as spoilage organisms in tropical crustacean shellfish 81 as spoilage organisms in vacuum-packed dried fish 111 growth and survival in cured. adverse condition on poorly preserved fish 110 EC grading scheme. as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45-6 pathogen profile 227-9 Coated fish products. of prepared fish products 44 Escherichia coli. growth and survival in crabmeat 84 growth and survival in cured. source of azaspiracids 159 source of DSP toxins 148-9. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. of fish – different processes 94 Dun. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. detection methods 148 implicated in amnesic shellfish poisoning 146 in crustacean shellfish 87 toxicology 147 Dried fish 93-121 Dry salting 97-8 Dry/wet salting 99 Drying. forms for purchase 80 Crustacean shellfish 79-92 Cured fish 93-121 Cyclic imines 160-1 detection methods 162 origins and distribution 161 toxicology 162 Degradation. to detect NSP toxins 155 ELISA kit (Biosense). to determine fresh fish spoilage 11 Eels.

of prepared fish products 44 Gambiertoxins. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. of fish at sea 5 Fatty fish. causing pink condition on fish surface 110 in fish sauce 127 Handling. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. of fresh fish spoilage 911 Evisceration. fermented fish product 126 Green cured split cod 97. as source of ciguatoxins 156 Garlic. as source of inulin in lactic acid fermentation in fish products 134 Garos. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. 102 Gymnodimines. for evaluation of fish quality 9 Eurotium. 130-5 Fish-spoilage bacteria 12 Fishtester. source of PSP toxins 142 Gyoshoyu. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. concerning crustacean shellfish 87-8 European Grading Scheme. as autochthonous bacterial flora of fish 3 in molluscan shellfish 60 spoilage organism in oysters 65 Food hygiene legislation 189-224 Food of animal origin. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. fish sauce 123 HACCP. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. mould causing spoilage of dried fish 110 Evaluation. definition of terms 176-7 implementation and review of plan 184-6 in fish and seafood product manufacture 175-88 seven stages of system 17784 Halobacterium spp. to measure fish skin resistance 10 Flavobacterium. of fish – effects on microflora 5-6 Hepatitis (non-A. processing 35 Fish component. preserved by marinating 101 Gravlaks. member of cyclic imine group 161 Gymnodinium catenatum. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. specific EU hygiene rules 196-224 Fresh fish. non-B) viruses.FISH AND SEAFOOD EU legislation. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes.

concerning crustacean shellfis 88 Jellied eels. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. for microbial inactivation in fish products 127. as source of botulism 136 International legislation. definition 1 Legislation. 100-1 Masmeen 107 Matjes. for fresh and frozen fish (Table) 20 Micrococcus spp. preserved by salt-boiling 100 Karenia brevis. smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 .INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. outbreak caused by gravid 115 Lobster. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. as autochthonous bacterial flora of fish 3 Lean fish. preserved by salt-boiling 100 Ionising radiation. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured. food hygiene 189-224 Lingulodinium polyedrum. 135 I-zushi. hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. to reduce rate of fish spoilage 12 Ikan kaju. as source of yessotoxins 152 Lipid content. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. as autochthonous bacterial flora of fish 3 as spoilage organisms in oysters 65 as spoilage organisms in tropical crustacean shellfish 81 in fish sauce 127 Microflora. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. preserved by marinating 101 Microbiological criteria. enabled by carbohydrate source in fish products 130. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. in fish fillet 1 Lipid oxidation.

growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . associated with consumption of fermented fish products 137 Oxidative rancidity. fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P. smoked and dried fish – effects of processing 96-108 of fermented fish – effects of processing 127-35 of fish – effects of processing 5-9 of ingredients of chilled/frozen prepared fish products – effects of processing 32-42 of molluscan shellfish – effects of processing 60-5 Modified-atmosphere packaging 6-9 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee guidance 7 of fresh chilled products 42 Molluscan shellfish 53-77 commercial products 54-9 definitions 53-9 Moraxella.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay. caused by lipid oxidation 11 Oysters. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. as TTX producer 145 Norovirus. smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. of crustacean shellfish – effects of processing 81-3 of cured. shigelloides. reduction in smoking process 108 Norcardiopsis dassonvillei. in crustacean shellfish 867 Norwalk virus. 153 for detection of LST toxins 151 for detection of NSP toxins 155 for detection of pinnatoxins and pteriatoxins 165 for detection of PSP toxins 144 for detection of spirolides 164 for detection of tetrotoxin 145 Mussels. for detection of azaspiracids 160 for detection of ciguatoxins 157 for detection of DA 148 for detection of DSP toxins 150.

contact details 241-2 Prorocentrum spp. in fish fillet 1 Proteus. spoilage organism in oysters 65 Protoceratium reticulatum. as component for seafood ready meals 40 Pathogen profiles. 81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 . smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. 163. 69-70 Polycyclic aromatic hydrocarbons. for detection of PSP toxins 144. reduction in smoking process 103. 161. as cause of histamine fish poisoning 15 as spoilage organism in chilled products 43 as spoilage organism in fresh fish 14-5 Pickling 94. preserved by salt-boiling 100 Pink. 152-3 Penicillium spp. 197 Plesiomonas. pathogen profile 230 in molluscan shellfish 5960. detection methods 165 origins and distribution 165 toxicology 165 Pissala. 100-1 Pindang. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation.INDEX Parasites. as source of DSP toxins 148-9 Protein content. 108 Polypaecilum pisce. as source of yessotoxins 152 Protoperidinium cassipes. growth and survival in chilled and frozen prepared fish products 46-7 growth and survival in chilled and frozen raw fish 15-9 growth and survival in crustacean shellfish 84-7 growth and survival in cured. in molluscan shellfish 60 Pseudomonas spp. as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. smoked and dried fish 115 Pasta. as source of toxins 159. effects on microflora of fish 5-9 Professional bodies. 145-6 definitions 28-30 non-seafood ingredients 32 Processing. fermented fish product 124 Plankton. growth and survival in chilled and frozen raw fish 19 growth and survival in crustacean shellfish 86 growth and survival in cured. planktonic source of azaspiracids 159 Pseudomonadaceae. causing spoilage of dried fish 110-1 Photobacterium phosphoreum. as component for seafood ready meals 40 Prawns. mould causing spoilage of dried fish 110 Potato. 225-39 Pathogens. causing spoilage in vacuum-packed dried fish 111 Pseudomonads. adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164.

36 Seafood. for microbial inactivation in fish products 127. flow diagram (Fig. 83 254 . as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 growth and survival in cured. in molluscan shellfish 60 Rice. as carbohydrate source in lactic acid fermentation in fish products 134 as component for seafood ready meals 40 RIDASCREEN. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products. for evaluation of fish quality 9 Radiation. vacuum packing 105 Salmonella spp. detection methods 165 member of cyclic imine group 164 origins and distribution 165 toxicology 165 Puffer fish.) 41 ingredients preparation. producers of brevetoxins 154 Raw fish. as source of tetrotoxin 144-5 Putty fish. processing and effects on microflora 60-1 Shewanella.) 41 Recipe fish products 38-40 Rhodotorula rubra. processing 34. 135 Rakefisk. flow diagram (Fig. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. preserved in saturated brine 99 Sauce. 100 Salting 97-9 definitions 93-4 Sardines. chilled and frozen 1-25 Recipe dish. hazard in salting of fish 97 Salt concentration. description and uses 58-9 Scampi. as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. to measure fish skin resistance 10 Salmon. resulting from poorly salted cod 110 Pyrodinium. detected in cod 115 Pteriatoxins.FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries. growth and survival in chilled and frozen raw fish 19 Pseudoterranova dicipiens. antibody-based kit to detect STX 144 RT Freshmeter. fermented fish product 126 Raphidophytes. processing 35 Shellstock. importance in lactic acid fermentation in fish products 130 importance in salting. source of PSP toxins 142 Quality Index Method. for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops. assembly and packaging. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. fermented fish product 126 as source of botulism 136 Rakørret. smoking and marinating of fish 96 Salt-boiling 94. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova.

fish sauce 123 Shrimps. 13. role in fish degradation 10. in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. 101 Storage. 14 Torry Scale. grading of 31 hot-smoking 107 US legislation. processing and effects on microflora 63-4 Skin resistance. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. in molluscan shellfish 60 Tuna. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru. as indicator of fresh fish spoilage 11 Toxins. fish and shellfish 141-74 Trade associations. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. smoked and dried fish 114 pathogen profile 232-3 pathogenic microorganisms in molluscan shellfish 59-60 potential for growth in pasta component of seafood ready meals 40 Stockfish. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 . 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. contact details 241-2 Trematode infections foodborne 137 Trichosporon. of crustacean shellfish 80. as measure of fish degradation 10 Sliming. concerning crustacean shellfish 89 Vacuum packaging. as contaminant in scampi 36 as contaminant in enrobing process 37 as hazard in dry salting of fish 89 growth and survival in chilled and frozen prepared fish products 46-7 growth and survival in crustacean shellfish 84 growth and survival in cured.INDEX Shigella spp. of fish after capture 5 Surstrømming. fermented fish product 126 Taste panel evaluation. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. for evaluation of fish quality 9 Torrymeter. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO.

152-3 256 . as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. of shellstock 61. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in fermented fish 136 growth and survival in molluscan shellfish 66-67 pathogen profile 234-5 Vibrio vulnificus. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 67 pathogen profile 235-6 Vibrionaceae. processing for seafood recipe meal centres 39 Vibrio spp. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. 68 Vibrio cholerae non-01. as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. 62 Yersinia spp. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80.

Master your semester with Scribd & The New York Times

Special offer for students: Only $4.99/month.

Master your semester with Scribd & The New York Times

Cancel anytime.