Leatherhead Food International Leatherhead Food International

MICROBIOLOGY HANDBOOK FISH AND SEAFOOD
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

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CONTRIBUTORS
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
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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

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

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6 Published Microbiological Criteria 3.7 References 4.6 Pathogens: Growth and Survival 1.2 Initial Microflora 4.7 References 3.6 Published Microbiological Criteria 4.3 Processing and its Effects on the Microflora 1. 3.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.8 References MOLLUSCAN SHELLFISH 3.1 Introduction 2.5 Pathogens: Growth and Survival 3.2 Initial Microflora 1.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.4 Spoilage 1.1 Definitions 4.4 Spoilage 4.1 Definitions 1.8 Further Reading CRUSTACEAN SHELLFISH 4.1 Definitions 3. CHILLED AND FROZEN RAW FISH 1.6 Pathogens: Growth and Survival 2.7 Published Microbiological Criteria 1.3 Processing and its Effects on the Microflora 3. 4.5 Spoilage 2.2 Definitions 2.5 Pathogens: Growth and Survival 4. vii .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 2.4 Spoilage 3.3 Processing and its Effects on the Microflora 4.2 Initial Microflora 3.4 Processing and its Effects on the Microflora 2.5 Factors Affecting Fresh Fish Spoilage 1.3 Initial Microflora 2.8 References 1.7 Published Microbiological Criteria 2.

1 Introduction 8.5.4 Spoilage 6.5 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 6.6 References 6.9 Azaspiracids 7.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.8 Ciguatoxins 7. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9. 852/2004 on the General Hygiene of Foodstuffs 191 9.6 Lipophilic Shellfish Toxins (LST) 7.8 Further Reading FERMENTED FISH 6.1 Definitions 5.2 Definitions 8.10 Further Reading 223 viii .8 Guidance 222 9.3 Regulation (EC) No.3 Tetrodotoxin 7.5 Regulation (EC) No.5 Pathogens: Growth and Survival 6. 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 9.6 Regulation (EC) No.11 Conclusion 7.7 References 5.7 Food Hygiene (England) Regulations 2006.3 Processing and its Effects on the Microflora 5.4 Amnesic Shellfish Poisoning (ASP) Toxins 7. EU FOOD HYGIENE LEGISLATION 189 9.4 Spoilage 5.4 Regulation (EC) No. 8. 9.2 Initial Microflora 5.6 Published Microbiological Criteria 5. 7.I.9 Other Relevant Legislation 223 9. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9. SMOKED AND DRIED FISH 5. CURED.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.1 Introduction 189 9.5 Pathogens: Growth and Survival 5.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.10 Cyclic Imines 7. S.3 Stages of a HACCP Study 8. 14 (Hygiene requirements specific to the UK) 221 9.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.7 Further Reading FISH AND SHELLFISH TOXINS 7.2 Legislative Structure 190 9. 2006 No.4 Implementation and Review of the HACCP Plan 8.1 Introduction 7.3 Processing and its Effects on the Microflora 6.2 Initial Microflora 6.1 Definitions 6.

2 Clostridium botulinum 10.7 Staphylococcus aureus 10.4 Listeria spp.5 Plesiomonas 10.6 Salmonella spp.8 Vibrio cholerae 10.3 Clostridium perfringens 10.10 PATHOGEN PROFILES 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .10 Vibrio vulnificus 10. 10.1 Aeromonas spp. 10. 10.9 Vibrio parahaemolyticus 10.

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

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

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

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

Fish flesh is dominated by the abundance of white muscle in relatively short segments. and excludes molluscs and Crustacea.2 . and is mostly interspersed between the muscle fibres. typically fluctuates between 0. it shows a typical season pattern especially in migratory species such as herring or mackerel. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. Fish fillets are a poor source of carbohydrates.5% (1). Protein content in fish fillet varies typically from 16 .1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills.21%. In addition. crustaceans. which can be up to 67%. ‘fish’ also refers to the flesh of such animals used as food. The lipid fraction is the component showing the greatest variation. migratory patterns.1. typically 3 and 15% of total weight. In this chapter. while fatty fish have fat cells distributed along their bodies. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1.20%. Fish can be divided into fatty and lean fish. The connective tissue content of fish is also lower than that found in meat. In addition. giving it its characteristically flaky structure. This super class of vertebrates includes all the bony and cartilaginous finfish. lean fish are those fish that store most of their fat in the liver. 1 . and spawning season. Fish fillet composition can vary significantly within the same species due to feed intake. However. but typically present a low fat percent when compared to other meats. offering less than 0. Muscle composition and structure of fish also differ from those found in other meat. and other forms of aquatic animal as part of their ‘fish’ definition. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. most fish contain omega 3-fatty acids and other essential nutrients. fins and a streamline body. The lipid content. “fish” will be used for fresh and seawater finfish. CHILLED AND FROZEN RAW FISH Associate Prof. respectively (1). Fish are an important part of a healthy diet since they contain high quality protein.

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

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

II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.106 cells/g. 105 . Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.35 °C 8 . 4 .102 101 .109 cells/g) * Some sources considered L. 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. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.26 °C 10 . stream water.19 µg/kg bodyweight from < 102 . vulnificus) 5 . estuarine environments.1 . and mud Ubiquitous. birds. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 .103 High (107 .14 . Plesiomonas shigelloides 3 .I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00.37 °C High for most species.37 °C Unknown Unknown TABLE 1. However. In addition. sewage.106 cells/g (exception: V. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. Toxin levels 0. Escherichia coli Yersinia enterocolitica 101 . numbers of pathogenic bacteria in raw fish tend to be low. and risk associated with the consumption of seafood is low (2. 4).0.FISH AND SEAFOOD TABLE 1.>106 Salmonella spp.

1. However. and their re-use will increase the crosscontamination of other fish with such microorganisms. chilled brine or refrigerated seawater at -2 °C. cortisone levels increase during prolonged stress and can alter the fillet quality. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating. The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. In addition. has an effect on the amount of stress and physical damage that the fish will suffer during capture. ropes. After capture. decks. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside. Increasingly. The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea. This contrasts greatly with the meat industry. and the killing process is designed to minimise stress. In eviscerated fish. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria. or traps.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. human hands and clothing. offering better control of fish health prior to.3.3 1. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating. and conditions of storage. During capture and storage. in which the bacterial load 5 . in recent decades. icing or freezing. mackerel or herring. and especially when fish is stored on board for longer periods. however. the cuttings provide areas of exposed flesh that are open to microbial attack. with the production of off-odours. hook and line. with fish such as tuna and cod being more commonly eviscerated than sardines. handling and processing Wild finfish are usually caught by net. If evisceration is carried out at sea. aquaculture practices have been expanding worldwide. Physical damage such as loss of scales. The length of time that set nets have been in the water or the time trawlers’ nets are towed. finfish will almost invariably come into contact with nets. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. and during harvest. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. in which the health of each animal can be assessed prior to slaughter. These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans.1 Processing and its Effects on the Microflora Capture. digestion and off-flavours in uneviscerated fish. Fish may be eviscerated prior to storage at sea . potential contaminants added during handling and processing.a practice that may have both advantages and disadvantages. with very little control over the condition of the fish at the time of death or the duration of the killing process. and spoilage. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms. boxes and/or baskets. birds and soil.

As with all foods. In the absence of oxygen. not part of the normal flora of the meat animals (8). allowing more growth of spoilage organisms. These containers are also used for displaying the fish during auction at the quayside. by actively altering the composition of the 6 . 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. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. 7). MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. 1.3. fish have a higher post mortem pH. and to limit the loss of quality (5). from water activity (aw) through to physical damage. use of effective detergents and disinfectants. In MAP. and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO). By vacuum packing foods. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. cleaning of the work environment. they are usually process contaminants. and minimising handling will all reduce microbial cross-contamination. but isolate the food in an environment different from the bulk atmosphere. However. Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. Physical barriers not only protect from physical damage.). Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. in time. Anaerobic atmospheres have less effect on fresh fish shelf life. Packaging changes the intrinsic and extrinsic parameters affecting a product. The atmospheric conditions surrounding a product may be passively or actively altered. but if applied properly should extend the life of the product.2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. ferric ion (Fe3+)). nitrogen. and of the quality of the water used for ice and to wash fish. will result in fully anoxic conditions. These changes can be deleterious. if there is some oxygen demand from the product. however. What is more. oxygen etc. Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics.FISH AND SEAFOOD can be substantial. partial fermentation of sugars occur leading to lower pH. a reduction in the oxygen tension is achieved which. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. for example. careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. often in the absence of adequate refrigeration.

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

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

Loss of freshness and spoilage cannot be separated as processes. or for that matter evaluation of the taste of the fish. B. so lower carbon dioxide concentrations may have better preservative effects. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). and metabolic activities of microorganisms present in the fish. and the flesh turns from bright and elastic to dull and soft (1). to brown and slimy with an intense off-odour.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). these are summarised in Table 1. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis.4 1. the gills from pink and shiny. reactions caused by activities of the fish’s own enzymes.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. Over time. 18) and are probably the most widely used. A.4. 1. The eyes turn from convex and clear to concave and opaque. and this pattern is the basis of freshness grading schemes. as traditional spoilage patterns of the external organs are typically very distinct.III.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. smell and 9 . When working with fillets. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids). texture. Taste panel evaluation of cooked fish may be used to score or grade colour. TABLE 1. Degradation of whole fresh fish stored in ice generally follows a set pattern. Various methods for whole fish freshness evaluation by a trained panel have been proposed. 17. the skin turns from iridescent to dull and bleached with bacterial slime. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . C (C=reject) Significantly.CHILLED AND FROZEN RAW FISH off-odours. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. with no smell.

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

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

5. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. For fresh fish iced immediately after capture and continually stored in ice. Also. biochemical and microbiological reactions taking place within and on the fish. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. the slower the spoilage processes proceed (within limits). All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. The reasons for the differences between tropical. but also crucial for the assurance of consumer safety. The sum of these changes may result in a different spoilage profile.and temperate-water fish spoilage rates may be many. The factors are interactive and cannot be completely isolated. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 .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. Fish spoil as a result of the chemical. careful temperature control during storage is not only important in terms of quality loss.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).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. a minority are often associated with the majority of the spoilage. 1. The concept of specific spoilage organisms (SSOs) is not new.FISH AND SEAFOOD 1.5 1. 1. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. For example. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. For fresh fish. the results of altering one parameter may have far-reaching consequences on others. by altering the gas atmosphere (extrinsic).5. 34).5. 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. the lower the temperature of storage. 5). The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks.

its ability to reduce TMAO to TMA. Members of the genus are able to produce pigments. The importance of Sh. It is able to grow as fast as. 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. 1. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. They are generally characterised as Gram-negative rods. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine.3. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. These organisms are described as potential spoilers. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. During the 1970s. obligate respiratory bacteria. produce hydrogen sulphide (H2S) from cysteine. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. oxidase-positive.5. putrefaciens growth may be used as indicators of spoilage (30). it is often isolated as about 1 . which produced the characteristic spoilage compounds of the fish (30. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. It is also present in fresh water. motile with polar flagella. As previously noted.CHILLED AND FROZEN RAW FISH started in the 1940s. mediated spoilage is characterised by ‘fruity’. (9) and they are considered SSO of iced freshwater fish (35). The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp. i.10% of the total flora of fresh fish from temperate marine waters. the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. or faster than the rest of the flora of ice-stored fresh marine fish (9). and may play some role in the spoilage of freshwater fish. on fish are diverse and in many cases species-specific. aldehydes. 35. Sh.3.1 Pseudomonas spp. Pseudomonas spp. It is thus an important spoilage organism of gadoid fish such as cod. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. putrefaciens spoilage of fish is due to its biochemical action on muscle.2 Shewanella putrefaciens Sh. frozen) fish products. 36).5. therefore. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s. catalase-positive. processed (e. more especially. 13 . ‘oniony’ and ‘faecal’ odours from the production of ketones. the spoilage of a product may be strongly influenced by the conditions under which the product is held.e. plus other characteristic compounds of the species responsible for spoilage.g. although only since the late 1960s has it been realised that specific metabolites of Sh. 35). 37).

Shewanella waksmanii. putrefaciens growth has been collected (12. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). putrefaciens lies in its chemotaxis: the bacterium. they are heterogeneous. This wide-ranging respiratory capability is thought to be unique (39). Shewanella affinis. 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. 41). mesophilic strains of Sh. Research has shown that approximately 107 cfu/g of Ph. putrefaciens has been studied. the bacteria have a significant problem to overcome . Sh. Shewanella oneidensis. whereas >108 cfu/g Sh. 38). phosphoreum has also been identified as responsible for histamine fish poisoning. Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection. like all members of the Pseudomonadaceae. Shewanella olleyana. In terms of its taxonomy.5. unlike many other motile bacteria. and Shewanella profunda were described. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39). Later. putrefaciens strains are characterised as microaerophiles or anaerobes. does not show carbon-source chemotaxis. Ph. owing to its cell size and activity. 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. but Sh. In fact. 1.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). at the time of rejection. baltica has been identified as the main H2S producer in cod during cold storing (37). The prevalence of the bacterium in so many environments. Initially.3. and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles.how to reduce a molecule that they are not able to take into their cells efficiently. additional psychrotrophic species such as Shewanalla baltica. although. Shewanella frigidimarina. In 50% N2/50% CO2 MAP cod. Shewanella gelidimarina. Sh. putrefaciens were required. it is strictly respiratory. It is also able to produce hydrogen sulphide and a range of other off-odour compounds. Shewanella livingstonensis. and recent taxonomic description using modern molecular methods bifurcated this species. putrefaciens was not present in such numbers (23. Shewanella denitrificans. with several new species being described. and Shewanella aquimarina. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). phosphoreum were required for 50% of taste panellists to reject a sample.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. Ph. Another distinct feature of the physiology of Sh. putrefaciens were identified as Shewanella algae. The ferric reductase activity of Sh. phosphoreum sufficient to cause spoilage was found. a population of Ph. as the ferric iron is insoluble. 41). Sh. This type of intoxication occurs when bacteria convert the histidine 14 .

types B. thermosphacta is a well-characterised psychrophilic spoilage organism of meat.6 Pathogens: Growth and Survival Endogenous chemicals. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). 5) reports that fresh fish consumption has never been shown to cause human botulism. thermosphacta did not produce off-flavours until 2 . 1. C1. bacteria and higher parasites all present some risk associated with fish consumption (1. 6). Fewer still are capable of growing on fresh fish. However. Adult disease is caused by the production of the botulinum toxin. they do not produce significant product spoilage). 5).3 °C (Group II) and these are non-proteolytic (i. D. algal toxins. it is bacterial risks that increase after capture of the fish. human viruses.e. Of 404 intoxication outbreaks of type E C. Review of outbreak data suggests fresh fish to be safe. Growing evidence suggests a role for B. this is probably due to spoilage occurring before toxin elaboration. 5. thermosphacta in the spoilage of some MAP fish. age. botulinum recorded up to 1963.6. 1. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. Types A.5. The heterogeneity of the types (A. Acetate production has been reported as a good indicator of spoilage by this organism. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions.CHILLED AND FROZEN RAW FISH present in fish into histamine. B. F and G) is well documented (44). E and F include bacteria with minimum growth temperatures of approximately 3. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. lightly preserved fish products are associated with botulism. Recent studies have investigated the dominance of B.1 Clostridium botulinum C.3 days after having reached 108 cfu/g (42). When inoculated into VP corned beef and sliced ham. B. these risks are substantially reduced (1. botulinum. especially type E botulinum intoxication. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. C2. B. where a particular 15 . a convenient but diverse species of bacterium with a number of different types. including antisera. E. (and very rarely F and G) have been reported to cause human disease. Some Ph. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). 1.4 Brochothrix thermosphacta and lactic acid bacteria B.3. previous exposure and access to supportive treatment. 75% (with a 35% mortality rate) occurred in Japan. Of these. E. but with proper cooking. presents a potential risk. Huss (1.

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

shigelloides are encountered. however. shigelloides is similar to Aeromonas spp. 5. Although the psychrophiles are unable to grow at body temperature. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. their potential to produce disease-causing exotoxin in food has not been fully examined (48). Some isolates are true psychrophiles. listeriosis cases have been reportable to the CDC. cytotoxic. infection. They are aquatic organisms with an epidemiology that is yet to be fully understood. coli on isolation media (50). possibly due to under-reporting because of their likeness to E. V. cholerae O1.20 °C. in its habitat.4 Listeria monocytogenes L. Again.6. Despite this. Since 2000. are present on fish and can grow to significant numbers during storage (1. which is often fatal (>50%). though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. Similar problems as those encountered for Aeromonas spp. monocytogenes has been well documented as a foodborne human pathogen. others are psychrotrophic mesophiles (49).6.000 17 . and proper cooking will substantially reduce risk. sodium channel blocking and haemolytic activity (48). It is known that Aeromonas spp. Chill storage may not eliminate growth.CHILLED AND FROZEN RAW FISH disorders). 1. proper chill storage will eliminate any risk associated with this organism. 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. The major virulence factor appears to be the production of toxins with enterotoxic.3 Aeromonas Aeromonas spp..55 cases per 100. there is no risk from the consumption of properly handled and cooked fresh fish. and especially Aeromonas hydrophila. Most cases have been sporadic rather than associated to large outbreaks. but proper cooking should substantially reduce risk of infection. P. In the US. and especially estuarine environment. 17). but the bacterium is known to survive and grow in the shallow marine. and fish and shellfish are probably the major reservoirs for the organism (50). with minimum growth temperatures around 0 °C and optima of 15 . There is no risk associated with the consumption of properly chilled and cooked fresh fish. L. 1. is historically associated with faecally contaminated water. in assessing the risk of fishborne infection by P. the causative agent of cholera. monocytogenes incidence has been between 0.26 to 0. are associated with human diarrhoeal illnesses.

there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. at risk groups . 1.6. full risk analysis is not possible (52).. 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. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54).FISH AND SEAFOOD persons. foetuses. Photobacterium spp. Being a psychrotroph. 1. there is a possibility for growth of the organism on chill-stored fish.2002 (55)) and is also common worldwide. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . 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. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. it is not apparent to the consumer and it cannot be destroyed by cooking. Like other toxins. alcoholics.48 minutes in salmon have been reported (53). D60 of 1. The organism poses a serious risk in chilled products not cooked before consumption. Lightly preserved products.pregnant women.98 minutes in cod and D60 of 4.. and Lactobacillus spp. but cooking will significantly reduce the risk of infection. Clostridium spp. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. some Vibrio spp. especially pickled fish. but more especially by Morganella morganii. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. 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. It is regularly isolated from seafoods (51). 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. leads to the production of histamine. but proper cooking will reduce any risk. neonates. If fish are not properly refrigerated.. Infections are mostly associated with the consumption of raw or 18 .5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. Klebsiella pneumoniae and Hafnia alvei. 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.6. Fortunately.

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

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

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

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

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

. Cann D. International Journal of Food Microbiology.. Journal of Food Science. 6. in Fish as Food. Trolle G. Labuza T.M. 317-21... Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches.. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. Ravn Jørgensen B.H. Aberdeen. 271-85. 13-32. 24 . Picard G. 1984.A. 35. International Journal of Food Microbiology. Taoukis P. Sumner J. Villemure G. McMeekin T. 15. 1. 1.2 Spoilage Shewan J. 65-72. 295-307. 741-50. Fisheries and Food. Hobbs G. 1989. The Microbiology of Seawater Fish. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. 1997. Akiba M. Huss H. Food and Agriculture Organisation. Eds.L. 1988. Journal of Food Protection. Shinano H.. Magno-Orejana F. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures.9. Bulletin of the Faculty of Fisheries (Hokkaido University). International Journal of Food Microbiology. 51. 4.E.P. Smith G. Lancaster.. Academic Press. 1989. 1984. Rome. Jones Y.D. 1985. Ed. 1986. Huss H.H. Gibson D. Dalgaard P. Ross T. Florida. Application of predictive microbiology to assure the quality and safety of fish and fish products. Handling and processing of scad. Gibson D. Estimation of the bacteriological quality of fish by automated conductance measurements. 127-34. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. Torry Research Station of the Ministry of Agriculture. 1. Huss H. 73-89. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Technomic Publishers. in Torry Advisory Note No 93. Fisheries and Food... Cho Y. Houston N.. 55. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review..FISH AND SEAFOOD 1. Torry Research Station. Torry Research Station of the Ministry of Agriculture..9.. Shahidi F.D.. 1992.. Kitts D..H.). in Seafood Science and Technology. Ed.. Olley J. Aberdeen. Microbiological quality and shelf life prediction of chilled fish. Ogden I. Gram L. 1961. Food and Agriculture Organisation of the United Nations. Borgstrom G.C. 317 (Suppl. Changes in microflora and chemical compounds in mackerel stored by partial freezing. Simard R.9 1. in FAO Fisheries Report No.1983.M.1 Further Reading Processing Anon.G.M.S. 1992.... 62-70. International Journal of Food Microbiology.

. http://eur-lex. Jacobsen M. 1993.H. Klausen N. 1992.9.. Liston J.. 65-72. Effect of culture age. 20023. Otero A. International Journal of Food Microbiology. in Quality Assurance in the Fish Industry. Garcia M. Sala F. Gram L. 1987. 322-6.. Applied and Environmental Microbiology.J.. International Journal of Food Microbiology. Elsevier Science.3 Pathogens Condon S. 59. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. 137-46. 4. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. 1987. Huss H. Huss H.K.eu/LexUriServ/LexUriServ. 1997. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. 2197-203.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A.. 5.L. Gram L..M. Modified atmosphere packaging of fish and fish products. in Fish processing technology. Blackie Academic & Professional. 72. Trolle G. London.9. Eds.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. Huss H. A rapid method for detection of histamine producing bacteria.CHILLED AND FROZEN RAW FISH Einarsson H. 25 .R.H. 1992.europa. Amsterdam.H. 479-88. Ed. Journal of Applied Bacteriology. 1.. 1. Hall G.

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

and these are increasingly being presented with an ever-expanding range of accompaniments. and this has produced a greater availability and diversity of products in the marketplace. but should be equivalent to cold-water products. They are typically flown into export markets. Listeria spp. At the same time. molluscs.. Microbiological hazards include Salmonella. where the preparation has already been carried out. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP). often using the simplicity of a microwave oven. they are usually of a better standard than species such as tilapia. shellfish and molluscs. including freshwater fish.FISH AND SEAFOOD Recent trends in the market for food and seafood products. They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. 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.. 2. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein. This includes all categories of seafood including trout and other freshwater species. and the consumer only has to heat (cook) the product. Seafood is the term used descriptively to include all categories of product. prawns. 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. which can be farmed. Doubts have been raised over the microbiological status and handling practices of reef-caught fish. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish. have shown the increasing importance of convenience foods. etc. and pathogenic Vibrio spp. and are well known for their diet consisting of waste materials. Shelf life on ice. but these are now finding favour as products in their own right. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. The expected shelf life attainable for this format is usually up to 8 days. rather than frozen.2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. Prepared natural fillets are fillets of fish without any additional coating. in particular. as they tend to lose colour and sheen after defrosting. the processing industry has developed a wide range of value added products. Nevertheless. Frozen fillets may have a surface layer of 28 . they are usually supplied filleted with head on. from both wild capture and aquaculture sources. or in MAP depends on catch conditions and treatment.

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

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

Tuna that does not meet the highest of grades based on colour. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). and have emphasised the origin and sustainability of the fish as key marketing points. The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. storage. will the saprophytic flora start to invade the surface flesh (1. sensory assessment of freshness etc. fat content. and processing conditions such as dehydration or poor filleting practices. including seasonal variation. species (Bluefin>Bigeye>Yellowfin>Albacore). i. Grading is based on size of fish (>25 kg). or visible damage from gaff marks (4). remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. Only after death. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. as a result there is an increase in spoilage rate thereby reducing the product shelf life. 6). The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna.I. A primary role in fish spoilage has been assigned to this organism.3. Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. Contamination will. 2). and intestines of the fish. Dalgaard et al. skin. searing or bar marking). handling or preparation stage. which may become a focus of contamination deep within the muscle. In summary. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. is not used for sushi or other high risk (and high value) products. whilst gaping is also dependent on a range of other factors that effect the muscle composition. Recent public opinion has favoured the use of dolphin. After capture and during any subsequent processing.e. and pathogenic flora associated with seafood. the microbial flora of seafood is associated with the gills. product handlers. 31 . it may be used for other products or markets depending on the product specification. For a summary of background information on the hazards associated with fisheries products and ingredients. on the whole.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. see Table 2.3 2. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. and ingredients added to the final product. The edible tissue of some species of fish are more susceptible to gaping than others.and turtlefriendly long lining to the less discriminatory netting.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. and the potentially toxic by-products of histamine-producing bacteria (5.

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

but it also poses a risk of cross-contaminating other products undergoing tempering. Where the raw materials are frozen. 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.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. Rapid controlled cooling of heat-processed components will help to minimise spoilage.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. For some components. depending on the size of the fillet 33 . Band-sawn materials or fish dice are normally promptly further processed. and potential opportunities for growth of foodpoisoning bacteria. 2. 2.3 Preparation of fish component of chilled and frozen recipe dishes 2.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. It is recommended that the principles of HACCP are applied to novel processes. but will also need to ensure that the optimum shelf life can be safely obtained. as the full complexity of the processes used for specific products cannot be covered in a general text.4. there are a number of distinct methods used. up to -5 °C so as to improve the cutting and handling properties during processing.2. to evenly raise the temperature of the entire frozen product from -20 °C or lower. and contamination from equipment to clean fish muscle. but it is assumed that any band sawing or dice process will introduce heat from blade friction. defined holding time at a chill temperature (0 – 3 °C). line extensions and process reviews.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material.1 . this may be an incidental effect. Generic examples have been used to illustrate only the major points.4. the core temperature. Figures 2. 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. as a heat process is often required for functional or organoleptic reasons. Examples of tempering techniques include the use of heated air. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface.3. or the use of microwave technology. Effects on the microflora are minimal during tempering (7). All have the same intention.4.

The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). frozen in a nitrogen blast freezer and weighed into sacks for later processing. by mincing and mixing with a binding agent (as described in section 2.1.1B . On arrival at the processing plant. after landing. The “blown” meat is graded.1A . drained and then refrozen for storage until required for further processing. 2.4. It is then size-graded.7 days.Fish Cakes). 2. quickfrozen.1.3. Fig.3. may not rise above 0 ºC during processing. 2.Scampi. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. the scampi is washed in potable water to remove foreign bodies.Shaped Fillets.2.1 (2. Raw material preparation stages for chilled or frozen coated fish products 34 . are placed into plastic boxes and covered in ice to preserve their condition. 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.1. Frozen cores are usually batter-enrobed. chilled or sold chilled on defrost with an expected shelf life of 6 . with the coating heat-fixed.1C .FISH AND SEAFOOD processed. 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 2.4.2).

2. 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. and water is added to rehydrate the components. The fish component of the cake will. as the physical structure of the material has been destroyed. Therefore. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. or sold as a chilled product after thawing. Fish cake cores may be batter-enrobed. but. additional spoilage organisms may be introduced..CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.g. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. are handled and stored as any other food product. quickfrozen.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. Waste material produced from this type of processing is often reused in the third category (fish cakes).block-sawn dust. The mix is shaped and formed ready for packing or for enrobing. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. spoil more rapidly than the dry ingredients. chilled. iii) Fresh/fillet/minced fish block. with the coating heat-fixed.3. those that use frozen blocks of whole fillets.off-cuts or belly flaps. or those where the trimmings. fish fingers). because of its nature. once the dry components have been rehydrated. more seriously. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. It is for this reason that all by-products from primary processing. such as aerobic spore formers and Staphylococcus spp.3. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . with an expected shelf life of 6 .1. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. material will need to be transferred swiftly to a chiller. that are to be reused for other foods. or promptly processed to avoid further quality loss. This is especially the case with block-sawn dust. ii) Fish-filleting waste .7 days.4. unless it is poorly handled.2 Shaped products This type of product falls into two categories. pathogens such as Salmonella and Listeria. from potato flake. 35 . The remaining dried ingredients are mixed with the fish.4.1.

and if required. potentially. 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. lactobacilli from the milk powder and flour. or sold as a chilled product after thawing. a.2 illustrates the typical stages of enrobing or breading. e. Natural fillets d. chilled. starch. Products are preformed and presented to the process (either three. perfringens could. quickfrozen. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. rusk or crumb. such as Listeria and Vibrio spp. with an expected shelf life of 6 .7 days. e. a characteristic flora will develop from the batter components. fish cakes Portions or fillets may be batter-enrobed.e. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. IQF/pre-formed fillets c. milk powder and spices. Initially. Batter contains water. i.g. which typically consists of flour. which can occasionally be isolated from protein-handling processes. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. A large variety of product formats and types can be batter-enrobed. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. is the addition of a pre-dust. and may include milk powder. and to carry flavourings where required. absorb any excess moisture on the surface of the fish portion.4 Battering/enrobing process Figure 2.4. spices. and possibly Staphylococcus spp.or fivestage) frozen or chilled. as well as ensuring that there is minimal contamination from human processors. vibrios and other fishassociated spoilage organisms. from flour. Bacillus spp. Over time. which is product-dependent and not always required. Spore-forming anaerobes such as C. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process.FISH AND SEAFOOD In all of the above examples. some of this initial flora 36 . Tempered band-sawn materials b. egg. Breaded or battered fish fillets are fully heat-processed before consumption. and Enterobacteriaceae from flour.. because of the intensive handling required during processing. raising agents. with the coating heat-fixed. If a crumb coating is required. rapidly multiply if strict temperature control was not maintained. any excess is removed with an air knife. The function of this is to improve the adhesion of subsequent coatings. 2.g. flour. The first stage. Fish products.

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

pasta or potato. 2. and must not show evidence of disease. Fig. The vegetables must withstand processing.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. but they are supplied without a source of carbohydrate e. 38 .1 ºC for no more than 3 hours is recommended (1). Figure 2. There are two main categories.5. A temperature no greater than 10 ºC for 12 hours cumulatively or 21. Flow diagram outline for recipe dish ingredients preparation 2.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes.4. rice. The protein source can be fish fillet or block. to produce a specific product. as this could introduce off-flavours and reduce shelf life.2. canned fish may be used in their preparation owing to it having wider availability and low wastage. For example.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products.g.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe.4. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7).3.

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

and then cooled slowly allowing germination and re-growth.4. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. This is a particular issue especially where products are heat treated to eliminate vegetative cells. 11). should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. a heat process in excess of 73 ºC will be required to rehydrate this component fully. Placing such products in a standard chill store to cool down could result in condensation. cereus). if subjected to temperatures between 20 and 40°C. If the sauce contains modified starches for freeze/chill stability. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. Potato toppings made from flake or powder will normally have little associated spoilage flora. For this reason all products. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. An incident involving Staph. and an increased risk of contamination from L. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. but temperature-abused pasta can also spoil rapidly by mould growth. a process equivalent to at least a 5D Listeria kill will be necessary.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. Products or dried ingredients contaminated with either spores of aerobic bacteria (e.4. cereus. Appropriate handling facilities or equipment will be required to protect the product. On hydration.g.) to the product (12. Sauces can be multi-component preparations or more simple preparations. Typically. Sauces or marinades are added to fish to broaden appeal or to accompany components. spore-formers can proliferate if adequate control of temperature is not maintained. aureus toxin is well documented (10. B. where composite meals are heat-treated. 40 . with the range always expanding as new tastes and cuisines are developed. such as butter garnish with black pepper. Alternatively. rely on the use of microbiologically sound ingredients.6.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. especially in those products that have a vegetable component or rice accompaniments.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. 13). or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. can germinate and rapidly multiply. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. cereus are well known and are still a common issue in the UK (9). the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. 2.

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

1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).4. Flow diagram outline assembly and packaging of recipe dish 2.FISH AND SEAFOOD Fig. botulinum to grow. which may restrict either microbial growth or enzyme activity.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth.4.7. unless the temperature is strictly controlled to below 5 °C throughout the supply chain.2 Modified-atmosphere packaging Originally developed for the red meat industry.7. 2.5. generally carbon dioxide and nitrogen. These include: 2.2. as it dissolves in the surface moisture to form a weak acid. The oxygen-free conditions will however allow other anaerobic bacteria such as C. 42 . The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. the method replaces air in the sealed packaging with a defined mixture of gases. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues.4. and tuna loins are packed by placing the product in a plastic pouch and removing all the air.

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

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

soil. Additional hazards from marine bio-toxins. cereus. Cases quoting contamination of seafood with organisms such as S.6.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish.104/ g should be treated with some concern (26). with pathogenic microorganisms or viruses. histamine intoxication. especially molluscan filter feeders. Incidents in such diverse materials as cream. potato. 27). Vibrio spp. Therefore. Hazards attributable to the non-seafood components are listed below and this information.1 Bacillus cereus This is a common contaminant of dust. are dealt with elsewhere in this book. in some cases. botulinum type E and Vibrio parahaemolyticus (25)..6. air. permit rapid growth of B. 45 . Counts of between 103 . farmed or free-living crustacea (30). typhi.3 Clostridium perfringens A study examining C.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. water. low oxygen or anaerobic conditions. etc. any risk of C.e. and Campylobacter are recorded in filter feeding molluscs such as mussels. In addition. 13. A number of other Bacillus spp.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. spices and herbs and vegetable shoots have been reported (12.6. may be of benefit within any risk assessment for chilled and frozen prepared fish products. botulinum when held in favourable conditions for growth (28) i. perfringens contaminating seafood will depend on the source of raw material. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. or a heat shock followed by holding at a temperature suitable for growth. These are C. and many raw or processed foods (26). along with previous discussion. Inappropriate holding times and temperatures will. and this issue may be of greater consequence for warmwater. Germination of spores requires favourable conditions. have been linked with food poisoning and should also be considered. 2. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). 2. agricultural run-off or sewage can contaminate in-shore species. and some effects related to product type and time to toxin production have been noted. 2. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage.

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

aureus enterotoxins and B.CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph. (36). although due to long analysis times required. are not mandatory. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40). as published by for example. 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.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts. pasta. etc. and as such the monitoring is for verification purposes rather than control. pastry. The following tables (Table 2. but these may be eliminated by a subsequent heat-treatment stage. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). 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. and are a recognised means of providing data on manufacturing standards.III based on IFST Guidelines (39)) provide example guidelines for seafood products. For instance. such as batter mixes. 2.7 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. with respect to the number and types of ingredients used. It should be remembered that 47 . but provide the industry with an indication of the microbial level at which action is required to ensure product safety. Microbial guidelines. although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”. . Microbiological specifications do not necessarily relate to product safety. and not to final products. the fresh product is often consumed before the results are obtained. 2.7 Published Microbiological Criteria Due to the complexity of many processed food products. and their association with similar materials. it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur. unless the criteria is for a pathogenic organisms where only non detection is acceptable. Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems.II and 2. 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.6. cereus emetic toxin are often confused owing to similar symptoms.

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

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

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

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

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6 litres. clams. In most species. completely encloses the body. whelks. snails. S. P. Jr.1. Plankton and algae form the bulk of their diet from seawater. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. Pelecypods have little need for locomotion. Pelecypoda. cockles. Box 3012 Bothell WA 98041-4421 United States of America 3. Other species of shellfish such as crabs. Food and Drug Administration 22201 23rd Dr. Gastropoda. Scaphopoda. O. chitons. the shell. Pacific Regional Laboratory Northwest U. oysters and clams can process about 3.3. which includes oysters.7 litres of water per hour. where it is eventually swallowed and digested. viruses. The term shellfish in this context is limited to oysters. respectively. 4). the foot is compressed to form a muscular spade for digging and the head is greatly reduced. composed of two valves hinged together dorsally. abalones. and 2. the adductors. The phylum consists of five classes: Amphineura.1 Definitions General features of the mollusc The phylum Mollusca. lying well within the mantle cavity. In these forms.1 3. clams. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. and Cephalopoda. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). and shrimp. octopi. oysters. mussels. scallops. cockles and scallops. Many microorganisms ingested by shellfish survive the digestive process (3. can hold the shells tightly shut against enemies (1).E. The class Pelecypoda includes all of the bivalves such as clams. cockles. Special tracts of cilia move this mucus toward the mouth. they also trap bacteria. while 53 . The class having species of economic and public health importance is the Pelecypoda. and scallops. lobster. mussels. squid. MOLLUSCAN SHELLFISH Carlos Abeyta. In the process of filtering water. slugs.S. chemical contaminants and other impurities on the mucus of the gills. Strong muscles. mussels. For example.

Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific . 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 . 3.South America Pacific – Japan Quahaug (also known as hard clam and hard shell clam) Coquina clam Hen clam Butter clam (also called the Washington clam) Mogal clam Little neck clam (also called rock Cockle Pacific littleneck) Atlantic – North American/Europe Atlantic – Europe/North America Japan Atlantic/PacificNorth America Donax spp.North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp.1. Clams are benthic and live burrowed in the mud.FISH AND SEAFOOD commercially valuable. are not as vulnerable to contamination through pollution of their beds as the bivalves. many species of bivalves are edible (Table 3. dwelling on the bottom or attached to structures in the water column.2 International commercial products Globally.I). but maintain contact with the water by means of a siphon tube (5). The habitat of oysters and mussels are sessile. and usually at higher concentrations than found in their habitat. The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3. Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 .

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.

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

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

by interlocking tufts of hair-like cilia. in which the individual branches. Smoked: meats may be canned in edible oil. scallops possess a single large adductor muscle. Salted: cooked meats bottled in brine. Semi-preserves: cooked meats with vinegar-acidified brine and spices. cooked meats packed in dry salt for transport. vinegar solution. also with other ingredients. Unlike other sessile bivalve molluscs.FISH AND SEAFOOD 3.) Dried: peeled. In some species. flattened ear or wing at the hinge area. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. The familiar marine mussel is asymmetrical. 58 . and held apart.2.2. boiled. are united. Canned: cooked meats. The posterior halves of the valves contain a large adductor muscle that holds the valves closed. Fresh: cooked meats. Fresh: shelled meats (often only the adductor muscle and the roe are eaten. or packed with jelly. or filaments. with the posterior ends of the shells (valves) broad and rounded. The adductor muscle is the only edible part of the animal. Internally. 3. gutted. The common scallop is about 5 cm long. mussel in butter sauce. mussel paste. scallops can swim or leap by a series of rapid closings and openings of the valves. and the anterior end smaller and pointed.1. The shells are fluted and round with a broad. sauces. Bait: used live extensively for baiting fish hooks for line fishing. brine. smouldered and afterwards dried (Japan). in mussel liquor. They have filibranch gills. Live: whole with shells. 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 a radially ribbed shell with the edge undulated and swim by opening and closing the valves.1.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms.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.

Live: in shell. Whatever contaminants are in the water will eventually get into the shellfish (7). Raw and partially cooked shellfish. Cockles range from about 1 to 15 cm in length and have two valves of equal size. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding.2.MOLLUSCAN SHELLFISH Frozen: shelled meats. they accumulate pathogenic microorganisms from polluted waters. also bottled in brine. 3. Salted: meats either lightly or heavily dry salted.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves. in sauce. The microflora are dependent upon a number of factors such as season. salinity. pH. 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. depending on length of journey. and the environment. 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 . are vectors of disease transmission (8). Canned: in brine. Because molluscan shellfish are filter feeders. and butter. Vinegar cured: bottled in malt vinegar after brining.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. which affects the temperature. mostly in oysters. Fresh: meats removed from the shell by boiling. clams and mussels. nutrient concentration and pollutants of the water column. Canned: in own juice. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms. 3.1.

Moraxella spp. Campylobacter coli Bacillus cereus Hepatitis non-A. 11. the initial flora composition and concentration of bivalves are highly variable. Other microorganisms found in lesser numbers are the Achromobacter spp. observed microscopically. but have never been cultured (14. For example. Pseudomonas spp..3.1. spirochetes of the genus Cristispira and Saprospira. mechanical harvesters. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. and yeast cells of Rhodotorula rubra and Trichosporon spp..3. clams.1. salinity. whole or in part.1 Processing and its Effects on the Microflora Oysters.1. from which one or both shells have been removed.. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. 10. In summary. Vibrios. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. Acinetobacter spp. or by 60 .2. rakes. shellstock (live) which is shellfish in the shell..1 and 3. nutrients.1 Shellstock 3. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora.. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. Bacillus spp. and shucked shellfish that is shellfish. and Aeromonads. 15). (9. At harvest.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. Flavobacterium spp. mussels. There are microorganisms. Enterococci spp. The largest groups found are the Pseudomonads..3. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves.000 /g. They are termed viable non-culturable microorganisms. 3. 3. 12. Temperature.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. Water temperatures affect the concentrations of microflora in the water. 13).3 3. that have never been cultured on laboratory media. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. and cockles The processing of bivalves takes two forms. responding to the environment in which they are harvested. Alcaligenes spp. dredges... Micrococcus spp..

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

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

If shellfish are not reasonably clean at the time of processing. 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). Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab.2 Effects of microflora during processing The same considerations as previously discussed. whereas cotton become wet and 63 . washing and culling shellfish prior to storage is essential. shellstock is subjected to contamination when stored on the floor in standing water. After washing of meats.3.2). It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. and fish. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. Thus. thus contributing to high bacterial counts in the final product (28). the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank.1. Due to the nature of the shucking operation. In addition. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. clothing becomes very soiled. lobsters. 3.2. for the effects of microflora during the processing of shellstock.2 Shucked shellfish 3.1. 3.2.1 Processing description The processing of shucked shellfish is limited to washing of shellstock.3. To remove the shellfish attached to the remaining valve. Under these conditions if the shuckers enter the packing room. 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.3. Shuckers routinely use either cotton or rubber gloves during the shucking operation. Containers are placed in packing boxes for shipping at refrigerated temperatures. are applied to the shucking operation of shellfish. or subjected to splash from foot traffic. and packing shellfish into containers (Figure 3. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported. the finished product may become contaminated.1. Studies have shown the use of rubber gloves reduces the contamination level. the tank is opened through a drain gate and chute. through which the washed meats are discharged for rinsing and then packed in containers. In the blower-washer. removal of shellfish (shucking).MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. washing.

within one hour. and sanitised before each filling.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). Fig 3. 3. 64 . which may lead to contamination of product. Shucked meats are thoroughly drained. aprons. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7.2 ºC or less. 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. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. Shucking containers are rinsed clean with running water.2. and gloves help prevent the contamination of shellfish. There is a tendency to store such articles on the shucking benches or in packing rooms. 2. Separate rooms or lockers for storage of clothing. 4.

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

nausea. where outbreaks occur regularly. only a small percentage of strains in the marine environment are potentially pathogenic (45). Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). widespread in both polluted and unpolluted waters. It is a common marine bacterium. during processing. there is a recent report of low levels of V. parahaemolyticus multiplies rapidly at temperatures above 20 ºC.1. They can be introduced to the shellfish from the environment or added during processing.e. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). Major outbreaks are associated with the warmer months. approximately 23 cfu/g. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. Diseases caused by Vibrios are enteric in nature. and to the point of sale. shellstock and packing of shellfish in jars) (29). and estimates based from a hospital study show less than 0.5. although non-hemolytic strains have been associated with illness (43. prevention of cross-contamination. Between May and September 1997. and harvesting of shellfish in the cooler months.1 Vibrio parahaemolyticus V. 44). One of 250 cases reported involved shucked products with low levels. there were over 250 cases of illness on the West Coast of North American extending from California to Canada.48 hours with abdominal pain. from harvesting to finished product (i. lot and retail outlet. Sporadic cases are frequent along the coast of the United States. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration.FISH AND SEAFOOD 3. coastal areas). headache and fever. ranging from epidemic cholera to sporadic cases of diarrhoea. diarrhoea. of V.000 population/year (46). parahaemolyticus from shellfish is low. These organisms can be present in the shellfish from the point of harvesting. The incidence of V.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. Fortunately. parahaemolyticus. and very common in Japan. V. parahaemolyticus possibly producing illnesses (40). Products involved were from the same processor. This bacterium is present in all the major processing steps of a shellfish plant. parahaemolyticus causes gastroenteritis lasting 24 . 3. however. V. 41). 66 . 3.5 cases/100. primarily in brackish or saltwater.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish.5.

Hazards from this bacterium can be controlled by thorough cooking of shellfish. Control of V. diabetes. 57. vulnificus. 53. In one study. Onset of symptoms occurs in 48 hours and can be severe. 49). V. 54. or immunosuppression.0. cholerae. It is one of the most severe foodborne infectious diseases.4 . 46.8 case/100. these individuals are advised not to eat raw shellfish. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. chills. 48). Again. 3.5. Primarily. lasting 6 . like V. 55. cholerae non-01 (51). parahaemolyticus the estimated annual incidence rate is very low. vulnificus are fever.5. 58).II (8. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). Control of oyster-associated 67 . and rapidly become inactive at low storage temperatures.3 Vibrio cholerae non-01 Illness caused by V. vulnificus is by rapid refrigeration during warm weather months. abdominal cramps. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. the organism has been found in the Gulf of Mexico. or primary septicaemia.MOLLUSCAN SHELLFISH Control of V. nausea. parahaemolyticus growth in shellfish meats is temperature dependent. Death has been reported to occur within 36 hours from onset.1.7 days.48 hours of onset. All isolates are considered pathogenic. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. and preventing cross contamination once the product is cooked. The occurrence of V. V. parahaemolyticus. 56). cirrhosis. Inadequate refrigeration will accelerate the growth of V.000 population/year (8. High-risk individuals. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. It causes diarrhoea. and nausea within 24 . and vomiting. in the range of 0. gastroenteritis. They are sensitive to cold. 46. however. fever. 3. cholerae non-01 is ubiquitous in the estuarine environment. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. leukaemia. which have been implicated in shellfish-related disease outbreaks (31). Oysters are the most hazardous food associated with V.1. those who have liver disease. The bacterium is an estuarine species that causes wound infections.2 Vibrio vulnificus The symptoms of V. V. The organism attaches to the intestine and release of a toxin is suspected. 23 of 24 major USA-West Coast estuaries were positive for V. are particularly susceptible to primary septicaemia. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). with a fatality rate of 50% for individuals with septicaemia. Healthy individuals are susceptible to gastroenteritis. cholerae non-01 has been associated with septicaemia.

acidity and heat. There are several other Vibrio spp.5. and shock.5. associated with shellfish-borne illness outbreaks. cholerae non-01 illnesses is by cooking seafood thoroughly. They are common marine bacteria. dehydration. e.5 Other Vibrios spp. is the same as previously discussed for the non-01 strains. In addition.FISH AND SEAFOOD V. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American.1. however. and nausea. In developing countries.g. mimicus. drying.10% carbon dioxide for optimal growth conditions. headache and fever. 61. 60). respectively. diarrhoea. parahaemolyticus and V. The frequency of infection of these organisms is no different from other Vibrios. cholerae 01 in molluscan shellfish. including V. salinity. undercooked. 3. these organisms cause gastroenteritis lasting 24 . particularly in oysters. 3. jejuni requires reduced levels of oxygen (3 . nausea.S. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. Illnesses are associated with the warmer months. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. preventing crosscontamination of cooked seafood. The control of V. V. In general. V.2 Campylobacter jejuni C. Gulf coast environment. Illness can include abdominal cramps. Control of infection from these organisms is no different from the other Vibrios. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59).48 hours with abdominal pain. hollisae. and the transmission of the disease by consumption of raw. It causes diarrhoea. V. cholerae 01. abdominal pain. Death may occur after severe fluid and electrolyte loss. (8. 62.5. and illness lasts 7 . Onset of disease is 6 hours to 5 days. 63). or cross-contaminated shellfish (57. Onset of illness is from 3 to 5 days after ingestion of contaminated food. and V. 3. There is evidence of strains of V. Poor sanitation and contaminated water supplies will spread the disease. vomiting.5%) and 2 . processors should know the product source. low pH. imported product from a country experiencing an epidemic. widespread in both polluted and unpolluted waters. the pathogenic severity is lower than V.1. fever. 68 . fluvialis. nausea. The presence of these strains appears to be in low numbers and non-virulent (59). and southern Asia including India and Bangladesh. vulnificus. cholerae 01 establishing in the U.10 days. cholerae 01 is transmitted by faecal contamination of food or water.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools.

although their role as a pathogen is highly controversial and circumstantial.500 cells/gram of Campylobacter may cause illness (64. V. C. Human feeding studies suggest that about 400 . 81. jejuni does not multiply in shellstock at refrigerated temperatures. 66). they survive well at refrigerated temperatures. shigelloides (79. ranging from 0. hollisae Infective dose varies with host susceptibility and virulence of the strain. infection occurred among participants of a firemen's banquet. control of C. cholerae 01 and non-01. 84. hydrophila group (75. fluviallis. V. 85). and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. The seasonality of 69 . vulnificus. jejuni were recovered. jejuni begins in the harvest of shellstock from approved waters. 80) are ubiquitous in the marine environment. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. have been isolated from shellfish beds throughout the world (67. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. Recently. 76. 78) and P. V. 65. 73). 69. 68.3. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). 82. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. 77. however.II Molluscan shellfish associated outbreaks caused by Vibrio spp. parahaemolyticus. of Incidents 4 279 0 1 0 No.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. V. 72. whereas at 10 ºC Campylobacter survive only for 5 days (71). and their immediate proper refrigeration.4 to 114 cells. 83. however. To reduce the risk of shellfish-borne campylobacteriosis. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. of Cases 4 362 0 1 0 a Includes V.3 Other organisms of concern 3. 70. 3. This finding is significant in that C. 71). C. and V.5. In one case.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. Campylobacter appear to survive well in oysters during storage.5.MOLLUSCAN SHELLFISH TABLE 3. mimicus. Campylobacter spp. from Pacific shellstock oysters purchased from a retail grocery store (74). Although. They have been implicated in limited shellfishassociated gastroenteritis cases (8. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters.

and where indicated are intended to be used for official control purposes. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. Note: Although molluscan shellfish are not included in the selected high risk food. Control of these organisms in shellfish products is limited. aureus C. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. 2 Source (90).600. Test Standard Plate Counts E. FDA/ Administrative Guidelines. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. coli Salmonella V. hydrophila group stored at refrigeration temperatures. 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.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. coli Enterotoxigenic E.000 to 3. jejuni V.0001 negative for the presence3 Under review4 Source (89). suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. 87) of the outgrowth of A. cholerae V. non-dairy frozen desserts. Similar results were seen by other investigators (86. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. prepared sandwiches. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. vulnificus in molluscan shellfish.000 cfu/g1 <230/100 g2 <1000 per gram.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. 70 . being isolated more often during the warmer month. They are easily destroyed with mild cooking temperatures. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. FDA/Administrative Guidelines 3 Source (91). 3. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. soy products. parahaemolyticus Staph. Caution should be exercised by individuals with haematological malignancies (88).000/gram (29). dried milk/whey drink powders. In one study.

.N. A typhoid fever epidemic caused by carrier bootlegging oysters.. 419-25. 1995.B. Nauman R. 2.J.R.. 19-34. Eds. 1981. Applied and Environmental Microbiology. 447-52.H. Infectious diseases associated with molluscan shellfish consumption.D. American Journal of Public Health. Ward D. 755-76. in Fishery Bulletin 1964 No.7 1. Applied and Environmental Microbiology. 1975. Lovelace T.F. 46. Jr. 104-9. Colwell R. Rowse A. 42. Cook D... 3. in General Zoology.S. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. U. 8. in Microbiology of Marine Food Products.R. 544-8. Galtsoff P. 29. 30. 1982. Veliger.. Tubiash H. Thompson C. 33643.. Journal of Milk and Food Technology. 9. Wiley-Blackwell. Philadelphia. 1983. 6. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. 71-6. 13. 1940. Tall B.E. 1994.R. Rippey S. Smith F. 44. Vanderzant C. 105-8. Organisation for Economic Co-operation and Development. American Journal of Hygiene. Ed. 10. 1936. Colwell R.W. Bacterial flora of the market oyster.S.L. Hackney C. 82-7. in Multilingual Dictionary of Fish and Fish Products. 1970. Villee C. 6. Applied Microbiology. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). Walker W.. 231-51. Fish and Wildlife Service. 12. Jr. 72-8. 64. 14. 33-6. Schmidt C. USA. Journal of Food Protection.A.. 1960. 245-7. Ray S. 633-40. American Journal of Public Health. Eds. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas).M. 11. Microbiology of bivalve molluscan shellfish.R. 71 . Galtsoff P. Effects of harvesting waters and storage conditions on yeast population in shellfish. water and sediment from Galveston Bay. 13. Hood M.E.S. Fleet G. Journal of the Mississippi Academy of Sciences. 1973. Viability and release Salmonella charity and Escherichia coli from oyster faeces. Pfeifer D.MOLLUSCAN SHELLFISH 3. 36. 8..W. Proceedings of the National Shellfisheries Association. Government Printing Office. Microbiological characteristics of Dungeness crab (Cancer magister). 16. Old H.R. W.. References The Phylum Mollusca.R. 15. Cook D. Clinical Microbiology Review. 17.S. Fate of enteric bacteria in estuarine sediments and oyster faeces.K. The American Oyster.A. 1984. 1964.. Liston J. 4. 7. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. Bernard F.K. 7. New York. 1968. 58. 1926. Biology of the oyster in relation to sanitation.C. 1991. 26.A.C. Lee J. Van Nostrand Reinhold. Applied Microbiology. Eliot C. 1963. Gill S. Saunders Company.. Washington D. 30. 5.

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

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

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

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

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

R. Government Printing Office ..8 Further Reading Ahmed F.C. Aspen Publishers Inc. Sherlock C.W. Gaithersburg. Ed. Food and Drug Administration. in U. U. Washington D. 63-8. 560.C.600 (CPG 7119. Gould G. Washington D.S. Clams.C. Program 7303. mussels. 90. 1995. Chapter 5: Foods. Washington D.. U. Inc. Food and Drug Administration. Ed. Lancaster..C.L. oysters.S. 3. Food and Drug Administration.S. 1987.. National Academy Press. 77 .. Baird-Parker T. in National Shellfish Sanitation Program Manual of Operations. Food and Drug Administration. 1991. Part I Sanitation of Shellfish Growing Areas. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. Food and Drug Administration Compliance Policy Guides. U.S.S. Food and Drug Administration. in U.A.S.W. 1995.. Ed.. 7.1987. 1997.Pathogen monitoring of selected high risk foods (FY 88/89).E. Food and Drug Administration Compliance Program Guidance Manual.H. U. 89. and HACCP: A Global Focus.S. fresh or frozenadulteration by bacteriological contamination Sec. The Microbiological Safety and Quality of Food. Quality Control. U. Burdge D. Technomic Publishing Co. U. Smith J. Slavin J. Fish Inspection. Martin R. Collette R.S. Pittsurgh. Food and Drug Administration. 91. Food and Drug Administration. Food and Drug Administration.M.030. 2000.E..12).. Seafood Safety. Lund B.MOLLUSCAN SHELLFISH 88.

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and available in 0. lobsters and crabs have ten legs. peeled product. (deep-water shrimp). Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. and are usually aquatic.5 and 1. and frozen in 1 and 2. Peeled ready-to-cook shrimp are raw. with additional water to form a glaze. Peeled and cooked shrimp are ready-to-eat after controlled thawing. CRUSTACEAN SHELLFISH Linda Nicolaides. lobsters. Shrimps. crabs.5 and 1. 79 . with shells and heads on or off are usually packed.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda.0 kg polythene bags. Crayfish.. crayfish and shrimp (1). Pandalus borealis (deep-water shrimp).g. the front pair ends in claws.Ph.. Parapeneopsis spp. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. Such products are usually only partially cooked before freezing. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). M. Species eaten from tropical waters include Penaeus spp. e. They possess a segmented body.5 kilo pack (boxes or bags). FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4.4. jointed limbs and a chitinous exoskeleton.0 kg polythene bags. and Trachypenaeus spp. packed and frozen in 0.. Pandalus montagui and Crangon crangon (inshore shrimp).

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

whether GHP are in place. Pseudomonas spp.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. The rate of spoilage will be a function of time and temperature. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected.3. chilled 81 . bacterial contamination and spoilage are inevitable. especially if they are stored in ice or iced water. or transported directly to processing plants.for example. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters.. however. The claws are usually bound together to prevent the animals fighting and causing damage to each other. 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. As with finfish.1). lobsters and crayfish are harvested from cages or pots containing bait.3 Processing and its Effects on the Microflora Crabs. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . They are then taken to market alive in either holding tanks. peeled and cooked. In comparison shrimps die quickly after capture. 4. A large proportion of shrimps are produced by aquaculture. Shrimps can be processed whole. These animals can survive out of water if the surrounding environment is kept cool and damp. 4.10 days for temperate water species (3). Shrimp might be further processed with the shell on. Hence. In contrast. or with the shell and central vein removed. uncooked prawns and shrimp is presented in Figure 4.. gills and thorax. the genera associated with tropical species include Enterobacteriaceae.1. Alcaligenes. corynebacteria. When the shrimp arrive at the processing plant they are washed and then sorted into size. thus. glazed and frozen (see Figure 4. Micrococcus spp. and Achromobacter spp. chilled sea water. Vibrio spp. graded according to size. so they are either sold on the local market or are processed in local processing plants. with “head on” or “head off” whereby the edible tail is removed from the head. or are found in brackish water around the shores. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. or cooked..1 Prawns and shrimps An example of the process used for preparing frozen. from the time of capture and processing. up to 16 days for tropical species compared with 8 . they are washed. After capture and transport to the processing plant. Shrimp will then be glazed and frozen in a raw state. the rate of deterioration of the crustacean shellfish by these organisms is reduced. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots.

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

viruses or parasites. An offensive odour is also produced. putrefaciens and Pseudomonas spp. If boiled. and have a sweetish. or packaged and processed. indicative of protein degradation and the growth of the principal spoilage bacterium Sh.CRUSTACEAN SHELLFISH 4. Once removed and cleaned. contaminating microflora may include enteric bacteria. there is no indication of life. they include Sh. In comparison. bacteria of public health significance. 4. whilst boiled crustaceans have a pleasant slightly sweet smell. bright and dry appearance. the joint between the tail and carapace is open and the limbs hang loosely. 4. wet and ‘soapy’ or ‘jammy’ to the touch. with the growing demand for ready-to-eat products from crab and lobster meat. The shell will be soft. Heat is produced if the shrimps decompose in a confined space. With spoiled crabs and lobsters.4. putrefaciens. dull and sticky. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. slightly iodine smell. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. or in hermatically sealed cans that have undergone an appropriate heat treatment. spoiling prawns and shrimps have shells that are soft. However. 4.2 Cooked crabs and lobsters As indicated earlier. since crabs and lobsters are traditionally consumed immediately after cooking. and a strong ammoniacal odour is produced. the meat is either repacked and sold in the shells.3. If the tail drops. immediately prior to boiling. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail.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.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. for crabs. there is no associated spoilage problem. the tightness of limbs will indicate whether they were killed by immersion in boiling water or.4. 83 . If hygienic procedures are not followed. The shells have a crisp.

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

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

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. parahaemolyticus. jejuni has not. Astroviruses and non-A. vulnificus is an invasive and lethal pathogen associated with wound infections. whilst 5.8% of samples contained C. 4.6 Campylobacter jejuni and Campylobacter coli C.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans.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. However.30 MPN/g) (27). they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). hydrophila has been isolated from prawns (29). hydrophila is able to grow well at refrigeration temperatures (0 .5.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. however. However. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. a marine halophile. Hepatitis A virus presents the most serious hazard. Quantitative levels were below limits of detection in all cases (<0. coli. or freezing at a temperature of -23 °C for 7 days (30. Of these. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. 4. although A.7 Aeromonas hydrophila and Aeromonas sobria A. V. e. but has the potential to cause fatal foodborne illness (26). V. Caliciviruses.5. Parasitic infections are controlled by either cooking infected material prior to consumption.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States.5. hydrophila and A. occurs in crustaceans from warm environments. been implicated in a case of foodborne infection associated with crustaceans. Japan. non-B Hepatitis viruses (31). A. 31. It is more commonly implicated in countries where crustacean shellfish are eaten raw. 4. particularly trematodes (6). to date. Norovirus. the potential hazard exists as C. South America. 4.g. in a survey carried out in the United States between 1977 and 86 .5. 32).

although it is one of the European Union (EU) legislated allergens.5. The risk of such hazards is far higher with fin fish and molluscan shellfish. 4. 4. Provisions laid down concerning the use of additives would also apply to these types of products. Norwalk viruses can also withstand heating to 60 °C. 4. consistent with that in the stomach. which results in a persistent and apparently permanent loss of short-term memory (36). No foodborne incidents have been attributed to this source to date. 38. which could be transferred to the meat during unskilled removal from the shell (35). 2073/2005.CRUSTACEAN SHELLFISH 1981.5.11 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. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). They can survive low pH levels.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. Selected parts from this regulation is presented below.6. 4. The toxin causes amnesic shellfish poisoning (ASP) when consumed. or chlorine concentrations similar to those found in drinking water. 87 . 39).1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. from farm to fork (37. 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. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. A summary of microbiological criteria for crustacean shellfish is listed below. are included in the Commission Regulation (EC) No.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. only 0.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). produced or imported into the European Union (EU).

FISH AND SEAFOOD

4.6.1.1 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 4.6.1.1.1 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. 4.6.1.1.2 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

88

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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. 4.6.2.1 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.

References
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.

3.

4.

89

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.

19.

20.

21.

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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.

27.

28. 29.

30.

31. 32.

33.

34. 35. 36.

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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.

38.

39.

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.

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

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

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

is employed. When a milder brine. all using salting steps. sometimes the salt crystallises onto the surface of the fish. a 90º or above brine (237.0 .0 158.4% salt (Table 5. it is important to understand the concept of salt concentration.0 g of salt per litre of brine). TABLE 5.3 Processing and its Effects on the Microflora Owing to the wide variety of products. % 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.I).4 52. it causes swelling because the fish absorbs water from the salt solution (1). When working with salting. it is impossible to establish standardised processing stages in the curing of fish. and salt content. Different brine strengths are required according to the curing processes and/or the type of product. a 0º brine saturation means pure water. Thus.2 g of salt per litre of brine).6 g of salt or more per litre of brine) may cause salt burn in this way. smoking or marinating processes. At 18 °C a 100º brine is produced when the solution contains 26.6 264. The strength of the solution is described as degrees of saturation (brineometer degrees).2 105.FISH AND SEAFOOD 5. When a 100º brine is used. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. about 50º (132.4 184. only the most common processes can be described here. or salt in the water phase. it is important to state the temperature.8 79. the fish is usually salted with a 70 . Therefore.80º brine (184. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.8 211.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. processes and producers’ private recipes. Salt concentration means the percentage of salt in the water contained in the fish. For smoking salmon.6 132. The dissolution of salt in water is dependent on the temperature. This should be distinguished from the salt content. Salt in the water phase links the two conditions of moisture and saltiness.2 237. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat.8 211. etc.

‘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. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts.CURED. 5.0. and such delays. and salt can speed this oxidation process. when the air humidity is high. and essentially free from iron and copper (3). This condition is known as ‘putty fish’ (4).1 Salting Salting of fish for preservation purposes has been used since pre-history.1.1 Dry salting / Kench curing Usually.35% and 0. for which there is a high demand in Mediterranean countries.15 . mainly in tropical conditions. The most common fish used in the northern hemisphere is cod (Gadus morhua).3. which consequently inhibits the diffusion of the water from the bulk of the product to the surface.1. See Figure 5. SMOKED AND DRIED FISH 5.15% calcium and magnesium. The salt used should be of food-grade quality. and a ‘putty’ fish product is also obtained (1).3. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. outgrowth of halophilic bacteria may occur. This is because fatty fish turn rancid quickly. Salting can be done in a number of different ways. For a light salting.1 ppm and 10 ppm. Also. can be dried further to produce ‘klipfish’ or ‘bacalão’. but the concentration of copper and iron must be kept under 0. the fish is restacked with fresh salt after about a week. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. 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. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. On the other hand. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). the fish is mixed with one part of salt per eight parts fish. are recommended to help make the product whiter. However. Usually it is followed by other preservation techniques such as drying and/or smoking. low in calcium and magnesium. respectively (1). When pure salt is used. lean white fish is used for this purpose. having a short section at the end of the tail. respectively. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. caught in North Atlantic waters. if very pure sodium chloride is used for curing purposes. ‘Green cured’ split cod with about 60% of the backbone removed. however. it tends to lead to a slight yellowing and softening of the product. are likely to cause hydroscopicity of the product and. addition of levels of 0. and for a heavy salting a ratio of 1:3 salt:fish is used (1). 97 . The calcium and magnesium impurities. and the salt has penetrated throughout the fish (3). which include calcium and magnesium. producing characteristic pink patches.

Processing of dried salted cod 5.90 are rarely achieved.5. Where fish are salted in their whole form. The greatest barriers to salt penetration are the skin and scales. Food-poisoning organisms.1.2 Wet salting or brining As referred to above. eviscerated and/or split). such as Staphylococcus aureus could survive and grow at such water activities in the product. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole.3. so that the risk of spoilage is high. it appears that water activities much less than 0.1. when a light brine is used. Only small fish can be salted whole. the process is mainly for flavouring purposes prior to the use of other preservation processes. For large fish it is advisable to split it before salting. although toxin production would be unlikely (4). Fig. such as 98 .FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat.

sardines.78 2.10 1.86 2.78 6.91 2.76 99 .85 3.25 5.30 2.11 3.3.0 3.5 4.56 6.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) 2.23 8. with pressure being applied to the top of the container to help the brine to drain away.46 4.05 7.54 5.26 4. Histamine formation is of importance when using scombroid and some clupeidean species. 5.61 3.51 5.49 4. with a shelf life of about 4 weeks (water activity (aw) about 0.76 4.44 2. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.3 days. The fish are kept immersed in their own brine near saturation point for several days.98 6. headed and gutted. they can be stored at 28 °C.66 5.85 3.35 4.04 5. Anchovies are the main fish used for these purposes. The main objective of the brining method of salting fish is to limit air penetration during processing.98 6.98 5.76 4.74 9.26 2.76 4. the liquid that comes out of the fish forms a brine.26 3. the fish enzymes play an important role by maturing or ripening the product.5 3.90 (1). Histamineforming bacteria do not generally grow at water activities below 0.II.47 6.CURED.51 3. SMOKED AND DRIED FISH smoking.23 2.70 6.06 4. the brine is then replaced over the fish.96 1.41 6.88 5.23 2. herrings and other fatty fish can be used.41 5. can be cured in a saturated brine solution for 6 days and. There are several variations of this process.76 4.03 4.00 3.33 5. Fresh sardines. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .03 2.26 1.70 3.78) (1).16 7.09 8.60 2.0 4. Here.5 3. This is why it is the preferred method for fatty fish.44 2.19 3.1. For preservation purposes.25 5. but as it is placed in containers. See Table 5.5 10. after being drained and pressed at 465 kg/m2 for 15 hours.84 1.45 3. limiting the amount of rancidity that can develop.61 3.26 7.99 2.64 4.76 4.23 3.11 9.11 4.09 8.65 2. but mackerel. TABLE 5. the fish is dry-salted. thus.73 1.17 5.22 6.80 5.95 3. For example.41 4.41 3. the product obtained is called gaspé-cured (5). if split fish is dry-salted.66 5.17 4.0 2.3 Dry / wet salting In this process. a saturated brine is used. and a red burgundy colour will develop in the product. and can be collected and replaced again over the fish.

After this. salt.4% of salt) for distribution and sale (1). In a second step. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. then packed in barrels with a 70º brine. because of the salt content of the product. outgrowth of bacterial spores after processing may occur (4). Ceviche. However.FISH AND SEAFOOD 5.7%) for as long as 3 weeks.3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. A much firmer product is obtained when the fish is salted prior to acidification.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. even if the product is hermetically sealed. and 6% salt. and lasts for 3 . The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten.30 minutes. Another variation uses sour cream instead of the second brine step. containing 1 kg of salt for every 25 kg of fish. ‘ikan kaju’. they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. usually flavoured with onions and hot chillies. usually requiring refrigeration below 10 °C for longer-term storage. is made of different varieties of fish. As referred to above. and spices) is added to the brine.5% 120grain distilled vinegar. Once cooled. They will have a salt content >6% (w/w) in the water phase. All these products are ‘semi-preserves’. they are packed for consumption within 2 weeks at chill temperature (6). drying and salt-boiling steps until a hard translucent block is obtained (4). There is epidemiological evidence that these products have been the cause of some 100 . Other preservation techniques can be used for extending the shelf life of these products. herring or other fatty species are used. underlines the need for careful control (4). whereas the latter method preserves the fish for only a few days.2 months. a Peruvian product. up to 1 . Usually. has a shelf life of several years. and packed (3). Traditional jellied eels are a product where pieces of fish are cooked in boiling water. flavouring (sugar. the occasional implication of pindang in cases of foodborne illness. but it has further smoking. Herring can also be cured for several days in an 80 to 90º brine with 2. The first process can give a longer shelf life. until the flesh is soft. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is.3. and even death. which is dipped into a boiling brine for 10 .6 days without refrigeration (1). Afterwards. or can be boiled by being placed in a basket. giving a cooked appearance to the product. only the nonspore-forming organisms are eliminated in the process and. Another salt-boiled Indonesian product.3. placed in a marinade of lime juice. or a pH <5 and they are stabilised for a limited period. 5. The fish can be soaked in a high concentration of vinegar (about 4 .2% acetic acid and 2 . The lime juice plays the role of the vinegar. it can be repacked in milder brine (1 .

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

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

Maintaining the temperatures of pyrolysis between 425 and 200 °C. sprats. oysters. such as Escherichia coli. Top-quality fish must always be used. Blends of various woods can be formulated to produce sawdust with specific flavours. In the mechanical kiln.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. softwood colours the product quickly. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10).3.5 Smoking Smoking today. 5. The quality of the smoke is related to the type of wood used. is mainly a process for flavouring fish rather than a preservation technique. Hardwoods are considered better because of the milder flavour they impart to the food. There are many fish species that are used for smoking purposes . making full description impossible here. and the use of smoke generated by superheated steam or liquid smoke. tuna. cod roes. botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc.CURED. It can be used for cold or hot smoking. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns. clams. SMOKED AND DRIED FISH TABLE 5. Staph. aureus (toxin) C. mackerel. aureus and Saccharomyces cerevisiae (10).salmon. cod. The smoking procedures vary widely. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish. using electrostatic filtration of the smoke. eel. mussels and many others from tropical countries. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. However. trout. haddock. swordfish. Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . Aroma and flavour are a blend of smoke components (3). white mangrove and abura has an inhibitory effect on some microorganisms. herring. as referred to above. It has been reported that the smoke from mahogany.

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

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

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

moulding it into long thick sections. a very expensive and fashionable smoked product in the Netherlands. are usually hot-smoked after being immersed in 80º brine or dry-salted (6).CURED. white fish about 8 days. Katsuobushi. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. 5. which means that any bacterial contamination and growth after the heat treatment is a serious hazard that must be controlled by Good Manufacturing Practices (GMP) and factory hygiene/sanitation after the heat treatment (7). 18). Aspergillus ostianus. The product is removed from the drying cycle. and many closely related Aspergilli (Aspergillus ochraceus. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. synthetic smoke flavours. Aspergillus oryzae. including Eurotium repens and Eurotium rubrum. (Penicillum cyclopium and Penicillium puterellii) (19). and substances unconnected with smoke but with a smoky flavour or smell.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output.5. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. Boiled or steamed clams. Aspergillus tamarii) and several Penicillium spp. apart from the improved appearance. Vacuum packaging retards the onset of rancidity when fatty fish is used. are used. Natural smoke extracts. including ‘umami’ (16.6 hours and smoked for 3 . The classical liquid smoke is 107 . 17. Growth of these moulds on the tuna results in the extraction of moisture. 21. similar to masmeen. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. Eels. When refrigerated at 3 °C. such as yeast derivatives. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). fatty fish will remain in good condition for 6 days. are usually 80º brined for 5 . 23). Hot-smoked products made from white fish will usually keep better than those from fatty fish. but seems. Mackerel.3½ hours using the typical temperature set for hot-smoking referred to above (12). A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. chilled or freeze-thawed. cooking in brine. 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). they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). rewetted and remoulded to give the desired shape and consistency. break down of fats and the moderation of smoke flavour.3. They will keep well frozen. 22. resulting in a characteristic flavour (20. In terms of safety. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). then hot smoking and finally sun drying it. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. but mechanical drying is used instead of sun drying. They are generally not cooked prior to consumption. to have no further advantage over other packing methods for other fish (12).

both of which are considered potential carcinogens. Fresh fish spoilage may be measured by 108 . Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine.03 μg/kg of benzopyrene. The European Scientific Commitee for Food. drying. 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. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). When liquid smoke was used. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). nutritional neutrality and antioxidant activity (27). Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. or smoke extracts in organic solvents.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. smoking. Liquid smoke was considered to provide advantages of safety.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. 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. and less than 0. According to some recent research.3.6 to 2. injecting it into the fish flesh.8% and still provide protection for 7 days (25). or applying it as an atomised spray within a modern automatic smoke kiln. the salt level could effectively be reduced from 4. 5. 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.5. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. consistency of flavour and colour. applying it as a dip after brining. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. It has been shown that liquid smoke can reduce the formation of PAHs (3).06 μg/kg of benzoanthracene in the final foodstuff (28). 5. In experiments comparing traditionally smoked fish with liquid smoke. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). so as to impart an unnatural red colouration to the flesh.4 Spoilage Brining. 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).

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

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

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

Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms.variable.FISH AND SEAFOOD TABLE 5. + to +++ strength of preservative effect 4 Post-process contaminants 112 .IV Spoilage of salted. Dun.not used/unchanged. boiled. halophila Halophilic spoilage organisms Halophilic spoilage organisms.W. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. LAB. 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. sebi and B. marinated and smoked fish Spoilage controlled by pH Heat Ant1 Ref2 Process Product Aw Probable spoilage organisms None Fresh fish spoilage organisms Halophilic spore formers and PPC4 Halophilic spore formers and PPC Halophilic organisms: Pink-Halococcus spp. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted. . and Halobacterium spp.

when present in the raw material. botulinum. but if the fish products are stored at temperatures <10 °C. with this change. 9 and 8 of the outbreaks.97 0.5-10 4. In the United States.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. popular demand for healthy foods has led to a change in the tastes preferred by the public and.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13).5 5. the products were heavily salted.CURED. Since neither the smoking nor the drying parts of this curing process are particularly severe.5 5.97 0. and as botulism is a serious hazard in foods. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth.0-9. botulinum type E and production of toxin.5 4. types A and B are not affected by 3 . salted and pickled products accounted for 10. botulinum Staph. Before. the importance of cold-chain integrity must be emphasised. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C.5-10. under optimal conditions. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13).86 0. The presence of salt in cured products has a great effect on the growth of the bacteria. respectively. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. The concentration required to prevent growth at room temperature can vary from as low as 3.5% to 5%.5 0-11 3. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E.5-8. smoked and dried. the non-proteolytic type E strains can grow in up to 5% sodium chloride.61 Organisms Most Pseudomonadaceae C. aureus Yeasts Moulds Lactic acid bacteria pH 5. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. sodium 113 . It has been shown that. could not grow and produce toxin. botulinum type E. although the combination of the two factors.5-8.75 0. pressure on the traditional methods of curing. SMOKED AND DRIED FISH TABLE 5.5. the fermented products being the most dangerous group (7).8 0.8 1. However. botulinum type E Staph. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). and C.9 0.

5.86 and 10% sodium chloride.4.4 Listeria monocytogenes The psychrotrophic L. if storage temperatures are over 10 °C. It is a halotolerant bacterium able to grow at water activities as low as 0. 5. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. throat and skin. 5.5% salt in the water phase. if it contains sodium nitrite ≥100 ppm. related to the consumption of a raw smoked fish meal.0 . if the person handling the product is a carrier of Staph. However. when the bacterial load of the product is lower. Sliced smoked fish products. 5. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . as they require more handling. Once the product is contaminated.or cold-smoked fish should have at least 3. aureus. more importantly. gills and guts as well as in several lightly preserved fish 114 . the organism can survive most of the light curing processes. but. the human nose. coli all occur on fish products as a result of contamination from animal/human reservoirs.FISH AND SEAFOOD nitrite can be added to the brine but this is not permitted in EU countries because nitrites might lead to formation of the carcinogenic nitrosamines. The organism is mesophilic with a minimum growth temperature of 7 °C. growth and toxin formation may occur. 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.3 Enterobacteriaceae The pathogens Salmonella. In foodstuffs. As they are poor competitors. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and.30% of the population positive for enterotoxin-producing strains (41). are particularly susceptible to contamination and. 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.94 (7). they will grow rapidly (7). it should contain not less than 3. Infection by Shigella sonnei has increased in the UK. if recontamination occurs and storage conditions are favourable. the organism may contaminate the product. As with other organisms. but requires higher temperatures for toxin production (>10 °C). has been reported (42).5.5.5.0% salt in the water phase (30). Shigella and E. but. aureus growth. Thus. after processing. 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. Usually. The FDA established that hot. the risk of infection can be eliminated by proper cooking. and the minimum pH for growth is 4.

have been shown to have an anti-listerial activity in many food products. a number of fish products. the CT 3162-FAIR program. the only concern to public health is when eating raw or uncooked fish products. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. do not survive 1 min at 60 °C. gravad fish. which.514. Of microbiological concern in the northern hemisphere are two parasites . when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). or nematodes. Some LAB.92 . most of them are of little concern to public health. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism.8 . favours growth of psychrotrophic LAB. Pseudoterranova dicipiens. such as ‘lightly preserved’ fish products.9. monocytogenes pathogenic for only specific segments at risk. pH 4.6 and water activities below 0.5. Thus. and in CT95-1207FAIR). monocytogenes in levels at or above 102 per gram (46). which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48).95 (45). both being detected in cold-smoked salmon.the round worm Anisakis spp. Its limiting conditions for growth are 0 . are considered unsafe.12%. SMOKED AND DRIED FISH products such as cold-smoked fish. a round worm.2. from freshwater fish. which means that the process must conform to a level of ‘no detectable L. widely used as starter cultures in food processes.5 Parasites Despite the presence of parasites in fish being very common. cold-smoked fish. This includes matjes herring. and other local traditional products (7). Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. and the tape worm Diphyllobothrium spp. They have complicated life cycles. salt levels of 8 . the USA established ‘zero tolerance’ for the organism.CURED. has also been detected in cod (7). from seawater fish. Two outbreaks in the United States have been reported to be linked to salmon (10). which considered L. Unlike some European countries. in combination with curing agents and refrigerated storage. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. monocytogenes’ in 25 g of the finished product.45 °C. marinades (ceviche) (43) and gravad fish (44). lightly salted caviar. 5. The use of LAB in fish preservation is a topic that has been studied in projects (UP. a short period of freezing either of the raw material or of 115 . researchers are looking for other ways of preventing the pathogen’s occurrence or growth.0. Until 1997. 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. which include intermediate hosts. The round worms. in the EU Fisheries R&D-FLAIR program. there were no reported outbreaks related to cured fish. As they are heat-sensitive.

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

. Introduction to Fish Technology. Flick G. Leatherhead. Handling and Processing Eels.. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. Ed.E. Fernandes R. USA..CURED.H. Urner Barry Publications Inc. Torry Advisory Note No. Marinades. monocytogenes Salmonella spp. Wareing P.F.7 1. Huss H. Hall G. 8. Flick G. 9. 1982. Birnie K. parahaemolyticus Vibrio vulnificus C.. Bannerman A. Catching. 120-38. Wareing P. 1993. London.F. 56. Martin R. 6. Eds. Food spoilage fungi.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. 1994. Regenstein C. 7. FAO Fisheries Technical Paper. 254-82 5. 82 (revised). coli (ETEC) L.M. SMOKED AND DRIED FISH TABLE 5. New York. LT or ST positive Presence of the organism Presence of the organism Positive for enterotoxin or ≥104 (Most Probable Number) Presence of toxigenic 01 or non-01 ≥ 1 x104 (Kanagawa positive or negative) Presence of the organism Presence of viable spores or vegetative cells in products that will support their growth. Smoked. Leatherhead Publishing. 1991. 1979. cured and dried fish. 1997. Eds. 1984. Moody M. 334. except in the viscera of dungeness crab. The Smoked and Cured Seafood Guide. McLay R. 4.. Horne J.. Torry Advisory Note No. Chapman & Hall. Doré I.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. Horner W. 37(revised). Preservation of fish by curing (drying. salting and smoking).M. * Staph. 3. or presence of toxin 0. in The Seafood Industry. Fernandes R. in Micro-Facts: The Working Companion for Food Microbiologists. Hot Smoking of Fish. 117 . 3273. New York. 2. in Fish Processing Technology. Assurance of Seafood Quality. 381-406.A. Van Nostrand Reinhold. Torry Advisory Note No. 2007. botulinum * Limit of tolerance 1x 103 ETEC/g. aureus * Vibrio cholerae V. References Regenstein J. Van Nostrand Reinhold.W. 1990.

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

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

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

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

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

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

g. cencalok burong-isda. e. 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. pa-chao. som-pa-keng pekasam. burong-dalag.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi.12 months 10% raw sugar added. mam-chao. mam-pa-kor. burong-bangus pla-ra.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. pa-khem.FERMENTED FISH TABLE 6.8 months Main dish TABLE 6. funa-suchi. shrimps. crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 . mam-seeing sikhae som-kay-pa-eun. tai-suchi phaak. burong-ayungi. e. crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 . pla-som.g.3 months Main dish Tai-pla Bowels 1:3 6 . ayu-sushi. pla-chao. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps. som-pa. som-fak/ som-fug 125 Malaysia Philippines Thailand .4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 .

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

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

Patis (Condiment) 128 . 6.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 .12). 6. Fig. Belacan (Condiment) Fig.2.1.

To produce semi-solid fish pastes as opposed to fish sauces. In fish sauce production. Ika-Shiokara (side dish) Fig. eviscerated fish are sometimes used to slow the autolytic process. however. 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.FERMENTED FISH Fig. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. the 129 .6.4. 14). 6. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are.3.

6 – 6. Figure 6.5. for example.V and Figures 6. 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. In contrast to free-swimming fish.5). filtered and bottled. Some examples of products and their formulation are presented in Table 6. which is eventually separated from the residual fish tissues.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.3. 6. snack) 130 . particularly glutamate. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. 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. volatile fatty acids and nucleotides. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens.8. Fig. 6.

NE 10 . north-east.FERMENTED FISH TABLE 6. Fig. 3:1:0.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0.6.03 Preparation time 6 . the prefix 'pla' is replaced by 'kung'. NE Formulation Fish (whole or pieces): salt: roasted rice. In these cases. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. C NE. side dish) 131 .12 months Shelf life Use 1 . S.7 days 5 .12 months Main dish 2 Pla-jao C.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 .5-1. C NE. N.6.4 are also produced using shrimps. Kung-Chom (staple food. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. N.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. C. south of Thailand. north. NE.12 days 3 . N. N S 5 . central.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .2-4 Fish (pieces): salt:khaomark (fermented rice).

Red burong isda 132 . Phillipines Journal of Food Science and Technology. typically in the range 2 . Generally. these products are salted at lower levels than the fish sauces and pastes..salt and lactic acid. their growth and metabolism are nonetheless slowed. If acidity is a desired sensory property of the product.VI).1 The use of higher salt levels results in a longer production phase but a better keeping quality product. then this will take longer to develop than if lower salt levels are used. 2. It also indicates that. 106) Fig. while they are able to grow at high salt levels.7. of the two factors contributing to shelf life .3.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. (from Arroyo et al. giving a reduced rate of acid generation. 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 6.6. The LAB isolated from these products are those with more marked salt tolerance but. the salt is of far greater importance in determining the product’s long-term stability. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6.10%. This is readily explained by the susceptibility of LAB to elevated salt concentrations.2. but there is considerable variation in this.

1 4.9. 106) Fig.0 Shelf life 3 weeks 6 . 133 .8 .5.8.5.4.4.3 .9 .12 months 1 .5.7 .FERMENTED FISH (from Arroyo et al. Philippines Journal of Food Science and Technology.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 2.4 .6.5 3.6.8 pH 4.5.0 .2 4.2 7.8 .17.1 .6.5 .9 4.8 2.6 3. White balao-balao TABLE 6.3 5.9 4.0 .5 .0 .2 4..3 years 1 .5.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16). 2.

In several products.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.3.5) despite other factors. Where they are used. a common ingredient in these products. For example. alternatively.FISH AND SEAFOOD 6. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. inulin. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. such as salt content.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. 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. Garlic. Addition of carbohydrate will.VII).1 . a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. also lower the unit cost of the product and this is probably a significant factor in many cases. they increase the amount of soluble sugars produced and thus the range of LAB that can grow.4.2.9) compared with the white product (pH 4.0 . This results in a more active lactic fermentation and a lower pH. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. of course. being similar (18. 21). 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 . although the final population of LAB may be similar. 19). Although the ability to ferment starch is not widespread in the LAB. When added as boiled or roasted rice alone. Rice is the most common carbohydrate source used. the rice may be added in a partially saccharified form such as kao-mark (Thailand). also serves as a valuable source of the more readily fermented carbohydrate. TABLE 6. 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).3.

around 12%. is something of an exception since millet is used along with garlic as the carbohydrate source. in turn. lead to an increase in pH.5.9%.2 at the end of fermentation. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). will not reliably exclude air. Results of a product survey were consistent with this. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products.5:9:3. The mixture is allowed to ferment for 2 .7 up to 6. reporting the product’s pH to range from 4. the Korean fermented fish product. markedly extending the shelf life (25). It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others.2. presumably correlating with the salt content. 6. Excessive growth and acid production by the LAB could render products too sour for acceptability. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. allowing other spoilage organisms to grow. but they are likely to behave similarly to lactic fermented products produced from other commodities.7 to below 4. In a successful fermentation. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. The growth of yeasts and moulds could. 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). There is little published information on the factors that limit the shelf life of the low-salt products. such as wrapping in banana leaves. 135 . The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. the level of salt employed is on the high side for fish/salt/carbohydrate products. and there is a marked regional variation in the preferred type.8 . One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. the pH drops from 6.FERMENTED FISH Sikhae. Generally. Clearly. 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. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage.3 weeks at around 20 °C.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). Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet.3 days. Traditional packing techniques used with lactic fermented fish products. thereafter declining to about 4. which ranged from 7.5 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.17.

fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. During fermentation. viable numbers will probably decline quite rapidly. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. and consumed without cooking. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. but we are unaware of any reports of this happening. although these limits apply when all other factors are optimal (26). Several measures have been suggested for reducing the risk of botulism. fish products are the most common source of botulism. 6. After capture. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. In the fish sauces and pastes. outbreaks of 136 . are Vibrio parahaemolyticus and Clostridium botulinum. under conditions inimical to growth and high ambient storage temperatures. 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. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. The bacterial pathogens most commonly associated with the living environment of fish. In Japan.8 and at salt levels as high 10%. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. typically 30 °C. Because of the severity of the illness it causes. There is little in the way of documented information associating fermented fish products with foodborne illness. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. V. however. 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. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27).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.87. parahaemolyticus has been reported growing down to a pH of 4. which accounted for 88% of outbreaks in the period 1952 . and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. As in Norway.

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

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

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

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

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

7. dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). Gymnodinium catenatum. even crustaceans such as lobsters. Many of these species are extremely common and are found throughout the world in a variety of cold. 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. all of which contain a tetrahydropurine skeleton (Figure 7. Not all the toxins found in shellfish extracts are microalgal products.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.FISH AND SEAFOOD has been produced (9). Around twenty PSP toxins are known. can become contaminated with these toxins. In addition. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). neoSTX and Gonyautoxins (GTX) 1-4. a great variety of shellfish from all latitudes. and a review on the risk assessment of marine toxins has also been produced (14). Consequently.g. 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. 7.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium.2. Saxitoxin Group instead of the Paralytic Shellfish Poisons). A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. and a tropical species Pyrodinium bahamense (16).1). 142 . The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). some are the result of biotransformation processes by the filter-feeding host. with intoxications occurring in many countries even today (15). PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. All PSPs degrade rapidly in alkaline conditions. and another. The Ncarbamoyl sulfates are readily hydrolysed to STX. temperate. and sub-tropical waters and habitats (15).

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

7. Taiwan. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). Two approaches have been taken here.2. Thailand. 24) (Figure 7. other derivatives have been reported. a pre-column method has received Association of Analytical Communities (AOAC) approval (18).2). TTX most commonly occurs in many species of puffer fish. and over ten analogs are known (22. a criticism has been that such methods are time consuming. Most intoxication events have occurred in South East Asia. China.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. After some development and various trials. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. However it remains as the monitoring method of choice in many countries. Alternative HPLC methods have subsequently reduced this to a single HPLC run. 23. specifically 144 . or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. and has also been evaluated by a number of EU laboratories. including Japan. While the chemical detection methods are most promising for the future. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. The mouse bioassay has been in place for decades and the method has been standardised.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). Since its discovery.8 µg/g) of shellfish or seafood tissue. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). The latter test is accepted by the United States Food and Drug Administration (US FDA) for use in the United States National Shellfish Sanitation Program. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). 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. and while simple and relatively inexpensive to operate. and islands of the South Pacific. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. 15. 17).FISH AND SEAFOOD 7.3. as well as other parts. 7.

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

but 146 . and while promising.FISH AND SEAFOOD column oxidation to the fluorescent derivatives. need further development (24). Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. a polar glutamate agonist. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. However this method has been criticised for being complex. but it is known that such congeners are less toxic than DA itself (30).4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). 7.3.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. 7.4. Structure of domoic acid and three naturally occurring photoisomers 7. 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. Immunoassay methods have also been investigated. Currently 12 isomers of DA have been reported (see Figure 7.3 for examples). but all of them are considerably less toxic than DA itself (29). Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28).

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

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

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

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.3 days. and a detection limit of 100 µg/kg has been reported (12). A regulatory level of 80 .FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). Due to the lipophilic nature of the toxins. vomiting. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. 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. even with the minimal of clean up steps (48). nausea. which can lead to an underestimation of the total DSP content in a sample. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . Since the late 1970s. Human symptoms include diarrhoea. various European countries. 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.5.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. and while this provides critical seafood safety information.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. 7. and abdominal pain. DSP episodes have been reported worldwide and are included in many monitoring programs (47). 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. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. DSP-Check is a commercially 150 . DSP episodes have been reported from Japan. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. it does not provide information on the DSP toxin profile. 7. New Zealand and Chile. 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). 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). In recent years.5. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. and DTX-2.

6. 7. 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. 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 and 7. However. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). 7. Fig.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).5. In yet another approach. Within this group. 58). for many years these compounds were included in the DSP toxin complex. and another rapid antibodybased test kit for DSP toxins has also been reported (53). over 20 PTXs are known (56) and more than 30 YTXs have been reported (57.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. with good agreement with the mouse bioassay and LC–based methods (54. 55). 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. Indeed. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . with a claimed detection limit of 20 µg/kg. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. Some representative examples of both groups of toxins are shown in Figures 7.

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

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

Fibrocapsa japonica. and off the coast of New Zealand (65).7.1 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. and cockles (65. the same toxins have been reported to be produced by three genera of Raphidophytes.7. was chemically characterised as PbTx-2 (67). 7. A complicating factor both in terms of establishing the profile and toxicity of brevetoxin-contaminated shellfish has been the discovery of the so-called “brevetoxin metabolites”. 66).FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida. 154 . They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. which are found in Japanese waters (66). and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf. Interestingly. whelks. Many of these species are associated with massive fish kills. These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). brevetoxins are found in mussels. and one of the brevetoxin metabolites BTX-B2 7. Structure of the two parent brevetoxins. verruculosa off the mid Atlantic coast of the U. 66). In the Gulf of Mexico. PbTx-1 and PbTx-2. namely Chatonella marina. Fig.S. but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65. and Heretosigma akashiwo. oysters.

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

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

FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. Another symptom that can develop over time is extreme fatigue. Globally. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . 75. The onset of symptoms can occur rapidly within 24 hours. although not all CTXs are equipotent. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. barracudas.76). These include paresthesias (numbness and tingling of the extremities). 7. which can be confused with chronic fatigue syndrome. 7. groupers. and it is estimated that ingestion of as little as 0. sharks. and in fact CTX poisoning is often only identified after consumption of contaminated fish. and although the gastronomical problems may dissipate within a day or so. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. there are neither standards nor an official testing program. 78). weeks or even months (74 . where cold objects feel hot and vice versa. Even when suspect samples are available. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. which can return following the consumption of alcohol and certain foods such as oily fish (74.1 µg of ciguatoxin can result in human illness (74. and there are suggestions that such incidents are increasing and spreading worldwide (74). though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. Like the brevetoxins. which in turn are consumed by carnivores such as snappers.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. and the sensation of loose teeth (77). 75). The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. Other symptoms include a metallic taste. arthralgia. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. all of which can be linked to the effect of CTX on excitable membranes. 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. Local island populations attempt to avoid poisoning by catching only smaller fish. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. only trace amounts of CTXs can cause illness following consumption of contaminated fish. and temperature sensation reversal.8.3 Detection methods As a consequence of their potency. myalgia. and moray eels.8. the neurological symptoms can persist for days. pruritus. Patients have reported symptoms lasting weeks and even months.

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

3 days. Following the original episode. oysters (Crassostrea gigas. clams (Tapes philipinarum. 90). and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. scallops (Pecten maximus). In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. but current data indicates that unlike the DSP compounds. Structure of the three principal azaspiracids found in shellfish 7. cassipes (83). and to date this has been the only source from which these toxins have been found at toxic levels. and to date no long-term effects have been recorded (83.9. 88. Donax spp. which can persist for 2 .). and most recently in a crustacean (Cancer pagurus). as in other cases of shellfish toxins. and off the coast of Morocco (83. diarrhoea and stomach cramps. 7. cockles (Cerastoderma edule). Mytilus galloprovincialis). which are ingested by P. There has been considerable debate over the source of azaspiracids. The bivalves affected include mussels (Mytilus edulis. 84. 7. Scandinavia. AZA occurrence has been reported by several European countries including France. cassipes produces AZAs in culture. AZA toxins do 159 .9. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). 89). Ostrea edulis).1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. vomiting. While this result is not necessarily in debate. these molluscs were never considered to be the biogenetic source. it has been impossible to establish that P.FISH AND SHELLFISH TOXINS Fig.9. and some time after the original episode in Ireland. However. Ensis siliqua. These symptoms are not unlike those experienced following the consumption of DSP toxins.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. Portugal.

7. but all contain a spirocyclic or cyclic imine function within their structure. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. though the possibility exists that they may inhibit other phosphatases. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. which are particularly amenable to detection by positive ion electrospray mass spectrometry.9.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. and ASP. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. However. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . 7. 85). AZAs also display cytotoxicity towards various mammalian cell lines (83.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2.10 Cyclic imines These toxins are represented by several distinct structural types. 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. and closely related ptierotoxins (Figures 7. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action.12). Using the same approach. Once again. 91. PSP. 92). 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). Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. and necrosis of epithelial cells on the microvilli (91). The mouse bioassay has been used to monitor shellfish extracts. parenchymal cell pyknosis in the pancreas. Based on IP minimum lethal dose data. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. the spirolides. and erosion and bleeding in the stomach. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). the pinnatoxins.10 – 7. 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. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. 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. 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. abnormalities in the thymus and spleen.

it was proposed that a dinoflagellate identified as Gymnodinium spp.. At the time of the initial observation of gymnodimine.10. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . 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. selliformis (97). but they are included here for completeness. Subsequent taxonomic studies revised this again to a new dinoflagellate K. Some time later. two additional congeners. 7. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis).FISH AND SHELLFISH TOXINS health.10.10. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin. 95). cockles. they are sometimes referred to as emerging toxins of uncertain human health concern. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94.1 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. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. Structure of the cyclic imine gymnodimine 7. surf clam and New Zealand abalone. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7.520 Da (94. 161 .1. Pacific oyster.10). 95). Fig. In only two cases (gymnodimines and spirolides) has a planktonic source been identified. scallops. 7.1 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.

Based on this diminished oral toxicity. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode.900 µg/kg). The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. and lack of responsiveness to stimuli.1.3 Detection methods The characteristic “fast-acting. The time lapse between ingestion and death is between 5 and 15 minutes. 7.11). and this is the detection method of choice. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 .10.FISH AND SEAFOOD 7. 95) (Figure 7. Most cyclic imines lack an extended chromophore.100). abdominal breathing. Reduction of the imine group eradicated the toxicity. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. and so spectroscopic detection by UV methods is not favoured. They were originally discovered in extracts of mussels (M. almost a dozen derivatives in total have been identified to date (94. 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 . 95).2 Toxicology A review of the toxicology of all cyclic imines has been published (98). edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay.10. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds.1. 7. 50%) of 96 µg/kg. 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. Oral administration of the toxin to mice by gavage resulted in about an 8. paralysis of the hind quarters and eventually complete immobility. together with a lack of evidence of human intoxication. However. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. There is a rapid onset of symptoms including respiratory distress. Whether isolated from dinoflagellate cells directly or from shellfish tissue. it has been stated that regulation against gymnodimine is not required (98). finishing with 100% methanol. In particular. A later study using purified toxin established an LD50 dose (Lethal Dose. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. Since their original discovery which resulted in the characterisation of spirolides A-F (99.10.

Fig. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada.10. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). Structural differences among the various spirolides result from variations in the number of methyl groups and the sequence of spiroketal rings 6-5-5 in one group and 6-6-5 in the other. by gavage (87 . southern Ireland. or by oral ingestion (381 .2. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins.11. those spirolides possessing the vicinal dimethyl grouping (see Figure 7. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . Structure of four spirolide toxins A-D 7. 7. the western coast of Scotland. All data reported here are for desmethyl spirolide C.2 Toxicology The spirolides are the most studied group of the cyclic imines. fasted mice). in fjords in Denmark and Norway. fasted mice).166 µg/kg.2.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). While oral toxicity is considerably less than by IP injection. 7.11.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. Interestingly.8 µg/kg). 95).10. the gulf of Maine.707 µg/kg. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94.

While the mechanism of action remains to be clearly defined. A number of methods following this procedure have been described (94.12). Fig. which includes two diastereomers. To date.B contain a spiro imine moiety similar to that found in the spirolides.10. and in fact these compounds share a number of other structural features.2.FISH AND SEAFOOD (100). consistent with modulation of acetyl cholinergic receptors (103).3 Detection methods As discussed for gymnodimine. 95. 7. 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. However. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. Structure of the pinnatoxin group and the closely related pteriatoxins 164 .12. 7. the mouse bioassay can be used to detect these fast acting toxins. four members of the group have been identified – two of them being the C-34 diastereomers. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. 102).3 Pinnatoxins and pteriatoxins Pinnatoxins A . experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C.10. Three members of this group have been reported. 7. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry.

1 Origins and distribution Shellfish of the genus Pinna. but this remains to be conclusively established (106). while pinnatoxin D was essentially inactive. 99%) values for pinnatoxins A-C are in the range (22 . This is generally a result of increased monitoring and continually improving detection technologies and methods. 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. which occupy tropical waters around Okinawa and the south east coast of China.11 Conclusion There has been considerable progress in the detection of known marine toxins. However. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105).3. With increasing knowledge of the chemistry of toxins. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins.3. 7.100 µg/kg). Only the acute toxicity in mice following IP injection has been reported. While this is recognised as a real possibility for a wide range of toxins. There is strong circumstantial evidence to suspect that these toxins. which in many cases are now found to be more widespread than previously believed. 7. Coupled with this. to date action or 165 .most likely a dinoflagellate. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other.3. have long been suspected of causing human illness after ingestion. The LD99 (Lethal Dose. there is no compelling evidence to suggest that they have resulted in human toxicity (98).10. new toxin groups such as the azaspiracids and cyclic imines have also been reported. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures.3 Detection methods No formal detection methods for these compounds have been reported.10. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. and more advanced means of detection. 7. The LD99 dose for the pteriatoxins is reported to be in the range (8 .180 µg/kg).2 Toxicology There is little information describing the toxicology of these compounds (98).10. arise from an algal source . like the gymnodimines and spirolides. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). Despite this progress.FISH AND SHELLFISH TOXINS 7.

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

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.

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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.

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8. HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
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
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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.

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operation or stage in the food chain including raw materials. 8. procedures. 7. 6.A point.The application of methods. ingredient and packaging buyers. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. Determine the CCPs. from primary production to final consumption. quality assurance or control. food microbiology. food chemistry or engineering. Establish the critical limit(s). An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available.Obtaining evidence that the elements of the HACCP plan are effective. 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. Figure 8. in addition to monitoring to determine compliance with the HACCP plan. Verification . Establish a system to monitor control of the CCP. The team should also include 177 . 3.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . tests and other evaluations.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. 5. Conduct a hazard analysis. 8. procedure. 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 procedures for verification to confirm that the HACCP system is working effectively. Establish documentation concerning all procedures and records appropriate to these principles and their application. Each stage is discussed below in detail. 2.1 Assemble HACCP team HACCP requires management commitment of resources to the process.1 is a flow diagram illustrating this 12-stage logic sequence. Validation . hygiene managers.3. 4.

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

processing conditions (e.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.). It is also important to identify the consumer target groups. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. including practices onboard fishing vessels.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora. heat treatment. pH. physical and chemical structure (including water activity (aw). etc.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.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e.1. may need to be considered specifically.g. etc. In addition. such as children or the elderly.3. storage and distribution conditions and instructions for use. packaging.2 Describe the product It is important to have a complete understanding of the product. there may be species-specific hazards related to certain fish and shellfish. is a cooking process required?).3. fermentation. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . 8.3. The description should include information such as the product name. naturally present on fish from warm waters (> 15 °C). aquaculture drug residues and potentially pathogenic Vibrio spp. or in aquaculture operations. freezing. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. contaminated fish and shellfish should not be used. composition. marine toxins associated with particular fish and shellfish species. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation.3.). These factors are likely to have a significant effect on some potential hazards. such as enteric viruses in shellfish. 8. Vulnerable groups of the population. which should be described in detail. However. If the processing does not include a kill step. 8.g. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. 8. Expertise on such inherent hazards.3. shelf life.

3. conduct a hazard analysis. ingredient specifications. 8. The operation should be observed at all stages. survival or multiplication of pathogenic microorganisms. and identify any measures to control identified hazards The HACCP team should list all hazards that may reasonably be expected to occur at each step in the production process.6 List all potential hazards associated with each step.3. because the hazard analysis and decisions regarding CCPs are based on these data.FISH AND SEAFOOD product. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. It is essential that the diagram is accurate. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. it is essential that the details of each step are fully appreciated and recorded. 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 should also include details of CCP steps. Ideally. If HACCP studies are applied to proposed new process lines or products. and therefore a detailed description of the procedure applied must be available. or circumstances of any reclaim or rework activity. However. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects.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. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. features of equipment design. For example. once determined. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. It may include details of the factory and equipment layout. time/temperature data.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. 8. cleaning and hygiene procedures and storage conditions. 180 ● ● .4.3. the qualitative and/or quantitative evaluation of the presence of hazards. 8.

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

it is necessary to specify both time and temperature for a thermal process.FISH AND SEAFOOD 8. structured approach to decision making. but for which no control exists at that step or any other.7. chemical. If a significant hazard has been identified at a step where control is required for safety. 8. 8.1 Fish and seafood Again.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. More than one critical limit may be defined for a single step.3. 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 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.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. Where possible. given the enormous variety of fish products and processes in use. For example. application of the decision tree should be flexible.3. it is not possible to generalise on likely CCPs.3. then the process must be modified to include a control measure. However. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. and the reader is referred to the appropriate product chapter. biological or sensory parameters. it can be said that effective control measures are likely to include the following: • • • • • 8. critical limits should be specified and validated for each CCP.3. Some examples relevant to fish products are: • • • Time and temperature limits for pasteurised and cooked products Temperature limits for chilled fish Brine concentration in cured fish 182 . Criteria used to set critical limits must be measurable and may include physical.8.2) to provide a logical. and its use may not always be appropriate. However.

HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE

• • • 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
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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.

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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

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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.

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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.

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

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

24.1991.9. 190 • • . J. packaging. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin.B. 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. as last amended by Regulation (EC) No 806/2003.7. Subsequently. storing. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. 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. transportation.1991. as last amended by Regulation (EC) No 806/2003.1992.9. manufacturing.15). 41). 7. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004.2 Legislative Structure From 1 January 2006. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. distribution. handling and offering for sale or supply to the consumer.1). and sets out what those enforcing the provisions have to do. 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. p. p. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. European Communities L 268. p. A number of more detailed implementing and transitional measures have been adopted at EC level. processing. 24. N. 9. 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. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No.

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. national legislation in the form of a Statutory Instrument (S.g. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. There are some exemptions. the legislation is directly applicable law. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production.11) The EU hygiene regulations apply to all stages of food production including primary production.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. of stock farming. right through to the implementation of procedures based on HACCP principles.1993.I. 9. setting offences. Additionally the requirements of EC Regulation 853/2004 must be complied with. 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. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. 191 . 1 . for example. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. and binding in its entirety on all member states from the date of entry into force.) in England. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. 2006 No. food storage that is for private or domestic consumption. penalties and powers of entry. Although the regulations have the force of law. 2073/2005. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No.I. with primary production. is required to give effect to the EU regulations. As regulations. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. and equivalent legislation in Scotland. 852/2004. p. e. Wales and Northern Ireland). Primary products are defined as products of primary production including products of the soil. revocation of existing implementing legislation.7. etc.14. domestic preparation or handling.3 Regulation (EC) No. which will be covered later in this chapter. 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 Food Hygiene (England) Regulations 2006 (S. Wales and Northern Ireland. and of hunting and fishing. 19.

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9.3.1

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: 9.3.2.1 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.

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Additionally, there are requirements for provision of adequate lavatories, basins, ventilation, lighting and drainage.

9.3.2.2 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.

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9.3.2.3 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.

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9.3.2.4 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.

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9.3.2.5 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.

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9.3.2.6 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.

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9.3.2.7 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.

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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.

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9.3.2.8 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. 9.3.2.9 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.

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9.3.2.10 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 9.3.2.11 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). 9.3.2.12 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.

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9.3.3

Registration

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:

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

4.2 Form of the identification mark (i) The mark must be legible and indelible. 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. EK. and the characters easily decipherable and be clearly displayed. the new mark must show the approval number of the establishment where this occurred. EÜ.FISH AND SEAFOOD 9. Section I of Regulation (EC) 853/2004 are given as follows: 9. 198 . Extracts of the requirements of Annex II.2.1 Identification marking An identification mark needs to be applied following the requirements of Annex II.2. EY. EB or WE.4. EK. 9. please refer to Regulation (EC) 853/2004. EC. EG. the code would be UK.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. (iv) If the identification mark is applied in an establishment within the Community. For the UK.4. EO. Section I of Regulation (EC) 853/2004. the mark must be oval shaped and include the abbreviation CE.1. the mark must show the approval number of the establishment. the food business operator may apply the same identification mark to both types of food. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not.2. (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. ES. a new mark must be applied to the product. (iii) Additionally. EF. (ii) However.1.4. (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. In this case. for other codes. when a product's packaging and/or wrapping is removed or is further processed in another establishment.2 Fishery product requirements 9.

the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . its wrapping or the packaging.3 Method of marking (i) The mark is to be applied directly to the product. the colourings used must be EC authorised colours. it is sufficient to apply the mark to the exterior of that package only. a) requirements for establishments. wrapping or packaging in another establishment. With the exception of Chapters I and II. echinoderms.1. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. as amended. Extracts of the requirements of Annex III.e. Chapter IV. C and D. specifically relating to fishery products are given below.LEGISLATION 9. Chapter III. the wrapping or the packaging. Therefore. The requirements of this section accompany those in Regulation (EC) 852/2004. This section does not apply to bivalve molluscs. Section VIII of Regulation 853/2004. (iii) In the case of liquid. processing.4. it applies to those animals when not placed on the market live. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. reference should be made to the actual Regulation. Parts A.2. Part A and Chapter V apply to retail.2. including vessels. they would need to be obtained following Section VII on live bivalve molluscs. 9. and fishery products carried in bulk. granulate and powdered products of animal origin carried in bulk. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. i. and are intended for further handling. or be printed on a label affixed to the product. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. tunicates and marine gastropods when placed on the market live. (v) When the mark is applied directly to products of animal origin.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. the mark may be applied to the outer surface of the container or packaging. (ii) For products of animal origin that are placed in transport containers or large packages. for full requirements.4. the approval number of the establishment and the abbreviation for the community where appropriate.

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

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

the product must be kept at no more than -9 °C. 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. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. The brine must not be a source of contamination. c) Fishery products. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption. Handlers may use spiked instruments to move large fish or fish that might injure them. must be chilled or.4. other than those kept alive.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. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. chilled and wrapped. where fishery products are displayed for sale. 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 . e) In cases where whole fish intended for canning is frozen in brine. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above.Requirements during and after landing 9. B. b) Factory vessels on board which crustaceans and molluscs are cooked.2.2. Hygiene requirements Hygiene requirements refer to the following: a) Parts of vessels used for storage of fishery products must be cleaned and maintained in a good state with no contamination. if this is not possible. provided no damage occurs to the flesh. Chapter II . do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. (ii) In the case of food business operators responsible for auction and wholesale markets etc.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. landed as soon as possible.

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

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

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

and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing.2.4. 206 .2. Fresh. (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. Molidae.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.2. and when fishery products are wrapped on board fishing vessels. 9.7 Chapter VII . The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption. thawed unprocessed fishery products.4. food business operators must ensure that wrapping material is not contaminated and is stored in such a way that it is not exposed to contamination and is easy to clean where necessary. In the case of whole frozen fish in brine intended for the manufacture of canned food. E. it should be kept at no more than -9 °C. Fishery products contaminated with parasites may not be placed on the market for human consumption. 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. Diodontidae and Canthigasteridae.2.Storage of fishery products (i) Fresh fishery products. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum.FISH AND SEAFOOD D. prepared and processed fishery products belonging to the family Gempylidae.6 Chapter VI . Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. 9.

(c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. 2074/2005. species belonging to the Gadidae family. species belonging to the Merlucciidae family.Transport of fishery products The above points for Chapter VII apply in relation to transport. provided that the journey is short and the competent authority allows this.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. melt water must not remain in contact with the products. 9. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. Helicolenus dactylopterus. 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. (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 . (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. 9.4.2. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. The temperature of frozen fishery products must not fluctuate above -15°C. Sebastichthys capensis.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.). 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.2.2. (ii) If fishery products are kept under ice.8 Chapter VIII .4.

please refer directly to the regulation. and point 3 of Chapter VII applies to retail. reference should be made to the actual 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.4. 9.3. (b) In the case of other operations. where the raw material complies with points (a). Live bivalve mollusc requirements - 9. specifically relating to bivalve molluscs are given below. for full requirements. The copy of 208 . Extracts of the requirements of Annex III. as amended. they supplement the requirements of Annex II to that Regulation. Section VII of Regulation 853/2004. VIII and IX. Chapter IX applies to pectinidae harvested outside those areas. The section is split into 9 chapters for which a summary is provided as follows: Chapters I to VIII apply to animals harvested from production areas that the competent authority has classified in accordance with Regulation (EC) No 854/2004.4. they supplement the requirements of Annex I to that Regulation. VI. this Section also applies to live echinoderms. The information required in the registration document is given in point 4 and includes the health status of the production area. shellfish species etc. taken from at least three different points and mixed together by grinding. 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.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. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. (b) and (c) of Part B(1) of that Chapter. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. Chapters V. however.3 The regulation details specific hygiene requirements for bivalve molluscs.FISH AND SEAFOOD Regulation (EC) No 853/2004. The sample must consist of about 100 g of flesh. tunicates and marine gastropods. Apart from the provisions on purification.1 Chapter I .

Hygiene requirements for the production and harvesting of live bivalve molluscs A. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. Requirements for production areas (i) The competent authority classes (classed A. 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. 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.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. Transport needs to allow sufficient drainage for best survival conditions. 209 . (ii) Provided the requirements of chapter V are met.Class A means they can be placed directly on the market . C.2 Chapter II . processing or relaying. collection from: . B. B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with. 9. poles or other fixed means are to identify the boundaries of the sites.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market .LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority.4.3.

B. and there are requirements for tanks and water storage containers. the purification system to allow them to resume /maintain filter-feeding activity. 800 micrograms per kilogram. requirements if there are many batches of live bivalve molluscs. Requirements for dispatch centres Handling should not result in contamination to any degree. 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.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements.4.3. the levels of marine biotoxins should not exceed the following limits.5 Chapter V .3 Chapter III .4 Chapter IV . (iv) yessotoxins. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. 210 .3.Hygiene requirements for purification and dispatch centres A.4. 160 micrograms of okadaic acid equivalents per kilogram. for: (i) paralytic shellfish poison (PSP).4. Before dispatch.3. 9.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. containers and a label to identify purification. 1 milligram of yessotoxin equivalent per kilogram. Requirements for purification centres There are requirements for live bivalve molluscs to be washed. dinophysistoxins and pectenotoxins together.FISH AND SEAFOOD 9. (ii) amnesic shellfish poison (ASP).

9. Registration document requirements also apply. 9.7 Chapter VII .4.3. by way of derogation from directive 2000/13/EC.3. Also. dispatch centre or processing establishment.1). The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.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. Finally.4. the species of bivalve mollusc and date of packaging (at least the date and month) are required.3.6 Chapter VI . (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. must be waterproof.4.2. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards. they cannot be re-immersed in or sprayed with water. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.3. 160 micrograms of azaspiracid equivalents per kilogram.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. 211 .4.9 Chapter IX . 9.8 Chapter VIII . Requirements of Chapters III and IV are to be complied with. (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.4.Identification marking and labelling In addition to general identification marking (contained in 9. the label.LEGISLATION (v) azaspiracids. including the identification mark.

5. In this regulation. or - - 212 . Therefore the health standards for fishery products as previously stated in Annex III. Fishery products in particular must comply with the requirements of Annex III of the regulation. echinoderms. 9. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. 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. Chapter V of Regulation 853/2004 apply. chemical. (ii) fishery products not complying with the requirement concerning biotoxins. 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. 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. This refers to official controls or production and placing on the market of fishery products. or (iii) bivalve molluscs. Section VIII.FISH AND SEAFOOD 9. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. they derive from: (i) poisonous fish.5 Regulation (EC) No. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic.

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

5. 2073/2005. 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. boundaries or class of a production area or its closure. opening or closing a production area.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. E. F.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004.5. 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. the competent authority is to close the production area. 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. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. which has applied since 1 January 2006.2 Chapter III .2.6 Regulation (EC) No. 9. 9. Procedures for re-opening a closed production area are stated D. establishes microbiological criteria for a range of foods. 214 . 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. 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.FISH AND SEAFOOD C. 9.

applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. and the legislation applies to all businesses involved in food production and handling.4 referring to fishery products. Two kinds of criteria have been established: food safety criteria. 9. based on the absence. volume.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. 9. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product.II. per unit(s) of mass.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria.I. ensuring that foods being placed on the market do not pose a risk to human health. The criteria are outlined in Table 9. with chapter 2. presence or number of microorganisms. area or batch. The relevant criteria are outlined in Table 9. a batch of foodstuffs or a process. 215 .6.6.LEGISLATION The aim of this legislation is to complement food hygiene requirements. and/or on the quantity of their toxins or metabolites.

monocytogenes.3 Ready-to-eat foods unable to support the growth of 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. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 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 1.2 Ready-to-eat foods able to support the growth of L. monocytogenes L. other than those intended for infants and for special medical purposes EN/ISO 11290-1 Products placed on the market during their shelf life Before the food has left the immediate control of the food business operator.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life .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.TABLE 9.

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. coli (15) 1.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1.25 Live bivalve molluscs and live echinoderms.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. tunicates and gastropods 9 (18) 2 E.27 Fishery products which have undergone enzyme maturation treatment in brine.17 Live bivalve molluscs and live echinoderms. manufactured from fish species associated with a high amount of histidine (17) Histamine .

Clupeidae. products with a shelf-life of less than five days shall be automatically considered to belong to this category. wine. Single samples may be taken at retail level. Malle P..1.25. A pooled sample comprising a minimum of 10 individual animals. cider. Coryfenidae. m = M. products heat treated in their final package). Valle M. 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... — — bottled or packed waters. excluding sprouted seeds. 43-9. — bread. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002.92. sugar. AOAC Internat. Dervin C. honey and confectionery. Pomatomidae.0 and aw ≤ 0. Other categories of products can also belong to this category. 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. This criterion shall apply to products before they have left the immediate control of the producing food business operator. Scombresosidae.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. 2. 79. 1996. monocytogenes. shall not apply.1 . Engraulidae. Particularly fish species of the families: Scombridae. Malle P. when recontamination is not possible after this treatment (for example. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter.. including cocoa and chocolate products. according to which the whole batch is to be deemed unsafe. when he is not able to demonstrate. Bouquelet S. Bouquelet S. 5 This criterion shall apply if the manufacturer is able to demonstrate. The most recent edition of the standard shall be used. soft drinks. beer. to the satisfaction of the competent authority. Products with pH ≤ 4. spirits and similar products. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. that the product will not exceed the limit 100 cfu/g throughout the shelf life. biscuits and similar products. products with pH ≤ 5.4 or aw ≤ 0. E. uncut and unprocessed vegetables and fruits. J. Duflos G. References: 1. Assay of biogenic amines involved in fish decomposition. to the satisfaction of the competent authority. live bivalve molluscs. coli is used here as an indicator of faecal contamination. c = number of sample units giving values between m and M For point 1. subject to scientific justification.94.. — fresh. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 .

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

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. 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 .TABLE 9.

— acceptable.7 Food Hygiene (England) Regulations 2006. if one or more of the values observed are > M or more than c/n values are between m and M. Hot-held foods (food having been cooked or reheated and is for service or on display for sale) must not be kept below 63 °C. if all the values observed are ≤ m. 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.I. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health. and the rest of the values observed are ≤ m.II The limits given refer to each sample unit tested. which overrides the 8 °C requirement. 2006 No. — unsatisfactory. 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. For example. Schedule 4 of the Food Hygiene (England) Regulations 2006.7. The regulations prescribe a chilled food holding temperature of 8 °C or less. — acceptable. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. 2006 No. and the rest of the values observed are ≤ m. The regulations provide for defences in relation to upward variations of the 8 °C temperature.1 In the U. 14 details temperature control requirements for foods in general. had not been 221 . if a maximum of c/n values are between m and M.LEGISLATION 1 n = number of units comprising the sample. S. 2 Interpretation of the test results relating to Table 9. 9.I. the defendant will need to prove that the food was on service or display. — unsatisfactory.K. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. The defendant may be required to produce well-founded scientific proof to support his claims. with chill holding tolerance periods. if all the values observed are ≤ m. The test results demonstrate the microbiological quality of the process tested. E. 14 (Hygiene requirements specific to the U.K) Temperature control requirements 9. tolerance periods for chill-holding of foods and hot-holding variations. S.

K. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. 2006 No. 2006 No.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 .FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. For other food. 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. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions. 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. 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. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety.I.S. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. 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.gov. and was kept at above 8 °C or the recommended temperature. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S.food.I. was kept at above 8 °C or the recommended temperature for an unavoidable reason. or. for a limited period consistent with food safety. 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. including: Shelf life: where the minimum durability or 'use by' indication is required according to Regulation 20 or 21 of the Food Labelling Regulations 1996 (form of indication of minimum durability and form of indication of ‘use-by date’) the period up to and including that date. S. Alternatively. 9. 3). such as that below.8 Guidance In the U. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing.

2076/2005.4. 2076/2005. 206). Regulation(EC) No. Regulation (EC) No. 22) as amended by Regulation (EC) Nos.4. 852/2004 is now set out in a Corrigendum (OJ No. 854/2004 is now set out in a Corrigendum (OJ No. Regulation (EC) No. 2074/2005.12.6. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 30. namely.2004. L226. 14. 3) as amended by Regulation (EC) No. 62) as amended by Regulation (EC) No. 55).6. 2074/2005 and Regulation (EC) No. 223 . Regulation (EEC) 2136/89. The revised text of Regulation (EC) No.2004. The revised text of Regulation (EC) No. L226. L139. 25. 853/2004 is now set out in a Corrigendum (OJ No. The revised text of Regulation (EC) No. 22. 30. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41.4. 30.2004. L338.6. L226. 25. as amended. L139. these are not be discussed here. Regulation (EC) No. L283.2004.2006. Regulation (EC) No.10. 25. 2074/2005. 2076/2005. laying down common marketing standards for preserved sardines. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 2076/2005. 1663/2006. 83) as amended by Regulation Nos 882/2004. 2074/2005.9 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. 1).10 Further Reading Regulation (EC) No. 2075/2005 and Regulation (EC) No. Regulation (EC) No. 2073/2005 on microbiological criteria for foodstuffs (OJ No. L139. 9. 1688/2005. 1.2004. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. 1662/2006.2004.2005. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. 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. Commission Regulation (EC) No.LEGISLATION 9. 1019/2008 and as read with Regulation 2073/2005. as read with the corrigenda at OJ No. 1441/2007.

22.2001. 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 854/2004.10. 22. 22) Commission regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products as amended by Commission Regulation (EC) No 1792/2006 of 23 October 2006 (OJ L 278.FISH AND SEAFOOD Commission Regulation (EC) No 2074/2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC)No 882/2004 of the European Parliament and of the Council. (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. 23.12. 6) 224 .1.2000.2005. 21. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004.12. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.2005.

5. Morphology Gram-negative rods. and 2.1. a 10 to 1.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.1.1 Aeromonas spp.1.0 x 1.5 pH Under otherwise optimal conditions. At 5 °C. Aeromonas has the potential to grow over a pH range of ca 4 .0 . and D-values at 45 and 51 °C have been reported as 29.1. The optimum growth temperature is 28 °C (2). PATHOGEN PROFILES 10.5.5 µm 10.1.45 °C (1). At pH 4. 10.1 10.10 (3).2 Oxygen requirements Facultative anaerobe.000-fold increase in number is observed after one week in food systems (1).1.3 . under optimal incubation temperatures. Some death will occur at this pH if the temperature is 4 °C. 10.3.5 minutes. respectively (1). The minimum pH value for growth is thought to be pH 5. no growth is observed (1).3 minutes.3 Temperature Aeromonas grows at temperatures of 2 . 225 . 0. 10.10.

1 Morphology Gram-positive. 6).7 .4 x 1. 226 .2. which may be inactivated by heating at 80 ºC for 20 . and is sensitive to > 4. botulinum is an obligate anaerobe. being capable of slow growth and toxin production at low temperatures . forming at least seven different toxins.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 .30 minutes. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth.2 Clostridium botulinum Seven different types of C.22. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C. B.2. All C.even as low as 3 ºC (7). F are the types that are responsible for most cases of human botulism (5. E and. 10.2.2 Oxygen requirements Although C.0 µm. but the interior of the food may do so. or at 90 ºC for a few seconds. There are four main groupings of the organism. types B and F can be either.3 Temperature All strains of C. 0.2% sodium chloride (NaCl) (1). but the spores of this organism are more so.5 . Group I will not grow at temperatures of 10 ºC or less. 10.FISH AND SEAFOOD 10. to a lesser extent. at 85 ºC for 5 minutes. A to G. 10.2.2.4 Heat resistance The vegetative cells of C.5% NaCl (4). botulinum is prevented.1. and groups I and II are those responsible for the botulism cases. botulinum are known. 10. botulinum grow reasonably well in the temperature range of 20 45 ºC. botulinum are not particularly heat resistant. 10. Types A. Thus an aerobically packed product may not support the growth of the organism on the surface. All type A strains are proteolytic and type E strains are usually non-proteolytic. spore-forming rods. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. but Group II strains are psychrotrophic. botulinum types produce heat labile toxins.

In commercial situations. These values have been established under carefully controlled laboratory conditions.1 Clostridium perfringens Morphology Gram-positive spore-forming rods. pH and temperature.3 . 10. vacuum packed. parcooked meals with pH value > 5.2. they may survive mild heat treatments (70 . and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. perfringens . These concentrations correspond to limiting aw of 0.6 and pH 5. The organism will grow well in the centre of meat or poultry dishes.94 for Group I and 0. 11)).0.21 minutes) (9). particularly by cooking. 10. grow on the surface of foods unless they are vacuum or gas-packed. therefore.10. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I.0 µm.0 and aw > 0. Note also that if humectants other than salt are used.3 10.2.PATHOGEN PROFILES 10. botulinum depends on solute.9 x 2. botulinum spores The most heat resistant spores of Group I C.1. 10. respectively (8.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. 10. 227 .2. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains.is an obligate anaerobe. where oxygen levels are reduced.97 for Group II (5).5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4.g.0 .2 Oxygen requirements C. safety margins must be introduced to allow for process variability. 0. the minimum aw for growth may be lower.97) (10.3.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. It will not. 9).3. However.like other clostridia . botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0.7 Characteristics of C.

Recorded heat 228 .10 ºC. perfringens is not tolerant of low water activities.3. As in the case of other factors limiting the growth/survival of this organism. etc. Under otherwise ideal conditions.3 Temperature The most significant characteristic of C. While some growth can occur at 50 ºC. perfringens is not a tolerant organism with respect to pH. type of solute. 10. perfringens has the potential ability to grow within the temperature range 15 .8% inhibit growth of most C. perfringens.it is destroyed by heating at 60 ºC for 10 minutes (14.3. 15. The lowest aw recorded to support the growth of C. will survive greater extremes of pH (and aw) (12. 18). (It is unusual for spores to be formed in foods after the growth of this organism (14)). 10. 13). perfringens appears to be 0. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. perfringens in vacuum packed beef (18). It grows best at pH values between 6 and 7 (the same pH as most meats).5 minutes (17). perfringens in roast beef at 60 ºC is 14. lower concentrations may be effective in combination with other factors.3. the limits for aw are affected by temperature. Its optimum temperature for growth is 43 .3. In addition. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. perfringens strains.5 pH C. 16).97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. perfringens spores The spores of C. Salt concentrations of 6 .4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C.7 Characteristics of C. however.45 ºC. depending on strain and other conditions. vegetative cells die rapidly (12). 10. Spores.6 Aw/Sodium chloride C. 0 . perfringens can vary quite considerably in their heat resistance. At cold temperatures. 10.3. 13). although C. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures.FISH AND SEAFOOD 10.93 and 0. The D-value for C. pH. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12.50 ºC. Some studies indicate that the presence of 3% NaCl delays growth of C.3. and their heat resistance is also affected by the heating substrate. the enterotoxin is not heat resistant .

4 10.5 pH The ability of Listeria to grow at different pH values (as with other bacteria) is markedly affected by the type of acid used. being potentially capable of growing . monocytogenes is reported to be 45 ºC (20).2. however.19 minutes.4. 0. using hydrochloric acid as an acidulant).0.6 . it is not a sporeformer.3 . The upper temperature limit for the growth of L. and where acetic acid is used as an acidulant. D-values for L. pH < 5. Under ideal conditions.1 Listeria spp.0 µm. respectively (21). 60 and 65 ºC were reported as 10.4.4 . the lowest limit for growth is likely to be considerably higher .3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. Morphology Gram-positive short rods. -0. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.2 has been suggested as the lowest working limit (22). However. growth at low temperatures can be very slow. Its optimum growth temperature. requiring days or weeks to reach maximum numbers.4. so can be destroyed by pasteurisation. the organism is able to grow at pH values well below pH 5 (pH 4. and temperature.23. is between 30 and 37 ºC.64.2 Oxygen requirements Aerobe or microaerophilic.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17.98 and 0. 229 . monocytogenes is not a particularly heat resistant organism. however.0 minutes for heat resistant spores to 1. 10.3 is the lowest value where growth has been recorded. 10. In foods.especially at refrigeration temperatures.5 . 10.2.4. 10.albeit slowly .4 ºC is probably a more likely minimum for growth in foods (19). 1.at refrigeration temperatures down to.4 Heat resistance L. or even below 0 ºC.4.8 minutes for heat sensitive spores (12). monocytogenes in crawfish tail meat at 55.5 x 0. 10.

10. Morphology Gram-negative short rods.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.97.1 Salmonella spp.0 .8 .1 Plesiomonas Morphology Gram-negative.7 x 1.6. The bacterium may survive for long periods at aw values as low as 0. peritrichous flagella.2 Oxygen requirements Facultative anaerobe. 10.6 10.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24). but has been shown to be able to grow at an aw level of 0. at salt levels found in foods (10 .5.90.8.83 (20). 10.5 10. 0. 10.0 µm.5 . 10.0. monocytogenes is quite tolerant of high NaCl/low water activities.5.3 Temperature Plesiomonas can grow in the temperature range of 8 .1. or even grow. 230 .5. 10. 25).4.5 . It is likely to survive. 0.6 Aw/Sodium chloride L.5 pH Plesiomonas has a pH range for growth of pH 4.5.FISH AND SEAFOOD 10.5 (24.0 x 1 . It grows best at aw of ≥ 0.3.5. 10.3 µm.45 ºC.12% NaCl or more). 25).5. the optimum being 30 ºC (23).

231 . strains vary in their ability to withstand heating. 10. Salmonella senftenberg 775W is about 10 . temperature and pH.4% NaCl. Salmonellae are quite resistant to drying.0. or possibly 0. Furthermore. although salt tolerance increases with increasing temperature (28). pasteurisation and equivalent treatments will destroy the organism under normal circumstances.PATHOGEN PROFILES 10.37 ºC. The optimal pH for Salmonella growth is between 6. therefore. Growth is slow at temperatures below about 10 ºC.945.6.6. some strains are able to grow at temperatures as low as 4 °C (26).5 .2 Oxygen requirements Facultative anaerobe. The growth of Salmonella is generally inhibited by the presence of 3 .6 Aw/Sodium chloride Where all other conditions are favourable.6.5.6. a heat resistant organism. under otherwise ideal conditions.3 Temperature Salmonellae can grow in the temperature range of 7 .7.8 .93 (as reported in dried meat and dehydrated soup). and with an appropriate acid.48 ºC. depending on the type of acid and temperature.20 times more heat resistant than the average strain of Salmonella at high aw (27). However. Salmonella has the potential to grow at aw levels as low as 0.4 Heat resistance Salmonella is not a spore-forming organism.5. substrate.9.6. with a z-value of 4 . and appropriate heat treatments must be determined experimentally for low-aw foods. D60 ºC values normally range from about 1 to 10 minutes.5 pH Salmonella has a pH range for growth of pH 3. Salmonellae are quite resistant to freezing. Some death will occur at pH values of less than about 4. depending on serotype. However. the optimum being 35 . high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. 10. 10. 10.5 ºC. It is not.

It can survive well at low temperatures.7. depending on aw. fat content. In meat macerate. with an optimum of 35 .20 minutes.0. aureus is more limited under anaerobic than aerobic conditions. Staph. Under most circumstances.37 ºC (29). The limits for toxin production are also narrower than for growth. aureus viability.5). 10. aureus than citric acid).0 . aw.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH.3 Temperature Under otherwise ideal conditions. the D60 ºC value is 2 .1 Staphylococcus aureus Morphology Gram-positive cocci. stressed cells can also be less tolerant of heating. Most strains of staphylococci can grow within the pH range 4.9 µm diameter. 232 .). however. 10. Freezing and thawing have little effect on Staph.7.5 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. 31).7. As is the case with other bacteria.7 10. 10. 0. aw and temperature (sensitivity to acid increases with temperature).7.2 to 9.FISH AND SEAFOOD 10. under otherwise ideal conditions (29. The following relate to limits for growth only. NB: The growth of Staph.7. aureus can grow within the temperature range 7 – 48. but may cause some cell damage (30).2 Oxygen requirements Facultative anaerobe.5 ºC.3 (optimum 7. 10. the organism is heat sensitive and will be destroyed by pasteurisation.7 .7. etc.

0 (optimum 7.2 Oxygen requirements Facultative anaerobe. but certain strains can grow in 20% NaCl.7. 10.7.86 is the generally recognised minimum in foods (32). pH.5) (29.8. 10. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. strain.99 little or no toxin production in anaerobically packed foods. In general. 0.PATHOGEN PROFILES 10.0 .43 ºC (35).99 aerobically under otherwise optimal conditions.6 Aw/Sodium chloride Staph.8.7. 10. and solute. atmosphere. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant. In general. an aw of 0.8.0.6 µm 10. 233 . 31) between 0.7 Limits permitting toxin production 10 .2. with a temperature range for growth of 10 .2 . 34).9.8 x 1. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. Staph. * dependent on temperature. An effect of increasing salt concentration is to raise the minimum pH of growth.5 .10% NaCl.1 Vibrio cholerae Morphology Gram-negative curved rods. It can grow over the aw range 0.83 . aureus can grow in 7 . Staphylococci are more resistant to salt present in foods than other organisms.4 .3 Temperature The optimum temperature for growth has been reported as 30 ºC. However.8 10.87 and > 0.>0.

10. 10.9. cholerae may persist on frozen fish for >180 days (36).FISH AND SEAFOOD Also. V. 10. it has a pH range for growth of pH 5 .6 (35. under otherwise favourable conditions. respectively. 234 .0.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.8.97 .65 minutes (35). declining in number during storage.0% NaCl (35).5 ºC). It can grow in the presence of 0.4. with an optimum of 0.9. 38).2 Oxygen requirements Facultative anaerobe. 10. 37.9.8. parahaemolyticus is easily destroyed by drying. with optimum at 7. cholerae in crab meat homogenate at 60 ºC is 2.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. but not acid conditions.984 (35). The D-value for V. are 5 and 43 ºC.998.6 Aw/Sodium chloride V.6.4 Heat resistance V. and it is sensitive to refrigeration temperatures (0 . there is evidence that V. cholerae is sensitive to heat at temperatures of >45 ºC.5 pH This organism is tolerant of high pH values. cholerae can grow over the aw range 0.9. but only moderately sensitive to freezing (40).1 . with an optimum of 37 ºC (39). 10.8.9 10. 10.

its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.2 Oxygen requirements Facultative anaerobe. vulnificus is heat sensitive. with a temperature range for growth of 8 .8 .0.5 pH The pH range for growth is 4. parahaemolyticus are not heat resistant.29 minutes. 10.3 Temperature The optimum temperature for growth has been reported at 37 ºC.1 Morphology Gram-negative curved rods. parahaemolyticus will grow at aw levels as low as 0.4% (39.PATHOGEN PROFILES 10.5 minutes. with an optimum pH of 7.11.9. but the optimum concentration is 2 . and are readily destroyed by cooking. V. 10.43 ºC (42).10. The D-values at 55 ºC for clam and crab homogenate are 0.6 .9.10. parahaemolyticus is unable to grow in the absence of salt.922.4 Heat resistance V. The organism can survive for long periods at freezer temperatures (42). respectively (39). respectively.6 (41). 41).9. 10. and 2.10. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).6 Aw/Sodium chloride V. 235 . depending on solute. The optimum concentration of NaCl equates to aw of approximately 0.4 Heat resistance V. 41).98 (35.8.02 . 10.10.5% NaCl. 10.10 Vibrio vulnificus 10. 10.

FISH AND SEAFOOD 10. Faris A.H. Clostridium botulinum. ASM Press. Smith J.10. 5-19...M. Marcel Dekker. Montville T.M. Cliver D. New York. Murrell K.H. Eds. Great Britain. 329-49. Peck M.6 Aw/Sodium chloride V. Blackie.D. Kovacek K.0% NaCl.. 1994. Beuchat L. Kim J. Notermans S. Gorham J. Food Control. 8. 2001. Aeromonas. Volume 5: Microbiological Specifications of Food Pathogens.D. Dodds K. in Food Microbiology: Fundamentals and Frontiers. 10.. London.. Palumbo S.. Hui Y.. 1994. 2.R. Johnson E.H. New York.W. Marcel Dekker Inc.. Eds. Dodds K. Washington D. Clostridium botulinum. Austin J. 2005. Eds. 1-19.5. with an optimum pH of 7.A. 6 (1). in Foodborne Disease Handbook. 1993.8 (42).W. Eds.. References International Commission on Microbiological Specifications for Foods. Volume 1. Ed. Aeromonas hydrophila group. Stelma G. 4.11 1.5 pH The pH range for growth is 5 . Doyle M. in Clostridium botulinum: Ecology and Control in Foods. 6. Aeromonas hydrophila.H. Doyle M. Marcel Dekker. Foegeding P.. 111-89. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’. Novak J... Caister Academic Press.L. with an optimum of 2.5 . in Clostridium botulinum: Ecology and Control in Foods.10.E.R.10. Clostridium botulinum.. in Foodborne Bacterial Pathogens. 9. Bhunia A. 279-303.. Fratamico P.W. 383-408. in Microorganisms in Foods. Hui Y.P. vulnificus grows over the range 0. Marcel Dekker. 1989. 57-63. in Foodborne Pathogens. New York. 1995... Marcel Dekker. Marcel Dekker.N. Bier J.M. Eds. Aeromonas species. Dodds K. Juneja V.. Betts G. Marcel Dekker.L. Clostridium botulinum and Clostridium perfringens.D.O. in International Handbook of Foodborne Pathogens. International Commission on Microbiological Specifications for Foods. Cliver D.. New York. in Foodborne Bacterial Pathogens.. New York. 3. Eds. 97-131. 11.L. Volume 1: Diseases Caused by Bacteria. Hauschild A. in Foodborne Disease Handbook. 2003..O. Doyle M.D.P. New York. 121-76. 10. Ed. Principles of control. Gorman J.N.L. Miliotis M... 1989. Hauschild A. Potential hazards associated with REPFEDS. 5.H. Lund B. Ed. Hauschild A. Stelma G. Murrell K.R.W.W. Diseases Caused by Bacteria.. New York. Microbiology and Molecular Biology. 7.K. Eds. 1-27.P.C. 236 . 1993. 10. 357-67. Gaze J.J.H. Abeyta C.5% (42). 1996.

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

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

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

FISH AND SEAFOOD 240 .

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.org.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.defra.gov.uk or v.org.org.seafish.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 Web site: www.uk Web site: www.uk Web site: www.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.ifm.gsi.scotland.gov.org/ .CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: cawoods@denholmseafoods.gov.holt@ifm.gov.gsi.co.

dk Web site: www.ifpea@eircom.dardni.dk Web site: www.suite. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.is Web site: www.dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .gov. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.is/ 242 .org/fishery/ European Association of Fish Producers’ Organisations H.ie/templates/homepage.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.bim. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo.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.no Web site: www.htm Irish Fish Processors and Exporters Association 25 Kincora Avenue Clontarf Dublin 3 Eire Ireland Tel: + 353 1 833 7882/ 02770249 Fax: + 353 1 833 7882 Email: tormg.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: post@fiskeriforskning. Andersens Boulevard 37.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: matis@matis.eapo.fiskehandlerne.C.gov.uk/index/fisheriesfarming-and-food.uk Web site: www.matis.fao.difta.fiskeriforskning.FISH AND SEAFOOD Department of Agriculture and Rural Development.dk Danmarks Fiskehandlere H.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. 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.org Web site: www.com Web site: www.

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

se Web site: www.who.codexalimentarius.who.uk/scieh.co.int Web site: http://www.uk/index.de 244 .co.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.int Web site: www.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/en/ WHO Regional Office for Europe Scherfigsvej 8 DK-2100 Copenhagen Ø Denmark Tel: + 45 39 171717 Fax: + 45 39 171818 Email: postmaster@euro.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.int/foodsafety World Health Organization (WHO) Headquarters CH .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.who.asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.int/ SIK .who.sofht.euro.scot.scot. Web site: www.net/web/indexen.sik.nhs.uk Web site: www.uk Web site: www.euro.org Web site: www.se/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: fao-who-cc@bgvv.nhs.int.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.org. jsp Food Safety 10 .hps.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: foodsafety@euro.sfam.

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

FISH AND SEAFOOD 246 .

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

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

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. 106 Contacts 241-5 Conway method. to detect NSP toxins 155 ELISA kit (Biosense). 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. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. for detection of DA 148 ELISA kit (DSP-Check). of prepared fish products 44 Escherichia coli. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). 152 Diphyllobothrium. to determine fresh fish spoilage 11 Corynebacteria. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. 149-50. processing 334 Cockles. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . hot-smoking 107 ELISA assay. as autochthonous bacterial flora of fish 3 Crabs. growth and survival in crabmeat 84 growth and survival in cured. of fish – different processes 94 Dun. 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. 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.INDEX pathogen profile 226-7 Clostridium perfringens. adverse condition on poorly preserved fish 110 EC grading scheme. smoked and dried fish 114 Enzymic spoilage. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. as contaminants in scampi 36. source of azaspiracids 159 source of DSP toxins 148-9. to determine fresh fish spoilage 11 Eels. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp.

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

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

reduction in smoking process 108 Norcardiopsis dassonvillei.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. caused by lipid oxidation 11 Oysters. of crustacean shellfish – effects of processing 81-3 of cured. for detection of azaspiracids 160 for detection of ciguatoxins 157 for detection of DA 148 for detection of DSP toxins 150. in crustacean shellfish 867 Norwalk virus. associated with consumption of fermented fish products 137 Oxidative rancidity. shigelloides. as TTX producer 145 Norovirus. 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. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. 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.

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

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

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

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

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