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


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

vi .

2 Initial Microflora 1.7 Published Microbiological Criteria 2.6 Published Microbiological Criteria 3.7 References 4.8 References MOLLUSCAN SHELLFISH 3.3 Processing and its Effects on the Microflora 1.1 Definitions 3.8 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 2.7 References 3.6 Pathogens: Growth and Survival 2.6 Pathogens: Growth and Survival 1.4 Spoilage 3.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.4 Processing and its Effects on the Microflora 2.2 Initial Microflora 4.2 Initial Microflora 3. 4.4 Spoilage 1.1 Introduction 2.3 Initial Microflora 2. vii .5 Spoilage 2.2 Definitions 2.5 Pathogens: Growth and Survival 4.4 Spoilage 4.7 Published Microbiological Criteria 1. CHILLED AND FROZEN RAW FISH 1.6 Published Microbiological Criteria 4.5 Pathogens: Growth and Survival 3.8 Further Reading CRUSTACEAN SHELLFISH 4.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.1 Definitions 1.8 References 1.1 Definitions 4.3 Processing and its Effects on the Microflora 3.5 Factors Affecting Fresh Fish Spoilage 1.3 Processing and its Effects on the Microflora 4. 3.

7 References 5.6 References 6. S.2 Definitions 8. CURED.I. 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 9.4 Spoilage 6.7 Further Reading FISH AND SHELLFISH TOXINS 7.8 Further Reading FERMENTED FISH 6.9 Azaspiracids 7. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.8 Guidance 222 9.1 Introduction 8.6 Lipophilic Shellfish Toxins (LST) 7. 852/2004 on the General Hygiene of Foodstuffs 191 9. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.5.6 Regulation (EC) No.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.1 Introduction 189 9.4 Regulation (EC) No.3 Processing and its Effects on the Microflora 6.3 Processing and its Effects on the Microflora 5. 14 (Hygiene requirements specific to the UK) 221 9.5 Regulation (EC) No.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7. 9.10 Further Reading 223 viii .1 Definitions 5.2 Legislative Structure 190 9.4 Implementation and Review of the HACCP Plan 8. 8.5 Pathogens: Growth and Survival 6.3 Regulation (EC) No.1 Definitions 6.9 Other Relevant Legislation 223 9.6 Published Microbiological Criteria 5.3 Stages of a HACCP Study 8.4 Spoilage 5.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.8 Ciguatoxins 7.3 Tetrodotoxin 7.10 Cyclic Imines 7.2 Initial Microflora 5.11 Conclusion 7.4 Amnesic Shellfish Poisoning (ASP) Toxins 7. EU FOOD HYGIENE LEGISLATION 189 9. 2006 No.1 Introduction 7. 7.7 Food Hygiene (England) Regulations 2006.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.5 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. SMOKED AND DRIED FISH 5.2 Initial Microflora 6.5 Pathogens: Growth and Survival 5.

11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .1 Aeromonas spp. 10.8 Vibrio cholerae 10.4 Listeria spp.6 Salmonella spp.10 PATHOGEN PROFILES 10.2 Clostridium botulinum 10.10 Vibrio vulnificus 10.3 Clostridium perfringens 10.9 Vibrio parahaemolyticus 10.5 Plesiomonas 10.7 Staphylococcus aureus 10. 10. 10.


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

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

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

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

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

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

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

102 101 .I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. and risk associated with the consumption of seafood is low (2. and mud Ubiquitous.106 cells/g.1 .0. 4 .19 µg/kg bodyweight from < 102 .37 °C Unknown Unknown TABLE 1. Plesiomonas shigelloides 3 . Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. estuarine environments. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp.106 cells/g (exception: V. stream water. birds.FISH AND SEAFOOD TABLE 1.26 °C 10 . 105 .1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.37 °C High for most species. sewage. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria. numbers of pathogenic bacteria in raw fish tend to be low.103 High (107 . 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. However.>106 Salmonella spp. Escherichia coli Yersinia enterocolitica 101 . Toxin levels 0. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 .35 °C 8 .14 . 4). In addition.109 cells/g) * Some sources considered L. vulnificus) 5 .II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.

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

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

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

Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. and the use of a HACCP plan. The addition of high levels of carbon dioxide to these fish. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. as already stated. from both a microbiological and a chemical standpoint. 3. In the United Kingdom. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16).g.g. grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. Only the highest-quality fish. Although all fish species need to be treated separately. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. The growth of these bacteria is reduced by high carbon dioxide concentrations but. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). but is recognised as a drawback of carbon dioxide application. 4. this problem occurs to a lesser extent with meat. 5. 2.3 °C at all points from packaging onwards. It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. For other less fatty marine fish. greater advantages are seen. The problem of water loss in MAP fish is common. 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). For fatty fish. adequate product labelling in respect of storage temperature. use of high-quality raw fish. at all times.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13). trimethylamine and sulfides). should be used. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. however. botulinum. 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. owing to the dissolution of cellular structures. Subsequent storage must be at or below 3 °C. TMAO) that can be reduced to produce 8 . adequate shelf life and cooking requirements. These conditions include storage of the product at ≤ 3. however. The replacement gas should contain at least 40% carbon dioxide. may result in unacceptable colour and textural change and hence little shelf life extension (11).

reactions caused by activities of the fish’s own enzymes. with no smell.4 1. C (C=reject) Significantly. texture. TABLE 1. Various methods for whole fish freshness evaluation by a trained panel have been proposed.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. smell and 9 . loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. 18) and are probably the most widely used. the skin turns from iridescent to dull and bleached with bacterial slime. and this pattern is the basis of freshness grading schemes. Over time. Loss of freshness and spoilage cannot be separated as processes. B. so lower carbon dioxide concentrations may have better preservative effects. and metabolic activities of microorganisms present in the fish. When working with fillets. 17. 1.4.III. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation.CHILLED AND FROZEN RAW FISH off-odours. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids). The eyes turn from convex and clear to concave and opaque. A. to brown and slimy with an intense off-odour. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). these are summarised in Table 1.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. the gills from pink and shiny. or for that matter evaluation of the taste of the fish. and the flesh turns from bright and elastic to dull and soft (1). as traditional spoilage patterns of the external organs are typically very distinct.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). Degradation of whole fresh fish stored in ice generally follows a set pattern. Taste panel evaluation of cooked fish may be used to score or grade colour. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 .

texture becomes soft and watery or hard and dry. or simply a waste product present due to bioaccumulation (22). IMP. However. Generally. all marine fish use/have TMAO (21). possibly slightly metallic. By measuring the concentrations of the six components.5 (18.FISH AND SEAFOOD taste of cooked fish. an intense odour compound. These processes are strongly influenced by the physical conditions of storage. 19). From being flaccid. The process of ATP breakdown is used as an indicator of fish freshness (24). the Torrymeter. is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20.8 to 6. little flavour. cabbage-like. an osmoregulator. this will generally correlate with the length and severity of rigor. thus. the conductance generally increases. To a varying extent. or its remaining shelf life (1). 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. bacterial reduction of TMAO to TMA. the pH drop of the muscle is correspondingly less. During the initial storage. 23). After death. 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. ADP. Because fish tend to have relatively little residual glycogen compared with mammals. the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). ii) Neutral taste. which increases from 0% towards 100% with time. As the fish degrades. post-rigor values are typically in the range of pH 5. After death. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical. HxR and Hx) gives a quantity (K-value. biochemical and microbiological actions. sulphurous and/or rancid smells. After some time. fruity and/or bitter off-flavours. RT Freshmeter or Fishtester). sea-weedy taste. the Eh of fish muscle remains relatively high (20). iv) Enhancement of the spoilage characteristic of phase iii. the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. AMP. rigor resolves. iii) Traces of sour. a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. as little endogenous reduction of TMAO occurs (18). 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. an anti-freeze. development of sickly sweet. chemical. reduction in ATP and increase 10 . rigor mortis is the first noticeable change in the fish. TMAO permits a high Eh to remain in the muscle tissue. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. %). spoiled and putrid.g. the muscles relax again and the fish returns to a flaccid state. ammoniacal.

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

For example. the lower the temperature of storage. The concept of specific spoilage organisms (SSOs) is not new.5. the results of altering one parameter may have far-reaching consequences on others. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity.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). 1. 34). by altering the gas atmosphere (extrinsic).5 1. 5). The sum of these changes may result in a different spoilage profile. For fresh fish iced immediately after capture and continually stored in ice. Also.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. careful temperature control during storage is not only important in terms of quality loss. a minority are often associated with the majority of the spoilage. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks.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.5. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. the slower the spoilage processes proceed (within limits). All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. For fresh fish. The factors are interactive and cannot be completely isolated. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH.and temperate-water fish spoilage rates may be many. biochemical and microbiological reactions taking place within and on the fish. 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. Fish spoil as a result of the chemical. 1. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. but also crucial for the assurance of consumer safety.5. 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.FISH AND SEAFOOD 1.

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

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

1. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. age. botulinum recorded up to 1963. B. types B. Recent studies have investigated the dominance of B. where a particular 15 . 5) reports that fresh fish consumption has never been shown to cause human botulism. Some Ph. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. a convenient but diverse species of bacterium with a number of different types. Review of outbreak data suggests fresh fish to be safe. botulinum. Growing evidence suggests a role for B. previous exposure and access to supportive treatment. E. lightly preserved fish products are associated with botulism. B. 1.6 Pathogens: Growth and Survival Endogenous chemicals. Adult disease is caused by the production of the botulinum toxin. (and very rarely F and G) have been reported to cause human disease. thermosphacta did not produce off-flavours until 2 . but with proper cooking. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. bacteria and higher parasites all present some risk associated with fish consumption (1. 6).3 °C (Group II) and these are non-proteolytic (i. this is probably due to spoilage occurring before toxin elaboration. Acetate production has been reported as a good indicator of spoilage by this organism. algal toxins. However. 75% (with a 35% mortality rate) occurred in Japan. C1. C2. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43).5. E. When inoculated into VP corned beef and sliced ham. 5. 1. D. they do not produce significant product spoilage). these risks are substantially reduced (1.CHILLED AND FROZEN RAW FISH present in fish into histamine. presents a potential risk.1 Clostridium botulinum C. thermosphacta in the spoilage of some MAP fish.3. it is bacterial risks that increase after capture of the fish. Types A.6. The heterogeneity of the types (A. 5). Of 404 intoxication outbreaks of type E C.4 Brochothrix thermosphacta and lactic acid bacteria B. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. Of these. human viruses.e. F and G) is well documented (44). Fewer still are capable of growing on fresh fish. E and F include bacteria with minimum growth temperatures of approximately 3. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). including antisera.3 days after having reached 108 cfu/g (42). B. especially type E botulinum intoxication. Huss (1.

which was probably contaminated from an uncooked food source or an excreting foodhandler. and when in contact with contaminated water (no CDC reported cases from fish. 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. sushi) (5). salted or pickled) fish have also been associated with botulism. and proper cooking (boiling for 1 minute. the risk posed to the consumer of fresh fish. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. smoked. France. V. Proper chilling. The bacterium causes primary septicaemia. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . 1978 . A unique clone of V. Generally. parahaemolyticus cases have increased across the world.g. accounted for the majority of cases (1.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. 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. USA. is small. Other lightly or semi-preserved (e. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. This strain has been responsible for 50 to 80% of all V. Japan. storage and preparation (11). again. The first report of the organism being a foodborne agent was with shirasu. parahaemolyticus outbreaks. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47).g. whether stored aerobically or in modified-atmospheres. and North America. the majority of outbreaks are caused by consumption of raw fish products (e. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling.2 Vibrio parahaemolyticus and other vibrios V. which can cause human gastroenteritis. Despite the ubiquitous nature of type E C. I-sushi. Russia. Currently. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). parahaemolyticus is a marine organism. Only about 1% of marine isolates produce a thermostable haemolysin. Southeast Asia. 11). parahaemolyticus O3:K6 is responsible for many of the recent V. a Japanese boiled and semi-dried sardine dish. there is no specific guideline that describes a minimum level of V. Since 1996.6. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. V. vulnificus is associated with warm marine and estuarine waters. which is believed to be required for virulence. were associated with finfish between 1978 and 1998 (46). parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. 1. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile.2005).3 °C) will eliminate all risk. Proper temperature control (<3. due to the cultural preference for raw fish dishes. including epidemics in India. 5). In Japan. the majority are associated with products not cooked immediately before consumption.

which is often fatal (>50%). but proper cooking should substantially reduce risk of infection. possibly due to under-reporting because of their likeness to E.. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. Similar problems as those encountered for Aeromonas spp. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. Since 2000. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. Chill storage may not eliminate growth. shigelloides is similar to Aeromonas spp. V. In the US.000 17 .26 to 0. Again. monocytogenes has been well documented as a foodborne human pathogen. with minimum growth temperatures around 0 °C and optima of 15 .3 Aeromonas Aeromonas spp.4 Listeria monocytogenes L. Most cases have been sporadic rather than associated to large outbreaks. P. 5.55 cases per 100. there is no risk from the consumption of properly handled and cooked fresh fish. Although the psychrophiles are unable to grow at body temperature. monocytogenes incidence has been between 0. and especially estuarine environment.20 °C. infection.CHILLED AND FROZEN RAW FISH disorders). and proper cooking will substantially reduce risk.6. are associated with human diarrhoeal illnesses. their potential to produce disease-causing exotoxin in food has not been fully examined (48). are present on fish and can grow to significant numbers during storage (1. in assessing the risk of fishborne infection by P. listeriosis cases have been reportable to the CDC. cytotoxic. the causative agent of cholera. L. others are psychrotrophic mesophiles (49). cholerae O1. sodium channel blocking and haemolytic activity (48). is historically associated with faecally contaminated water. There is no risk associated with the consumption of properly chilled and cooked fresh fish.6. proper chill storage will eliminate any risk associated with this organism. and fish and shellfish are probably the major reservoirs for the organism (50). The major virulence factor appears to be the production of toxins with enterotoxic. and especially Aeromonas hydrophila. 1. but the bacterium is known to survive and grow in the shallow marine. Some isolates are true psychrophiles. coli on isolation media (50). 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. 1. Despite this. shigelloides are encountered. It is known that Aeromonas spp. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. 17). however. They are aquatic organisms with an epidemiology that is yet to be fully understood. in its habitat.

full risk analysis is not possible (52). Klebsiella pneumoniae and Hafnia alvei. leads to the production of histamine.. D60 of 1..FISH AND SEAFOOD persons. Like other toxins. 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. If fish are not properly refrigerated. there is a possibility for growth of the organism on chill-stored fish. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. It is regularly isolated from seafoods (51). foetuses. but proper cooking will reduce any risk. alcoholics. it is not apparent to the consumer and it cannot be destroyed by cooking.6. neonates.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses.98 minutes in cod and D60 of 4. Lightly preserved products. 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.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. Being a psychrotroph. The organism poses a serious risk in chilled products not cooked before consumption.pregnant women.6. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54).2002 (55)) and is also common worldwide. at risk groups . 1. some Vibrio spp. 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. 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. but cooking will significantly reduce the risk of infection. Fortunately. especially pickled fish. 1. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6).. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. 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. Infections are mostly associated with the consumption of raw or 18 . and Lactobacillus spp. Photobacterium spp. Clostridium spp.

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

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

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

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

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

Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere.G. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Simard R. Huss H. Borgstrom G.1983. 73-89. Ed. 1992. Application of predictive microbiology to assure the quality and safety of fish and fish products.S. Technomic Publishers. Aberdeen. Changes in microflora and chemical compounds in mackerel stored by partial freezing. 1986. Handling and processing of scad.C.M. 317-21. Lancaster. 127-34. Gram L. International Journal of Food Microbiology. Estimation of the bacteriological quality of fish by automated conductance measurements. McMeekin T. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. Taoukis P. Food and Agriculture Organisation of the United Nations. Magno-Orejana F. 317 (Suppl. 6.. Gibson D.1 Further Reading Processing Anon. Smith G. International Journal of Food Microbiology.. Ravn Jørgensen B. Akiba M. Microbiological quality and shelf life prediction of chilled fish.D. Sumner J. Food and Agriculture Organisation. 1. Torry Research Station of the Ministry of Agriculture. International Journal of Food Microbiology. Bulletin of the Faculty of Fisheries (Hokkaido University).. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Eds.). Hobbs G. in Fish as Food. Journal of Food Protection. Academic Press.9. Gibson D. Fisheries and Food.M.2 Spoilage Shewan J. Torry Research Station... Ogden I. 1961. Torry Research Station of the Ministry of Agriculture. 15. 1984. Dalgaard P. Olley J... 1989. 4. 741-50.. Huss H. 65-72. 1988. The Microbiology of Seawater Fish... Huss H. in Torry Advisory Note No 93.9 1. Jones Y. 13-32. Ed. Cho Y. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review.E. Shahidi F.M.FISH AND SEAFOOD 1. Cann D. Kitts D. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches. 271-85.. 1.P.. 24 . 295-307. in FAO Fisheries Report No. Studies on the microbiological ecology of mackerel stored by the method of partial freezing.L.D.. 1992. 1989. Labuza T. 1. 62-70.. Trolle G. Villemure G. Florida.. 51. Picard G. 1984. International Journal of Food Microbiology. 1985.H. Ross T.. in Seafood Science and Technology. Rome. 1997.9..H. Shinano H..H. 55.. Fisheries and Food.A. Houston N. Aberdeen. Journal of Food Science. 35..

..europa. Huss H.L. 322-6. 1992. 5.R. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. Blackie Academic & Professional.. A rapid method for detection of histamine producing bacteria. Amsterdam.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. 1992. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. Gram L. Huss H. in Quality Assurance in the Fish Industry. 72. Sala F.CHILLED AND FROZEN RAW FISH Einarsson H. 2197-203.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products.H..M. in Fish processing technology. http://eur-lex. 25 .K. 1997. Otero A... International Journal of Food Microbiology. 1. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. Applied and Environmental Microbiology.. Journal of Applied Bacteriology. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. 1993. Modified atmosphere packaging of fish and fish products. International Journal of Food Microbiology. 4.9. Elsevier Science.J.H. Gram L. 479-88. 65-72. Eds. Hall G. Liston J. Huss H. 20023.H. Klausen N. Trolle G. Ed. 1987. 1. Jacobsen M. 59. 1987. London.3 Pathogens Condon S. Effect of culture age.eu/LexUriServ/LexUriServ. Garcia M.9.. 137-46.


the exceptions being Sushimi (high quality raw fish). Sushimi tuna). CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr. and crustacea such as shrimp and crabs. emphasising the high quality of the seafood and the gourmet nature of the prepared products. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e.2. Few seafood products are sold ‘ready-to-eat’. 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. Even though heating prior to consumption will eliminate pathogenic microbes. it does not necessarily apply to the toxins that may have been produced.).g. although recent developments in improved supply chains.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. or could be disguised by sauces. This is no longer the case with many processed products now available in stores. The majority of seafood products produced require a cooking stage before consumption. smoked (hot) mackerel. The same used to be said for the difference between fresh and frozen products. smoked (cold) salmon. smell etc. colourings etc. but still acceptable and meeting all safety criteria. 27 .g. seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. 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. due to the early cessation of spoilage. colour of gills. products of lesser quality. 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. clarity of eyes. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2.

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

in VP or through mail order. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. the ingredients premixed. mackerel. mackerel.g. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product. VP products) because of the high salt content acting as a barrier to spoilage. salmon. The expected shelf life of these products is up to 8 days (or longer for some chilled. haddock. natural fillets of fish such as herring. or pre-packed in MAP or vacuum packaging (VP). Added colours can be included with the brine solution to meet specific requirements for products e.g. or pre-packed frozen with added glaze. 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. This process in effect cooks the product and reduces or eliminates microbial contaminants. 29 . which may.2.5%. resulting in a product that is ready-to-eat without any further cooking. be flavoured with herb dips or peppercorns.g. annatto with smoked haddock. The result is fish with a typical residual salt content of 1. Some products may be salted for flavour only (e. with a minimum residual salt content of 3. Hot-smoked fish are prepared from brined. 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. and salmon. sawdust with other woods adjusted for flavour profile and burn duration). cod). A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes.5% in the aqueous phase. Fresh fillets are sold from wet fish counters. etc.5 . The shelf life for MAP fillets is typically up to 5 days. which is approximately 30% (or 1 . swordfish steaks). Fish steaks can be cut from a natural fillet or loin (e.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. and the product then shaped and heat-processed to cook the product before being chilled. 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. Cold smoking is essentially a flavouring process. in addition. tuna steaks) or band-sawn from a frozen block (e. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products..g.2 days) longer than that of non-prepacked fish. but may be increased (up to 1 month) by packaging methods like MAP or VP. Again. a protective glaze may be added to the frozen steak. Pâtés are prepared with raw fish. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. They are sold from wet fish counters.

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

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

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

Generic examples have been used to illustrate only the major points.4.4. Where the raw materials are frozen. Examples of tempering techniques include the use of heated air. as the full complexity of the processes used for specific products cannot be covered in a general text. Figures 2. the core temperature. 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. Effects on the microflora are minimal during tempering (7). 2. For some components. depending on the size of the fillet 33 . Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. there are a number of distinct methods used.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. Band-sawn materials or fish dice are normally promptly further processed. but will also need to ensure that the optimum shelf life can be safely obtained.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.3 Preparation of fish component of chilled and frozen recipe dishes 2.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. to evenly raise the temperature of the entire frozen product from -20 °C or lower.4. but it is assumed that any band sawing or dice process will introduce heat from blade friction.2. or the use of microwave technology. 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. up to -5 °C so as to improve the cutting and handling properties during processing. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. but it also poses a risk of cross-contaminating other products undergoing tempering.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. All have the same intention. as a heat process is often required for functional or organoleptic reasons. this may be an incidental effect. and potential opportunities for growth of foodpoisoning bacteria. It is recommended that the principles of HACCP are applied to novel processes. defined holding time at a chill temperature (0 – 3 °C). Rapid controlled cooling of heat-processed components will help to minimise spoilage. and contamination from equipment to clean fish muscle. line extensions and process reviews. 2.3.1 .

with the coating heat-fixed. 2. may not rise above 0 ºC during processing.1B .2). Fig.3.2. by mincing and mixing with a binding agent (as described in section 2.1 (2. It is then size-graded. The “blown” meat is graded. drained and then refrozen for storage until required for further processing. after landing.Fish Cakes).Shaped Fillets. Raw material preparation stages for chilled or frozen coated fish products 34 .1A . the scampi is washed in potable water to remove foreign bodies.1. 2.7 days.4.1. 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. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). 2. On arrival at the processing plant. Frozen cores are usually batter-enrobed. chilled or sold chilled on defrost with an expected shelf life of 6 . Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. The processing of three examples types are described in Figure 2.FISH AND SEAFOOD processed.1C . The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.Scampi.3.4. quickfrozen.1. frozen in a nitrogen blast freezer and weighed into sacks for later processing.

It is for this reason that all by-products from primary processing. those that use frozen blocks of whole fillets. or promptly processed to avoid further quality loss.3. with the coating heat-fixed. spoil more rapidly than the dry ingredients. quickfrozen. iii) Fresh/fillet/minced fish block.1. as the physical structure of the material has been destroyed. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . or sold as a chilled product after thawing. Fish cake cores may be batter-enrobed. with an expected shelf life of 6 . Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. are handled and stored as any other food product.2 Shaped products This type of product falls into two categories. 35 . Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. Therefore.g. or those where the trimmings. more seriously. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. from potato flake. additional spoilage organisms may be introduced. The remaining dried ingredients are mixed with the fish.off-cuts or belly flaps. but.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. 2.7 days. material will need to be transferred swiftly to a chiller.3. such as aerobic spore formers and Staphylococcus spp.4. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. This is especially the case with block-sawn dust.4. and water is added to rehydrate the components. because of its nature.1. The fish component of the cake will. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish.block-sawn dust. fish fingers).. unless it is poorly handled. Waste material produced from this type of processing is often reused in the third category (fish cakes). that are to be reused for other foods. The mix is shaped and formed ready for packing or for enrobing. pathogens such as Salmonella and Listeria. ii) Fish-filleting waste .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. once the dry components have been rehydrated. chilled. 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.

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

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

There are two main categories.4. canned fish may be used in their preparation owing to it having wider availability and low wastage.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe. to produce a specific product. pasta or potato.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes.2. 2.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers.3. The vegetables must withstand processing. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). The protein source can be fish fillet or block. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products.g. as this could introduce off-flavours and reduce shelf life. A temperature no greater than 10 ºC for 12 hours cumulatively or 21. and must not show evidence of disease.4.1 ºC for no more than 3 hours is recommended (1). Fig. Flow diagram outline for recipe dish ingredients preparation 2. rice.5. Figure 2.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. For example. but they are supplied without a source of carbohydrate e. 38 .

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

Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. 2. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. spore-formers can proliferate if adequate control of temperature is not maintained. 13). 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. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. where composite meals are heat-treated. rely on the use of microbiologically sound ingredients.g. and an increased risk of contamination from L. An incident involving Staph. can germinate and rapidly multiply. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. If the sauce contains modified starches for freeze/chill stability.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. cereus are well known and are still a common issue in the UK (9). cereus. a heat process in excess of 73 ºC will be required to rehydrate this component fully. such as butter garnish with black pepper. if subjected to temperatures between 20 and 40°C. For this reason all products. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. and then cooled slowly allowing germination and re-growth. but temperature-abused pasta can also spoil rapidly by mould growth.4. 40 . Sauces or marinades are added to fish to broaden appeal or to accompany components. with the range always expanding as new tastes and cuisines are developed. Placing such products in a standard chill store to cool down could result in condensation. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. Appropriate handling facilities or equipment will be required to protect the product. cereus). On hydration. especially in those products that have a vegetable component or rice accompaniments. B. Products or dried ingredients contaminated with either spores of aerobic bacteria (e. aureus toxin is well documented (10. Typically. This is a particular issue especially where products are heat treated to eliminate vegetative cells.) to the product (12. Sauces can be multi-component preparations or more simple preparations.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. Alternatively.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. 11). Potato toppings made from flake or powder will normally have little associated spoilage flora.4.6. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC.

herbs. and handling of processed sauce can be less rigorous than. The format uses flavour compounds . 13).4. flour products. for instance. Figures 2. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. Marinade-type products are becoming more popular. 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. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. that of a dairy-based sauce. 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.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. respectively.2. Without barriers. 41 .5 illustrate the recipe dish ingredients preparation and assembly and packaging.4 and 2. etc. spices. 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.

The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. Flow diagram outline assembly and packaging of recipe dish 2. The oxygen-free conditions will however allow other anaerobic bacteria such as C.4.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. These include: 2.7. which may restrict either microbial growth or enzyme activity.FISH AND SEAFOOD Fig. the method replaces air in the sealed packaging with a defined mixture of gases. 42 . unless the temperature is strictly controlled to below 5 °C throughout the supply chain. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. as it dissolves in the surface moisture to form a weak acid. 2. botulinum to grow.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).4.4. generally carbon dioxide and nitrogen.5.7. and tuna loins are packed by placing the product in a plastic pouch and removing all the air.2. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues.2 Modified-atmosphere packaging Originally developed for the red meat industry.

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

Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme.. As a note of interest.. The type and nature of the enzymes and their activity will depend on the bacterial species. However. The conditions and freezing process most favourable for good-quality products will. freezing also has a protective role in improving the safety of raw materials by. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp.5. Serratia spp.g. producing characteristic off-flavours. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24). botulinum. which would warn the consumer of the potentially unsafe nature of the product. By retaining the intrinsic spoilage flora of seafood. and Aerobacter spp. Bacillus spp.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. within dairy products are well known. for example. 2. the shelf life of products is limited by the natural spoilage process described earlier. auto-oxidation of fats/oils). where the variety of components and ingredients used in manufacture is varied and complex. Products that are developed with an extended shelf life beyond the 6 . also be those that have the least effect on the viability of microorganisms (21). released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15).3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10).. Still. This reinforces the need for prompt processing and temperature control of ingredients and finished products. enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage. For example. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. 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. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e. Pseudomonas spp. however.FISH AND SEAFOOD 2. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. 44 .7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C.5. eliminating nematodes in products that do not have a heat treatment step. These have been proved to impart off-flavours. this should also take into account shelf life of product components used in the assembly of the final product.

air. have been linked with food poisoning and should also be considered. botulinum when held in favourable conditions for growth (28) i.6. may be of benefit within any risk assessment for chilled and frozen prepared fish products. and this issue may be of greater consequence for warmwater. potato. permit rapid growth of B. Hazards attributable to the non-seafood components are listed below and this information. 13. Cases quoting contamination of seafood with organisms such as S. 27).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. These are C. and some effects related to product type and time to toxin production have been noted.6. farmed or free-living crustacea (30).104/ g should be treated with some concern (26). any risk of C. soil. etc. Incidents in such diverse materials as cream. cereus.6. Counts of between 103 . or a heat shock followed by holding at a temperature suitable for growth. perfringens contaminating seafood will depend on the source of raw material. histamine intoxication. Germination of spores requires favourable conditions. agricultural run-off or sewage can contaminate in-shore species.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. especially molluscan filter feeders.1 Bacillus cereus This is a common contaminant of dust. and many raw or processed foods (26). Vibrio spp.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. typhi. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. water. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). A number of other Bacillus spp. Therefore. Additional hazards from marine bio-toxins. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. 45 .. 2. are dealt with elsewhere in this book. and Campylobacter are recorded in filter feeding molluscs such as mussels. In addition. botulinum type E and Vibrio parahaemolyticus (25).e. spices and herbs and vegetable shoots have been reported (12. with pathogenic microorganisms or viruses. in some cases. Inappropriate holding times and temperatures will. along with previous discussion. 2. 2.3 Clostridium perfringens A study examining C.

L.6. L. However. Chilling may help Shigella to survive. at the present time. when introduced or inoculated into sterile components of chilled prepared recipe dishes. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. and this is believed to be due to a tolerance to brine used in the smoking process (32). 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).6. the restrictions on growth rate seen with seafood will not apply. An acceptance has now developed in the UK of the presence at low levels of L. which is not affected by any subsequent cooking process. Rates of contamination with smoked seafood are much higher than those with unsmoked products. These include inadequate refrigeration and preparing foods far in advance of planned service. and will therefore cause “food poisoning” even in a product that is cooked before consumption. but under the appropriate conditions. 46 .5 Shigella spp. aureus enterotoxins in foods. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces.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). if any other potentially pathogenic organisms. Sources of contamination are many because of the widespread distribution of this organism in the environment. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). 2. It is unclear. monocytogenes as a contaminant in foods (31). 2. infected food handlers. Shigella spp.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. growth may occur in foods (26). monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. will colonise factory environments in a similar fashion except for possibly Vibrio spp. 2.6.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). These conditions will allow the production of the toxin. indigenous or other. but. and inadequate cooking or holding within the temperature growth range (27). aureus (35).

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

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. batter etc) should also be consulted when establishing the specifications for raw materials and final product. L. coli GMP* <102 Maximum 104 TABLE 2. TABLE 2.g. aureus V. Maximum= Levels at end of stated shelf life 48 . 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.FISH AND SEAFOOD the microbial criteria for other products (rice. fish fingers.III IFST Guidelines for processed foods .fish products. pasta. parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E. monocytogenes B. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp.II IFST Guidelines for part-cooked foods – e.

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

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

McCarthy S.K. 60 (4). 1999.eu/LexUriServ/ LexUriServ. Institute of Food Science and Technology. 34. Evans M.G.2. 50 (9). Shigella as a foodborne pathogen. Smith J. Development and Use of Microbiological Criteria for Foods. Journal of Food Protection. 372-6. 37. Woodhead Publishing Ltd. Principles and Practices for the Safe Processing of Foods.chilledfood. Shapton N. London. 1991. Psychrotrophic Bacteria in Foods: Disease and Spoilage. 1997. Chilled Food Association. http://www.co. CRC Press.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology. Wolfe Publishing. 39.A.uk/cday/posters2008/npd6. 1998. Microbiological Criteria. 35. Campden & Chorleywood Food Research Association. Journal of Food Protection.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. Kraft A. Gloucestershire. 2006.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. London.campden. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33.H.pdf Shapton D. British Retail Consortium.europa. 788-801. Cambridge. 38. Foodborne Pathogens an Illustrated Text. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. 41. Varnam A. 1992. Phil Voysey...A. 51 . 36. U. http://www. 1987.A. U. 40.K.F.L. http://eur-lex. Chilled Food Association.


lobster. clams. can hold the shells tightly shut against enemies (1). Jr. whelks. Special tracts of cilia move this mucus toward the mouth. For example. Many microorganisms ingested by shellfish survive the digestive process (3. In these forms.3.1 Definitions General features of the mollusc The phylum Mollusca. scallops. S. mussels. mussels. In the process of filtering water. cockles and scallops. Other species of shellfish such as crabs.E. while 53 . The digestion process requires less than 2 hours in actively feeding adult bivalves (2). Pelecypoda. snails. clams. O. which includes oysters. respectively. Pelecypods have little need for locomotion. Food and Drug Administration 22201 23rd Dr. Plankton and algae form the bulk of their diet from seawater. the shell. and Cephalopoda. Box 3012 Bothell WA 98041-4421 United States of America 3.7 litres of water per hour.S. cockles. composed of two valves hinged together dorsally. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. where it is eventually swallowed and digested. P. Pacific Regional Laboratory Northwest U. they also trap bacteria. The phylum consists of five classes: Amphineura. Gastropoda. chemical contaminants and other impurities on the mucus of the gills. oysters. Scaphopoda.1 3. and 2. octopi. cockles. chitons. The term shellfish in this context is limited to oysters. slugs. lying well within the mantle cavity. and shrimp. MOLLUSCAN SHELLFISH Carlos Abeyta. In most species. 4). Strong muscles. The class Pelecypoda includes all of the bivalves such as clams. squid. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. abalones. oysters and clams can process about 3. the adductors. mussels.1. completely encloses the body.6 litres. The class having species of economic and public health importance is the Pelecypoda. and scallops. viruses. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells.

3. The habitat of oysters and mussels are sessile. Clams are benthic and live burrowed in the mud.FISH AND SEAFOOD commercially valuable. The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3. Whatever contaminants are in the water will eventually get into the shellfish.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. Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific .1. many species of bivalves are edible (Table 3. Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 . dwelling on the bottom or attached to structures in the water column. but maintain contact with the water by means of a siphon tube (5).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.2 International commercial products Globally. and usually at higher concentrations than found in their habitat.I).North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp.

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.

and other names.1 Clam Clam is the common name of a large group of often edible. Buried clams leave tubes. extended above the surface to maintain water currents needed for respiration and feeding. mostly marine bivalve molluscs.1. geoduck. The muscular foot is used to burrow in mud or sand. Different species are variously called quahog.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. hard-shell or soft-shell clam. 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. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint. littleneck. A mantle tissue encloses the body of the clam. or siphons. Most clams are only a few centimetres in their maximum dimensions. cherrystone. but 56 .2. laterally compressed.

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

smouldered and afterwards dried (Japan). vinegar solution.) Dried: peeled. 3. In some species. Unlike other sessile bivalve molluscs. The adductor muscle is the only edible part of the animal. mussel paste.1. Salted: cooked meats bottled in brine. scallops possess a single large adductor muscle. mussel in butter sauce. Smoked: meats may be canned in edible oil. The shells are fluted and round with a broad. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). They have filibranch gills. The common scallop is about 5 cm long. Live: whole with shells. brine. in which the individual branches. Fresh: cooked meats. flattened ear or wing at the hinge area.1. sauces. or filaments. boiled. Canned: cooked meats. in mussel liquor. Bait: used live extensively for baiting fish hooks for line fishing.2. The posterior halves of the valves contain a large adductor muscle that holds the valves closed.FISH AND SEAFOOD 3.2. 58 .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. The familiar marine mussel is asymmetrical. Internally. are united. Semi-preserves: cooked meats with vinegar-acidified brine and spices. scallops can swim or leap by a series of rapid closings and openings of the valves. or packed with jelly.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. and the anterior end smaller and pointed. cooked meats packed in dry salt for transport. with the posterior ends of the shells (valves) broad and rounded. gutted. also with other ingredients. by interlocking tufts of hair-like cilia. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves. Fresh: shelled meats (often only the adductor muscle and the roe are eaten. and held apart.

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

There are microorganisms. 3.. Campylobacter coli Bacillus cereus Hepatitis non-A. In summary.1 and 3. For example. or by 60 . but have never been cultured (14. mechanical harvesters. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. and shucked shellfish that is shellfish. clams.. Water temperatures affect the concentrations of microflora in the water. Temperature. 15). that have never been cultured on laboratory media. the initial flora composition and concentration of bivalves are highly variable. and Aeromonads.2.3 3. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. Enterococci spp. Micrococcus spp. Flavobacterium spp. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. The largest groups found are the Pseudomonads. whole or in part.1 Processing and its Effects on the Microflora Oysters. 10. shellstock (live) which is shellfish in the shell.3.. Moraxella spp.1. Alcaligenes spp.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. from which one or both shells have been removed. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. responding to the environment in which they are harvested.1 Shellstock 3. Vibrios. 11..3. 13). and yeast cells of Rhodotorula rubra and Trichosporon spp. (9.. nutrients. 12. They are termed viable non-culturable microorganisms. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100.3. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves.000 /g. and cockles The processing of bivalves takes two forms. rakes. observed microscopically. At harvest. dredges. Bacillus spp.1.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. mussels. Other microorganisms found in lesser numbers are the Achromobacter spp. Pseudomonas spp...1. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. 3. Acinetobacter spp.. spirochetes of the genus Cristispira and Saprospira.. salinity.

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

Generally.2 ºC (45 ºF) or below (19). with prolonged storage the microbiological quality of the bivalve decreases. If long delays occur before reaching the processing facility and/or refrigeration. Harvest conditions are another factor that influences the growth of bacteria in shellstock. 26). 17). there is evidence that in the absence of adequate refrigeration. 24. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. Over a period. The washing of sediment and detritus soon after harvesting will prevent further contamination. 22). Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. At harvest. and temperature storage of shellstock to point of sale.3. shellstock are not refrigerated on the harvest boat. 21. Enterobacter. 25). Washing shellstock helps prevent quantities of sediment and bacteria being mixed with.1. however. handling. for a considerable length of time. The most important factor in controlling bacterial growth in shellstock is temperature. Generally. shellstock are placed in refrigerated trucks for transport or placed in wet storage. some pathogenic organisms will survive and multiply in shellfish (20. 16. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. 23. vulnificus. hydrophila (18). coli. the bacterial counts in the meats will increase. Furthermore. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. including V. Klebsiella. when it reaches the processing facility. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . Water used for shellstock washing should be of good sanitary quality. without adequate refrigeration. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. to avoid possible contamination of the shellstock. are prevented. An outbreak of infectious hepatitis in Sweden in 1956. At this temperature the multiplication of faecal coliforms (E. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. and the sanitary practices applied during harvesting and shipping. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. and Citrobacter species) and Vibrios. with the exception of A. synthetic seawater for the purpose of de-sanding and salt uptake. bivalves tightly close their shells thus trapping water and microflora. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. and thus contaminating the shucked meat at the point of sale.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock.1.FISH AND SEAFOOD 3.

In the blower-washer. the finished product may become contaminated. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. Shuckers routinely use either cotton or rubber gloves during the shucking operation. 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. or subjected to splash from foot traffic.2 Shucked shellfish 3.2 Effects of microflora during processing The same considerations as previously discussed.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation.2).1 Processing description The processing of shucked shellfish is limited to washing of shellstock. If shellfish are not reasonably clean at the time of processing.1. lobsters. Thus.2. Due to the nature of the shucking operation. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. and packing shellfish into containers (Figure 3. for the effects of microflora during the processing of shellstock. the tank is opened through a drain gate and chute. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. and fish. shellstock is subjected to contamination when stored on the floor in standing water.3.2. washing and culling shellfish prior to storage is essential. 3. clothing becomes very soiled. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station.3.1. 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). Studies have shown the use of rubber gloves reduces the contamination level. are applied to the shucking operation of shellfish. whereas cotton become wet and 63 . After washing of meats. through which the washed meats are discharged for rinsing and then packed in containers. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported. In addition.1. Under these conditions if the shuckers enter the packing room. removal of shellfish (shucking). To remove the shellfish attached to the remaining valve. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. Containers are placed in packing boxes for shipping at refrigerated temperatures. thus contributing to high bacterial counts in the final product (28). washing.3.

and gloves help prevent the contamination of shellfish.2. Separate rooms or lockers for storage of clothing. within one hour. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. 4.2 ºC or less. There is a tendency to store such articles on the shucking benches or in packing rooms. Shucking containers are rinsed clean with running water.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). Shucked meats are thoroughly drained. 3. which may lead to contamination of product. aprons. and sanitised before each filling. 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. 2. Fig 3. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. 64 .

Controls that should be applied include using clean packing equipment. and yeasts. Micrococcus. blowers and other utensils that have cracked. in oysters. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. Alcaligenes. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. spoilage will occur. streptococci. clean gloves. Shucked shellfish are an excellent medium for the growth of bacteria. 31. skimmers. at the turn of the century. As described previously. 34. Glycogen. abundant gas production. particularly shucked meats. Flavobacterium. Achromobacter and Flavobacterium levels decrease during storage. 36). In summary. Proteus. These organisms may play a significant role in the spoilage processes. and proteolysis (10). 3. 33. All equipment coming in contact with the shellfish such as colanders. This process changes the bacterial profile to microorganisms favouring low pH. Shucked shellfish are held and transported at temperatures of 7.2 ºC or less. particularly oysters stored at 7 ºC. The spoilage process in shellfish may be divided into three stages: increase of acidity. For example. Much of the work has been done with oysters and clams. 35. where there is a breakdown of temperature control or prolonged storage of shellfish. anywhere during the process of shellfish from harvesting to the point of sale. Pseudomonas. Lactobacilli seem to be the predominant microflora in shellfish. these include lactobacilli. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. 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. The environment has an effect on the bacterial population found in shellfish. Glycogen is hydrolysed by the initial flora found in shellfish.8% (37). a carbohydrate. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. and packing the shellfish in a timely manner.2 ºC. The initial flora found in shellfish is the major factor of spoilage in bivalves. whereas bacterium such as Pseudomonas. Shellfish.47 to 6. Temperature abuse will result in high bacterial counts. shucking pails. 32). thus producing acid and gas (38). is stored in the bivalve as a source of energy. are an excellent medium for growth of bacteria. glycogen levels range from 0. rough. 65 . Earlier studies.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. showed that organisms such as Achromobacter. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7.

V. 3. during processing. there is a recent report of low levels of V. 3. lot and retail outlet. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. Sporadic cases are frequent along the coast of the United States. prevention of cross-contamination. One of 250 cases reported involved shucked products with low levels.1 Vibrio parahaemolyticus V.FISH AND SEAFOOD 3. Diseases caused by Vibrios are enteric in nature. parahaemolyticus multiplies rapidly at temperatures above 20 ºC.5. coastal areas). Between May and September 1997. diarrhoea. parahaemolyticus from shellfish is low. and harvesting of shellfish in the cooler months. however. 66 . of V. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. These organisms can be present in the shellfish from the point of harvesting. ranging from epidemic cholera to sporadic cases of diarrhoea. approximately 23 cfu/g. although non-hemolytic strains have been associated with illness (43.5. and to the point of sale. from harvesting to finished product (i. Fortunately. only a small percentage of strains in the marine environment are potentially pathogenic (45). 44). headache and fever. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). parahaemolyticus causes gastroenteritis lasting 24 . Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). 41). Major outbreaks are associated with the warmer months. where outbreaks occur regularly. and very common in Japan. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. parahaemolyticus possibly producing illnesses (40). The incidence of V.48 hours with abdominal pain. Products involved were from the same processor. shellstock and packing of shellfish in jars) (29). parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. parahaemolyticus. They can be introduced to the shellfish from the environment or added during processing. It is a common marine bacterium. V.1. This bacterium is present in all the major processing steps of a shellfish plant. and estimates based from a hospital study show less than 0. nausea.e.5 cases/100. widespread in both polluted and unpolluted waters.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. primarily in brackish or saltwater.000 population/year (46).

cholerae non-01 is ubiquitous in the estuarine environment. abdominal cramps.II (8. vulnificus. Hazards from this bacterium can be controlled by thorough cooking of shellfish. cholerae non-01 has been associated with septicaemia. diabetes. cholerae non-01 (51). and preventing cross contamination once the product is cooked. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. cirrhosis. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. which have been implicated in shellfish-related disease outbreaks (31). Control of oyster-associated 67 . Onset of symptoms occurs in 48 hours and can be severe. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. Healthy individuals are susceptible to gastroenteritis. 56). Again. parahaemolyticus. parahaemolyticus growth in shellfish meats is temperature dependent. parahaemolyticus the estimated annual incidence rate is very low.8 case/100. those who have liver disease. 46. the organism has been found in the Gulf of Mexico. vulnificus is by rapid refrigeration during warm weather months. in the range of 0. 55. cholerae. 46.5. V. Death has been reported to occur within 36 hours from onset. chills. or immunosuppression.5.1. nausea. It causes diarrhoea.000 population/year (8. fever. leukaemia. 3. are particularly susceptible to primary septicaemia. and rapidly become inactive at low storage temperatures. In one study. The organism attaches to the intestine and release of a toxin is suspected. All isolates are considered pathogenic.0. 48). Oysters are the most hazardous food associated with V. however. and vomiting. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50).MOLLUSCAN SHELLFISH Control of V.3 Vibrio cholerae non-01 Illness caused by V. with a fatality rate of 50% for individuals with septicaemia. or primary septicaemia.1.7 days. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). The bacterium is an estuarine species that causes wound infections. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. 23 of 24 major USA-West Coast estuaries were positive for V. Primarily. 49). gastroenteritis. like V. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. these individuals are advised not to eat raw shellfish. 58). and nausea within 24 .2 Vibrio vulnificus The symptoms of V. They are sensitive to cold. 3. V. It is one of the most severe foodborne infectious diseases. The occurrence of V. High-risk individuals.48 hours of onset. lasting 6 . 53. 54.4 . Inadequate refrigeration will accelerate the growth of V. 57. vulnificus are fever. Control of V. V.

Control of infection from these organisms is no different from the other Vibrios.10 days. Poor sanitation and contaminated water supplies will spread the disease.FISH AND SEAFOOD V. Illness can include abdominal cramps.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. cholerae 01 is transmitted by faecal contamination of food or water. and illness lasts 7 . or cross-contaminated shellfish (57. headache and fever. and the transmission of the disease by consumption of raw.10% carbon dioxide for optimal growth conditions. 62. It causes diarrhoea. imported product from a country experiencing an epidemic. jejuni requires reduced levels of oxygen (3 . cholerae non-01 illnesses is by cooking seafood thoroughly. Onset of disease is 6 hours to 5 days. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59).2 Campylobacter jejuni C. cholerae 01 establishing in the U. respectively. 60). including V. these organisms cause gastroenteritis lasting 24 . It is considered a fragile and sensitive organism to environmental stresses such as oxygen. V. nausea. There are several other Vibrio spp. Onset of illness is from 3 to 5 days after ingestion of contaminated food.1.5 Other Vibrios spp. 61. dehydration. In developing countries. acidity and heat. fluvialis. In general.5. and shock. vomiting. 3. mimicus. vulnificus. and nausea. drying. low pH. Death may occur after severe fluid and electrolyte loss. salinity. particularly in oysters.5%) and 2 . 63). the pathogenic severity is lower than V. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. hollisae.g. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. V. however. abdominal pain. 3. fever. nausea. The presence of these strains appears to be in low numbers and non-virulent (59). preventing crosscontamination of cooked seafood. (8.48 hours with abdominal pain. There is evidence of strains of V. associated with shellfish-borne illness outbreaks. widespread in both polluted and unpolluted waters.1. and southern Asia including India and Bangladesh.5. They are common marine bacteria. and V. undercooked. 3.S. Illnesses are associated with the warmer months. The frequency of infection of these organisms is no different from other Vibrios. cholerae 01. The control of V. is the same as previously discussed for the non-01 strains. processors should know the product source. e. Gulf coast environment. In addition. parahaemolyticus and V. 68 . V. cholerae 01 in molluscan shellfish.5. diarrhoea.

whereas at 10 ºC Campylobacter survive only for 5 days (71).3 Other organisms of concern 3.3. however. 77. 85). 80) are ubiquitous in the marine environment. 78) and P. Campylobacter appear to survive well in oysters during storage. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. 73). of Incidents 4 279 0 1 0 No. vulnificus. 70. V. 72. jejuni were recovered. jejuni begins in the harvest of shellstock from approved waters. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. control of C.MOLLUSCAN SHELLFISH TABLE 3. from Pacific shellstock oysters purchased from a retail grocery store (74). C. 66). Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). To reduce the risk of shellfish-borne campylobacteriosis. V. 76. parahaemolyticus. 69.II Molluscan shellfish associated outbreaks caused by Vibrio spp. have been isolated from shellfish beds throughout the world (67.5. although their role as a pathogen is highly controversial and circumstantial.500 cells/gram of Campylobacter may cause illness (64. of Cases 4 362 0 1 0 a Includes V. mimicus. hydrophila group (75. and their immediate proper refrigeration. 82. The seasonality of 69 .1 Aeromonas hydrophila group and Plesiomonas shigelloides A. 81. In one case. shigelloides (79. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. V. 3. 71). They have been implicated in limited shellfishassociated gastroenteritis cases (8. they survive well at refrigerated temperatures. 83. ranging from 0. Campylobacter spp. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. 68. jejuni does not multiply in shellstock at refrigerated temperatures.5. 84. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. Human feeding studies suggest that about 400 . Recently.4 to 114 cells. and V. 65. This finding is significant in that C. fluviallis. however. hollisae Infective dose varies with host susceptibility and virulence of the strain. infection occurred among participants of a firemen's banquet. C.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. cholerae 01 and non-01. V. Although.

suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. aureus C. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. FDA/Administrative Guidelines 3 Source (91). action is considered on a case-by-case basis taking into account all the factors associated with the specific situation.000 cfu/g1 <230/100 g2 <1000 per gram. In one study. FDA/ Administrative Guidelines. Caution should be exercised by individuals with haematological malignancies (88). 70 . Test Standard Plate Counts E.600.000 to 3. Similar results were seen by other investigators (86. prepared sandwiches. 2 Source (90). coli Salmonella V. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. 3. cholerae V. Note: Although molluscan shellfish are not included in the selected high risk food. 87) of the outgrowth of A. 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. vulnificus in molluscan shellfish. Control of these organisms in shellfish products is limited. soy products. They are easily destroyed with mild cooking temperatures. being isolated more often during the warmer month. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. and where indicated are intended to be used for official control purposes. jejuni V.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide.000/gram (29). FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. dried milk/whey drink powders. coli Enterotoxigenic E.0001 negative for the presence3 Under review4 Source (89).000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. non-dairy frozen desserts. parahaemolyticus Staph.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. hydrophila group stored at refrigeration temperatures. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures.

Old H... Fish and Wildlife Service. 16. 10. Tall B. Biology of the oyster in relation to sanitation.R. 633-40. 46. 64. Viability and release Salmonella charity and Escherichia coli from oyster faeces. 33643. in Fishery Bulletin 1964 No. Hackney C. W. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). 245-7.. 1995. Rowse A. 1936. Smith F. Fleet G. Clinical Microbiology Review. 4. Government Printing Office. 1964. Hood M. 1960. USA. 1991.F. 2.N.A. Jr. Proceedings of the National Shellfisheries Association.. Microbiological characteristics of Dungeness crab (Cancer magister). 1994. Fate of enteric bacteria in estuarine sediments and oyster faeces. Tubiash H. 19-34. Villee C. 72-8.S. Applied and Environmental Microbiology. Organisation for Economic Co-operation and Development.C. 12. Ray S. Saunders Company. 30.D.W. 71 . Rippey S. Schmidt C. Veliger. American Journal of Public Health. 447-52.K. Effects of harvesting waters and storage conditions on yeast population in shellfish. Journal of Food Protection.K. 9.. 1984. water and sediment from Galveston Bay. 104-9. Ward D. Cook D. in Multilingual Dictionary of Fish and Fish Products. 42. Bacterial flora of the market oyster.E. 17.MOLLUSCAN SHELLFISH 3. Cook D. 6. 26. in Microbiology of Marine Food Products. 1926.H. 105-8. Galtsoff P. 13. 544-8. 29. Wiley-Blackwell.C. Microbiology of bivalve molluscan shellfish. The American Oyster.R. Journal of the Mississippi Academy of Sciences. 8. Colwell R.S.R. Bernard F. Eds. 11. Van Nostrand Reinhold. Applied Microbiology. Colwell R. Applied and Environmental Microbiology. Washington D. 36. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves.E. in General Zoology. References The Phylum Mollusca... Philadelphia. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters.. 3. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). 71-6. 1975. 755-76.A.. 82-7. Pfeifer D. U. 1968. 419-25.S.B. Thompson C. 58.R. Jr.. 231-51.J. 15.7 1. Galtsoff P. 30. Walker W. 44.R. Infectious diseases associated with molluscan shellfish consumption. Lee J. Ed. New York.M. 1963. 1970.. American Journal of Hygiene. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. 1981. Applied Microbiology. 7. 33-6.R.W. 13. 7. Eliot C. Lovelace T. 1982. Liston J. American Journal of Public Health. Nauman R.. A typhoid fever epidemic caused by carrier bootlegging oysters. 6. 14. 1940. 8. Gill S. 1973. Vanderzant C. 1983.A.L.. 5. Journal of Milk and Food Technology.S. Eds.

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

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

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

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

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

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


and are usually aquatic. Pandalus borealis (deep-water shrimp). Peeled and cooked shrimp are ready-to-eat after controlled thawing. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. Pandalus montagui and Crangon crangon (inshore shrimp).. jointed limbs and a chitinous exoskeleton. They possess a segmented body. Parapeneopsis spp..4. crayfish and shrimp (1). M. the front pair ends in claws.5 and 1.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. Peeled ready-to-cook shrimp are raw. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. e. and Trachypenaeus spp. Species eaten from tropical waters include Penaeus spp. lobsters and crabs have ten legs.0 kg polythene bags. packed and frozen in 0. with shells and heads on or off are usually packed.5 kilo pack (boxes or bags). with additional water to form a glaze.5 and 1. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi).0 kg polythene bags.Ph. lobsters. Shrimps. Crayfish.g. CRUSTACEAN SHELLFISH Linda Nicolaides. and frozen in 1 and 2. Such products are usually only partially cooked before freezing. (deep-water shrimp). peeled product. 79 . and available in 0. crabs. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4..

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

Shrimp might be further processed with the shell on. bacterial contamination and spoilage are inevitable.1 Prawns and shrimps An example of the process used for preparing frozen. thus. A large proportion of shrimps are produced by aquaculture.3 Processing and its Effects on the Microflora Crabs. Hence. Vibrio spp. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. peeled and cooked. These animals can survive out of water if the surrounding environment is kept cool and damp. corynebacteria. especially if they are stored in ice or iced water. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . up to 16 days for tropical species compared with 8 . Shrimp will then be glazed and frozen in a raw state.. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. or transported directly to processing plants. graded according to size. however. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails.for example. In comparison shrimps die quickly after capture. chilled 81 . 4. After capture and transport to the processing plant. The rate of spoilage will be a function of time and temperature.. Alcaligenes. Shrimps can be processed whole. or are found in brackish water around the shores. they are washed. glazed and frozen (see Figure 4.3. Pseudomonas spp. uncooked prawns and shrimp is presented in Figure 4. from the time of capture and processing.1. Micrococcus spp. As with finfish. whether GHP are in place. They are then taken to market alive in either holding tanks. lobsters and crayfish are harvested from cages or pots containing bait. or cooked.10 days for temperate water species (3). or with the shell and central vein removed. chilled sea water. 4. the genera associated with tropical species include Enterobacteriaceae. and Achromobacter spp. The claws are usually bound together to prevent the animals fighting and causing damage to each other. with “head on” or “head off” whereby the edible tail is removed from the head. 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.. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. gills and thorax. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots.1). In contrast. When the shrimp arrive at the processing plant they are washed and then sorted into size. so they are either sold on the local market or are processed in local processing plants. the rate of deterioration of the crustacean shellfish by these organisms is reduced.

frozen uncooked prawns 82 . Flow diagram . 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. After cooking. Good Manufacturing Practice (GMP). including GHP. Fig. spices and cooking batter.1. should be followed to ensure that the product is not re-contaminated with microorganisms present in the processing environment or from the workers. 4. Breaded products introduce an added risk of spore forming bacteria. and packed to meet the customer’s specification.FISH AND SEAFOOD frozen.

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

However.5% of frozen raw shrimps from different countries (13). and usually for both. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L.1 Pathogens: Growth and Survival Salmonella spp. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. although efficient cooking will quickly eliminate it (6).5. monocytogenes in cooked ready-to-eat foods. fish fingers. Although foodborne illness had not been associated with the product. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). 5. as well as for Salmonella. in 24. even if GMP is followed.5. 16). The presence of L. a common pond fertiliser used in aquaculture. 6. later surveys in 1990 recovered the pathogen from retail raw. 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. Following this. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). The above results prompted the FDA to develop a compliance programme for the pathogen.5 4. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. monocytogenes was absent from 40 samples of cooked prawns. shrimps and cockles sold (17). nearly 30 years elapsed before the potential hazard of L. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. 7).FISH AND SEAFOOD 4. Salmonella cannot be completely removed from raw shrimp during processing. in domestic/imported shrimps (15). Further surveys identified Listeria spp.cooked prior to consumption. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . 4.2 Listeria spp. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. Hence. An import alert was put in place in June 1987. animal manure. monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). cooked and smoked fish. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . shrimp and shellfish (11). Moreover.

cholerae is divided into two groups. To date. although contamination by types A and B may occur from the environment. Once the competing microflora has been removed by a heat treatment. The main source of contamination is the hands of processing personnel. therefore. cholerae.CRUSTACEAN SHELLFISH old. e. pregnant women and the immuno-compromised. as part of the natural bacterial flora. pasteurisation and cooking. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. However. 85 .5. 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. the pathogen can grow at temperatures as low as 3 °C. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. aureus on their hands.5. Vibrio cholerae.4 Clostridium botulinum C. 24. the causative agent of cholera. 4.5 Vibrio spp. 4.3 Staphylococcus aureus Staph. 22) as well as from undercooked crustacea when harvested from contaminated waters (23.5. V. which can become colonised by V. 25). The former. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. Crustaceans have been linked with food intoxications attributed to Staph. botulinum type E is commonly associated with fish and fishery products. no incidence of foodborne illness caused by L.g. especially in products that have been vacuum packaged. Control should be achieved by promoting GHP and staff training programmes. was associated with seafood consumption. aureus is not a member of the intrinsic flora associated with crustacean shellfish. whilst boils and cuts are also common sources of the pathogen. The pathogen has resistant spores. particularly crabmeat (18. Crabs are covered in a chitin exoskeleton. aureus because of the ubiquity of this organism in the processing environment. Researchers have demonstrated that inoculated shrimp supported toxin production by C. monocytogenes has been attributed to crustaceans. cholerae O1 and V.70% of healthy individuals carry toxin-producing strains of Staph. An epidemic that began in South America in 1991. combined with the degree of handling of these types of product. 4. is widely distributed in aquatic environments. 19). cholerae non-O1. V. about 50 . GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. so is able to survive heat treatments that eliminate vegetative cells. botulinum type E at 10 °C but not at 4 °C (20).

5. 4. 32). hydrophila has been isolated from prawns (29).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.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. a marine halophile. Parasitic infections are controlled by either cooking infected material prior to consumption. been implicated in a case of foodborne infection associated with crustaceans. Of these. jejuni has not. parahaemolyticus. particularly trematodes (6).30 MPN/g) (27). coli. but has the potential to cause fatal foodborne illness (26). the potential hazard exists as C. e. 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. Hepatitis A virus presents the most serious hazard. 31.7 Aeromonas hydrophila and Aeromonas sobria A. however.5. 4. or freezing at a temperature of -23 °C for 7 days (30.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. It is more commonly implicated in countries where crustacean shellfish are eaten raw. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. vulnificus is an invasive and lethal pathogen associated with wound infections.5. However. V. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. Norovirus.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. hydrophila and A. hydrophila is able to grow well at refrigeration temperatures (0 . 4. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. V. Japan. Astroviruses and non-A. However. Quantitative levels were below limits of detection in all cases (<0. occurs in crustaceans from warm environments. non-B Hepatitis viruses (31). whilst 5. A. South America.5. although A. in a survey carried out in the United States between 1977 and 86 .6 Campylobacter jejuni and Campylobacter coli C.8% of samples contained C. Caliciviruses. 4.g. to date.

10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. The toxin causes amnesic shellfish poisoning (ASP) when consumed. although it is one of the European Union (EU) legislated allergens.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). Selected parts from this regulation is presented below. 4. 87 .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.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive.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. 4. 2073/2005. 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. only 0. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. are included in the Commission Regulation (EC) No.5. which results in a persistent and apparently permanent loss of short-term memory (36). Norwalk viruses can also withstand heating to 60 °C. produced or imported into the European Union (EU).6. They can survive low pH levels. The risk of such hazards is far higher with fin fish and molluscan shellfish. 4. No foodborne incidents have been attributed to this source to date. 38. consistent with that in the stomach.5. or chlorine concentrations similar to those found in drinking water. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). 4. A summary of microbiological criteria for crustacean shellfish is listed below. which could be transferred to the meat during unskilled removal from the shell (35). 39).CRUSTACEAN SHELLFISH 1981. from farm to fork (37.

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.


dependent on the country or the type of product. where the salt replaces moisture in the fish flesh. This can be called ‘kench’ curing and is mainly used for non-fatty white fish. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. fish is immersed for a few hours in a relatively low concentration brine. drying. Salting is both a method of preserving fish. The processes take anywhere from a few hours to a few days or even months. and a preliminary operation to some smoking. The fish is headed. Fish curing is practised worldwide. split and opened out flat. and the extracted moisture is allowed to drain away. and salting is done by stacking the fish in layers. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. Usually. and marinating processes. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5.5. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. SMOKED AND DRIED FISH Dr. CURED.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. both to preserve and to create a variety of new products. 93 . rather than for preservation itself. It includes the salting. drying and smoking processes. and is sometimes known as ‘pickling’. alternating fish layers with a layer of salt. Wet salting or brining. their combinations and/or variations. and before significant spoilage takes place. Fish can be salted in three different ways or their combination: Dry salting.

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

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

smoking or marinating processes.8 211. Salt in the water phase links the two conditions of moisture and saltiness. TABLE 5. it is impossible to establish standardised processing stages in the curing of fish. it is important to understand the concept of salt concentration. is employed.3 Processing and its Effects on the Microflora Owing to the wide variety of products. The dissolution of salt in water is dependent on the temperature. Thus. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. For smoking salmon. the fish is usually salted with a 70 .4 184. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. % 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. a 0º brine saturation means pure water. or salt in the water phase.0 g of salt per litre of brine). Different brine strengths are required according to the curing processes and/or the type of product. This should be distinguished from the 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) 26.8 79.0 .FISH AND SEAFOOD 5.6 g of salt or more per litre of brine) may cause salt burn in this way.I). The strength of the solution is described as degrees of saturation (brineometer degrees).6 132. it causes swelling because the fish absorbs water from the salt solution (1). At 18 °C a 100º brine is produced when the solution contains 26. about 50º (132. When working with salting. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. it is important to state the temperature.2 g of salt per litre of brine). etc.2 105.0 158. When a milder brine.2 237.4 52. Salt concentration means the percentage of salt in the water contained in the fish. When a 100º brine is used. sometimes the salt crystallises onto the surface of the fish.4% salt (Table 5. only the most common processes can be described here. a 90º or above brine (237. Therefore.80º brine (184. processes and producers’ private recipes. all using salting steps. and salt content.8 211.6 264.

addition of levels of 0. and such delays.35% and 0.1 Salting Salting of fish for preservation purposes has been used since pre-history. This is because fatty fish turn rancid quickly. The most common fish used in the northern hemisphere is cod (Gadus morhua). it tends to lead to a slight yellowing and softening of the product. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. 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. are recommended to help make the product whiter. low in calcium and magnesium. See Figure 5. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. ‘Green cured’ split cod with about 60% of the backbone removed. which include calcium and magnesium. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). Also. For a light salting. mainly in tropical conditions.3.CURED. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. the fish is restacked with fresh salt after about a week. and salt can speed this oxidation process. ‘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.1 ppm and 10 ppm. 97 . the fish is mixed with one part of salt per eight parts fish. Salting can be done in a number of different ways. outgrowth of halophilic bacteria may occur. when the air humidity is high.1. and a ‘putty’ fish product is also obtained (1). can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. producing characteristic pink patches. On the other hand. However. respectively.1 Dry salting / Kench curing Usually. and for a heavy salting a ratio of 1:3 salt:fish is used (1). Usually it is followed by other preservation techniques such as drying and/or smoking. 5.15 .0. caught in North Atlantic waters. SMOKED AND DRIED FISH 5.15% calcium and magnesium. This condition is known as ‘putty fish’ (4). The calcium and magnesium impurities.3. and the salt has penetrated throughout the fish (3). respectively (1). The salt used should be of food-grade quality. can be dried further to produce ‘klipfish’ or ‘bacalão’. having a short section at the end of the tail.1. if very pure sodium chloride is used for curing purposes. are likely to cause hydroscopicity of the product and. but the concentration of copper and iron must be kept under 0. for which there is a high demand in Mediterranean countries. and essentially free from iron and copper (3). however. lean white fish is used for this purpose. When pure salt is used.

3. For large fish it is advisable to split it before salting.2 Wet salting or brining As referred to above. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. such as Staphylococcus aureus could survive and grow at such water activities in the product. Only small fish can be salted whole. Processing of dried salted cod 5. eviscerated and/or split). Food-poisoning organisms. it appears that water activities much less than 0. Fig.1.1. the process is mainly for flavouring purposes prior to the use of other preservation processes. Where fish are salted in their whole form. such as 98 . so that the risk of spoilage is high.5.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. The greatest barriers to salt penetration are the skin and scales. although toxin production would be unlikely (4). when a light brine is used.90 are rarely achieved.

limiting the amount of rancidity that can develop.98 6.16 7.65 2.78) (1).49 4.1.47 6.98 5.5 4. and a red burgundy colour will develop in the product.44 2. the liquid that comes out of the fish forms a brine.33 5.78 6.76 4.85 3.0 4.66 5.3. There are several variations of this process. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.35 4.88 5.II. Histamine formation is of importance when using scombroid and some clupeidean species. headed and gutted.51 3. For example.04 5.0 3.41 3.66 5. Here. but mackerel. 5.73 1.84 1.70 3.56 6.5 3.23 2.17 4.05 7. but as it is placed in containers. and can be collected and replaced again over the fish.76 4.76 4.09 8.64 4. a saturated brine is used.11 3. the fish is dry-salted.98 6. if split fish is dry-salted. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.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) 2.44 2.90 (1).54 5.22 6.00 3. with pressure being applied to the top of the container to help the brine to drain away. The main objective of the brining method of salting fish is to limit air penetration during processing.11 9.03 4. sardines.26 2.41 6.5 10.61 3.06 4.86 2.99 2.91 2.76 4. the fish enzymes play an important role by maturing or ripening the product.78 2.80 5.0 2.41 5.41 4. the brine is then replaced over the fish. The fish are kept immersed in their own brine near saturation point for several days.09 8.25 5. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .25 5.51 5. TABLE 5.46 4.85 3. See Table 5.3 Dry / wet salting In this process. can be cured in a saturated brine solution for 6 days and.23 3.76 4.61 3.17 5.23 8.03 2.26 3.74 9.5 3.30 2. SMOKED AND DRIED FISH smoking. they can be stored at 28 °C.CURED. For preservation purposes.60 2. Fresh sardines.10 1.76 99 . herrings and other fatty fish can be used.26 7.95 3.26 1. the product obtained is called gaspé-cured (5). thus. Histamineforming bacteria do not generally grow at water activities below 0.23 2. after being drained and pressed at 465 kg/m2 for 15 hours. Anchovies are the main fish used for these purposes. This is why it is the preferred method for fatty fish.45 3.11 4.96 1.70 6.3 days.26 4. with a shelf life of about 4 weeks (water activity (aw) about 0.19 3.

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

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

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

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

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

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

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

The production method is similar to masmeen. 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). (Penicillum cyclopium and Penicillium puterellii) (19). SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. resulting in a characteristic flavour (20. a very expensive and fashionable smoked product in the Netherlands. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). to have no further advantage over other packing methods for other fish (12).CURED. Aspergillus ostianus. 23). but seems. When refrigerated at 3 °C. The classical liquid smoke is 107 . is a boiled smoked fish product that is used in Japanese cooking to impart flavour. Katsuobushi. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. They are generally not cooked prior to consumption. 22. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). Eels. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. Mackerel. chilled or freeze-thawed. but mechanical drying is used instead of sun drying. moulding it into long thick sections. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. break down of fats and the moderation of smoke flavour. including Eurotium repens and Eurotium rubrum. are usually hot-smoked after being immersed in 80º brine or dry-salted (6).3. are used. Growth of these moulds on the tuna results in the extraction of moisture. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. Vacuum packaging retards the onset of rancidity when fatty fish is used. 21.6 hours and smoked for 3 . Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). Hot-smoked products made from white fish will usually keep better than those from fatty fish.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. white fish about 8 days. The product is removed from the drying cycle. Aspergillus tamarii) and several Penicillium spp.5. are usually 80º brined for 5 . 5. 17. apart from the improved appearance. cooking in brine. rewetted and remoulded to give the desired shape and consistency. fatty fish will remain in good condition for 6 days.3½ hours using the typical temperature set for hot-smoking referred to above (12). Aspergillus oryzae. 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). and substances unconnected with smoke but with a smoky flavour or smell. In terms of safety. then hot smoking and finally sun drying it. Natural smoke extracts. synthetic smoke flavours. similar to masmeen. 18). Boiled or steamed clams. They will keep well frozen. such as yeast derivatives. including ‘umami’ (16. and many closely related Aspergilli (Aspergillus ochraceus.

smoking.6 to 2. 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. The European Scientific Commitee for Food. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. injecting it into the fish flesh. so as to impart an unnatural red colouration to the flesh. and less than 0. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. drying. 5. According to some recent research.3. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). Fresh fish spoilage may be measured by 108 .8% and still provide protection for 7 days (25). 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. 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. 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). spraying is preferable because it is easiest to control flavour and acceptable with this method (3).03 μg/kg of benzopyrene.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. consistency of flavour and colour.4 Spoilage Brining. or smoke extracts in organic solvents. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25).06 μg/kg of benzoanthracene in the final foodstuff (28). Liquid smoke was considered to provide advantages of safety. 5. When liquid smoke was used. In experiments comparing traditionally smoked fish with liquid smoke.5. or applying it as an atomised spray within a modern automatic smoke kiln. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. applying it as a dip after brining. It has been shown that liquid smoke can reduce the formation of PAHs (3). both of which are considered potential carcinogens. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. nutritional neutrality and antioxidant activity (27). established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0.

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

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

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

W.variable. + to +++ strength of preservative effect 4 Post-process contaminants 112 . halophila Halophilic spoilage organisms Halophilic spoilage organisms. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. 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. 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. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. sebi and B. Dun.not used/unchanged. and Halobacterium spp.FISH AND SEAFOOD TABLE 5. LAB. . boiled.

respectively. popular demand for healthy foods has led to a change in the tastes preferred by the public and. the fermented products being the most dangerous group (7).97 0.5-8.5-8. 9 and 8 of the outbreaks. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E.0-9. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). the non-proteolytic type E strains can grow in up to 5% sodium chloride. In the United States. Before. but if the fish products are stored at temperatures <10 °C.75 0. aureus Yeasts Moulds Lactic acid bacteria pH 5. and as botulism is a serious hazard in foods. botulinum type E and production of toxin. under optimal conditions. botulinum Staph. could not grow and produce toxin. the importance of cold-chain integrity must be emphasised. smoked and dried. pressure on the traditional methods of curing. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14).86 0. the products were heavily salted.97 0. It has been shown that. Since neither the smoking nor the drying parts of this curing process are particularly severe. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). with this change. botulinum type E.5-10 4. when present in the raw material. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39).5 0-11 3. However. The concentration required to prevent growth at room temperature can vary from as low as 3. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0.5% to 5%.8 0. types A and B are not affected by 3 .8 1.5 5.9 0.5-10. although the combination of the two factors.5.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. sodium 113 .CURED. SMOKED AND DRIED FISH TABLE 5. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. The presence of salt in cured products has a great effect on the growth of the bacteria.5 5. and C. salted and pickled products accounted for 10.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. botulinum. botulinum type E Staph. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. Examination of 165 outbreaks of botulism caused by fish products showed that smoked.5 4.61 Organisms Most Pseudomonadaceae C.

gills and guts as well as in several lightly preserved fish 114 .4 Listeria monocytogenes The psychrotrophic L.5. As with other organisms. As they are poor competitors. the organism may contaminate the product. Usually.30% of the population positive for enterotoxin-producing strains (41). as they require more handling.5% salt in the water phase.4. Thus. but. but requires higher temperatures for toxin production (>10 °C). Once the product is contaminated.or cold-smoked fish should have at least 3. after processing. throat and skin. the human nose. it should contain not less than 3.3 Enterobacteriaceae The pathogens Salmonella. they will grow rapidly (7). if recontamination occurs and storage conditions are favourable.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. if it contains sodium nitrite ≥100 ppm.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. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. Infection by Shigella sonnei has increased in the UK. and the minimum pH for growth is 4. are particularly susceptible to contamination and. related to the consumption of a raw smoked fish meal. Sliced smoked fish products. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . 5. has been reported (42). It is a halotolerant bacterium able to grow at water activities as low as 0. aureus growth. the organism can survive most of the light curing processes. 5. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. aureus. if storage temperatures are over 10 °C.5.86 and 10% sodium chloride. but. 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. In foodstuffs. coli all occur on fish products as a result of contamination from animal/human reservoirs. more importantly. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). An outbreak of Shigella in an Italian cruise ship.0 . 5. if the person handling the product is a carrier of Staph. the risk of infection can be eliminated by proper cooking. The FDA established that hot. growth and toxin formation may occur. The organism is mesophilic with a minimum growth temperature of 7 °C. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. they do not grow well in the raw material.5. However.0% salt in the water phase (30). Shigella and E.94 (7).5. when the bacterial load of the product is lower.

widely used as starter cultures in food processes. or nematodes.5 Parasites Despite the presence of parasites in fish being very common. and the tape worm Diphyllobothrium spp. do not survive 1 min at 60 °C. Of microbiological concern in the northern hemisphere are two parasites . from freshwater fish. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. and in CT95-1207FAIR). Two outbreaks in the United States have been reported to be linked to salmon (10). SMOKED AND DRIED FISH products such as cold-smoked fish. Unlike some European countries.5. a short period of freezing either of the raw material or of 115 . Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. such as ‘lightly preserved’ fish products. which means that the process must conform to a level of ‘no detectable L.CURED. and other local traditional products (7). pH 4. in the EU Fisheries R&D-FLAIR program. The round worms.95 (45). salt levels of 8 . 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. Its limiting conditions for growth are 0 . which include intermediate hosts. Some LAB.514. the only concern to public health is when eating raw or uncooked fish products. a round worm. are considered unsafe. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. They have complicated life cycles.2. The use of LAB in fish preservation is a topic that has been studied in projects (UP. from seawater fish.6 and water activities below 0. 5. most of them are of little concern to public health.12%. the CT 3162-FAIR program. monocytogenes’ in 25 g of the finished product. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. lightly salted caviar. monocytogenes in levels at or above 102 per gram (46). a number of fish products. Thus. favours growth of psychrotrophic LAB.9.the round worm Anisakis spp.0. has also been detected in cod (7). which. monocytogenes pathogenic for only specific segments at risk. This includes matjes herring. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). which considered L. Until 1997. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). gravad fish. Pseudoterranova dicipiens. the USA established ‘zero tolerance’ for the organism. As they are heat-sensitive.45 °C. both being detected in cold-smoked salmon. cold-smoked fish. there were no reported outbreaks related to cured fish. have been shown to have an anti-listerial activity in many food products.92 .8 . marinades (ceviche) (43) and gravad fish (44). in combination with curing agents and refrigerated storage.

monocytogenes) (cfu/g unless stated) <106 106 . salami. (not L. Acceptability is based on appearance. perfringens B. coli (total) Listeria spp. 5. 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.<103 N/A 100 . smell. cholerae Vibrio parahaemolyticus L. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E.<100 >104 >100 N/A N/A <20 20 .<100 200 .VI Guidelines for the microbiological quality of ready-to-eat smoked fish. texture.<104 100 .<104 20 .<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.<107 ≥107 N/A <100 <20 100 . cereus and Bacillus subtilis# <20 <20** <20 <10 20 . soft cheese. and the levels or absence of indicator organisms or pathogens. E. not detected in 25 g Campylobacter spp.<100 20 .<104 104 .<100 10 .<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . the organism should be identified # N/A denotes not applicable 116 . † Guidelines for aerobic colony counts may not apply to certain fermented foods. If the Bacillus counts exceed 104/g. monocytogenes Staph.FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7).<100 20 . coli O157 & other VTEC Vibrio. These foods fall into category 5.100 >100 N/A Pathogens Salmonella spp. ‡ On occasions some strains may be pathogenic. and unpasteurised yoghurt. for example. aureus C.6 Published Microbiological Criteria TABLE 5.

. 1994. 1991. coli (ETEC) L. * Staph. Marinades. Preservation of fish by curing (drying. 2. Regenstein C. 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. 1982. Leatherhead.M. 56. McLay R..F. 381-406. Wareing P. 3.H. Smoked. botulinum * Limit of tolerance 1x 103 ETEC/g. Torry Advisory Note No. 1979. in Fish Processing Technology. salting and smoking).. in The Seafood Industry. cured and dried fish.A. The Smoked and Cured Seafood Guide. New York. Horner W.W. Urner Barry Publications Inc. References Regenstein J. 4. Hall G. Food spoilage fungi. FAO Fisheries Technical Paper. parahaemolyticus Vibrio vulnificus C. Van Nostrand Reinhold. Moody M. Fernandes R. Wareing P. Torry Advisory Note No. Horne J. 37(revised).M. except in the viscera of dungeness crab.. 117 . 120-38. aureus * Vibrio cholerae V. 9. 6. Bannerman A. USA. 254-82 5.CURED. Catching. 1997. New York. Handling and Processing Eels. Introduction to Fish Technology.F. Fernandes R. SMOKED AND DRIED FISH TABLE 5. Doré I. Ed. in Micro-Facts: The Working Companion for Food Microbiologists. 8. 2007. 3273. 1984. London. Eds. monocytogenes Salmonella spp. 7. Hot Smoking of Fish. Van Nostrand Reinhold.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.E. Leatherhead Publishing. or presence of toxin 0.7 1. Martin R. 334. Birnie K.. Assurance of Seafood Quality. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. Huss H. 1990. Eds.. Chapman & Hall. Flick G. 82 (revised). 1993. Torry Advisory Note No. Flick G.

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

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

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

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


1. koji. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. producing the fish sauces and pastes. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. Essentially. they are exported to European. FERMENTED FISH Prof. Fermented fish products are largely confined to east and south-east Asia. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6.1 Definitions As applied to fish products. typically in the range 15 .25% and are used mainly as a condiment. Carbohydrates are added to some fish sauces during their preparation. 6. For example. the most important of all fermented fish products.6.I. though some are produced elsewhere. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. The fish sauces are produced on the largest scale.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. added during preparation. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. in economic terms. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. The relative importance of these two activities depends on the formulation of the product. North American and other markets and are. the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. there are numerous variants of some common themes and a host of local names used to describe them. they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. lactic fermentation can play a significant role. As with most traditional products that are still produced principally on a cottage industry or domestic scale. whereas in those where a carbohydrate source such as rice or sugar is also added. 123 . Those to which only salt is added tend to be mainly autolytic.

6.II.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. a variety of products can be produced within a single country. and the more widely known Worcestershire sauce a very distant relative. salty taste.III. Fermented fish products are of very minor importance outside Asia. This is well exemplified by some of the fish/salt products of Thailand described in Table 6. to products analogous to other lactic fermented foods such as salami.FISH AND SEAFOOD TABLE 6. made from whole fish in France. made from fish livers in Greece. where the bacterial production of lactic acid is a major feature. A selection of Asian products is presented in Table 6.1. 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.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. and pissala. In Europe. These have long since fallen from use. who had adopted them from the Greeks. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. 124 . although garos. may be direct descendants. 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.

pla-chao. tai-suchi phaak. cencalok burong-isda.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. burong-dalag. som-pa. ayu-sushi.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi.3 months Main dish Tai-pla Bowels 1:3 6 . som-pa-keng pekasam. pa-khem. crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 .12 months 10% raw sugar added. mam-seeing sikhae som-kay-pa-eun. mam-chao. mam-pa-kor.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 . e. burong-bangus pla-ra.FERMENTED FISH TABLE 6. pla-som. e.8 months Main dish TABLE 6. 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. burong-ayungi. funa-suchi. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps. pa-chao. crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 .g. shrimps. som-fak/ som-fug 125 Malaysia Philippines Thailand .g.

relatively mild-tasting product. especially in fish/salt products. in a survey of fermented fish products in north-east Thailand. There are also more heavily fermented products. which employ lower levels of salt than Asian products. fermented trout.the flooded rice fields. The diversity of fish used in fermented products (see. in Norway and surstrømming. A modern account of their production. These are often described as fish/salt products. the most popular varieties of which are rakørret. Penaeus indicua Many of the fish used in fermentation are uneviscerated.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. states that sugar is often included with the salt. which has been increasing in popularity over the last 50 years. malt or flour (1). TABLE 6. rakefisk or surfisk. and the initial microflora is likely to be considerably larger in these cases.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. Table 6. For example. Here. paddy ponds beside rice fields used for collecting fish when the field has dried up. and a large local freshwater reservoir (2). 6. and old descriptions of their production often refer to the addition of whey. or buried salmon. freshwater fish are the usual raw material. 126 .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. and counts ranging from below 104 /g to in excess of 107 have been reported. however. is a traditional. and their microflora tends to reflect their local environment more than that of marine species.FISH AND SEAFOOD In Northern Europe. some fermented fish products remain popular. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. made from herring. it was found that several different sources were used for the fish . for example. Norwegian gravlaks. 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 Sweden.

Mucor.3. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns.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.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. Other halophilic species have been identified from fish/salt products. Bacillus spp. and studies 127 . Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that.FERMENTED FISH Where other ingredients are used. These preparations contain a mixed microflora of moulds and yeasts. which included nam-pla and patis. in many cases. Similar results were seen with patis.74 or below. It also demonstrated their significant proteolytic activity during the first month of fermentation. principally species of the genera Rhizopus. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. angkak (Philippines and Thailand) and look-pang (Thailand) are used. or boiled or roasted rice. they can contribute to the initial microflora. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). 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. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). (3. traditional starter culture preparations such as koji (Japan). however. 4). More recently. mean that microorganisms have been generally held to play little or no role in the process that follows. When this is a simple carbohydrate source such as sugar. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). sufficient to saturate the water phase present and produce an aw of 0. were found to be the predominant isolates. In some cases. 6. Endomycopsis and Saccharomyces . Hansenula. either directly or as the partially saccharified rice they produce. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5).4. however. the Philippine fish sauce. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter.3 6. and the anaerobic conditions that prevail in the bulk of the material.1 – 6. Generally. after one month. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. 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. This. In another microbiological study of four fish sauces.

Fig. 6.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 . 6.12). Belacan (Condiment) Fig.1.2. Patis (Condiment) 128 .

In fish sauce production. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. 6. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are. the 129 . eviscerated fish are sometimes used to slow the autolytic process. Ika-Shiokara (side dish) Fig.FERMENTED FISH Fig.6. 14). however.4.3. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. thought to be largely the result of autolytic breakdown of the fish tissues. To produce semi-solid fish pastes as opposed to fish sauces.

filtered and bottled.3. Hoi-Malangpu-Dong (side dish.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. which is eventually separated from the residual fish tissues. particularly glutamate.5). Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. 6. Figure 6. 6. volatile fatty acids and nucleotides.V and Figures 6. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. snack) 130 .8. Fig. Some examples of products and their formulation are presented in Table 6. In contrast to free-swimming fish.5. 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. for example.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.6 – 6.

3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0.6. central. C NE. NE. the prefix 'pla' is replaced by 'kung'. north-east.2-4 Fish (pieces): salt:khaomark (fermented rice). N. 3:1:0. S. C.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. Kung-Chom (staple food. N S 5 .12 months Main dish 2 Pla-jao C.7 days 5 . C NE. side dish) 131 .10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 . NE Formulation Fish (whole or pieces): salt: roasted rice.03 Preparation time 6 .6. In these cases. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus.5-1.12 months Shelf life Use 1 . N.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.FERMENTED FISH TABLE 6. Fig. NE 10 .4 are also produced using shrimps.12 days 3 . south of Thailand.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 3:1:3:0.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . N. north.

7.6. The LAB isolated from these products are those with more marked salt tolerance but. 2. while they are able to grow at high salt levels. then this will take longer to develop than if lower salt levels are used. It also indicates that.salt and lactic acid.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. If acidity is a desired sensory property of the product.2. these products are salted at lower levels than the fish sauces and pastes. (from Arroyo et al. Red burong isda 132 . typically in the range 2 . 106) Fig.1 The use of higher salt levels results in a longer production phase but a better keeping quality product. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6.VI).10%. their growth and metabolism are nonetheless slowed.. but there is considerable variation in this. Phillipines Journal of Food Science and Technology. giving a reduced rate of acid generation. the salt is of far greater importance in determining the product’s long-term stability. of the two factors contributing to shelf life .3. This is readily explained by the susceptibility of LAB to elevated salt concentrations. 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. Generally.

0 Shelf life 3 weeks 6 .5.2 Philippines Journal of Food Science and Technology.17.5 3.9 4.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).12 months 1 .2 4.5.1 .0 .8 4. 2.8 .5 .3 . White balao-balao TABLE 6.8 pH 7.0 .5 .9 .4 .FERMENTED FISH (from Arroyo et al.5.0 .9.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.9.7 ..3 years 1 .3 5.9 4.8 . 106) Fig. 133 .6 3.4.

TABLE 6. This results in a more active lactic fermentation and a lower pH.5) despite other factors. a common ingredient in these products. Garlic.FISH AND SEAFOOD 6. Where they are used. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH . inulin.4.9) compared with the white product (pH 4. 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). although the final population of LAB may be similar. alternatively. For example. 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 . being similar (18. Addition of carbohydrate will. also serves as a valuable source of the more readily fermented carbohydrate. 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 (20.3. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. Although the ability to ferment starch is not widespread in the LAB. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. Rice is the most common carbohydrate source used. of course. When added as boiled or roasted rice alone. such as salt content. also lower the unit cost of the product and this is probably a significant factor in many cases. In several products. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. the rice may be added in a partially saccharified form such as kao-mark (Thailand).VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi.1 .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.VII). 21). 19).

and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). which ranged from 7. 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. Results of a product survey were consistent with this. In a successful fermentation. and there is a marked regional variation in the preferred type.2 at the end of fermentation. Excessive growth and acid production by the LAB could render products too sour for acceptability. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23).8 . but they are likely to behave similarly to lactic fermented products produced from other commodities.7 up to 6.7 to below 4. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7.17.5:9:3. 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). Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. Clearly. presumably correlating with the salt content. will not reliably exclude air. reporting the product’s pH to range from 4. The growth of yeasts and moulds could. Generally. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. lead to an increase in pH.2. in turn. 135 . 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.9%. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. the pH drops from 6.FERMENTED FISH Sikhae. There is little published information on the factors that limit the shelf life of the low-salt products. Traditional packing techniques used with lactic fermented fish products. markedly extending the shelf life (25).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). allowing other spoilage organisms to grow. 6.3 days. the Korean fermented fish product. is something of an exception since millet is used along with garlic as the carbohydrate source. such as wrapping in banana leaves. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. around 12%.5. thereafter declining to about 4.3 weeks at around 20 °C. the level of salt employed is on the high side for fish/salt/carbohydrate products.5 in the first 2 . the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases.

under conditions inimical to growth and high ambient storage temperatures. 6. Several measures have been suggested for reducing the risk of botulism. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. viable numbers will probably decline quite rapidly. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. are Vibrio parahaemolyticus and Clostridium botulinum. In Japan.8 and at salt levels as high 10%. 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. typically 30 °C. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. parahaemolyticus has been reported growing down to a pH of 4. 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. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. however. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). and consumed without cooking. There is little in the way of documented information associating fermented fish products with foodborne illness. After capture. V. As in Norway. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. outbreaks of 136 . In the fish sauces and pastes. although these limits apply when all other factors are optimal (26). but we are unaware of any reports of this happening. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. The bacterial pathogens most commonly associated with the living environment of fish. Because of the severity of the illness it causes. which accounted for 88% of outbreaks in the period 1952 . fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage.87. fish products are the most common source of botulism. During fermentation.

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

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

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

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

.. most notably the azaspiracids and the cyclic imines as well as some additional polyethers. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. While some of these toxins only occur in seafood from a restricted geographical region. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins. Wright. several scientific reports have described their efficacy and potential. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. Another book dealing with the clinical effects of a variety of poisons and toxins. Ph. Scientific and Cultural Organisation) has been updated (7).3) and some practical applications for detecting shellfish toxins have been reported (4.7. Nevertheless. FISH AND SHELLFISH TOXINS Jeffrey L. known and unknown. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. and a second updated edition dealing with the pharmacology. Since the original chapter in this series was published (6). C. physiology and detection of marine and freshwater toxins has also appeared (8). The manual on harmful marine algae and their products. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7. including the well known marine toxins 141 . An emerging technology is the use of immunosensors (1 . 5). a number of new toxins and toxin families have been identified. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. Although most of these antibody-based test kits have yet to receive international regulatory approval. Increasingly sophisticated instrumentation (e.g. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity.M.D.C. It remains to be seen if this approach gains momentum in the future. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins.

The Ncarbamoyl sulfates are readily hydrolysed to STX. Not all the toxins found in shellfish extracts are microalgal products.FISH AND SEAFOOD has been produced (9). even crustaceans such as lobsters. temperate. and sub-tropical waters and habitats (15). The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). with intoxications occurring in many countries even today (15). Gymnodinium catenatum. 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. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). dealing with the detection methods and regulatory requirements for many marine toxins has been published (12).1). All PSPs degrade rapidly in alkaline conditions. 7. can become contaminated with these toxins. Consequently. 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. and a tropical species Pyrodinium bahamense (16). PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. 142 .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. Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). Saxitoxin Group instead of the Paralytic Shellfish Poisons). and a review on the risk assessment of marine toxins has also been produced (14). In addition.g. and another. neoSTX and Gonyautoxins (GTX) 1-4. Around twenty PSP toxins are known. some are the result of biotransformation processes by the filter-feeding host. all of which contain a tetrahydropurine skeleton (Figure 7. Many of these species are extremely common and are found throughout the world in a variety of cold. a great variety of shellfish from all latitudes.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. 7.2.

and humans seem to be particularly susceptible. tongue.1. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water.2 Toxicology PSP toxins affect the central nervous system of mammals.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. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. and fingers within 5 . However. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7.2. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis. Human symptoms begin with tingling and numbness of the lips.30 minutes of consumption. this is never sufficient to render the seafood safe to consume. 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). 143 . 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. 7.

and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. 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. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis.2). including Japan. 15. TTX most commonly occurs in many species of puffer fish. Most intoxication events have occurred in South East Asia. and over ten analogs are known (22. and islands of the South Pacific. 23.FISH AND SEAFOOD 7. Alternative HPLC methods have subsequently reduced this to a single HPLC run. 17). or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0.8 µg/g) of shellfish or seafood tissue. However it remains as the monitoring method of choice in many countries.3. a pre-column method has received Association of Analytical Communities (AOAC) approval (18). 24) (Figure 7. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®).1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. 7. and while simple and relatively inexpensive to operate. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. After some development and various trials. other derivatives have been reported. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). Taiwan. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. a criticism has been that such methods are time consuming. While the chemical detection methods are most promising for the future. 7. Thailand. The mouse bioassay has been in place for decades and the method has been standardised. as well as other parts. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). and has also been evaluated by a number of EU laboratories. Since its discovery. specifically 144 . 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. Two approaches have been taken here. China.2.

TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. and even newts. Typically this has been accomplished using HPLC followed by post 145 . the blue-ringed octopus.2 Toxicology Like the PSP toxins. Fig. 25.FISH AND SHELLFISH TOXINS in the liver. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. and once again the mouse bioassay has been used to detect the presence of the toxin. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. nausea and vomiting. horseshoe crabs. it is served in speciality restaurants with qualified chefs to properly prepare the fish. where puffer fish of the genus Fugu are a delicacy. and in mild cases results in distal muscle weakness. 26). raising speculation that TTX is produced by a bacterial source. respiratory failure and coma. molluscs. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. All of the observed toxicity is secondary to the action potential blockade. An oral dose of 1 . 7. Since then. In Japan.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions.2. Severe poisoning causes paralysis and respiratory failure. Structure of tetrodotoxin 7. gonads and roe (24).3. 27).2 mg of purified toxin can be lethal. 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. hypotension. A review of TTX detection methods has been published (12. 23. and more recently by an actinomycete Norcardiopsis dassonvillei (26). The onset of symptoms usually occurs rapidly. 7. and blocks diffusion of sodium through the sodium channel. preventing depolarisation and propagation of action potentials in nerve cells.3.

4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA).3.3 for examples).4. Immunoassay methods have also been investigated.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). 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). Fig. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28). However this method has been criticised for being complex. 7. but 146 . and while promising. Currently 12 isomers of DA have been reported (see Figure 7. but all of them are considerably less toxic than DA itself (29). Structure of domoic acid and three naturally occurring photoisomers 7. but it is known that such congeners are less toxic than DA itself (30). both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. 7. a polar glutamate agonist.FISH AND SEAFOOD column oxidation to the fluorescent derivatives. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. need further development (24).

but any impairment of the blood brain barrier results in additional risk. 147 . southern and western portions of Ireland. disorientation and coma. and in locations around New Zealand (32. Indeed. and is mainly distributed in the blood. For example. Victims display a variety of symptoms including headache. Okinawa and more northerly parts of Japan.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. 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. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp.. To a great extent.FISH AND SHELLFISH TOXINS only three strains (P. multiseries. another diatom species Nitzschia navisvaringica. Penetration of the blood brain barrier is poor in healthy subjects. and clams. found in the Pacific around Vietnam. the Philippines. 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. and review of the toxicologic pathology of DA has been published (36). Recently. Another factor is the movement of the toxin through the food chain. 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. The oral absorption of DA is around 5 . Pseudonitzschia australis. 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.10% of the administered dose. as does impaired renal function which increases serum concentration and residence time of the toxin. cormorants and pelicans (34). nausea. has been reported as a medium-level producer of DA (33). The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. Levels of DA in the other strains are generally <1 pg/cell.39). seizures. scallops. and sea animals such as sea lions and even whales (35). Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. including persistent short term memory deficits (36 .4. the Mediterranean. which has been well documented in studies along the west coast of the US. 33). and this has led to ASP toxicity in birds. 7. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). 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.

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

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

All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. and while this provides critical seafood safety information. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. DSP episodes have been reported worldwide and are included in many monitoring programs (47). and abdominal pain. 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). even with the minimal of clean up steps (48). various European countries.5. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. 7. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). A regulatory level of 80 . vomiting.5. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. nausea. 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. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 .FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. In recent years. 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. which can lead to an underestimation of the total DSP content in a sample. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. 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. DSP episodes have been reported from Japan.3 days.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. 7. Human symptoms include diarrhoea. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. New Zealand and Chile. Since the late 1970s. 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). DSP-Check is a commercially 150 . and a detection limit of 100 µg/kg has been reported (12). Due to the lipophilic nature of the toxins. 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. and DTX-2.

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. 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. Within this group. In yet another approach. However. Some representative examples of both groups of toxins are shown in Figures 7. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57.5 and 7. Indeed. 7.6. for many years these compounds were included in the DSP toxin complex. with a claimed detection limit of 20 µg/kg. and another rapid antibodybased test kit for DSP toxins has also been reported (53). 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.5. 7.6 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). with good agreement with the mouse bioassay and LC–based methods (54. 55). they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). 58).FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . Fig.

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).6.6.6. Structure of the two parent yessotoxins. 57. eastern Canada. Japan. 7. 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. 7. the toxin content and profile may vary among different strains. New Zealand and South America (49. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. many parts of Europe and the Mediterranean. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. which may co-occur in blooms with other organisms. As is typical for many algal species.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). YTX and homo-YTX 7.FISH AND SEAFOOD Fig.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). 58). While 152 .

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

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

8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. nausea. this family of toxins has proven to be a challenge both to identify and monitor. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. 7. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. The mouse bioassay is used as the primary detection method. In this application. The mean time to onset of symptoms is around 3 hours. 69). which can result in a cascade of associated events in a cell (66). and diarrhoea. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73).2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. vertigo. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 .3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. and mammalian body fluids without pretreatment or dilution (72). shellfish homogenates.7. 7.7. with hot spots in the coral reef areas of the Caribbean and French Polynesia. and in severe cases seizure. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). the competitive ELISA method correlated most closely with the mouse bioassay results. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). with a mean duration of symptoms of around 1 day. The human health implications following exposure to NSPs have been reviewed (69). A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. 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.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). 66. Ingestion of brevetoxins results in a variety of symptoms including parathesia. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. While the mouse bioassay continues to be used. malaise. 7.

1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins. 7.8. A detailed comprehensive review of ciguatoxins has recently been published (74). Distribution through the food chain begins with 156 . lipid soluble polyethers. The diversity of benthic phytoplankton that exist on reefs further complicates the picture. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. a group of closely related polyethers that are considered the source of ciguatoxins. CTXs are large. green and brown seaweeds and also occur in sediments and coral debris. 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. complex. Fig.8). and only a few structures have been identified. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish.FISH AND SEAFOOD transferred through the food chain to the larger carnivores. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7.8. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. but more are suspected to exist.

sharks. Local island populations attempt to avoid poisoning by catching only smaller fish. weeks or even months (74 . but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. groupers. Even when suspect samples are available. Other symptoms include a metallic taste.3 Detection methods As a consequence of their potency.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. the neurological symptoms can persist for days. The onset of symptoms can occur rapidly within 24 hours.8. and temperature sensation reversal. 75). arthralgia.8. which can be confused with chronic fatigue syndrome. Globally. although not all CTXs are equipotent. which can return following the consumption of alcohol and certain foods such as oily fish (74. myalgia. Like the brevetoxins. there are neither standards nor an official testing program. and moray eels.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. only trace amounts of CTXs can cause illness following consumption of contaminated fish. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. 75. where cold objects feel hot and vice versa. and the sensation of loose teeth (77). The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. pruritus. Patients have reported symptoms lasting weeks and even months.76). Nevertheless CTX poisoning continues to be a threat to local and visiting populations. all of which can be linked to the effect of CTX on excitable membranes. and it is estimated that ingestion of as little as 0.1 µg of ciguatoxin can result in human illness (74. 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. 7. and there are suggestions that such incidents are increasing and spreading worldwide (74). which in turn are consumed by carnivores such as snappers. 78). though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. barracudas. These include paresthesias (numbness and tingling of the extremities). and although the gastronomical problems may dissipate within a day or so. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. and in fact CTX poisoning is often only identified after consumption of contaminated fish. Another symptom that can develop over time is extreme fatigue.

In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). followed by clean-up on silica and aminopropyl cartridges. The majority of these have been proposed based on LC/MS analytical data. at C-3 and C-23). Consequently. Acetone extracts of fish tissue are subjected to solvent partitioning. The original structure proposed revealed an entirely new class of shellfish toxin. Recently. and following synthesis of the molecule some time later. 158 . following consumption of contaminated mussels Mytelius edulis (82). another in vitro sodium channel-based assay. Usually the major AZA derivatives found in shellfish are AZA1 (50 .85). A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine. has been developed. Since the original discovery and identification of the toxin. and the corrected structure was published in 2004 (86) (Figure 7. Immunoassays may be an alternative detection method (79). and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. This is then subjected to LCMS/MS analysis. 84). Recent reviews of this new group of toxins have appeared (83. and only a few structures (AZA 1 .5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis.80%). which are based on an LC-MS/MS approach.9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland.30%) and AZA3 (5 . although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80).20%) present in lesser amounts (Figure 7. Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. and another dealing with the chemistry and detection methods has been published (85). Chemical detection methods are severely compromised by the lack of standards.9).9). almost 20 additional derivatives have been reported (83 .FISH AND SEAFOOD Lethal Dose (MLD) 0. high-throughput screen (74). with AZA2 (10 . minor revisions of a few stereocenters were made. 7.25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. which features characteristic fragment ions corresponding to sequential losses of three molecules of water. and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge.g. but this only has application in a clinical setting (81). only a few laboratories are able to undertake such analyses. specific for CTX activity.

oysters (Crassostrea gigas. Mytilus galloprovincialis). While this result is not necessarily in debate.9. 90).2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. Structure of the three principal azaspiracids found in shellfish 7.9. Scandinavia.3 days. 88. Ostrea edulis). In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. it has been impossible to establish that P. These symptoms are not unlike those experienced following the consumption of DSP toxins. cockles (Cerastoderma edule). as in other cases of shellfish toxins. clams (Tapes philipinarum. 84. cassipes produces AZAs in culture. and to date no long-term effects have been recorded (83. However. but current data indicates that unlike the DSP compounds. these molluscs were never considered to be the biogenetic source. Following the original episode. Ensis siliqua. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp.9. 89). 7. AZA toxins do 159 . and some time after the original episode in Ireland. There has been considerable debate over the source of azaspiracids. cassipes (83). 7.FISH AND SHELLFISH TOXINS Fig. and most recently in a crustacean (Cancer pagurus). The bivalves affected include mussels (Mytilus edulis. AZA occurrence has been reported by several European countries including France. diarrhoea and stomach cramps. which are ingested by P. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87).). vomiting. and to date this has been the only source from which these toxins have been found at toxic levels.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. scallops (Pecten maximus). which can persist for 2 . Donax spp. and off the coast of Morocco (83. Portugal.

but all contain a spirocyclic or cyclic imine function within their structure. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. 7. Once again. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. and necrosis of epithelial cells on the microvilli (91). 91.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. 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). 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. Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. The mouse bioassay has been used to monitor shellfish extracts. Based on IP minimum lethal dose data. Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. Importantly these pathological outcomes were reported as being different from those caused by DSP. and erosion and bleeding in the stomach. the spirolides. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum.10 – 7. 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. 7.10 Cyclic imines These toxins are represented by several distinct structural types. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. and ASP. However.9. PSP. 92). and closely related ptierotoxins (Figures 7. the pinnatoxins. abnormalities in the thymus and spleen. AZAs also display cytotoxicity towards various mammalian cell lines (83. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg).FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. 85). parenchymal cell pyknosis in the pancreas.12). Using the same approach. which are particularly amenable to detection by positive ion electrospray mass spectrometry. 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. though the possibility exists that they may inhibit other phosphatases.

two additional congeners.10. 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. 95). Subsequent taxonomic studies revised this again to a new dinoflagellate K. cockles. but they are included here for completeness. Structure of the cyclic imine gymnodimine 7. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). 161 . they are sometimes referred to as emerging toxins of uncertain human health concern. 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. Some time later. 7.520 Da (94.10.10). The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. In only two cases (gymnodimines and spirolides) has a planktonic source been identified.. surf clam and New Zealand abalone. Pacific oyster. Fig.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. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94.1.FISH AND SHELLFISH TOXINS health. it was proposed that a dinoflagellate identified as Gymnodinium spp. 7. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. 95). a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin.10. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . At the time of the initial observation of gymnodimine.

cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. Oral administration of the toxin to mice by gavage resulted in about an 8.1. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine.to10-fold reduction of toxicity (LD50 600 .2 Toxicology A review of the toxicology of all cyclic imines has been published (98). abdominal breathing. 7. Most cyclic imines lack an extended chromophore. 7. paralysis of the hind quarters and eventually complete immobility. In particular. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94.10.1. and so spectroscopic detection by UV methods is not favoured. 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. However. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds.FISH AND SEAFOOD 7. A later study using purified toxin established an LD50 dose (Lethal Dose.10.11). There is a rapid onset of symptoms including respiratory distress. Whether isolated from dinoflagellate cells directly or from shellfish tissue. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. 95) (Figure 7. and lack of responsiveness to stimuli.100). The time lapse between ingestion and death is between 5 and 15 minutes. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. together with a lack of evidence of human intoxication. Since their original discovery which resulted in the characterisation of spirolides A-F (99. 50%) of 96 µg/kg.10. They were originally discovered in extracts of mussels (M. 95). The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 .900 µg/kg).3 Detection methods The characteristic “fast-acting. Based on this diminished oral toxicity. finishing with 100% methanol. Reduction of the imine group eradicated the toxicity. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. and this is the detection method of choice. it has been stated that regulation against gymnodimine is not required (98).

FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). by gavage (87 . All data reported here are for desmethyl spirolide C. those spirolides possessing the vicinal dimethyl grouping (see Figure 7.11. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins.2. While oral toxicity is considerably less than by IP injection. the western coast of Scotland. 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.8 µg/kg). They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102).2. Fig. 7. Interestingly.10. 7.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. 95). southern Ireland. fasted mice).10. fasted mice). or by oral ingestion (381 . in fjords in Denmark and Norway. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 .166 µg/kg. Structure of four spirolide toxins A-D 7.2 Toxicology The spirolides are the most studied group of the cyclic imines.11. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada.707 µg/kg. the gulf of Maine. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 .

Fig. and in fact these compounds share a number of other structural features. 95. 7. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. 7. Three members of this group have been reported. four members of the group have been identified – two of them being the C-34 diastereomers. A number of methods following this procedure have been described (94. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry.3 Detection methods As discussed for gymnodimine.12).2. consistent with modulation of acetyl cholinergic receptors (103).10.3 Pinnatoxins and pteriatoxins Pinnatoxins A . 7. Structure of the pinnatoxin group and the closely related pteriatoxins 164 .B contain a spiro imine moiety similar to that found in the spirolides.FISH AND SEAFOOD (100). While the mechanism of action remains to be clearly defined. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7.12. the mouse bioassay can be used to detect these fast acting toxins. 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. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C. To date. which includes two diastereomers. 102).10. However.

100 µg/kg). arise from an algal source . but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. 7. The LD99 (Lethal Dose. but this remains to be conclusively established (106). While this is recognised as a real possibility for a wide range of toxins. and more advanced means of detection.3. With increasing knowledge of the chemistry of toxins. Coupled with this. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). while pinnatoxin D was essentially inactive.3. like the gymnodimines and spirolides. This is generally a result of increased monitoring and continually improving detection technologies and methods. 99%) values for pinnatoxins A-C are in the range (22 . has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. have long been suspected of causing human illness after ingestion.10.3. 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.FISH AND SHELLFISH TOXINS 7.11 Conclusion There has been considerable progress in the detection of known marine toxins. new toxin groups such as the azaspiracids and cyclic imines have also been reported. 7. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. However. which in many cases are now found to be more widespread than previously believed. there is no compelling evidence to suggest that they have resulted in human toxicity (98). to date action or 165 .1 Origins and distribution Shellfish of the genus Pinna. Only the acute toxicity in mice following IP injection has been reported.10.10. Despite this progress. The LD99 dose for the pteriatoxins is reported to be in the range (8 . There is strong circumstantial evidence to suspect that these toxins.3 Detection methods No formal detection methods for these compounds have been reported. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105).most likely a dinoflagellate.180 µg/kg). which occupy tropical waters around Okinawa and the south east coast of China. 7.2 Toxicology There is little information describing the toxicology of these compounds (98).

1267-74. 2008.A.. 2007. Kane M.. . in order to fully study the synergistic effects of multiple toxin types..R. 15871.12 1.M. 2007. Gobi J. 7. 121. Paris. 166 4. Campas M. Joint FAO/WHO/IOC activities to provide scientific advice on marine biotoxins (research report). Biosensors and Bioelectronics. Food and Agriculture Organisation of the United Nations. Lawley R. Even if they do not completely replace other methods such as the mouse bioassay or chemical analysis that require sophisticated chemical analytical instrumentation. Recent advancements in surface plasmon resonance immunosensors for detection of small molecules of biomedical. 179-99.V... thus significantly reducing the number of mouse or chemical analyses that have to be performed.E.. Food and Agriculture Organisation.C... Enzymatic recycling-based amperometric immunosensors for the ultrasensitive detection of okadaic acid in shellfish. Eberhart B-T. 8. The challenge facing such approaches is the variety of matrices that must be examined.C. Prieto-Simon B.. 3. Campas M. de la Igelsia P.M. Rome. and in particular the ruggedness of such methods. 24 (4). Pravda M. Shankaran D. 2008. Stevens R. Soelberg S.C. Wekell J.. 166-74. Gibbs P. O’Sullivan C... 2007. 9. Chinowsky T. Anderson D. 2002. While the mouse bioassay continues to be the mainstay of most monitoring programs. 6.. 1998. 884-95. Novel electrochemical immunosensors for seafood toxin analysis... Miura N. Boca Raton. Wright J. 52. CRC Press. 10. food. FAO Food and Nutrition Paper 80. 40. Botana L.. Guilbault G. Manual on Harmful Marine Microalgae. Seafood and Freshwater Toxins. Marty J-L. Talanta. 716-22.FISH AND SEAFOOD regulatory levels for toxins continue to be assessed only for individual purified toxins. but these are gradually being overcome. Fish and shellfish toxins. never mind the amounts required of several groups. UNESCO. 2. 1735-45..K. they can provide a valuable “yes/no” filter for shellfish and finfish extracts. Marine Pollution Bulletin.. Biosensors to detect marine toxins: Assessing seafood safety... 7. Leatherhead Food RA. in Microbiology Handbook: Fish and Seafood. Spencer S. Trainer V. Detection of toxin domoic acid from clam extracts using a portable surface plasmon resonance biosensor. Hallegraeff G. Sensors and Actuators B: Chemical. Marine Biotoxins.L. Toyofuku H.M. and environmental interest. Leatherhead.G.. Furlong C. Toxicon. Diogene J.. 5.M. 2006. Cembella C.D. 6. Eds.L.P. 72. 2004.M. 2003. In most cases this is due to the paucity of material that is usually available for any single group of toxins. Le Berre M.L. Harmful Algae. References Kreuzer M. an encouraging development is the use of antibody-based methods to detect the presence of marine toxins. Marty J-L.

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

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

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

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

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

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.


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

biological. it may be obtained from external sources. chemical and/or physical). If expert advice is not available on-site. Fig. 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.g.1.FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations. 8. Logic sequence for application of HACCP 178 .

3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. fermentation.3. aquaculture drug residues and potentially pathogenic Vibrio spp.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora. composition. packaging. 8. marine toxins associated with particular fish and shellfish species. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments.g.g. In addition. including practices onboard fishing vessels. Vulnerable groups of the population.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.3.3. may need to be considered specifically. These factors are likely to have a significant effect on some potential hazards. 8. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. storage and distribution conditions and instructions for use. freezing. It is also important to identify the consumer target groups. Expertise on such inherent hazards.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries.1. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. or in aquaculture operations. If the processing does not include a kill step.).3.4 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. processing conditions (e. The description should include information such as the product name. such as children or the elderly. naturally present on fish from warm waters (> 15 °C). heat treatment.3. However.). such as enteric viruses in shellfish. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation. which should be described in detail.3. 8. shelf life. etc. pH. is a cooking process required?). contaminated fish and shellfish should not be used. etc. there may be species-specific hazards related to certain fish and shellfish. 8. physical and chemical structure (including water activity (aw). It should provide an accurate representation of all steps in the production process from raw materials to the end179 .2 Describe the product It is important to have a complete understanding of the product.

3. conduct a hazard analysis. However. 8. It may include details of the factory and equipment layout. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. It is essential that the diagram is accurate. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. Ideally. survival or multiplication of pathogenic microorganisms.3.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. 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. Many fish-processing operations have relatively few steps and the flow diagrams appear simple.FISH AND SEAFOOD product.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. 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 therefore a detailed description of the procedure applied must be available. If HACCP studies are applied to proposed new process lines or products. it should also include details of CCP steps. 180 ● ● .6 List all potential hazards associated with each step. 8. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. 8. once determined.3. the qualitative and/or quantitative evaluation of the presence of hazards. because the hazard analysis and decisions regarding CCPs are based on these data. features of equipment design. time/temperature data. or circumstances of any reclaim or rework activity. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects.4. ingredient specifications. The operation should be observed at all stages. cleaning and hygiene procedures and storage conditions. For example.

One control measure may be relevant to several process steps. and there are an equally varied range of potential hazards associated with them. The incidence of psychrotrophic. Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. For example. botulinum. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage. and the potential growth of Listeria monocytogenes on smoked fish.6. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual. For example. since this may interfere with the analysis. the hurdle effect. Vibrio spp. The HACCP team should then determine what control measures exist that can be applied for each hazard. the number of consumers potentially exposed and their vulnerability.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. or may originate from polluted water or from post-capture contamination (e.g. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed.). to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards. where a hazard is repeated.3.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products. 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. Salmonella spp. and viruses). modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. Therefore it is not possible. or desirable. Note: it is important at this stage that no attempt is made to identify CCPs. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin. any food safety objectives or manufacturer’s food safety requirements. 8. Hazards introduced during processing of fish products depend very much on the characteristics of the process.g. Pathogens may be part of the resident microflora of the living animal (e. 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. 181 .

structured approach to decision making.FISH AND SEAFOOD 8. 8.3. it is not possible to generalise on likely CCPs.1 Fish and seafood Again.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. biological or sensory parameters. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively.3. Where possible. critical limits should be specified and validated for each CCP. Criteria used to set critical limits must be measurable and may include physical.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits.8. If a significant hazard has been identified at a step where control is required for safety. 8. chemical. then the process must be modified to include a control measure. and its use may not always be appropriate. and the reader is referred to the appropriate product chapter. it is necessary to specify both time and temperature for a thermal process.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. 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 . More than one critical limit may be defined for a single step. However.3.3. it can be said that effective control measures are likely to include the following: • • • • • 8. For example.7. given the enormous variety of fish products and processes in use. application of the decision tree should be flexible.2) to provide a logical. However. but for which no control exists at that step or any other. 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.


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

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

the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. European Communities L 268. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. A number of more detailed implementing and transitional measures have been adopted at EC level.1991. transportation.2 Legislative Structure From 1 January 2006. manufacturing. 190 • • . processing. 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 187.1992. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No.9. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. distribution. 7.9. as last amended by Regulation (EC) No 806/2003. handling and offering for sale or supply to the consumer. 41). p. 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.15). 24.7. 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. packaging.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. storing.1).1991. 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. 24. p. N. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. Subsequently. Regulation 854/2004 relates to the organisation of official controls on products of animal origin.B. and sets out what those enforcing the provisions have to do. as last amended by Regulation (EC) No 806/2003. p. 9. J.

as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. with primary production. 19.I. and equivalent legislation in Scotland. domestic preparation or handling.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175.7. 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. 9. and binding in its entirety on all member states from the date of entry into force. Wales and Northern Ireland. Primary products are defined as products of primary production including products of the soil. national legislation in the form of a Statutory Instrument (S. of stock farming. etc. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. revocation of existing implementing legislation. for example. 2006 No.I. is required to give effect to the EU regulations. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. Additionally the requirements of EC Regulation 853/2004 must be complied with. penalties and powers of entry.g. 191 .1993. e. 1 . As regulations. and of hunting and fishing.14.) in England. 2073/2005. right through to the implementation of procedures based on HACCP principles. There are some exemptions. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. food storage that is for private or domestic consumption. Although the regulations have the force of law. the legislation is directly applicable law. p. which will be covered later in this chapter. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. setting offences.11) The EU hygiene regulations apply to all stages of food production including primary production. The Food Hygiene (England) Regulations 2006 (S.3 Regulation (EC) No. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. 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. 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. 852/2004. 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:


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

(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. 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. EY. the code would be UK. EB or WE.1 Identification marking An identification mark needs to be applied following the requirements of Annex II.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. please refer to Regulation (EC) 853/2004. EK. In this case.FISH AND SEAFOOD 9. and the characters easily decipherable and be clearly displayed. Section I of Regulation (EC) 853/2004. EO. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. EG. For the UK. when a product's packaging and/or wrapping is removed or is further processed in another establishment. the mark must show the approval number of the establishment.4. (iv) If the identification mark is applied in an establishment within the Community. (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. a new mark must be applied to the product. (ii) However.4. the food business operator may apply the same identification mark to both types of food. EF. for other codes. EK.2 Fishery product requirements 9.1. EÜ. (iii) Additionally.2.4.2. the mark must be oval shaped and include the abbreviation CE. ES. Section I of Regulation (EC) 853/2004 are given as follows: 9. the new mark must show the approval number of the establishment where this occurred. EC. 9. 198 .2 Form of the identification mark (i) The mark must be legible and indelible.1. Extracts of the requirements of Annex II.4.2.

2. the approval number of the establishment and the abbreviation for the community where appropriate. reference should be made to the actual Regulation.1.4. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. Section VIII of Regulation 853/2004. This section does not apply to bivalve molluscs.4. Chapter III.2. Extracts of the requirements of Annex III. it is sufficient to apply the mark to the exterior of that package only. and fishery products carried in bulk.3 Method of marking (i) The mark is to be applied directly to the product.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. (v) When the mark is applied directly to products of animal origin. as amended. wrapping or packaging in another establishment. and are intended for further handling. processing. (ii) For products of animal origin that are placed in transport containers or large packages. its wrapping or the packaging.e. (iii) In the case of liquid. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . echinoderms. Chapter IV. including vessels. C and D.LEGISLATION 9. 9. With the exception of Chapters I and II. Parts A. they would need to be obtained following Section VII on live bivalve molluscs. granulate and powdered products of animal origin carried in bulk. Part A and Chapter V apply to retail. it applies to those animals when not placed on the market live. the wrapping or the packaging. the mark may be applied to the outer surface of the container or packaging. i. tunicates and marine gastropods when placed on the market live. for full requirements. The requirements of this section accompany those in Regulation (EC) 852/2004. specifically relating to fishery products are given below. the colourings used must be EC authorised colours. or be printed on a label affixed to the product. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. Therefore. a) requirements for establishments. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels.

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

and not more than 0 °C after 16 hours.LEGISLATION (i) Such vessels need to have holds. 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. tanks must incorporate devices that allow a uniform temperature throughout the tanks. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. 4. A temperature recording device must be present and placed where the temperature in the hold is the highest. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. Storage areas for the finished products also need to be sufficiently large and cleanable. Also. Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C. where necessary. to ensure that melt water does not stay in contact with the products. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. (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. (iii) In vessels equipped for chilling fishery products in cooled clean seawater. recording of temperatures. 201 . 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. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. 3. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. and allow the monitoring and. any waste processing unit needs to have a separate hold to store the waste. where necessary.

must be chilled or.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. chilled and wrapped. the product must be kept at no more than -9 °C. The brine must not be a source of contamination.2. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. where fishery products are displayed for sale. Handlers may use spiked instruments to move large fish or fish that might injure them. if this is not possible. B. Chapter II . 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 . Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. 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.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. and products washed thoroughly. c) Fishery products. e) In cases where whole fish intended for canning is frozen in brine.2. provided no damage occurs to the flesh. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption.4. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. 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. other than those kept alive. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. b) Factory vessels on board which crustaceans and molluscs are cooked.Requirements during and after landing 9. (ii) In the case of food business operators responsible for auction and wholesale markets etc. landed as soon as possible.

They should be wrapped. and where necessary. (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.3 Chapter III . B. part I. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. they must be stored under ice in appropriate facilities. 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.2. points 1 and 2. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. Fillets and slices must then be removed from work tables as soon as possible after preparation. handling fishery products A. C. Additionally. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed.Requirements for establishments.4. Chapter I. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. prepared or processed immediately after arrival at an establishment on land. dispatched.LEGISLATION consumption.2. (ii) Heading and gutting must be carried out hygienically. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. 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. C. 9. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . including vessels. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. packaged and chilled as quickly as possible after preparation. 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. 854/2004.

must accompany fishery products referred to above when placed on the market. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. cooling must continue until a temperature 204 .4 Chapter IV . D. 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. (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. (iii) sprat. (iii) A document from the manufacturer. If no other method of preservation is used. 9. (ii) mackerel.2.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. stating the type of process they have undergone. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . Once produced. (b) fishery products from the following species. (iv) (wild) Atlantic and Pacific salmon.Requirements for certain processed fishery products A.4. and whole fish must be gutted and washed beforehand.2. if the processing is insufficient to destroy nematode larvae.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 cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. raw materials must have no guts. except when supplied to the final consumer. and (c) marinated and/or salted fishery products. they must be cooled rapidly. or a stabilising treatment.

B. refining and purification steps before being placed on the market for the final consumer. pressing. 9. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport.2. 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). 2074/2005. and after this. 2073/2005. 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. B. and the raw material is processed within 36 hours after loading. 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. 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. However. C. the following conditions need to be met: A.5 Chapter V .2. separation. Shelling/shucking is to be done hygienically. (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.LEGISLATION approaching that of melting ice is reached. 2073/2005). raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. processing. centrifugation.4.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. 205 . 2074/2005).

Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. it should be kept at no more than -9 °C. 9.7 Chapter VII . and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice.2.4. In the case of whole frozen fish in brine intended for the manufacture of canned food.2. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae.Storage of fishery products (i) Fresh fishery products. Fresh. prepared and processed fishery products belonging to the family Gempylidae. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. The scientific name must accompany the common name on the label. Diodontidae and Canthigasteridae. and when fishery products are wrapped on board fishing vessels. 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. 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.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.4.6 Chapter VI . Molidae. thawed unprocessed 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. Fishery products contaminated with parasites may not be placed on the market for human consumption. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. 206 .FISH AND SEAFOOD D. E.2.2.

2074/2005. species belonging to the Merlucciidae family. 9.Transport of fishery products The above points for Chapter VII apply in relation to transport. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp.8 Chapter VIII .LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. Sebastichthys capensis. (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.4.2.2. 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. species belonging to the Gadidae family. provided that the journey is short and the competent authority allows this. 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. 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. The temperature of frozen fishery products must not fluctuate above -15°C. melt water must not remain in contact with the products. 9.). Helicolenus dactylopterus. (ii) If fishery products are kept under ice. (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 .4.

(b) and (c) of Part B(1) of that Chapter. Section VII of Regulation 853/2004. as amended. The information required in the registration document is given in point 4 and includes the health status of the production area. Chapters V. where the raw material complies with points (a). 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. Apart from the provisions on purification.4.3. shellfish species etc. VI. specifically relating to bivalve molluscs are given below. reference should be made to the actual regulation. they supplement the requirements of Annex II to that Regulation. Chapter IX applies to pectinidae harvested outside those areas.FISH AND SEAFOOD Regulation (EC) No 853/2004.3 The regulation details specific hygiene requirements for bivalve molluscs. Live bivalve mollusc requirements - 9. for full requirements. 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 854/2004.1 Chapter I . this Section also applies to live echinoderms. however. please refer directly to the regulation.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. they supplement the requirements of Annex I to that Regulation. 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. The copy of 208 . Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. Extracts of the requirements of Annex III. The sample must consist of about 100 g of flesh. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. VIII and IX. 9. taken from at least three different points and mixed together by grinding. tunicates and marine gastropods. (b) In the case of other operations.4. and point 3 of Chapter VII applies to retail.

Transport needs to allow sufficient drainage for best survival conditions. 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. 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. poles or other fixed means are to identify the boundaries of the sites. 209 .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. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. processing or relaying. collection from: .4. B.Class A means they can be placed directly on the market .Hygiene requirements for the production and harvesting of live bivalve molluscs A. 9. 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. Requirements for production areas (i) The competent authority classes (classed A. B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with.3.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market .2 Chapter II . (ii) Provided the requirements of chapter V are met. C.

3.3. Requirements for dispatch centres Handling should not result in contamination to any degree. Requirements for purification centres There are requirements for live bivalve molluscs to be washed.FISH AND SEAFOOD 9. for: (i) paralytic shellfish poison (PSP). and there are requirements for tanks and water storage containers. dinophysistoxins and pectenotoxins together.Hygiene requirements for purification and dispatch centres A. Before dispatch.4. (ii) amnesic shellfish poison (ASP). 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 9.4 Chapter IV .4. 210 . (iv) yessotoxins. the levels of marine biotoxins should not exceed the following limits.4. 9.3. 800 micrograms per kilogram. B.5 Chapter V . 1 milligram of yessotoxin equivalent per kilogram. requirements if there are many batches of live bivalve molluscs.3 Chapter III . containers and a label to identify purification. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. the purification system to allow them to resume /maintain filter-feeding activity.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. 160 micrograms of okadaic acid equivalents per kilogram.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements.

the label.1).7 Chapter VII .6 Chapter VI .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. Also. Finally. the species of bivalve mollusc and date of packaging (at least the date and month) are required. Requirements of Chapters III and IV are to be complied with. including the identification mark.4.3. 9. by way of derogation from directive 2000/13/EC. they cannot be re-immersed in or sprayed with water.LEGISLATION (v) azaspiracids. dispatch centre or processing establishment.3.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.4. 9.4. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.2.3. (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. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.4.8 Chapter VIII .4. Registration document requirements also apply. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.Identification marking and labelling In addition to general identification marking (contained in 9. 211 . 9. 160 micrograms of azaspiracid equivalents per kilogram.3. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.9 Chapter IX . must be waterproof.

general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given. 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. 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. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. 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 . The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. Fishery products in particular must comply with the requirements of Annex III of the regulation. Chapter V of Regulation 853/2004 apply.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. they derive from: (i) poisonous fish. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004. echinoderms. or (iii) bivalve molluscs. In this regulation.FISH AND SEAFOOD 9. Therefore the health standards for fishery products as previously stated in Annex III. (ii) fishery products not complying with the requirement concerning biotoxins. 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.5 Regulation (EC) No. Section VIII. chemical. This refers to official controls or production and placing on the market of fishery products.5. 9.

coli per 100 g of flesh and intravalvular liquid. A summary is given as follows: 9. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. In the remaining 10 % of samples. Regulation (EC) 854/2004. as previously stated. Also.1 Chapter II . three dilutions MPN test specified in ISO 16649-3. in 90% of the samples.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. Sampling plans and frequency are required. 000 E.2 This states official controls for the production and placing on the market of live bivalve molluscs.000 E. 213 . coli per 100 g of flesh and intravalvular liquid. The reference method for this analysis is the five-tube.2. live tunicates and live marine gastropods. chapter V is reinforced. 4. section VII. Live bivalve molluscs from Class B areas must not exceed.5. Annex II 9. - - - - B.5. live bivalve molluscs must not exceed 46. The reference method for this analysis is the five-tube. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. live echinoderms. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated.Official controls concerning live bivalve molluscs from classified production areas A. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. Live bivalve molluscs from Class C areas must not exceed 46. Classification of production and relaying areas: This section reinforces that specified in Annex III.600 E. coli per 100 g of flesh and intravalvular liquid. that microbiological criteria and chemical contaminants amongst other requirements are met.

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. 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. boundaries or class of a production area or its closure.6 Regulation (EC) No.2 Chapter III . E. F.2. opening or closing a production area.FISH AND SEAFOOD C.5. 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.5. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. 9.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. 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. 9. which has applied since 1 January 2006. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. 9. 2073/2005. Procedures for re-opening a closed production area are stated D. establishes microbiological criteria for a range of foods. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. 214 .

volume. presence or number of microorganisms. 215 .LEGISLATION The aim of this legislation is to complement food hygiene requirements.4 referring to fishery products.II. and the legislation applies to all businesses involved in food production and handling. based on the absence. Two kinds of criteria have been established: food safety criteria.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. and/or on the quantity of their toxins or metabolites. ensuring that foods being placed on the market do not pose a risk to human health. The relevant criteria are outlined in Table 9. with chapter 2.I. per unit(s) of mass. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. 9.6. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. a batch of foodstuffs or a process.6. area or batch.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. The criteria are outlined in Table 9. 9.

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.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. other than those intended for infants and for special medical purposes(4) (8) L. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.TABLE 9. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1.2 Ready-to-eat foods able to support the growth of 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. monocytogenes. monocytogenes 1.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. monocytogenes 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. manufactured from fish species associated with a high amount of histidine (17) Histamine .27 Fishery products which have undergone enzyme maturation treatment in brine.25 Live bivalve molluscs and live echinoderms. 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 1.17 Live bivalve molluscs and live echinoderms. tunicates and gastropods 9 (18) 2 E.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.

sugar. — bread. 1996. to the satisfaction of the competent authority. J. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. Dervin C.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample.25. Duflos G. cider. subject to scientific justification. Coryfenidae.. to the satisfaction of the competent authority. Bouquelet S. — fresh. Malle P. products with a shelf-life of less than five days shall be automatically considered to belong to this category.1 . In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. according to which the whole batch is to be deemed unsafe. monocytogenes. Engraulidae. Particularly fish species of the families: Scombridae.. References: 1. Bouquelet S. excluding sprouted seeds. 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. shall not apply. when he is not able to demonstrate. soft drinks. coli is used here as an indicator of faecal contamination. Assay of biogenic amines involved in fish decomposition. E. Valle M.1.4 or aw ≤ 0. beer. products heat treated in their final package). when recontamination is not possible after this treatment (for example. 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. A pooled sample comprising a minimum of 10 individual animals. m = M.94. wine. This criterion shall apply to products before they have left the immediate control of the producing food business operator. Products with pH ≤ 4. products with pH ≤ 5. 2. spirits and similar products. c = number of sample units giving values between m and M For point 1. that the product will not exceed the limit 100 cfu/g throughout the shelf life. Malle P. Clupeidae. biscuits and similar products. uncut and unprocessed vegetables and fruits. honey and confectionery. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 .0 and aw ≤ 0. AOAC Internat. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. Single samples may be taken at retail level.92. The most recent edition of the standard shall be used.. 5 This criterion shall apply if the manufacturer is able to demonstrate. live bivalve molluscs. Pomatomidae... including cocoa and chocolate products. Other categories of products can also belong to this category. 43-9. 79. Scombresosidae. — — bottled or packed waters.

82. monocytogenes in ready-to-eat foods able to support the growth of L. J. Salmonella in different food categories: — satisfactory. — unsatisfactory. if any of the values are > the limit. monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory.I The limits given refer to each sample unit tested. AOAC Internat. if all the values observed indicate the absence of the bacterium. if the presence of the bacterium is detected in any of the sample units. if the presence of the bacterium is detected in any of the sample units. — unsatisfactory. (1) 219 . L. 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. if all the values observed indicate the absence of the bacterium. — unsatisfactory. if all the values observed indicate the absence of the bacterium.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. The test results demonstrate the microbiological quality of the batch tested(1). L. where the limit refers to a pooled sample. the mean value observed is ≤ m 2. Histamine in fishery products from fish species associated with a high amount of histidine: — satisfactory. excluding live bivalve molluscs and live echinoderms. — unsatisfactory. if all the values observed are ≤ the limit. a maximum of c/n values observed are between m and M 3. if the following requirements are fulfilled: 1. L. — unsatisfactory. 1097-101. no values observed exceed the limit of M. if the presence of the bacterium is detected in any of the sample units. 1999. tunicates and gastropods in relation to testing E. 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. monocytogenes in other ready-to-eat foods: — satisfactory. Interpretation of the test results relating to Table 9. coli. 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.

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 2.4.1 Shelled and E.

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

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. S. 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. For other food. 9. for a limited period consistent with food safety. 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.S. and was kept at above 8 °C or the recommended temperature. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. or. 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. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing.gov. 2006 No. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. such as that below. 2006 No.I.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 .food. 3).I. was kept at above 8 °C or the recommended temperature for an unavoidable reason. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. 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.K. 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. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment.8 Guidance In the U. Alternatively. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions.

2075/2005 and Regulation (EC) No. 1). 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No.6. L338. 1.2005. 1662/2006. 2076/2005.LEGISLATION 9. 2076/2005. as read with the corrigenda at OJ No.4. 1019/2008 and as read with Regulation 2073/2005. 30. L283. namely. L139. 83) as amended by Regulation Nos 882/2004. 25. L226. Regulation (EC) No.12. 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.4. 3) as amended by Regulation (EC) No. Regulation (EEC) 2136/89.2004. L226.2006. 2076/2005.4.2004. 30. 2074/2005.6. 1688/2005. 223 .6. The revised text of Regulation (EC) No.10. 2073/2005 on microbiological criteria for foodstuffs (OJ No. 30. Regulation (EC) No. Regulation(EC) No.2004. The revised text of Regulation (EC) No. 2074/2005. 55). 2074/2005 and 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. 853/2004 is now set out in a Corrigendum (OJ No. L226. The revised text of Regulation (EC) No.2004. 2076/2005. 62) as amended by Regulation (EC) No. 22. 25. 25. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito.10 Further Reading Regulation (EC) No. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 854/2004 is now set out in a Corrigendum (OJ No. Commission Regulation (EC) No. laying down common marketing standards for preserved sardines.2004. 2074/2005. Regulation (EC) No. as amended. 9. these are not be discussed here. 22) as amended by Regulation (EC) Nos. Regulation (EC) No. L139.9 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. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 852/2004 is now set out in a Corrigendum (OJ No. 1441/2007. L139.2004. Regulation (EC) No. 206). 14. 1663/2006.

22. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.1.2001. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.2000.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.12. 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. 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. 21. 22. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. 6) 224 .2005.10.12. 23. (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 338.2005.

45 °C (1).1.0 . 10. The optimum growth temperature is 28 °C (2).0 x 1.1 Aeromonas spp.1. At 5 °C. Some death will occur at this pH if the temperature is 4 °C. 225 .3 .1 10.5 µm 10. no growth is observed (1). respectively (1). a 10 to 1. Morphology Gram-negative rods. Aeromonas has the potential to grow over a pH range of ca 4 . PATHOGEN PROFILES 10.5.3 minutes.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.10 (3). and D-values at 45 and 51 °C have been reported as 29.1. and minutes.5 pH Under otherwise optimal conditions. At pH increase in number is observed after one week in food systems (1).3 Temperature Aeromonas grows at temperatures of 2 . 10.5. under optimal incubation temperatures.2 Oxygen requirements Facultative anaerobe. The minimum pH value for growth is thought to be pH 5. 0. 10.1.

and groups I and II are those responsible for the botulism cases. All C. 0. All type A strains are proteolytic and type E strains are usually non-proteolytic. Types A.5 . but the interior of the food may do so.1 Morphology Gram-positive. and is sensitive to > 4. spore-forming rods. 10.3 Temperature All strains of C. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. Group I will not grow at temperatures of 10 ºC or less. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. 10.2. at 85 ºC for 5 minutes. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C. B. 226 .2. A to G. or at 90 ºC for a few seconds.even as low as 3 ºC (7). 6).2. Thus an aerobically packed product may not support the growth of the organism on the surface.2 Clostridium botulinum Seven different types of C. 10. botulinum types produce heat labile toxins.1.4 Heat resistance The vegetative cells of C.2. but the spores of this organism are more so.2% sodium chloride (NaCl) (1). being capable of slow growth and toxin production at low temperatures . botulinum are known.0 µm. 10. types B and F can be either. botulinum is an obligate anaerobe. botulinum are not particularly heat resistant. which may be inactivated by heating at 80 ºC for 20 .5% NaCl (4).22.7 . botulinum is prevented. to a lesser extent. There are four main groupings of the organism. botulinum grow reasonably well in the temperature range of 20 45 ºC. 10.4 x 1.FISH AND SEAFOOD 10.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 .30 minutes. but Group II strains are psychrotrophic.2 Oxygen requirements Although C. E and.2. F are the types that are responsible for most cases of human botulism (5. forming at least seven different toxins.

The organism will grow well in the centre of meat or poultry dishes.0.PATHOGEN PROFILES 10. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.3 10.21 minutes) (9). the minimum aw for growth may be lower. botulinum depends on solute. parcooked meals with pH value > 5. 10.2. pH and temperature.2 Oxygen requirements C.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. 9). vacuum packed.2. 10.97) (10. These concentrations correspond to limiting aw of 0. safety margins must be introduced to allow for process variability. grow on the surface of foods unless they are vacuum or gas-packed.97 for Group II (5).7 Characteristics of C. It will not. In commercial situations.6 and pH 5. Note also that if humectants other than salt are used.2. respectively (8. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. perfringens .0 µm. they may survive mild heat treatments (70 .g. 0. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. botulinum spores The most heat resistant spores of Group I C. However.like other clostridia .3. 227 .1.is an obligate anaerobe.3. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. where oxygen levels are reduced.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C.9 x 2. 10.10.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4.3 .0 and aw > 0.0 .1 Clostridium perfringens Morphology Gram-positive spore-forming rods.94 for Group I and 0. 11)). These values have been established under carefully controlled laboratory conditions. therefore. 10. particularly by cooking.

10. the enterotoxin is not heat resistant . perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. 13). although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. The lowest aw recorded to support the growth of C. perfringens strains. While some growth can occur at 50 ºC. perfringens appears to be 0. type of solute. perfringens in roast beef at 60 ºC is 14.3 Temperature The most significant characteristic of C.50 ºC. and their heat resistance is also affected by the heating substrate. however. As in the case of other factors limiting the growth/survival of this organism. Recorded heat 228 . perfringens can vary quite considerably in their heat resistance. 0 .it is destroyed by heating at 60 ºC for 10 minutes (14.3. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. The D-value for C. lower concentrations may be effective in combination with other factors. depending on strain and other conditions. perfringens in vacuum packed beef (18). perfringens spores The spores of C. At cold temperatures.45 ºC. 10. perfringens has the potential ability to grow within the temperature range 15 .7 Characteristics of C.3. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. Some studies indicate that the presence of 3% NaCl delays growth of C. will survive greater extremes of pH (and aw) (12. perfringens is not tolerant of low water activities. 13). although C. vegetative cells die rapidly (12).97 (glycerol and sucrose respectively) depending on the solute used to control aw (13.8% inhibit growth of most C. pH. Salt concentrations of 6 . the limits for aw are affected by temperature. In addition. etc. It grows best at pH values between 6 and 7 (the same pH as most meats).4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. 15.5 minutes (17).93 and 0. 10.6 Aw/Sodium chloride C. Its optimum temperature for growth is 43 .FISH AND SEAFOOD 10. perfringens is not a tolerant organism with respect to pH. 10.10 ºC. Spores.5 pH C.3. perfringens. (It is unusual for spores to be formed in foods after the growth of this organism (14)). 18).3.3. Under otherwise ideal conditions.3. 16).

or even below 0 ºC. The upper temperature limit for the growth of L.2 has been suggested as the lowest working limit (22). however.4. the lowest limit for growth is likely to be considerably higher .5 . being potentially capable of growing . D-values for L. -0. 10.0.0 minutes for heat resistant spores to 1.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic.4 .4 10.5 x 0. 60 and 65 ºC were reported as Heat resistance L.19 minutes.4. the organism is able to grow at pH values well below pH 5 (pH 4.2. monocytogenes is reported to be 45 ºC (20). However. and where acetic acid is used as an acidulant. 10. In foods.98 and 0. so can be destroyed by pasteurisation.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. respectively (21). 229 . 10.at refrigeration temperatures down to. requiring days or weeks to reach maximum numbers. Under ideal conditions. is between 30 and 37 ºC. Its optimum growth temperature.4. 10.8 minutes for heat sensitive spores (12).albeit slowly .3 is the lowest value where growth has been recorded. 1. monocytogenes is not a particularly heat resistant organism.2.23.5 pH The ability of Listeria to grow at different pH values (as with other bacteria) is markedly affected by the type of acid used.1 Listeria spp. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens. 10.4 ºC is probably a more likely minimum for growth in foods (19).6 .3 . pH < 5. and temperature.4. it is not a sporeformer. however.0 µm.2 Oxygen requirements Aerobe or microaerophilic. monocytogenes in crawfish tail meat at 55. Morphology Gram-positive short rods.especially at refrigeration temperatures. growth at low temperatures can be very slow. 0. using hydrochloric acid as an acidulant).

monocytogenes is quite tolerant of high NaCl/low water activities.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).2 Oxygen requirements Facultative anaerobe. or even grow.0 .6 10.83 (20). 25). at salt levels found in foods (10 .6.3 µm.5. 10. It is likely to survive.97.1 Plesiomonas Morphology Gram-negative.1 Salmonella spp.5 .5. 10.5.3. 0.5 pH Plesiomonas has a pH range for growth of pH 4.5. .5 (24. 25). peritrichous flagella.FISH AND SEAFOOD 10.45 ºC.0 µm.5 10.12% NaCl or more). 10. 230 .6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.0 x 1 . the optimum being 30 ºC (23).90. 0. The bacterium may survive for long periods at aw values as low as 0.6 Aw/Sodium chloride L.8 . 10.5. Morphology Gram-negative short rods. but has been shown to be able to grow at an aw level of 0. 10. It grows best at aw of ≥ 0.4. 10.0.3 Temperature Plesiomonas can grow in the temperature range of 8 .7 x 1.1.

9.6. Furthermore.5 .20 times more heat resistant than the average strain of Salmonella at high aw (27). 10. depending on serotype. Salmonellae are quite resistant to drying.2 Oxygen requirements Facultative anaerobe.945. However.5 pH Salmonella has a pH range for growth of pH 3. 10. substrate.5 ºC. under otherwise ideal conditions. The optimal pH for Salmonella growth is between 6. D60 ºC values normally range from about 1 to 10 minutes. temperature and pH. 231 . However. with a z-value of 4 . although salt tolerance increases with increasing temperature (28).6.6. It is not. Salmonella senftenberg 775W is about 10 . depending on the type of acid and temperature.37 ºC.5. and with an appropriate acid. Some death will occur at pH values of less than about 4. some strains are able to grow at temperatures as low as 4 °C (26).7. 10.0. strains vary in their ability to withstand heating. The growth of Salmonella is generally inhibited by the presence of 3 .6.PATHOGEN PROFILES 10.5. Salmonellae are quite resistant to freezing. a heat resistant organism.6.93 (as reported in dried meat and dehydrated soup). Salmonella has the potential to grow at aw levels as low as 0. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. therefore.48 ºC. 10.6 Aw/Sodium chloride Where all other conditions are favourable.3 Temperature Salmonellae can grow in the temperature range of 7 .4% NaCl. and appropriate heat treatments must be determined experimentally for low-aw foods. or possibly 0. pasteurisation and equivalent treatments will destroy the organism under normal circumstances. the optimum being 35 . Growth is slow at temperatures below about 10 ºC.8 .4 Heat resistance Salmonella is not a spore-forming organism.

Staph.0 . 10.5 ºC. It can survive well at low temperatures. aw.1 Staphylococcus aureus Morphology Gram-positive cocci. The following relate to limits for growth only. NB: The growth of Staph.7. but may cause some cell damage (30). 10.7 10. under otherwise ideal conditions (29. the organism is heat sensitive and will be destroyed by pasteurisation.20 minutes.3 Temperature Under otherwise ideal conditions. aureus than citric acid).0.7.37 ºC (29).3 (optimum 7. aureus is more limited under anaerobic than aerobic conditions.7.9 µm diameter.7. with an optimum of 35 .). Freezing and thawing have little effect on Staph.5). 232 .2 Oxygen requirements Facultative anaerobe.FISH AND SEAFOOD 10. however. 0. 10. aureus viability. aureus can grow within the temperature range 7 – 48. depending on aw. The limits for toxin production are also narrower than for growth.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.2 to 9. Under most circumstances.7 . 10. aw and temperature (sensitivity to acid increases with temperature). Most strains of staphylococci can grow within the pH range 4.7. the D60 ºC value is 2 . In meat macerate. etc. stressed cells can also be less tolerant of heating. As is the case with other bacteria.7. 31).4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. fat content.

* dependent on temperature.7. 34). 0.86 is the generally recognised minimum in foods (32). An effect of increasing salt concentration is to raise the minimum pH of growth. atmosphere. Staph. but certain strains can grow in 20% NaCl.5 . 10. aureus can grow in 7 .0 (optimum 7.7.2 .0.>0.2.2 Oxygen requirements Facultative anaerobe. However.8. pH.43 ºC (35). In general. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities. In general.8.0 . 233 .9.5) (29.99 little or no toxin production in anaerobically packed foods.8. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.83 . strain.8 x 1.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5.1 Vibrio cholerae Morphology Gram-negative curved rods.8 10.10% NaCl.87 and > 0. and solute. with a temperature range for growth of 10 .6 µm 10.PATHOGEN PROFILES 10.3 Temperature The optimum temperature for growth has been reported as 30 ºC. an aw of 0.4 .7.99 aerobically under otherwise optimal conditions.6 Aw/Sodium chloride Staph. 10.7 Limits permitting toxin production 10 . 31) between 0. 10. Staphylococci are more resistant to salt present in foods than other organisms. It can grow over the aw range 0.

9 10.97 . it has a pH range for growth of pH 5 . 10. but not acid conditions. declining in number during storage.FISH AND SEAFOOD Also. but only moderately sensitive to freezing (40). 10.0. there is evidence that V. parahaemolyticus is easily destroyed by drying.8.65 minutes (35). 234 . cholerae in crab meat homogenate at 60 ºC is 2. It can grow in the presence of 0. 10.8.6 Aw/Sodium chloride V. Temperature The minimum and maximum temperatures reported for growth in laboratory media. 37. and it is sensitive to refrigeration temperatures (0 . with an optimum of 37 ºC (39).4 Heat resistance V.984 (35).1 . cholerae can grow over the aw range 0. 38). cholerae may persist on frozen fish for >180 days (36). respectively. 10.998. are 5 and 43 ºC.5 pH This organism is tolerant of high pH values.6 (35.9. with optimum at 7. with an optimum of NaCl (35).9.5 ºC). The D-value for V. V. under otherwise favourable conditions.8.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods.2 Oxygen requirements Facultative anaerobe. cholerae is sensitive to heat at temperatures of >45 ºC. 10.

5% NaCl. 10. parahaemolyticus is unable to grow in the absence of salt. 10.2 Oxygen requirements Facultative anaerobe. The D-values at 55 ºC for clam and crab homogenate are 0. The organism can survive for long periods at freezer temperatures (42).9. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43). 41).922.6 Aw/Sodium chloride V. vulnificus is heat sensitive.6 (41). respectively (39).98 (35. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.4 Heat resistance V.11.10.4 Heat resistance V.10. respectively.10.43 ºC (42). and Morphology Gram-negative curved rods. 41).6 .5 pH The pH range for growth is 4. 10. 235 . but the optimum concentration is 2 . with a temperature range for growth of 8 . parahaemolyticus will grow at aw levels as low as 0. and are readily destroyed by cooking. The optimum concentration of NaCl equates to aw of approximately 0.9. 10.4% (39. 10.10 Vibrio vulnificus 10.02 . 10. parahaemolyticus are not heat resistant.3 Temperature The optimum temperature for growth has been reported at 37 ºC.5 minutes.8 .29 minutes. with an optimum pH of 7.9.PATHOGEN PROFILES 10. depending on solute. V.10.

10. Marcel Dekker Inc. 1-27. 279-303.... Marcel Dekker. ASM Press. Betts G. with an optimum pH of 7. Hui Y. New York... Murrell K. 2001. Abeyta C. Lund B. 111-89..O. Beuchat L. Great Britain. Bier J. 5. Eds. 4.FISH AND SEAFOOD 10. Clostridium botulinum.10. Eds.D. 2005. Aeromonas.P.H. Kim J. Ed. Murrell K. Gaze J. Hauschild A. 1-19. London. Food Control.R. Peck M. Potential hazards associated with REPFEDS. Faris A. Austin J.H. Aeromonas hydrophila group. Principles of control. Blackie. 5-19.D. Smith J.10. 357-67. 10. 2003. Dodds K.. Cliver D. 1993.H. Eds. Hui Y. Marcel Dekker. 57-63. Caister Academic Press. in Clostridium botulinum: Ecology and Control in Foods. 8.D.C. Marcel Dekker. 97-131. Eds.. International Commission on Microbiological Specifications for Foods. Aeromonas species.K. 7..H.H. 1993..H. Juneja V..6 Aw/Sodium chloride V. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.. Stelma G. Gorham J. 6. Eds..L. Volume 1. Hauschild A. in Foodborne Bacterial Pathogens. 1989.N. Bhunia A. in Microorganisms in Foods.8 (42). 236 .5 . Volume 5: Microbiological Specifications of Food Pathogens. Montville T. 1994. Eds.L. 1994. New York. Dodds K. Marcel Dekker. Microbiology and Molecular Biology.M.. with an optimum of 2. vulnificus grows over the range 0. 9. Palumbo S. Diseases Caused by Bacteria. New York.0% NaCl. 3.N. Cliver D. Ed.. 1995. in Foodborne Disease Handbook. Clostridium botulinum. Volume 1: Diseases Caused by Bacteria.10. Eds. 1996. Miliotis M. Stelma G..M. Foegeding P..P. Washington D. 10.11 1.. in Foodborne Bacterial Pathogens.M.. Gorman J. Marcel Dekker.E. Ed. New York.J. in Clostridium botulinum: Ecology and Control in Foods.P. Aeromonas hydrophila. Doyle M. Doyle M. 329-49. 2. References International Commission on Microbiological Specifications for Foods. Novak J. 121-76.. Hauschild A. in Foodborne Pathogens. in Food Microbiology: Fundamentals and Frontiers. in International Handbook of Foodborne Pathogens.L. Notermans S..R.O. New York.L. Kovacek K. New York.. Dodds K. New York.A.W. Doyle M. 11.5% (42).W. in Foodborne Disease Handbook. Clostridium botulinum.D.. Fratamico P. Marcel Dekker.W. Clostridium botulinum and Clostridium perfringens.5 pH The pH range for growth is 5 . 1989.5. 383-408.R.W. 6 (1). Johnson E.W.

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

Blackie. Marcel Dekker. in Foodborne Infections and Intoxications. 45 (3). 426-35. 642. 36.FISH AND SEAFOOD 27. in Foodborne Pathogens. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. Staphylococcal intoxications. Gustafson J. 217-64. Cambridge. 2005. 34.. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. New York. Salmonella. 8.A. Academic Press.M. 31. Volume 5.J. Tohyama Y. 273-84. 331-57.R.. Staphylococcal gastroenteritis. Staphylococcus aureus. Eds. 1983. D’Aoust J-Y... Wilkinson B. Bergdoll M. Food and Environmental Sanitation. Caister Academic Press.M. 38. 414-25. 2005.P.C. Crowcroft N. Ed. Japanese Journal of Experimental Medicine.J. Riemann H... London.. International Commission on Microbiological Specifications for Foods. Academic Press.A. Stewart G. R15764. Salmonellae. CDR Review.S. Applied and Environmental Microbiology. 32. Volume 5. 1990. Ed. Eds. Microbiological Specifications of Food Pathogens.. Blackie. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). in Foodborne Pathogens: Microbiology and Molecular Biology.M. Lee-Wong A. Staphylococcal Food Poisoning. Jay J. 523.P.D. Marcel Dekker.O. Vibrio cholerae. in Foodborne Bacterial Pathogens. 985-98. Bhunia A. International Commission on Microbiological Specifications for Foods. International Commission on Microbiological Specifications for Foods. New York. Blackie. Doyle M. in Modern Food Microbiology.C. 29. 2005. Springer Science.P. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures. 1996. in Understanding Pathogen Behaviour Virulence. Ed. International Commission on Microbiological Specifications for Foods. Oliver J. 1989. Ed. Warner R.O. London. Reed G. 1996. London. Eds.62. Dairy. 327-52.K. Cleland D. Doyle M. 543-68. Jay J. 30. Cliver D.L.M. Bergdoll M. 85-106. Cholera: current epidemiology. 327-445. in Foodborne Disease. London. 4 (13). 545-66. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems. 35. Cliver D. Wymondham. International Commission on Microbiological Specifications for Foods. 39. New York.A. 1996. Golden D. Microbiological Specifications of Food Pathogens. Volume 5: Microbiological Specifications of Food Pathogens. Golden D. Twedt R.S. 1993. Loessner M. 13 (11). 28. in Microorganisms in Foods. Loessner M.. Ed. 2005.S. 37. Smith J. 1930. 40. 1994. Ed. Ed. in Microorganisms in Foods. Woodhead Publishing Ltd. in Microorganisms in Foods. 238 . Stress Response and Resistance.. Fratamico P. Vibrio parahaemolyticus. 33. London. Griffiths M. Vibrio parahaemolyticus.H. 1989. International Commission on Microbiological Specifications for Foods.

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


uk Web site: www.gov.gsi.uk or v.gov.org.uk Web site: www.org.gov.ifm.org.defra.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.co.gsi.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.org/ .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.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.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.seafish.gov.uk Web site: www.holt@ifm.scotland.

bim.dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .FISH AND SEAFOOD Department of Agriculture and Rural Development.eapo.fiskehandlerne.ifpea@eircom.C.ie/templates/homepage. 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. Andersens Boulevard 37. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo.org/fishery/ European Association of Fish Producers’ Organisations H.fao.matis. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: matis@matis.gov.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.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. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.com Web site: www.is Web site: www.fiskeriforskning.uk/index/fisheriesfarming-and-food.dk Web site: www.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: post@fiskeriforskning.org Web site: www.dardni.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.dk Web site: www.uk Web site: www.dk Danmarks Fiskehandlere H.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.gov.suite.no Web site: www.difta.is/ 242 .

org Web site: www.uk Web site: www.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.csiro.co.seafood.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 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.org.gsi.ac.com Web site: www.cmar.org.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.uk Web site: www.hpa.org.dh.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.nz Web site: www.fdf.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.com/ 243 .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/en/index.co.ifr.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.gov.au Web site: www.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.bsi-global.gov.

euro.uk Web site: www.sofht.scot. jsp Food Safety 10 .uk/index.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.int/ SIK .hps.int.nhs.uk/scieh.euro.int Web site: http://www.who.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.sik.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.se Web site: www.who.nhs.int Web site: www.codexalimentarius.who.uk Web site: www.net/web/indexen.de 244 .sfam.org.asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.co.scot.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.who. Web site: www.who.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.int/foodsafety World Health Organization (WHO) Headquarters CH .org Web site: www.co.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.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: foodsafety@euro.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.

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


growth and survival in crustacean shellfish 86 Aeromonas sobria. planktonic source of spirolides in shellfish 163 Allergens. 102 Bacillus spp. 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 . preserved by dry/wet salting 99 Anisakis spp. 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. causing spoilage of dried fish 110. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. as autochthonous bacterial flora of fish 3 Aeromonas spp. 111 Astroviruses. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. 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. source of PSP toxins 142 Alexandrium ostenfeldii. to test microbiological criteria 19 Aeromonadaceae.INDEX Achromobacter spp. 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. in crustacean shellfish 87 Alteromonas spp. in crustacean shellfish 86-7 ATP. 69-70 Aeromonas hydrophila.

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

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.INDEX pathogen profile 226-7 Clostridium perfringens. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. source of azaspiracids 159 source of DSP toxins 148-9. hot-smoking 107 ELISA assay. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . to determine fresh fish spoilage 11 Corynebacteria. to determine fresh fish spoilage 11 Eels. growth and survival in crabmeat 84 growth and survival in cured. 106 Contacts 241-5 Conway method. smoked and dried fish 114 Enzymic spoilage. 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. 152 Diphyllobothrium. for detection of DA 148 ELISA kit (DSP-Check). as autochthonous bacterial flora of fish 3 Crabs. processing 334 Cockles. as contaminants in scampi 36. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. adverse condition on poorly preserved fish 110 EC grading scheme. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. 149-50. of fish – different processes 94 Dun. 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. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. 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. to detect NSP toxins 155 ELISA kit (Biosense).

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

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

fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. reduction in smoking process 108 Norcardiopsis dassonvillei. 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. 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 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. of crustacean shellfish – effects of processing 81-3 of cured.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. shigelloides. as TTX producer 145 Norovirus. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. caused by lipid oxidation 11 Oysters. 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. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay. in crustacean shellfish 867 Norwalk virus. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. 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.

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

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

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

as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. 62 Yersinia spp. 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.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. 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 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. of shellstock 61. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. 152-3 256 . as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01. 68 Vibrio cholerae non-01.

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.