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

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8 Further Reading CRUSTACEAN SHELLFISH 4.5 Pathogens: Growth and Survival 3. 4.8 References MOLLUSCAN SHELLFISH 3.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.3 Processing and its Effects on the Microflora 1.4 Spoilage 1.6 Pathogens: Growth and Survival 1.6 Published Microbiological Criteria 4.7 Published Microbiological Criteria 1. vii . CHILLED AND FROZEN RAW FISH 1.6 Pathogens: Growth and Survival 2.4 Spoilage 4.3 Processing and its Effects on the Microflora 4.3 Processing and its Effects on the Microflora 3.5 Factors Affecting Fresh Fish Spoilage 1.4 Spoilage 3.7 References 4.2 Initial Microflora 3.2 Initial Microflora 4.2 Definitions 2.4 Processing and its Effects on the Microflora 2.1 Definitions 3.6 Published Microbiological Criteria 3.1 Definitions 4.2 Initial Microflora 1. 3.1 Definitions 1.7 References 3.5 Spoilage 2.3 Initial Microflora 2.7 Published Microbiological Criteria 2.1 Introduction 2.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.5 Pathogens: Growth and Survival 4.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.8 References 1.

SMOKED AND DRIED FISH 5.8 Ciguatoxins 7.3 Regulation (EC) No.I.9 Azaspiracids 7.6 Lipophilic Shellfish Toxins (LST) 7. CURED.2 Definitions 8.4 Spoilage 5. 9.1 Definitions 6.3 Processing and its Effects on the Microflora 6. 8.4 Regulation (EC) No.1 Introduction 8.3 Stages of a HACCP Study 8. 14 (Hygiene requirements specific to the UK) 221 9.5 Pathogens: Growth and Survival 5.11 Conclusion 7.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.10 Cyclic Imines 7. EU FOOD HYGIENE LEGISLATION 189 9.2 Initial Microflora 6.4 Spoilage 6. 2006 No.6 Published Microbiological Criteria 5.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.7 Further Reading FISH AND SHELLFISH TOXINS 7. 852/2004 on the General Hygiene of Foodstuffs 191 9.3 Processing and its Effects on the Microflora 5.5. 7.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.7 Food Hygiene (England) Regulations 2006.2 Legislative Structure 190 9. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.4 Implementation and Review of the HACCP Plan 8.1 Introduction 189 9.2 Initial Microflora 5.5 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 6.1 Definitions 5.5 Regulation (EC) No.9 Other Relevant Legislation 223 9. S.3 Tetrodotoxin 7.5 Pathogens: Growth and Survival 6.7 References 5. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.6 References 6.10 Further Reading 223 viii .8 Further Reading FERMENTED FISH 6.1 Introduction 7.6 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 212 9.8 Guidance 222 9.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.

10.1 Aeromonas spp.9 Vibrio parahaemolyticus 10.4 Listeria spp.6 Salmonella spp.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .3 Clostridium perfringens 10.10 PATHOGEN PROFILES 10. 10. 10.10 Vibrio vulnificus 10.5 Plesiomonas 10.2 Clostridium botulinum 10.8 Vibrio cholerae 10.7 Staphylococcus aureus 10.

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

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

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

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

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

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

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

Plesiomonas shigelloides 3 . Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. sewage. numbers of pathogenic bacteria in raw fish tend to be low.0.26 °C 10 .FISH AND SEAFOOD TABLE 1.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. 105 .106 cells/g.19 µg/kg bodyweight from < 102 .II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . 4 .37 °C High for most species.106 cells/g (exception: V.103 High (107 . Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. and mud Ubiquitous.37 °C Unknown Unknown TABLE 1.14 . birds.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.109 cells/g) * Some sources considered L. In addition. and risk associated with the consumption of seafood is low (2. However. estuarine environments. Escherichia coli Yersinia enterocolitica 101 . 4). Toxin levels 0.102 101 . vulnificus) 5 .35 °C 8 .>106 Salmonella spp. stream water. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.1 . monocytogenes as part of the natural aquatic flora It is apparent from the above that there is potentially a very diverse range of organisms present on fish.

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

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

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

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

so lower carbon dioxide concentrations may have better preservative effects. When working with fillets. The eyes turn from convex and clear to concave and opaque. and this pattern is the basis of freshness grading schemes.CHILLED AND FROZEN RAW FISH off-odours. 18) and are probably the most widely used. as traditional spoilage patterns of the external organs are typically very distinct. the gills from pink and shiny.III.4 1.1 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). Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids). loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . 1. or for that matter evaluation of the taste of the fish. A.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. texture. the skin turns from iridescent to dull and bleached with bacterial slime. smell and 9 . with no smell. and the flesh turns from bright and elastic to dull and soft (1).4. Taste panel evaluation of cooked fish may be used to score or grade colour. Various methods for whole fish freshness evaluation by a trained panel have been proposed. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1).60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. reactions caused by activities of the fish’s own enzymes. to brown and slimy with an intense off-odour. Degradation of whole fresh fish stored in ice generally follows a set pattern. and metabolic activities of microorganisms present in the fish. TABLE 1. Over time. C (C=reject) Significantly. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. these are summarised in Table 1. 17. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. Loss of freshness and spoilage cannot be separated as processes. B.

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

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

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 application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks. For fresh fish. The reasons for the differences between tropical.5. but also crucial for the assurance of consumer safety. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 . the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. the results of altering one parameter may have far-reaching consequences on others.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. the lower the temperature of storage. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. The factors are interactive and cannot be completely isolated. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33.2 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). biochemical and microbiological reactions taking place within and on the fish. 1.5 1.5.3 Recognised specific spoilage organisms (SSOs) The degree of spoilage leading to sensory rejection of fish is partly dependent on the perception of the consumer. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. Fish spoil as a result of the chemical. Also. by altering the gas atmosphere (extrinsic). 1. The concept of specific spoilage organisms (SSOs) is not new. For example. 5).and temperate-water fish spoilage rates may be many.FISH AND SEAFOOD 1. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. careful temperature control during storage is not only important in terms of quality loss. the slower the spoilage processes proceed (within limits). The sum of these changes may result in a different spoilage profile. 34). a minority are often associated with the majority of the spoilage.5. For fresh fish iced immediately after capture and continually stored in ice.

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

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

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

I-sushi. The first report of the organism being a foodborne agent was with shirasu. the risk posed to the consumer of fresh fish.g.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. 5). Since 1996. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. 1978 .6. Other lightly or semi-preserved (e. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). due to the cultural preference for raw fish dishes. 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. and proper cooking (boiling for 1 minute. smoked. V. Southeast Asia. and when in contact with contaminated water (no CDC reported cases from fish. Despite the ubiquitous nature of type E C. France. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. A unique clone of V. is small. there is no specific guideline that describes a minimum level of V. which was probably contaminated from an uncooked food source or an excreting foodhandler. salted or pickled) fish have also been associated with botulism. parahaemolyticus is a marine organism.2 Vibrio parahaemolyticus and other vibrios V. The bacterium causes primary septicaemia. storage and preparation (11). vulnificus is associated with warm marine and estuarine waters. This strain has been responsible for 50 to 80% of all V. parahaemolyticus cases have increased across the world. Japan. Russia. accounted for the majority of cases (1. parahaemolyticus outbreaks. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. USA. again. 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. whether stored aerobically or in modified-atmospheres. Generally. including epidemics in India. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans.3 °C) will eliminate all risk. 1. Currently. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. the majority are associated with products not cooked immediately before consumption. were associated with finfish between 1978 and 1998 (46). parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47).g. the majority of outbreaks are caused by consumption of raw fish products (e. 11). parahaemolyticus O3:K6 is responsible for many of the recent V. Proper temperature control (<3.2005). Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. which can cause human gastroenteritis. and North America. In Japan. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . Only about 1% of marine isolates produce a thermostable haemolysin. which is believed to be required for virulence. a Japanese boiled and semi-dried sardine dish. sushi) (5). V. Proper chilling.

there is no risk from the consumption of properly handled and cooked fresh fish. Despite this. Chill storage may not eliminate growth. shigelloides are encountered. in its habitat.6. in assessing the risk of fishborne infection by P. cytotoxic. Since 2000. 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. P. however. 1.000 17 . with minimum growth temperatures around 0 °C and optima of 15 .CHILLED AND FROZEN RAW FISH disorders). coli on isolation media (50).. the causative agent of cholera. Although the psychrophiles are unable to grow at body temperature. is historically associated with faecally contaminated water. Again. In the US.3 Aeromonas Aeromonas spp. and especially Aeromonas hydrophila. shigelloides is similar to Aeromonas spp. infection. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis.20 °C. It is known that Aeromonas spp. listeriosis cases have been reportable to the CDC. monocytogenes has been well documented as a foodborne human pathogen. but proper cooking should substantially reduce risk of infection. Similar problems as those encountered for Aeromonas spp. their potential to produce disease-causing exotoxin in food has not been fully examined (48). and proper cooking will substantially reduce risk. Some isolates are true psychrophiles. which is often fatal (>50%). and fish and shellfish are probably the major reservoirs for the organism (50). are associated with human diarrhoeal illnesses. others are psychrotrophic mesophiles (49). cholerae O1. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. and especially estuarine environment. possibly due to under-reporting because of their likeness to E.26 to 0.55 cases per 100. are present on fish and can grow to significant numbers during storage (1. Most cases have been sporadic rather than associated to large outbreaks. 17). sodium channel blocking and haemolytic activity (48). no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. There is no risk associated with the consumption of properly chilled and cooked fresh fish. 1. They are aquatic organisms with an epidemiology that is yet to be fully understood. L. 5. The major virulence factor appears to be the production of toxins with enterotoxic. V. but the bacterium is known to survive and grow in the shallow marine. proper chill storage will eliminate any risk associated with this organism. monocytogenes incidence has been between 0. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation.6.4 Listeria monocytogenes L.

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

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

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

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

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

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

M. Cho Y.1983. Aberdeen.L. in FAO Fisheries Report No. Ravn Jørgensen B. Huss H. Kitts D. 1989. 62-70.P.S. Picard G. Akiba M.. The Microbiology of Seawater Fish..M..9. Rome.. Olley J. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Borgstrom G.H. 1. 295-307. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches..D. Estimation of the bacteriological quality of fish by automated conductance measurements. 1986. 1988. 1. 1984. International Journal of Food Microbiology... 51. 741-50.).1 Further Reading Processing Anon. Academic Press.9 1..D. 35. Technomic Publishers. 73-89. 1989... Ed.M. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review.C. 24 . Gibson D. Studies on the microbiological ecology of mackerel stored by the method of partial freezing.. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Eds. Hobbs G.. International Journal of Food Microbiology. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere.9. Food and Agriculture Organisation.2 Spoilage Shewan J. Journal of Food Science. 1992. Aberdeen. 6. 55.. Jones Y. Application of predictive microbiology to assure the quality and safety of fish and fish products. Sumner J. Bulletin of the Faculty of Fisheries (Hokkaido University). 127-34.E. Gram L. Taoukis P. 1984.G. Magno-Orejana F. Handling and processing of scad. Food and Agriculture Organisation of the United Nations. Torry Research Station of the Ministry of Agriculture.. Huss H. Ed.. in Fish as Food. Torry Research Station. McMeekin T. Florida. Shahidi F. Huss H... Ross T. Microbiological quality and shelf life prediction of chilled fish. Fisheries and Food. Fisheries and Food. in Torry Advisory Note No 93. International Journal of Food Microbiology. 317 (Suppl.A. Dalgaard P. 1992. Trolle G. 15.FISH AND SEAFOOD 1. 1. in Seafood Science and Technology. Ogden I. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. International Journal of Food Microbiology. 4. Lancaster. 271-85. 1997. Gibson D. 317-21. Cann D. Houston N. Villemure G. Changes in microflora and chemical compounds in mackerel stored by partial freezing. 1985.. Labuza T... Journal of Food Protection. 1961. Smith G. Shinano H. Simard R. 13-32.H.H. Torry Research Station of the Ministry of Agriculture. 65-72.

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

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

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

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. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. with a minimum residual salt content of 3.2 days) longer than that of non-prepacked fish. haddock. Again. be flavoured with herb dips or peppercorns. Added colours can be included with the brine solution to meet specific requirements for products e. Fish steaks can be cut from a natural fillet or loin (e. 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. Hot-smoked fish are prepared from brined. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes. the ingredients premixed. The expected shelf life of these products is up to 8 days (or longer for some chilled. swordfish steaks).2. which may.5%.g. The result is fish with a typical residual salt content of 1.5 . natural fillets of fish such as herring. or pre-packed frozen with added glaze.. 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. Pâtés are prepared with raw fish. which is approximately 30% (or 1 .5% in the aqueous phase. 29 .g. sawdust with other woods adjusted for flavour profile and burn duration). Some products may be salted for flavour only (e. The shelf life for MAP fillets is typically up to 5 days. in VP or through mail order. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. mackerel. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. in addition. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. This process in effect cooks the product and reduces or eliminates microbial contaminants. Cold-smoked natural fillets are prepared from a range of cold water fish including herring.g. resulting in a product that is ready-to-eat without any further cooking. cod). but may be increased (up to 1 month) by packaging methods like MAP or VP. etc. They are sold from wet fish counters. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product.g. tuna steaks) or band-sawn from a frozen block (e. VP products) because of the high salt content acting as a barrier to spoilage. salmon. mackerel. Cold smoking is essentially a flavouring process. a protective glaze may be added to the frozen steak. annatto with smoked haddock. Fresh fillets are sold from wet fish counters. or pre-packed in MAP or vacuum packaging (VP). and the product then shaped and heat-processed to cook the product before being chilled. and salmon.

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

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

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

defined holding time at a chill temperature (0 – 3 °C).CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material.3 Preparation of fish component of chilled and frozen recipe dishes 2. 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 the full complexity of the processes used for specific products cannot be covered in a general text. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface.3. Generic examples have been used to illustrate only the major points. but will also need to ensure that the optimum shelf life can be safely obtained. Examples of tempering techniques include the use of heated air. and contamination from equipment to clean fish muscle. and potential opportunities for growth of foodpoisoning bacteria. as a heat process is often required for functional or organoleptic reasons. up to -5 °C so as to improve the cutting and handling properties during processing. line extensions and process reviews. Rapid controlled cooling of heat-processed components will help to minimise spoilage. the core temperature.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. Where the raw materials are frozen. For some components. depending on the size of the fillet 33 . 2.4. but it is assumed that any band sawing or dice process will introduce heat from blade friction.4.1 . Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components.4.2. 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. All have the same intention.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. Effects on the microflora are minimal during tempering (7). this may be an incidental effect. there are a number of distinct methods used. to evenly raise the temperature of the entire frozen product from -20 °C or lower. Band-sawn materials or fish dice are normally promptly further processed. It is recommended that the principles of HACCP are applied to novel processes. or the use of microwave technology. 2.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. but it also poses a risk of cross-contaminating other products undergoing tempering.

2. It is then size-graded. after landing. The “blown” meat is graded.3. 2. the scampi is washed in potable water to remove foreign bodies.Shaped Fillets.1C .1A . Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. by mincing and mixing with a binding agent (as described in section 2.FISH AND SEAFOOD processed. may not rise above 0 ºC during processing. Fig. Raw material preparation stages for chilled or frozen coated fish products 34 .4.7 days. 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. with the coating heat-fixed. drained and then refrozen for storage until required for further processing. are placed into plastic boxes and covered in ice to preserve their condition. 2.1.1 Whole-fillet / scampi Scampi (Nephrops norvegicus).3.1.1. Frozen cores are usually batter-enrobed. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen. quickfrozen.1 (2.4. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product.2. The processing of three examples types are described in Figure 2. On arrival at the processing plant.2). chilled or sold chilled on defrost with an expected shelf life of 6 .1B . frozen in a nitrogen blast freezer and weighed into sacks for later processing.Fish Cakes).Scampi.

This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. 2. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. from potato flake.3.7 days.1.g. or promptly processed to avoid further quality loss.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. 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. or those where the trimmings. 35 . The fish component of the cake will. chilled. It is for this reason that all by-products from primary processing. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. Waste material produced from this type of processing is often reused in the third category (fish cakes). quickfrozen. Fish cake cores may be batter-enrobed. such as aerobic spore formers and Staphylococcus spp. once the dry components have been rehydrated.1. unless it is poorly handled. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss.3. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . pathogens such as Salmonella and Listeria.4. with the coating heat-fixed..off-cuts or belly flaps. spoil more rapidly than the dry ingredients. The mix is shaped and formed ready for packing or for enrobing. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish.block-sawn dust. but. material will need to be transferred swiftly to a chiller. Therefore. This is especially the case with block-sawn dust. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. iii) Fresh/fillet/minced fish block.4. that are to be reused for other foods. fish fingers). The remaining dried ingredients are mixed with the fish.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. are handled and stored as any other food product. additional spoilage organisms may be introduced. because of its nature. with an expected shelf life of 6 . ii) Fish-filleting waste . as the physical structure of the material has been destroyed. more seriously. those that use frozen blocks of whole fillets.2 Shaped products This type of product falls into two categories. or sold as a chilled product after thawing. and water is added to rehydrate the components.

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

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

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.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe.4. as this could introduce off-flavours and reduce shelf life. pasta or potato. to produce a specific product.3.4. rice.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). For example. and must not show evidence of disease.1 ºC for no more than 3 hours is recommended (1).3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes. canned fish may be used in their preparation owing to it having wider availability and low wastage. There are two main categories. The protein source can be fish fillet or block.g. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. Figure 2. A temperature no greater than 10 ºC for 12 hours cumulatively or 21. Flow diagram outline for recipe dish ingredients preparation 2.5. 2. The vegetables must withstand processing. Fig.2. 38 . but they are supplied without a source of carbohydrate e.

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

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

41 . The format uses flavour compounds . etc. 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. Materials prepared with regard to good hygienic practice need only be assembled in a tray or pouch before chilling or freezing in readiness for distribution. Marinade-type products are becoming more popular. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. that of a dairy-based sauce.5 illustrate the recipe dish ingredients preparation and assembly and packaging. respectively. for instance. Without barriers.4 and 2.herbs. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. the sauce component will spoil rapidly and the possibility of vegetative pathogens from post-process contamination or raw components growing to significant numbers cannot be ruled out. flour products. 13). and handling of processed sauce can be less rigorous than. suspended in an acidified sauce with an oil component. spices.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. Figures 2.4.2.

botulinum to grow. generally carbon dioxide and nitrogen. The oxygen-free conditions will however allow other anaerobic bacteria such as C. These include: 2. the method replaces air in the sealed packaging with a defined mixture of gases.4. 2.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth.2 Modified-atmosphere packaging Originally developed for the red meat industry. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow.FISH AND SEAFOOD Fig. Flow diagram outline assembly and packaging of recipe dish 2.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions). which may restrict either microbial growth or enzyme activity.2. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms.7.5. 42 .7.4. and tuna loins are packed by placing the product in a plastic pouch and removing all the air. as it dissolves in the surface moisture to form a weak acid.4. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. unless the temperature is strictly controlled to below 5 °C throughout the supply chain.

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

Pseudomonas spp. These have been proved to impart off-flavours. freezing also has a protective role in improving the safety of raw materials by. also be those that have the least effect on the viability of microorganisms (21). This reinforces the need for prompt processing and temperature control of ingredients and finished products.5..7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24). It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. Bacillus spp.FISH AND SEAFOOD 2.5. 44 . however. 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. producing characteristic off-flavours. the shelf life of products is limited by the natural spoilage process described earlier. which would warn the consumer of the potentially unsafe nature of the product. where the variety of components and ingredients used in manufacture is varied and complex. for example.. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e. The type and nature of the enzymes and their activity will depend on the bacterial species. By retaining the intrinsic spoilage flora of seafood. Serratia spp. within dairy products are well known. 2. As a note of interest. Still.. and Aerobacter spp.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. botulinum. For example. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. eliminating nematodes in products that do not have a heat treatment step. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). However. 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. auto-oxidation of fats/oils). enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage. The conditions and freezing process most favourable for good-quality products will.g. this should also take into account shelf life of product components used in the assembly of the final product.

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

growth may occur in foods (26).6. at the present time. 2. L. indigenous or other. Rates of contamination with smoked seafood are much higher than those with unsmoked products. if any other potentially pathogenic organisms. will colonise factory environments in a similar fashion except for possibly Vibrio spp. Sources of contamination are many because of the widespread distribution of this organism in the environment. It is unclear. but. and this is believed to be due to a tolerance to brine used in the smoking process (32). However. and inadequate cooking or holding within the temperature growth range (27).6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. 2. aureus enterotoxins in foods. 46 .5 Shigella spp. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. which is not affected by any subsequent cooking process. monocytogenes as a contaminant in foods (31). the restrictions on growth rate seen with seafood will not apply. and will therefore cause “food poisoning” even in a product that is cooked before consumption.6. Shigella spp.4 Listeria monocytogenes Interest in this organism has been characterised by a number of very serious incidents associated with a variety of product types (27). monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). 2. but under the appropriate conditions. infected food handlers. An acceptance has now developed in the UK of the presence at low levels of 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). These conditions will allow the production of the toxin. L. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. when introduced or inoculated into sterile components of chilled prepared recipe dishes.6. These include inadequate refrigeration and preparing foods far in advance of planned service. Chilling may help Shigella to survive. monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. aureus (35).

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

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

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

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

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

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In the process of filtering water. Box 3012 Bothell WA 98041-4421 United States of America 3. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food.E. and Cephalopoda. Pelecypods have little need for locomotion. clams. mussels. slugs. Gastropoda. and scallops. and shrimp. clams. where it is eventually swallowed and digested. Plankton and algae form the bulk of their diet from seawater. In these forms.S. Special tracts of cilia move this mucus toward the mouth. cockles. which includes oysters. Pacific Regional Laboratory Northwest U. abalones. Pelecypoda. MOLLUSCAN SHELLFISH Carlos Abeyta. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. lying well within the mantle cavity. can hold the shells tightly shut against enemies (1).3.1 3. octopi.7 litres of water per hour. they also trap bacteria. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). Jr. In most species.1 Definitions General features of the mollusc The phylum Mollusca. chitons. while 53 . squid. the adductors. viruses. O.1. Strong muscles. 4). For example. snails. whelks. The class having species of economic and public health importance is the Pelecypoda. mussels. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. The term shellfish in this context is limited to oysters. Scaphopoda.6 litres. cockles and scallops. chemical contaminants and other impurities on the mucus of the gills. The phylum consists of five classes: Amphineura. composed of two valves hinged together dorsally. mussels. oysters. oysters and clams can process about 3. S. The class Pelecypoda includes all of the bivalves such as clams. respectively. P. Food and Drug Administration 22201 23rd Dr. Many microorganisms ingested by shellfish survive the digestive process (3. scallops. lobster. and 2. Other species of shellfish such as crabs. the shell. cockles. completely encloses the body.

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

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. 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 .

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

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

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

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

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

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

Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. handling. The most important factor in controlling bacterial growth in shellstock is temperature. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. Furthermore. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. there is evidence that in the absence of adequate refrigeration. Generally. with prolonged storage the microbiological quality of the bivalve decreases. and the sanitary practices applied during harvesting and shipping. with the exception of A. however. for a considerable length of time. An outbreak of infectious hepatitis in Sweden in 1956. and thus contaminating the shucked meat at the point of sale. 25). Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. without adequate refrigeration. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). involving 691 cases was attributed to oysters contaminated in a wet storage area (27). including V. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. If long delays occur before reaching the processing facility and/or refrigeration. shellstock are not refrigerated on the harvest boat. shellstock are placed in refrigerated trucks for transport or placed in wet storage. 24. 23. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. to avoid possible contamination of the shellstock. hydrophila (18). and Citrobacter species) and Vibrios. some pathogenic organisms will survive and multiply in shellfish (20.2 ºC (45 ºF) or below (19). bivalves tightly close their shells thus trapping water and microflora. 21. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. Generally. coli.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. Enterobacter. Over a period. Water used for shellstock washing should be of good sanitary quality. Shellfish may contain higher levels of pathogens than are found in the water in which they grow.1. Klebsiella. are prevented.FISH AND SEAFOOD 3.3. and temperature storage of shellstock to point of sale. The washing of sediment and detritus soon after harvesting will prevent further contamination. the bacterial counts in the meats will increase. vulnificus. synthetic seawater for the purpose of de-sanding and salt uptake. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18.1. 16. At harvest. 22). At this temperature the multiplication of faecal coliforms (E. Harvest conditions are another factor that influences the growth of bacteria in shellstock. when it reaches the processing facility. 17). Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. 26).

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

and gloves help prevent the contamination of shellfish. 2. 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. 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. 64 . within one hour. 3.2. and sanitised before each filling. which may lead to contamination of product. Separate rooms or lockers for storage of clothing. Shucked meats are thoroughly drained. 4. Fig 3.2 ºC or less. aprons. There is a tendency to store such articles on the shucking benches or in packing rooms.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). 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.

35. Glycogen. Glycogen is hydrolysed by the initial flora found in shellfish. shucking pails. anywhere during the process of shellfish from harvesting to the point of sale. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. particularly shucked meats. particularly oysters stored at 7 ºC. and proteolysis (10). The spoilage process in shellfish may be divided into three stages: increase of acidity. In summary. are an excellent medium for growth of bacteria. these include lactobacilli. blowers and other utensils that have cracked. Micrococcus. These organisms may play a significant role in the spoilage processes. Shucked shellfish are an excellent medium for the growth of bacteria. where there is a breakdown of temperature control or prolonged storage of shellfish. clean gloves. The environment has an effect on the bacterial population found in shellfish. showed that organisms such as Achromobacter. Lactobacilli seem to be the predominant microflora in shellfish. a carbohydrate. 34.2 ºC. Flavobacterium. For example. in oysters. spoilage will occur. Temperature abuse will result in high bacterial counts.2 ºC or less. and yeasts. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. rough. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. at the turn of the century. Shucked shellfish are held and transported at temperatures of 7. abundant gas production. All equipment coming in contact with the shellfish such as colanders. 32). keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. and packing the shellfish in a timely manner. Pseudomonas. 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. Proteus. 3. 33. streptococci. whereas bacterium such as Pseudomonas. Alcaligenes. Shellfish. As described previously. is stored in the bivalve as a source of energy.8% (37). or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. The initial flora found in shellfish is the major factor of spoilage in bivalves. Controls that should be applied include using clean packing equipment.47 to 6. 36). 65 . thus producing acid and gas (38). Earlier studies. skimmers. Much of the work has been done with oysters and clams. 31. Achromobacter and Flavobacterium levels decrease during storage.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. glycogen levels range from 0. This process changes the bacterial profile to microorganisms favouring low pH.

5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. Diseases caused by Vibrios are enteric in nature. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). parahaemolyticus possibly producing illnesses (40). coastal areas). nausea.5 cases/100. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. Between May and September 1997. These organisms can be present in the shellfish from the point of harvesting. and estimates based from a hospital study show less than 0. lot and retail outlet.5. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. V. from harvesting to finished product (i. parahaemolyticus from shellfish is low. there is a recent report of low levels of V. although non-hemolytic strains have been associated with illness (43. headache and fever. ranging from epidemic cholera to sporadic cases of diarrhoea.1 Vibrio parahaemolyticus V. V.5. This bacterium is present in all the major processing steps of a shellfish plant. Fortunately. diarrhoea. prevention of cross-contamination. The incidence of V.48 hours with abdominal pain. They can be introduced to the shellfish from the environment or added during processing. 3. however.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. One of 250 cases reported involved shucked products with low levels. Sporadic cases are frequent along the coast of the United States. widespread in both polluted and unpolluted waters. and to the point of sale. during processing. 41). parahaemolyticus causes gastroenteritis lasting 24 . Products involved were from the same processor. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. of V. 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. It is a common marine bacterium. and very common in Japan.000 population/year (46). parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8.e. where outbreaks occur regularly.1. 3. and harvesting of shellfish in the cooler months. parahaemolyticus. 44). Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. shellstock and packing of shellfish in jars) (29). only a small percentage of strains in the marine environment are potentially pathogenic (45). approximately 23 cfu/g.FISH AND SEAFOOD 3. primarily in brackish or saltwater. 66 .

7 days. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. nausea. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). are particularly susceptible to primary septicaemia. or immunosuppression. in the range of 0. 56). those who have liver disease. cholerae non-01 (51). cholerae non-01 is generally a less severe gastroenteritis than that caused by V.II (8. or primary septicaemia. parahaemolyticus growth in shellfish meats is temperature dependent. 48). vulnificus. however. with a fatality rate of 50% for individuals with septicaemia. parahaemolyticus. Hazards from this bacterium can be controlled by thorough cooking of shellfish. V. Control of V. V. like V. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. and nausea within 24 . Control of oyster-associated 67 . cirrhosis. The bacterium is an estuarine species that causes wound infections.000 population/year (8.5. 3. vulnificus are fever. 58).3 Vibrio cholerae non-01 Illness caused by V.4 . gastroenteritis.1. Onset of symptoms occurs in 48 hours and can be severe.5. cholerae. 46. It is one of the most severe foodborne infectious diseases. 23 of 24 major USA-West Coast estuaries were positive for V. vulnificus is by rapid refrigeration during warm weather months. and preventing cross contamination once the product is cooked. which have been implicated in shellfish-related disease outbreaks (31). the organism has been found in the Gulf of Mexico. diabetes. cholerae non-01 is ubiquitous in the estuarine environment. leukaemia. Healthy individuals are susceptible to gastroenteritis. cholerae non-01 has been associated with septicaemia. 3. these individuals are advised not to eat raw shellfish. It causes diarrhoea. 53. Death has been reported to occur within 36 hours from onset. Primarily.0.1. In one study. fever. 54. and vomiting. The organism attaches to the intestine and release of a toxin is suspected. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). All isolates are considered pathogenic. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness.48 hours of onset. 46. Again. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. 49). 57. parahaemolyticus the estimated annual incidence rate is very low. lasting 6 . 55. Oysters are the most hazardous food associated with V.2 Vibrio vulnificus The symptoms of V. abdominal cramps.8 case/100. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. The occurrence of V. Inadequate refrigeration will accelerate the growth of V. and rapidly become inactive at low storage temperatures. chills. V.MOLLUSCAN SHELLFISH Control of V. They are sensitive to cold. High-risk individuals.

Death may occur after severe fluid and electrolyte loss. mimicus.10 days. Poor sanitation and contaminated water supplies will spread the disease.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. 3. Onset of illness is from 3 to 5 days after ingestion of contaminated food. acidity and heat. In developing countries. V. and V. fever. They are common marine bacteria.5. undercooked.10% carbon dioxide for optimal growth conditions. 63). 60). In general. cholerae 01 is transmitted by faecal contamination of food or water. abdominal pain. (8.5. diarrhoea. cholerae 01 in molluscan shellfish. Onset of disease is 6 hours to 5 days. In addition. however. salinity. and southern Asia including India and Bangladesh.1. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. the pathogenic severity is lower than V. dehydration. The control of V. Illness can include abdominal cramps.5%) and 2 . e.5 Other Vibrios spp. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. 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). hollisae. drying. particularly in oysters. or cross-contaminated shellfish (57. The presence of these strains appears to be in low numbers and non-virulent (59). Illnesses are associated with the warmer months. nausea. processors should know the product source. cholerae 01. 3. 62.5. associated with shellfish-borne illness outbreaks. is the same as previously discussed for the non-01 strains. V.FISH AND SEAFOOD V. jejuni requires reduced levels of oxygen (3 . headache and fever. widespread in both polluted and unpolluted waters. low pH. and shock. respectively. vomiting. cholerae non-01 illnesses is by cooking seafood thoroughly. It causes diarrhoea.48 hours with abdominal pain. Gulf coast environment.g. cholerae 01 establishing in the U. nausea. and illness lasts 7 . and nausea.S. including V. vulnificus. There are several other Vibrio spp. 3. 61. these organisms cause gastroenteritis lasting 24 . 68 .1. parahaemolyticus and V. There is evidence of strains of V.2 Campylobacter jejuni C. preventing crosscontamination of cooked seafood. imported product from a country experiencing an epidemic. The frequency of infection of these organisms is no different from other Vibrios. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. and the transmission of the disease by consumption of raw. V. fluvialis.

although their role as a pathogen is highly controversial and circumstantial. V. whereas at 10 ºC Campylobacter survive only for 5 days (71). however. jejuni begins in the harvest of shellstock from approved waters.5.500 cells/gram of Campylobacter may cause illness (64. 69. have been isolated from shellfish beds throughout the world (67. jejuni does not multiply in shellstock at refrigerated temperatures. vulnificus. from Pacific shellstock oysters purchased from a retail grocery store (74). 72. They have been implicated in limited shellfishassociated gastroenteritis cases (8. Although. of Incidents 4 279 0 1 0 No. 77. 84. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. V. infection occurred among participants of a firemen's banquet.4 to 114 cells. In one case. 71). mimicus. 81. V. and their immediate proper refrigeration. cholerae 01 and non-01. hollisae Infective dose varies with host susceptibility and virulence of the strain. they survive well at refrigerated temperatures. This finding is significant in that C.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. 70. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. 76. Campylobacter spp. The seasonality of 69 .5. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. C. 68. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. 83. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP).3. parahaemolyticus.II Molluscan shellfish associated outbreaks caused by Vibrio spp.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No.3 Other organisms of concern 3.MOLLUSCAN SHELLFISH TABLE 3. 80) are ubiquitous in the marine environment. Recently. Human feeding studies suggest that about 400 . jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. 65. of Cases 4 362 0 1 0 a Includes V. C. 3. 73). jejuni were recovered. 82. 85). Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. 66). and V. Campylobacter appear to survive well in oysters during storage. V. To reduce the risk of shellfish-borne campylobacteriosis. 78) and P. fluviallis. control of C. ranging from 0. however. shigelloides (79. hydrophila group (75.

70 . They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. 3. jejuni V.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. 87) of the outgrowth of A. They are easily destroyed with mild cooking temperatures. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. non-dairy frozen desserts. cholerae V. Control of these organisms in shellfish products is limited. Note: Although molluscan shellfish are not included in the selected high risk food. Similar results were seen by other investigators (86. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500.000 to 3. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. 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. FDA/Administrative Guidelines 3 Source (91). aureus C. and where indicated are intended to be used for official control purposes. Caution should be exercised by individuals with haematological malignancies (88). hydrophila group stored at refrigeration temperatures.000 cfu/g1 <230/100 g2 <1000 per gram. prepared sandwiches. coli Enterotoxigenic E. soy products. vulnificus in molluscan shellfish. FDA/ Administrative Guidelines. In one study. 2 Source (90). dried milk/whey drink powders. being isolated more often during the warmer month. parahaemolyticus Staph.0001 negative for the presence3 Under review4 Source (89).000/gram (29). 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. coli Salmonella V. Test Standard Plate Counts E.600.

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

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

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

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

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

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

1995.S. Food and Drug Administration. 1997.R. 3.C. oysters. Washington D. Slavin J..C.12). 1987.. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease.MOLLUSCAN SHELLFISH 88.S. Food and Drug Administration.C. 2000. Technomic Publishing Co.S.S. U. 89. 560.030. Ed. Inc. Martin R. U.. Lancaster. 91. 7. U.E.S. Gaithersburg.S. Ed. 1995.L. Baird-Parker T. Food and Drug Administration. 63-8. Smith J. Burdge D..H.C.S. Food and Drug Administration.A. Food and Drug Administration.8 Further Reading Ahmed F. in U. Food and Drug Administration Compliance Program Guidance Manual.W. Gould G. Food and Drug Administration.W. mussels. in U.. National Academy Press. Collette R. Lund B. U. Clams. Pittsurgh. Program 7303.600 (CPG 7119. Seafood Safety. Food and Drug Administration Compliance Policy Guides.1987. Food and Drug Administration.. Fish Inspection. Quality Control. Sherlock C. and HACCP: A Global Focus.S. Washington D. fresh or frozenadulteration by bacteriological contamination Sec. in National Shellfish Sanitation Program Manual of Operations. U.E.Pathogen monitoring of selected high risk foods (FY 88/89). U. Food and Drug Administration. Government Printing Office .M. 1991. Aspen Publishers Inc. 77 . Part I Sanitation of Shellfish Growing Areas. Chapter 5: Foods. Ed. 90.. The Microbiological Safety and Quality of Food... Washington D.

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and available in 0. They possess a segmented body.. packed and frozen in 0.5 kilo pack (boxes or bags). Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. Shrimps.0 kg polythene bags. and frozen in 1 and 2.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. peeled product.Ph. M.4. crayfish and shrimp (1). Peeled ready-to-cook shrimp are raw. Pandalus borealis (deep-water shrimp). Peeled and cooked shrimp are ready-to-eat after controlled thawing. crabs. Pandalus montagui and Crangon crangon (inshore shrimp). Species eaten from tropical waters include Penaeus spp. with shells and heads on or off are usually packed.. and Trachypenaeus spp.0 kg polythene bags. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. Parapeneopsis spp. and are usually aquatic. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. with additional water to form a glaze. jointed limbs and a chitinous exoskeleton.5 and 1. e. 79 .5 and 1. Crayfish..g. (deep-water shrimp). lobsters and crabs have ten legs. lobsters. the front pair ends in claws. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). Such products are usually only partially cooked before freezing. CRUSTACEAN SHELLFISH Linda Nicolaides.

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

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

including GHP.FISH AND SEAFOOD frozen. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour.frozen uncooked prawns 82 . After cooking. should be followed to ensure that the product is not re-contaminated with microorganisms present in the processing environment or from the workers. spices and cooking batter. Fig. 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. Breaded products introduce an added risk of spore forming bacteria. Good Manufacturing Practice (GMP). 4.1. Flow diagram .

4. there is no indication of life.CRUSTACEAN SHELLFISH 4. with the growing demand for ready-to-eat products from crab and lobster meat. bright and dry appearance. slightly iodine smell. With spoiled crabs and lobsters. putrefaciens. the meat is either repacked and sold in the shells. If boiled. If the tail drops. The shell will be soft.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. and a strong ammoniacal odour is produced. However. An offensive odour is also produced. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. there is no associated spoilage problem. immediately prior to boiling. If hygienic procedures are not followed. spoiling prawns and shrimps have shells that are soft. and have a sweetish. The shells have a crisp. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. the joint between the tail and carapace is open and the limbs hang loosely. 4.3. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. 4. since crabs and lobsters are traditionally consumed immediately after cooking.4. putrefaciens and Pseudomonas spp. bacteria of public health significance. or packaged and processed.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. In comparison.2 Cooked crabs and lobsters As indicated earlier. Heat is produced if the shrimps decompose in a confined space. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. whilst boiled crustaceans have a pleasant slightly sweet smell. wet and ‘soapy’ or ‘jammy’ to the touch. they include Sh. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. Once removed and cleaned. or in hermatically sealed cans that have undergone an appropriate heat treatment. dull and sticky. for crabs.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. 83 .4. contaminating microflora may include enteric bacteria. viruses or parasites.

However. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). 7). The above results prompted the FDA to develop a compliance programme for the pathogen. a common pond fertiliser used in aquaculture. although efficient cooking will quickly eliminate it (6). shrimps and cockles sold (17). 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. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . 16). The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) .1 Pathogens: Growth and Survival Salmonella spp. Although foodborne illness had not been associated with the product. later surveys in 1990 recovered the pathogen from retail raw. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. monocytogenes in cooked ready-to-eat foods. Moreover. Salmonella cannot be completely removed from raw shrimp during processing. 6. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. An import alert was put in place in June 1987. cooked and smoked fish. Following this.2 Listeria spp. monocytogenes was absent from 40 samples of cooked prawns.cooked prior to consumption. nearly 30 years elapsed before the potential hazard of L. in 24. even if GMP is followed. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). fish fingers. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals.5.5 4. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. as well as for Salmonella.FISH AND SEAFOOD 4. animal manure. 5. Further surveys identified Listeria spp. shrimp and shellfish (11). Hence. 4.5% of frozen raw shrimps from different countries (13). The presence of L. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11).5. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. and usually for both. monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). in domestic/imported shrimps (15).

which can become colonised by V. was associated with seafood consumption. V. 4. V. cholerae non-O1. The former. 24. An epidemic that began in South America in 1991.3 Staphylococcus aureus Staph. the causative agent of cholera. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. no incidence of foodborne illness caused by L. Vibrio cholerae. The pathogen has resistant spores. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. Once the competing microflora has been removed by a heat treatment. is widely distributed in aquatic environments. particularly crabmeat (18. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. aureus on their hands. 19).g. as part of the natural bacterial flora. although contamination by types A and B may occur from the environment. especially in products that have been vacuum packaged. the pathogen can grow at temperatures as low as 3 °C.5. Crabs are covered in a chitin exoskeleton. botulinum type E is commonly associated with fish and fishery products.70% of healthy individuals carry toxin-producing strains of Staph. 85 . 22) as well as from undercooked crustacea when harvested from contaminated waters (23. botulinum type E at 10 °C but not at 4 °C (20). 4. 4. combined with the degree of handling of these types of product.5. Crustaceans have been linked with food intoxications attributed to Staph. monocytogenes has been attributed to crustaceans. cholerae. 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. Researchers have demonstrated that inoculated shrimp supported toxin production by C. Control should be achieved by promoting GHP and staff training programmes.CRUSTACEAN SHELLFISH old. therefore. However. cholerae O1 and V. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. pasteurisation and cooking. e. To date.5 Vibrio spp. whilst boils and cuts are also common sources of the pathogen. about 50 . The main source of contamination is the hands of processing personnel. aureus because of the ubiquity of this organism in the processing environment. so is able to survive heat treatments that eliminate vegetative cells.5. pregnant women and the immuno-compromised.4 Clostridium botulinum C. 25). aureus is not a member of the intrinsic flora associated with crustacean shellfish. cholerae is divided into two groups.

Astroviruses and non-A.5. 31. whilst 5. Of these. Hepatitis A virus presents the most serious hazard. however. hydrophila and A. V. However. V.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans.5. A. been implicated in a case of foodborne infection associated with crustaceans.8% of samples contained C. 32). a marine halophile. in a survey carried out in the United States between 1977 and 86 .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.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States.5. Quantitative levels were below limits of detection in all cases (<0. the potential hazard exists as C. hydrophila is able to grow well at refrigeration temperatures (0 . they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). Japan. non-B Hepatitis viruses (31). but has the potential to cause fatal foodborne illness (26). Parasitic infections are controlled by either cooking infected material prior to consumption. Caliciviruses.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A.g. South America.6 Campylobacter jejuni and Campylobacter coli C. It is more commonly implicated in countries where crustacean shellfish are eaten raw. particularly trematodes (6). jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. Norovirus. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. although A.7 Aeromonas hydrophila and Aeromonas sobria A. 4. 4. jejuni has not.5. coli. vulnificus is an invasive and lethal pathogen associated with wound infections. hydrophila has been isolated from prawns (29). e. Contamination by this organism can be controlled by rapid chilling of crustaceans after harvesting and avoidance of time/temperature abuse during distribution and processing chains. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. 4. to date. or freezing at a temperature of -23 °C for 7 days (30.30 MPN/g) (27). parahaemolyticus. occurs in crustaceans from warm environments. However. 4.

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. 4. are included in the Commission Regulation (EC) No. only 0.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). 39). 4. 38. Selected parts from this regulation is presented below. 4. 4.CRUSTACEAN SHELLFISH 1981. Provisions laid down concerning the use of additives would also apply to these types of products. They can survive low pH levels. The toxin causes amnesic shellfish poisoning (ASP) when consumed. which results in a persistent and apparently permanent loss of short-term memory (36).1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. which could be transferred to the meat during unskilled removal from the shell (35).5. from farm to fork (37.6.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish.5. No foodborne incidents have been attributed to this source to date. although it is one of the European Union (EU) legislated allergens. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. Norwalk viruses can also withstand heating to 60 °C. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). produced or imported into the European Union (EU).6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. A summary of microbiological criteria for crustacean shellfish is listed below. 87 . 2073/2005. or chlorine concentrations similar to those found in drinking water. consistent with that in the stomach. The risk of such hazards is far higher with fin fish and molluscan shellfish. 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.

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

CRUSTACEAN SHELLFISH

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

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

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

a 0º brine saturation means pure water.8 211. Therefore.FISH AND SEAFOOD 5.4 184. When a 100º brine is used. all using salting steps. % 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.6 264. When working with salting. it is impossible to establish standardised processing stages in the curing of fish. only the most common processes can be described here. At 18 °C a 100º brine is produced when the solution contains 26.8 211.8 79.2 237.4 52.I). etc. The strength of the solution is described as degrees of saturation (brineometer degrees). TABLE 5.2 g of salt per litre of brine).0 g of salt per litre of brine). or salt in the water phase. processes and producers’ private recipes. Thus. the fish is usually salted with a 70 .0 . a 90º or above brine (237. about 50º (132. it is important to state the temperature.6 g of salt or more per litre of brine) may cause salt burn in this way. The dissolution of salt in water is dependent on the temperature.2 105. This should be distinguished from the salt content. is employed.6 132.3 Processing and its Effects on the Microflora Owing to the wide variety of products. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. When a milder brine. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. smoking or marinating processes.4% salt (Table 5. it causes swelling because the fish absorbs water from the salt solution (1).0 158.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. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. it is important to understand the concept of salt concentration.80º brine (184. Salt concentration means the percentage of salt in the water contained in the fish. Different brine strengths are required according to the curing processes and/or the type of product. For smoking salmon. sometimes the salt crystallises onto the surface of the fish. Salt in the water phase links the two conditions of moisture and saltiness. and salt content.

respectively. lean white fish is used for this purpose.3. however. but the concentration of copper and iron must be kept under 0. The calcium and magnesium impurities. outgrowth of halophilic bacteria may occur. and essentially free from iron and copper (3). for which there is a high demand in Mediterranean countries. respectively (1). The salt used should be of food-grade quality. SMOKED AND DRIED FISH 5. addition of levels of 0. 97 . 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. ‘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. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. and a ‘putty’ fish product is also obtained (1). if very pure sodium chloride is used for curing purposes. On the other hand. 5. caught in North Atlantic waters. When pure salt is used. the fish is mixed with one part of salt per eight parts fish. which consequently inhibits the diffusion of the water from the bulk of the product to the surface.1 Dry salting / Kench curing Usually. mainly in tropical conditions. ‘Green cured’ split cod with about 60% of the backbone removed. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. the fish is restacked with fresh salt after about a week. having a short section at the end of the tail. it tends to lead to a slight yellowing and softening of the product. and for a heavy salting a ratio of 1:3 salt:fish is used (1).3.15 . are likely to cause hydroscopicity of the product and.CURED. This is because fatty fish turn rancid quickly.15% calcium and magnesium.0. and such delays.1.35% and 0. producing characteristic pink patches. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. This condition is known as ‘putty fish’ (4). For a light salting. However. low in calcium and magnesium. when the air humidity is high. Salting can be done in a number of different ways. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1).1 ppm and 10 ppm. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. Also.1. See Figure 5. which include calcium and magnesium. The most common fish used in the northern hemisphere is cod (Gadus morhua). are recommended to help make the product whiter. and the salt has penetrated throughout the fish (3). and salt can speed this oxidation process. Usually it is followed by other preservation techniques such as drying and/or smoking.1 Salting Salting of fish for preservation purposes has been used since pre-history. can be dried further to produce ‘klipfish’ or ‘bacalão’.

Food-poisoning organisms. such as Staphylococcus aureus could survive and grow at such water activities in the product.1.5.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. Where fish are salted in their whole form. it appears that water activities much less than 0. such as 98 . when a light brine is used. Fig.90 are rarely achieved. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole.1.2 Wet salting or brining As referred to above. the process is mainly for flavouring purposes prior to the use of other preservation processes. Only small fish can be salted whole. although toxin production would be unlikely (4). For large fish it is advisable to split it before salting. eviscerated and/or split). The greatest barriers to salt penetration are the skin and scales. Processing of dried salted cod 5.3. so that the risk of spoilage is high.

26 7.26 2.96 1.60 2. and a red burgundy colour will develop in the product.49 4.90 (1).91 2. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 . headed and gutted.76 4.98 5.26 4.80 5.51 5.00 3. the brine is then replaced over the fish.03 2. with pressure being applied to the top of the container to help the brine to drain away.22 6.78 6. For preservation purposes.61 3.98 6.44 2.85 3. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.46 4.66 5.85 3.5 10.41 5.41 6.17 4.11 9. a saturated brine is used.51 3.44 2. limiting the amount of rancidity that can develop. TABLE 5.3 days. Anchovies are the main fish used for these purposes.56 6. The fish are kept immersed in their own brine near saturation point for several days. but mackerel.70 3.II. For example.5 3.41 3.86 2.95 3. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained. Histamine formation is of importance when using scombroid and some clupeidean species. the fish enzymes play an important role by maturing or ripening the product. There are several variations of this process.23 2. This is why it is the preferred method for fatty fish.98 6. The main objective of the brining method of salting fish is to limit air penetration during processing.74 9. thus.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 3.5 3.30 2.99 2.76 4.23 2.19 3.CURED.09 8. the liquid that comes out of the fish forms a brine.65 2.61 3.26 1. Here.47 6.26 3. the product obtained is called gaspé-cured (5). and can be collected and replaced again over the fish.09 8. See Table 5.04 5.16 7. Fresh sardines.3.06 4.41 4.76 4. after being drained and pressed at 465 kg/m2 for 15 hours.3 Dry / wet salting In this process. Histamineforming bacteria do not generally grow at water activities below 0.25 5.5 4. can be cured in a saturated brine solution for 6 days and.54 5. herrings and other fatty fish can be used.78) (1).05 7. sardines. they can be stored at 28 °C.88 5.33 5.0 2.17 5.76 4. 5.0 4.64 4.0 3.23 8.76 99 .03 4.66 5.76 4.45 3. if split fish is dry-salted.11 3.73 1.10 1.84 1. but as it is placed in containers.70 6. SMOKED AND DRIED FISH smoking.1. with a shelf life of about 4 weeks (water activity (aw) about 0.35 4. the fish is dry-salted.78 2.11 4.25 5.

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

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

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

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

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

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

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

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

established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). smoking. the salt level could effectively be reduced from 4.3. 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. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines.5. 5. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). applying it as a dip after brining. Fresh fish spoilage may be measured by 108 . In experiments comparing traditionally smoked fish with liquid smoke. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. 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. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves.06 μg/kg of benzoanthracene in the final foodstuff (28). spraying is preferable because it is easiest to control flavour and acceptable with this method (3). or smoke extracts in organic solvents. both of which are considered potential carcinogens. Liquid smoke was considered to provide advantages of safety. so as to impart an unnatural red colouration to the flesh. and less than 0. According to some recent research. 5.4 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. When liquid smoke was used. It has been shown that liquid smoke can reduce the formation of PAHs (3).03 μg/kg of benzopyrene. consistency of flavour and colour.8% and still provide protection for 7 days (25). nutritional neutrality and antioxidant activity (27). 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). injecting it into the fish flesh. 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.6 to 2. The European Scientific Commitee for Food. or applying it as an atomised spray within a modern automatic smoke kiln.4 Spoilage Brining. drying.

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

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

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

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

It has been shown that. botulinum Staph.75 0. with this change.5-10 4. and as botulism is a serious hazard in foods.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. The presence of salt in cured products has a great effect on the growth of the bacteria. However.CURED. botulinum type E Staph. 9 and 8 of the outbreaks. Before.5-8.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13).5 4. the non-proteolytic type E strains can grow in up to 5% sodium chloride. botulinum. when present in the raw material. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). but if the fish products are stored at temperatures <10 °C. and C.97 0. although the combination of the two factors. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth.5 5.0-9.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. under optimal conditions. the importance of cold-chain integrity must be emphasised.86 0.5% to 5%. salted and pickled products accounted for 10.8 1. In the United States. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). SMOKED AND DRIED FISH TABLE 5. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). the products were heavily salted. respectively.5.5 0-11 3.5-10. the fermented products being the most dangerous group (7).8 0.5 5.9 0. types A and B are not affected by 3 .5-8. pressure on the traditional methods of curing. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. smoked and dried. aureus Yeasts Moulds Lactic acid bacteria pH 5. sodium 113 . botulinum type E and production of toxin.61 Organisms Most Pseudomonadaceae C. could not grow and produce toxin. The concentration required to prevent growth at room temperature can vary from as low as 3. Since neither the smoking nor the drying parts of this curing process are particularly severe. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C.97 0. botulinum type E.

more importantly. after processing.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. but. Thus.86 and 10% sodium chloride. growth and toxin formation may occur. As with other organisms. if recontamination occurs and storage conditions are favourable. As they are poor competitors.4 Listeria monocytogenes The psychrotrophic L. The FDA established that hot.4. Sliced smoked fish products. they will grow rapidly (7). but requires higher temperatures for toxin production (>10 °C). the organism may contaminate the product. are particularly susceptible to contamination and.0% salt in the water phase (30). It is a halotolerant bacterium able to grow at water activities as low as 0.5. if the person handling the product is a carrier of Staph. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin.5% salt in the water phase. the organism can survive most of the light curing processes. coli all occur on fish products as a result of contamination from animal/human reservoirs. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . and the minimum pH for growth is 4. Once the product is contaminated.5. as they require more handling. but. An outbreak of Shigella in an Italian cruise ship. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. The organism is mesophilic with a minimum growth temperature of 7 °C.or cold-smoked fish should have at least 3.0 . Infection by Shigella sonnei has increased in the UK. 5. In foodstuffs. has been reported (42). However. they do not grow well in the raw material. 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. the human nose. the risk of infection can be eliminated by proper cooking. related to the consumption of a raw smoked fish meal. it should contain not less than 3.5. if it contains sodium nitrite ≥100 ppm.94 (7). Usually. 5. Shigella and E. throat and skin.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. when the bacterial load of the product is lower. aureus. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. 5. aureus growth.5.30% of the population positive for enterotoxin-producing strains (41). gills and guts as well as in several lightly preserved fish 114 . if storage temperatures are over 10 °C.3 Enterobacteriaceae The pathogens Salmonella. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13).

Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. As they are heat-sensitive.the round worm Anisakis spp. or nematodes. The use of LAB in fish preservation is a topic that has been studied in projects (UP. widely used as starter cultures in food processes.2.12%. from freshwater fish. a short period of freezing either of the raw material or of 115 . which. cold-smoked fish. 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. a number of fish products. and in CT95-1207FAIR). are considered unsafe.5 Parasites Despite the presence of parasites in fish being very common.5. marinades (ceviche) (43) and gravad fish (44). gravad fish. have been shown to have an anti-listerial activity in many food products. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. monocytogenes in levels at or above 102 per gram (46). 5. there were no reported outbreaks related to cured fish. such as ‘lightly preserved’ fish products. the USA established ‘zero tolerance’ for the organism. and the tape worm Diphyllobothrium spp. favours growth of psychrotrophic LAB. most of them are of little concern to public health. monocytogenes’ in 25 g of the finished product.6 and water activities below 0. has also been detected in cod (7).95 (45). Pseudoterranova dicipiens.0. in combination with curing agents and refrigerated storage. and other local traditional products (7). Its limiting conditions for growth are 0 . Of microbiological concern in the northern hemisphere are two parasites . the only concern to public health is when eating raw or uncooked fish products.92 .514. which means that the process must conform to a level of ‘no detectable L. pH 4. from seawater fish.9. Two outbreaks in the United States have been reported to be linked to salmon (10). do not survive 1 min at 60 °C. lightly salted caviar.CURED. which include intermediate hosts. Thus. salt levels of 8 . Until 1997. They have complicated life cycles. a round worm. The round worms. monocytogenes pathogenic for only specific segments at risk. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). which considered L. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. in the EU Fisheries R&D-FLAIR program. Unlike some European countries. Some LAB. This includes matjes herring. SMOKED AND DRIED FISH products such as cold-smoked fish. both being detected in cold-smoked salmon.8 . As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. the CT 3162-FAIR program. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47).45 °C.

perfringens B.<107 ≥107 N/A <100 <20 100 . These foods fall into category 5.6 Published Microbiological Criteria TABLE 5.<104 20 .<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 .<100 200 . If the Bacillus counts exceed 104/g. salami.<100 >104 >100 N/A N/A <20 20 . and the levels or absence of indicator organisms or pathogens.<104 100 . coli O157 & other VTEC Vibrio. smell.<100 20 . ‡ On occasions some strains may be pathogenic. E.100 >100 N/A Pathogens Salmonella spp.FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). aureus C. and unpasteurised yoghurt.<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.<104 104 . (not L. cereus and Bacillus subtilis# <20 <20** <20 <10 20 . monocytogenes) (cfu/g unless stated) <106 106 . 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. coli (total) Listeria spp. texture. Acceptability is based on appearance. not detected in 25 g Campylobacter spp. for example.<100 10 . 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E. 5.<100 20 . † Guidelines for aerobic colony counts may not apply to certain fermented foods. the organism should be identified # N/A denotes not applicable 116 . monocytogenes Staph.<103 N/A 100 . cholerae Vibrio parahaemolyticus L.VI Guidelines for the microbiological quality of ready-to-eat smoked fish. soft cheese.

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

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

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

Susuki E. Hurst A. Paranjpye R. Pelaez C. Journal of Food Protection.html Gilbert R.E... Amsterdam. 80.8 Further Reading Gibson D. Ed.. Journal of the Association of Food and Drug Officials. 863-6. Clostridium botulinum and its importance in fishery products. Huang Y. Ribeiro C. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects. Davies A. Tham W. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions. 1991.M.. 1992. 47.A. Gibbs P.cfsan. 833-41.. Advances in Food Research. Huss H. Incidence and growth of psychrotrophic Clostridium botulinum in food. Cann D.. Acta Veterinaria Scandinavia. Loncarevic S. Wooten R. Danielsson-Tham M.. Peterson M. Pathogenic microorganisms of importance in seafood.fda. Cured.gov/~comm/haccp4x5. Unnerstad H. 120 . 22. Communicable Disease and Public Health.. 3. Ericsson H. 163-7.N. Torry Advisory Note No. Hughes A. 55 (4).L. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus. Richards J.A.. in smoked and "gravad" fish.. 197-209. 1994. 45. Canadian Journal of Microbiology. 45 (9). in Quality Assurance in the Fish Industry. Donovan T. 35 (11) 2904-7..H. 135-85. Little C.. 2001... Hobbs G. 26 (4). Microbiological guidelines for some ready-to-eat foods sampled at the point of sale. Washington D. 1989. Journal of Food Protection.. 188 (3). Prevalence of Listeria monocytogenes and other Listeria spp. 1995. and smoked fish establishments good manufacturing practices.. Eklund M.. Fletcher R. Association of Food and Drug Officials.C.V. 1996.D. Zeitschrift für Lebensmittel-Untersuchung und –Forschung. 5 (1). Food and Drug Administration. Tham W. 1982. Food Control. 1976. 1980. Roberts D. 2000.. http://www. US FDA Centre for Food Safety & Applied Nutrition.. Jacobsen M. 1997. Journal of Clinical Microbiology..M. Eklow A.. Elsevier Science Publishers B. 37 (1). Eds. Liston J..S. Harrison M. 232-6. 1980.. 507-10. An outbreak of listeriosis suspected to have been caused by rainbow trout.. de Louvois J. Round Worms in Fish.FISH AND SEAFOOD 44.. Nye K..C. 58 (8).. Moral A. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon.R.A.J. Beltran A. Danielsson-Tham M-L. 13-8. Loncarevic S.. Persson I. Bolton F. Mentzing L-O.. Appendix 5: FDA and EPA Guidance Levels. 46. Pelroy G. Garren D. 5. 49-59.J. 5-7.W..W. 48. in Fish and Fishery Products Hazards and Controls Guide. US FDA Centre for Food Safety & Applied Nutrition. salted.

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

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

where the bacterial production of lactic acid is a major feature. who had adopted them from the Greeks. made from fish livers in Greece. although garos. These have long since fallen from use.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. made from whole fish in France. and pissala. a variety of products can be produced within a single country. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. to products analogous to other lactic fermented foods such as salami. This is well exemplified by some of the fish/salt products of Thailand described in Table 6. In Europe. salty taste.III. may be direct descendants. 6. A selection of Asian products is presented in Table 6.II. 124 . Fermented fish products are of very minor importance outside Asia. 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. and the more widely known Worcestershire sauce a very distant relative. 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.1.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred.FISH AND SEAFOOD TABLE 6.

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

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

however. and the anaerobic conditions that prevail in the bulk of the material.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. in many cases. Bacillus spp. This. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. angkak (Philippines and Thailand) and look-pang (Thailand) are used. More recently. (3. either directly or as the partially saccharified rice they produce. they can contribute to the initial microflora. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. Mucor. Endomycopsis and Saccharomyces . suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). These preparations contain a mixed microflora of moulds and yeasts. 6. 4). In some cases. after one month.4. mean that microorganisms have been generally held to play little or no role in the process that follows. or boiled or roasted rice. Generally. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). 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. When this is a simple carbohydrate source such as sugar. which included nam-pla and patis. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. the Philippine fish sauce.1 – 6.3 6. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. however. were found to be the predominant isolates. traditional starter culture preparations such as koji (Japan). its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). It also demonstrated their significant proteolytic activity during the first month of fermentation.FERMENTED FISH Where other ingredients are used. Other halophilic species have been identified from fish/salt products. and studies 127 .3. In another microbiological study of four fish sauces. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7).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. Hansenula. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. Similar results were seen with patis. sufficient to saturate the water phase present and produce an aw of 0. 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.74 or below. principally species of the genera Rhizopus.

Patis (Condiment) 128 . 6. Fig. 6. Belacan (Condiment) Fig.1.12).FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 .2.

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

6 – 6. Hoi-Malangpu-Dong (side dish. volatile fatty acids and nucleotides. particularly glutamate.8. for example.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. Fig. filtered and bottled. Figure 6. 6. In contrast to free-swimming fish.5). snack) 130 . 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.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. Some examples of products and their formulation are presented in Table 6.3. 6. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. which is eventually separated from the residual fish tissues.5.V and Figures 6.

S. Kung-Chom (staple food. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. N.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .12 months Main dish 2 Pla-jao C.7 days 5 . N.5-1. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. 3:1:0. C NE. north.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. central. side dish) 131 . north-east.12 days 3 .03 Preparation time 6 . NE Formulation Fish (whole or pieces): salt: roasted rice. C NE.6. In these cases. south of Thailand.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C. NE.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0.2-4 Fish (pieces): salt:khaomark (fermented rice).4 are also produced using shrimps. N S 5 .3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . N. NE 10 . C.FERMENTED FISH TABLE 6.6. Fig. the prefix 'pla' is replaced by 'kung'.12 months Shelf life Use 1 .

If acidity is a desired sensory property of the product. Generally.1 The use of higher salt levels results in a longer production phase but a better keeping quality product.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. 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. these products are salted at lower levels than the fish sauces and pastes. The LAB isolated from these products are those with more marked salt tolerance but.. It also indicates that.7. Red burong isda 132 .salt and lactic acid. their growth and metabolism are nonetheless slowed. of the two factors contributing to shelf life .3. giving a reduced rate of acid generation.10%. typically in the range 2 . but there is considerable variation in this. 2.2. then this will take longer to develop than if lower salt levels are used. Phillipines Journal of Food Science and Technology.VI).6. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. the salt is of far greater importance in determining the product’s long-term stability. while they are able to grow at high salt levels. This is readily explained by the susceptibility of LAB to elevated salt concentrations. (from Arroyo et al. 106) Fig.

VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.7 .9 4.6.6.6 3. 133 .8 .12 months 1 .2 4.4.5. 106) Fig.1 .0 .5 3.0 .9.FERMENTED FISH (from Arroyo et al. Philippines Journal of Food Science and Technology.8 2.2 7.2 4..2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).1 4.5 .9 .3 .5.6.5 .0 .5.3 5. White balao-balao TABLE 6.0 Shelf life 3 weeks 6 .4 .8 .4.8 pH 4.9 4.9.5.5.8. 2.17.3 years 1 .

alternatively.FISH AND SEAFOOD 6. such as salt content.3. TABLE 6. also lower the unit cost of the product and this is probably a significant factor in many cases.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. Addition of carbohydrate will. being similar (18. a common ingredient in these products.2. although the final population of LAB may be similar.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. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. This results in a more active lactic fermentation and a lower pH. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. Rice is the most common carbohydrate source used.5) despite other factors.1 .9) compared with the white product (pH 4. 19). also serves as a valuable source of the more readily fermented carbohydrate. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. In several products. Garlic. When added as boiled or roasted rice alone.3.VII). Where they are used. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. 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 . of course. the rice may be added in a partially saccharified form such as kao-mark (Thailand). 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).4. For example. 21). inulin. 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 (20.0 . Although the ability to ferment starch is not widespread in the LAB.

3 weeks at around 20 °C. 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. the pH drops from 6. the Korean fermented fish product. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. the level of salt employed is on the high side for fish/salt/carbohydrate products. thereafter declining to about 4.5:9:3.2 at the end of fermentation. is something of an exception since millet is used along with garlic as the carbohydrate source. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. will not reliably exclude air. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. In a successful fermentation. reporting the product’s pH to range from 4.8 . and there is a marked regional variation in the preferred type. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. such as wrapping in banana leaves.3 days. in turn. 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).7 up to 6. Generally. 135 . red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7.7 to below 4. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products.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). around 12%. markedly extending the shelf life (25).5 in the first 2 . allowing other spoilage organisms to grow. Traditional packing techniques used with lactic fermented fish products. Excessive growth and acid production by the LAB could render products too sour for acceptability.9%.FERMENTED FISH Sikhae. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. which ranged from 7. Results of a product survey were consistent with this. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. Clearly.5. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. 6. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. but they are likely to behave similarly to lactic fermented products produced from other commodities. The mixture is allowed to ferment for 2 . lead to an increase in pH. There is little published information on the factors that limit the shelf life of the low-salt products. presumably correlating with the salt content.17. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23).2.

outbreaks of 136 . however. Several measures have been suggested for reducing the risk of botulism. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. Because of the severity of the illness it causes. As in Norway. and consumed without cooking. viable numbers will probably decline quite rapidly. The bacterial pathogens most commonly associated with the living environment of fish. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. which accounted for 88% of outbreaks in the period 1952 . There is little in the way of documented information associating fermented fish products with foodborne illness. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. In the fish sauces and pastes. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. 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.87. although these limits apply when all other factors are optimal (26). under conditions inimical to growth and high ambient storage temperatures.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. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. V. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90.8 and at salt levels as high 10%. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. After capture. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). are Vibrio parahaemolyticus and Clostridium botulinum. In Japan. During fermentation. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. parahaemolyticus has been reported growing down to a pH of 4. 6. typically 30 °C. but we are unaware of any reports of this happening. such as soaking the fish at temperatures below 5 °C before fermentation and the use of starter cultures to ensure a rapid and reliable decrease in pH. fish products are the most common source of botulism.

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

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

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

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

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

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

Human symptoms begin with tingling and numbness of the lips. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. this is never sufficient to render the seafood safe to consume. and fingers within 5 . 7.1. tongue.30 minutes of consumption. 143 . Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7.2. However. and humans seem to be particularly susceptible. 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). 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.2 Toxicology PSP toxins affect the central nervous system of mammals.FISH AND SHELLFISH TOXINS STX GTX2 GTX3 GTX5 (B1) C1 C2 NEO GTX1 GTX4 GTX6 (B2) C3 C4 R1 H H H H H H OH OH OH OH OH OH R2 H H OSO3 H H OSO3 H H OSO3 H H OSO3 - R3 H OSO3 H H OSO3 H H OSO3 H H OSO3 H R4 H H H SO3 SO3 SO3 H H H SO3 SO3 SO3 - Fig.

Thailand. a criticism has been that such methods are time consuming. However it remains as the monitoring method of choice in many countries. Most intoxication events have occurred in South East Asia. Two approaches have been taken here. TTX most commonly occurs in many species of puffer fish. After some development and various trials. While the chemical detection methods are most promising for the future. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). and over ten analogs are known (22. 7. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. other derivatives have been reported. as well as other parts. Taiwan. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). Alternative HPLC methods have subsequently reduced this to a single HPLC run. 23.2). 7. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12.2. a pre-column method has received Association of Analytical Communities (AOAC) approval (18).3.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. and islands of the South Pacific. 15. 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. and has also been evaluated by a number of EU laboratories.FISH AND SEAFOOD 7.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. The mouse bioassay has been in place for decades and the method has been standardised. 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. Since its discovery. including Japan.8 µg/g) of shellfish or seafood tissue. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. 17). specifically 144 . 24) (Figure 7. China. and while simple and relatively inexpensive to operate. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation).1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes.

This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. 7. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. 7. 27).2. All of the observed toxicity is secondary to the action potential blockade. 26).3. In Japan. 23. An oral dose of 1 . where puffer fish of the genus Fugu are a delicacy. and blocks diffusion of sodium through the sodium channel. hypotension. and even newts.FISH AND SHELLFISH TOXINS in the liver. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. and once again the mouse bioassay has been used to detect the presence of the toxin. Since then. it is served in speciality restaurants with qualified chefs to properly prepare the fish. 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. gonads and roe (24). preventing depolarisation and propagation of action potentials in nerve cells. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. The onset of symptoms usually occurs rapidly. and more recently by an actinomycete Norcardiopsis dassonvillei (26). respiratory failure and coma. Structure of tetrodotoxin 7. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. 25.3.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. and in mild cases results in distal muscle weakness. A review of TTX detection methods has been published (12. molluscs. the blue-ringed octopus. Typically this has been accomplished using HPLC followed by post 145 . horseshoe crabs. Fig. raising speculation that TTX is produced by a bacterial source.2 Toxicology Like the PSP toxins.2 mg of purified toxin can be lethal. nausea and vomiting. Severe poisoning causes paralysis and respiratory failure.

7. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28).4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). need further development (24). Currently 12 isomers of DA have been reported (see Figure 7. and while promising.3 for examples).3. a polar glutamate agonist. Fig. but 146 .4.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). 7. 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). but all of them are considerably less toxic than DA itself (29). However this method has been criticised for being complex. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule.FISH AND SEAFOOD column oxidation to the fluorescent derivatives. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. Immunoassay methods have also been investigated. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. but it is known that such congeners are less toxic than DA itself (30).

Okinawa and more northerly parts of Japan. Recently. but any impairment of the blood brain barrier results in additional risk. it has been reported that cooking or steaming contaminated mussels merely has the effect of spreading the toxin to other tissues of the shellfish as a result of disruption of the digestive gland. For example. Pseudonitzschia australis. disorientation and coma. and is mainly distributed in the blood. as does impaired renal function which increases serum concentration and residence time of the toxin. When DA was identified as a new marine toxin it was quickly determined to be a glutamate agonist that binds to a sub-class of neuronal receptors called kainite receptors. and review of the toxicologic pathology of DA has been published (36). Another factor is the movement of the toxin through the food chain. multiseries. 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.4. including persistent short term memory deficits (36 .FISH AND SHELLFISH TOXINS only three strains (P. Victims display a variety of symptoms including headache. has been reported as a medium-level producer of DA (33). southern and western portions of Ireland. Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. nausea. and clams. Levels of DA in the other strains are generally <1 pg/cell. the Mediterranean. Penetration of the blood brain barrier is poor in healthy subjects. and this has led to ASP toxicity in birds. another diatom species Nitzschia navisvaringica. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). To a great extent. which has been well documented in studies along the west coast of the US.. 7. The oral absorption of DA is around 5 . Indeed. scallops.10% of the administered dose. the Philippines. found in the Pacific around Vietnam. seizures. 33). 147 . The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. cormorants and pelicans (34). 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. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. and sea animals such as sea lions and even whales (35). 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. and in locations around New Zealand (32.39).

and an immunosensor method has also been reported (4). However. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. and this is typically the method of choice. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). In the context of this chapter.FISH AND SEAFOOD 7. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). but just over 15 years ago a third toxin. produce a variety of other 148 . Dinophysis spp. The benthic Prorocentrum spp.. 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). To further complicate matters. as can LC/MS. although in the latter study the probes for Pseudonitzschia pungens. 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. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. and these methods have been reviewed (40. in Monterey Bay. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them.4. dinophysistoxin-1 (DTX-1). australis. the heterotrophic dinoflagellates Dinophysis spp. and dinophysistoxin-2 (DTX-2) (Figure 7.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). 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. California. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. Since then. Alternatively. 41). are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . 48. The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. DTX-2 was discovered in contaminated Irish shellfish (46). and consequently were often overlooked as the real source of the toxins.4). Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective.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. 7. though this method is most commonly used to analyse plankton and seawater (42).. 49).

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

Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. 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. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48).160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. 7. 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. Since the late 1970s.5. and DTX-2. and abdominal pain. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. Human symptoms include diarrhoea.5. A regulatory level of 80 . Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. nausea. This regulatory level includes the possibility of LSPs in the DSP fraction being tested.3 days. or indeed on the presence of other associated LSTs that can occur with DSP toxins and which result in a positive mouse bioassay result (vide infra). DSP episodes have been reported worldwide and are included in many monitoring programs (47). and while this provides critical seafood safety information. New Zealand and Chile. and a detection limit of 100 µg/kg has been reported (12). even with the minimal of clean up steps (48). vomiting.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. In recent years. both critical enzymes involved in many key metabolic pathways in mammalian cells (50) and consequently affect a host of other biological processes in a cell including tumor promotion. it does not provide information on the DSP toxin profile. 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).3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. 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. various European countries.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. Due to the lipophilic nature of the toxins. DSP-Check is a commercially 150 . 7. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. which can lead to an underestimation of the total DSP content in a sample. DSP episodes have been reported from Japan.

58). 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. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. 7.5 and 7. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). 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. Within this group. Indeed. for many years these compounds were included in the DSP toxin complex. In yet another approach. 7. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. However.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. 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. Some representative examples of both groups of toxins are shown in Figures 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). fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays.6. 55). As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . with a claimed detection limit of 20 µg/kg. with good agreement with the mouse bioassay and LC–based methods (54. and another rapid antibodybased test kit for DSP toxins has also been reported (53).

As is typical for many algal species. the toxin content and profile may vary among different strains. 58). 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). many parts of Europe and the Mediterranean. 7. 57.1 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).6. Structure of the two parent yessotoxins. Japan. New Zealand and South America (49. which may co-occur in blooms with other organisms.FISH AND SEAFOOD Fig. 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. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. eastern Canada.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. YTX and homo-YTX 7. While 152 .6.6. 7.

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

7. was chemically characterised as PbTx-2 (67). which are found in Japanese waters (66).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. Many of these species are associated with massive fish kills. PbTx-1 and PbTx-2.7. but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65. and off the coast of New Zealand (65). oysters. Interestingly.FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida. and Heretosigma akashiwo. They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. Structure of the two parent brevetoxins. brevetoxins are found in mussels. 154 . In the Gulf of Mexico. namely Chatonella marina. verruculosa off the mid Atlantic coast of the U. whelks. Fibrocapsa japonica. and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf.S. and one of the brevetoxin metabolites BTX-B2 7. 66). 7. Fig. These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). 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). and cockles (65. the same toxins have been reported to be produced by three genera of Raphidophytes.

and mammalian body fluids without pretreatment or dilution (72). malaise. this family of toxins has proven to be a challenge both to identify and monitor. 66. vertigo. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. the competitive ELISA method correlated most closely with the mouse bioassay results. shellfish homogenates. and diarrhoea. 7. The mouse bioassay is used as the primary detection method.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68).8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. and in severe cases seizure. 7.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. 69). The brevetoxin metabolites are reported to be less toxic than the parent compound (70). but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). While the mouse bioassay continues to be used. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish.7. The mean time to onset of symptoms is around 3 hours. 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 human health implications following exposure to NSPs have been reviewed (69). with hot spots in the coral reef areas of the Caribbean and French Polynesia. 7. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. which can result in a cascade of associated events in a cell (66). nausea. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 .3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. with a mean duration of symptoms of around 1 day.7. Ingestion of brevetoxins results in a variety of symptoms including parathesia. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. In this application. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65.

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

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

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

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

Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. and erosion and bleeding in the stomach. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. 7.9.12). Importantly these pathological outcomes were reported as being different from those caused by DSP. the pinnatoxins. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . though the possibility exists that they may inhibit other phosphatases. 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). 92).3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. abnormalities in the thymus and spleen. the spirolides.10 – 7. 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. 91.10 Cyclic imines These toxins are represented by several distinct structural types. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. parenchymal cell pyknosis in the pancreas. and ASP. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. Once again. AZAs also display cytotoxicity towards various mammalian cell lines (83.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. Using the same approach. However. and closely related ptierotoxins (Figures 7. but all contain a spirocyclic or cyclic imine function within their structure. 85). and necrosis of epithelial cells on the microvilli (91). 7. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. PSP. Based on IP minimum lethal dose data. 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. which are particularly amenable to detection by positive ion electrospray mass spectrometry. The mouse bioassay has been used to monitor shellfish extracts. 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.

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

The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. 50%) of 96 µg/kg. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. Most cyclic imines lack an extended chromophore.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). almost a dozen derivatives in total have been identified to date (94. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. 95) (Figure 7. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay.100).11). extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. 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.10. and this is the detection method of choice.FISH AND SEAFOOD 7.1. Whether isolated from dinoflagellate cells directly or from shellfish tissue. The time lapse between ingestion and death is between 5 and 15 minutes.900 µg/kg). and so spectroscopic detection by UV methods is not favoured. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. finishing with 100% methanol. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . 7.1.to10-fold reduction of toxicity (LD50 600 . This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. paralysis of the hind quarters and eventually complete immobility. Based on this diminished oral toxicity. In particular. There is a rapid onset of symptoms including respiratory distress. Reduction of the imine group eradicated the toxicity.10.3 Detection methods The characteristic “fast-acting. 95). together with a lack of evidence of human intoxication. 7. However. abdominal breathing. and lack of responsiveness to stimuli. Since their original discovery which resulted in the characterisation of spirolides A-F (99. Oral administration of the toxin to mice by gavage resulted in about an 8. A later study using purified toxin established an LD50 dose (Lethal Dose.10. They were originally discovered in extracts of mussels (M. it has been stated that regulation against gymnodimine is not required (98).

the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). Structure of four spirolide toxins A-D 7. fasted mice). the western coast of Scotland. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. Fig. 95). these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . While oral toxicity is considerably less than by IP injection. the gulf of Maine. 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.10.2. fasted mice).707 µg/kg. or by oral ingestion (381 . This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 .11.10. southern Ireland.166 µg/kg. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. 7. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. 7.8 µg/kg).2. All data reported here are for desmethyl spirolide C.2 Toxicology The spirolides are the most studied group of the cyclic imines. in fjords in Denmark and Norway. those spirolides possessing the vicinal dimethyl grouping (see Figure 7.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). by gavage (87 .11. Interestingly.

95. 7. four members of the group have been identified – two of them being the C-34 diastereomers.10.B contain a spiro imine moiety similar to that found in the spirolides. consistent with modulation of acetyl cholinergic receptors (103). Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. which includes two diastereomers.2.10. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. the mouse bioassay can be used to detect these fast acting toxins. 102). Fig.12.12).3 Detection methods As discussed for gymnodimine. 7. While the mechanism of action remains to be clearly defined. and in fact these compounds share a number of other structural features. Three members of this group have been reported.FISH AND SEAFOOD (100). experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. To date. 7. A number of methods following this procedure have been described (94.3 Pinnatoxins and pteriatoxins Pinnatoxins A . However. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . 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.

180 µg/kg). 7. Despite this progress. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. new toxin groups such as the azaspiracids and cyclic imines have also been reported. While this is recognised as a real possibility for a wide range of toxins. like the gymnodimines and spirolides.3. 7. and more advanced means of detection. but this remains to be conclusively established (106).10. there is no compelling evidence to suggest that they have resulted in human toxicity (98). 99%) values for pinnatoxins A-C are in the range (22 . while pinnatoxin D was essentially inactive. which occupy tropical waters around Okinawa and the south east coast of China. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). The LD99 (Lethal Dose. 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. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104).100 µg/kg). once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures.10.3. With increasing knowledge of the chemistry of toxins. to date action or 165 . The LD99 dose for the pteriatoxins is reported to be in the range (8 .11 Conclusion There has been considerable progress in the detection of known marine toxins.most likely a dinoflagellate.2 Toxicology There is little information describing the toxicology of these compounds (98).FISH AND SHELLFISH TOXINS 7.1 Origins and distribution Shellfish of the genus Pinna. have long been suspected of causing human illness after ingestion. There is strong circumstantial evidence to suspect that these toxins. which in many cases are now found to be more widespread than previously believed. However.3 Detection methods No formal detection methods for these compounds have been reported. 7.10. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. Only the acute toxicity in mice following IP injection has been reported. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. arise from an algal source . Coupled with this.3. This is generally a result of increased monitoring and continually improving detection technologies and methods.

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

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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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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6. ingredient and packaging buyers. procedures. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. 5. operation or stage in the food chain including raw materials. 8.3.The application of methods.A point. 3.1 Assemble HACCP team HACCP requires management commitment of resources to the process. 8. Establish the critical limit(s). 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. 7. 2. Determine the CCPs. Establish a system to monitor control of the CCP. 4. tests and other evaluations. The team should also include 177 . 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. food microbiology. quality assurance or control.Obtaining evidence that the elements of the HACCP plan are effective. Establish documentation concerning all procedures and records appropriate to these principles and their application. Validation .1 is a flow diagram illustrating this 12-stage logic sequence. procedure. Figure 8. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. Each stage is discussed below in detail.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. Verification . in addition to monitoring to determine compliance with the HACCP plan. Establish procedures for verification to confirm that the HACCP system is working effectively. food chemistry or engineering. hygiene managers. from primary production to final consumption.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . Conduct a hazard analysis.

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

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

The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. 180 ● ● . conduct a hazard analysis. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. because the hazard analysis and decisions regarding CCPs are based on these data. 8.3. once determined.6 List all potential hazards associated with each step. The operation should be observed at all stages. it is essential that the details of each step are fully appreciated and recorded. features of equipment design. time/temperature data.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. cleaning and hygiene procedures and storage conditions. 8. and therefore a detailed description of the procedure applied must be available. or circumstances of any reclaim or rework activity.FISH AND SEAFOOD product. If HACCP studies are applied to proposed new process lines or products. ingredient specifications. 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. and identify any measures to control identified hazards The HACCP team should list all hazards that may reasonably be expected to occur at each step in the production process. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. However. 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. For example. It is essential that the diagram is accurate.3. the qualitative and/or quantitative evaluation of the presence of hazards. survival or multiplication of pathogenic microorganisms.3. Ideally.4. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. it should also include details of CCP steps. 8.5 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.

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

and its use may not always be appropriate.2) to provide a logical. However. biological or sensory parameters.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. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. chemical. Criteria used to set critical limits must be measurable and may include physical. However. Where possible.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. given the enormous variety of fish products and processes in use. it is not possible to generalise on likely CCPs. critical limits should be specified and validated for each CCP. and the reader is referred to the appropriate product chapter. More than one critical limit may be defined for a single step.3. If a significant hazard has been identified at a step where control is required for safety.7.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8.3.FISH AND SEAFOOD 8. but for which no control exists at that step or any other.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. it can be said that effective control measures are likely to include the following: • • • • • 8. structured approach to decision making. application of the decision tree should be flexible.3. 8. 8.8. For example. 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 .1 Fish and seafood Again. then the process must be modified to include a control measure. it is necessary to specify both time and temperature for a thermal process.

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

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

Regulation 854/2004 relates to the organisation of official controls on products of animal origin. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. 9. packaging. transportation. as last amended by Regulation (EC) No 806/2003. as last amended by Regulation (EC) No 806/2003. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. distribution. 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. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. p.2 Legislative Structure From 1 January 2006. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No.1991.15).B. 24. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. manufacturing. A number of more detailed implementing and transitional measures have been adopted at EC level. Subsequently. 24.1991.7. handling and offering for sale or supply to the consumer. 7. processing. 41). 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. J. and sets out what those enforcing the provisions have to do. 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. p. European Communities L 268. 190 • • .1). N. p.1992. 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. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004.9.9. storing.

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. and binding in its entirety on all member states from the date of entry into force. penalties and powers of entry. 2006 No. etc.) in England. The Food Hygiene (England) Regulations 2006 (S. the legislation is directly applicable law. food storage that is for private or domestic consumption. As regulations.1993. domestic preparation or handling.11) The EU hygiene regulations apply to all stages of food production including primary production. is required to give effect to the EU regulations. right through to the implementation of procedures based on HACCP principles. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No.I. Wales and Northern Ireland. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs.g. 852/2004. 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. and equivalent legislation in Scotland. 1 . and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. p. and of hunting and fishing. Wales and Northern Ireland). setting offences. 2073/2005. of stock farming. national legislation in the form of a Statutory Instrument (S. which will be covered later in this chapter. There are some exemptions. Primary products are defined as products of primary production including products of the soil. Additionally the requirements of EC Regulation 853/2004 must be complied with.14. for example. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. revocation of existing implementing legislation. Although the regulations have the force of law. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No.I. 9. 191 . 19.3 Regulation (EC) No. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. e. with primary production. 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.7.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175.

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

1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. 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.2 Fishery product requirements 9. please refer to Regulation (EC) 853/2004. EK.2. and the characters easily decipherable and be clearly displayed. (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. EC. 198 .2 Form of the identification mark (i) The mark must be legible and indelible. the food business operator may apply the same identification mark to both types of food. (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.4. EO. EÜ. EY.1. EB or WE. EF. (iv) If the identification mark is applied in an establishment within the Community.4. 9. a new mark must be applied to the product. Section I of Regulation (EC) 853/2004.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. the new mark must show the approval number of the establishment where this occurred. ES. EG. for other codes. the mark must be oval shaped and include the abbreviation CE.4.1.4.2. (iii) Additionally. For the UK. the mark must show the approval number of the establishment. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not.2.FISH AND SEAFOOD 9. the code would be UK. when a product's packaging and/or wrapping is removed or is further processed in another establishment. Section I of Regulation (EC) 853/2004 are given as follows: 9. (ii) However. In this case. Extracts of the requirements of Annex II.

i.LEGISLATION 9. Extracts of the requirements of Annex III. With the exception of Chapters I and II. granulate and powdered products of animal origin carried in bulk. Section VIII of Regulation 853/2004.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. the mark may be applied to the outer surface of the container or packaging. The requirements of this section accompany those in Regulation (EC) 852/2004.1. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code.3 Method of marking (i) The mark is to be applied directly to the product. its wrapping or the packaging. for full requirements. specifically relating to fishery products are given below. (ii) For products of animal origin that are placed in transport containers or large packages. as amended. Chapter III. This section does not apply to bivalve molluscs. they would need to be obtained following Section VII on live bivalve molluscs. and are intended for further handling. Chapter IV. and fishery products carried in bulk. a) requirements for establishments. reference should be made to the actual Regulation. (v) When the mark is applied directly to products of animal origin. processing. it is sufficient to apply the mark to the exterior of that package only. the colourings used must be EC authorised colours. Part A and Chapter V apply to retail.e. tunicates and marine gastropods when placed on the market live. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 .2. including vessels. 9. the wrapping or the packaging. Parts A.2. the approval number of the establishment and the abbreviation for the community where appropriate. it applies to those animals when not placed on the market live. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels.4. Therefore. (iii) In the case of liquid. wrapping or packaging in another establishment.4. echinoderms. or be printed on a label affixed to the product. C and D.

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

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

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

handling fishery products A. part I.Requirements for establishments. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. Chapter I. they must be stored under ice in appropriate facilities. Additionally. (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. and where necessary. 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. Fillets and slices must then be removed from work tables as soon as possible after preparation. packaged and chilled as quickly as possible after preparation. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . including vessels. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed. 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. 854/2004. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices.LEGISLATION consumption. dispatched. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. 9.2. C. prepared or processed immediately after arrival at an establishment on land. points 1 and 2. C. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice.2. They should be wrapped.3 Chapter III . 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. B. (ii) Heading and gutting must be carried out hygienically. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels.4.

If no other method of preservation is used. if the processing is insufficient to destroy nematode larvae. raw materials must have no guts. and whole fish must be gutted and washed beforehand.Requirements for certain processed fishery products A. (iii) A document from the manufacturer. or a stabilising treatment. (ii) mackerel. cooling must continue until a temperature 204 . (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. they must be cooled rapidly.4.2. except when supplied to the final consumer. D. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. and (c) marinated and/or salted fishery products. stating the type of process they have undergone.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. (iii) sprat. must accompany fishery products referred to above when placed on the market. Once produced. 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. (b) fishery products from the following species. 9. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting.4 Chapter IV .2.20 °C in all parts of the product for at least 24 hours (this treatment must be applied to the raw product or the finished product): (a) fishery products to be consumed raw or almost raw. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. (iv) (wild) Atlantic and Pacific salmon. Requirements concerning parasites (i) The fishery products below must be frozen at no more than .

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

Diodontidae and Canthigasteridae.4. Fresh. 9. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. and when fishery products are wrapped on board fishing vessels.7 Chapter VII . 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. 206 . in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. 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.6 Chapter VI .2. In the case of whole frozen fish in brine intended for the manufacture of canned food. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. 9. it should be kept at no more than -9 °C. Molidae.2.4. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing.FISH AND SEAFOOD D.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. prepared and processed fishery products belonging to the family Gempylidae.2. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. Fishery products contaminated with parasites may not be placed on the market for human consumption. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. The scientific name must accompany the common name on the label.Storage of fishery products (i) Fresh fishery products.2. E. 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.

9.4. species belonging to the Gadidae family. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. Note: (i) Frozen fishery products do not need to be kept at a temperature of -18 °C if they are being transported from a cold store to an approved establishment to be thawed on arrival for the purposes of preparation and/or processing. provided that the journey is short and the competent authority allows this.2. Sebastichthys capensis. melt water must not remain in contact with the products.2. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. species belonging to the Merlucciidae family.).4.Transport of fishery products The above points for Chapter VII apply in relation to transport. The temperature of frozen fishery products must not fluctuate above -15°C. 2074/2005.3 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. (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 .2.8 Chapter VIII . The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. (ii) If fishery products are kept under ice. 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.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. 9. Helicolenus dactylopterus.

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

B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. 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 B means treatment in a purification centre or after relaying is done before it can be placed on the market .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.3.4.Hygiene requirements for the production and harvesting of live bivalve molluscs A. 209 . poles or other fixed means are to identify the boundaries of the sites. C. collection from: . Transport needs to allow sufficient drainage for best survival conditions. 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.Class A means they can be placed directly on the market . processing or relaying. Requirements for production areas (i) The competent authority classes (classed A. (ii) Provided the requirements of chapter V are met.2 Chapter II . B. 9. 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.

(ii) amnesic shellfish poison (ASP). 1 milligram of yessotoxin equivalent per kilogram. 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. Requirements for purification centres There are requirements for live bivalve molluscs to be washed.3.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. Requirements for dispatch centres Handling should not result in contamination to any degree.4. 9.3.4 Chapter IV .4.4.5 Chapter V .FISH AND SEAFOOD 9. the purification system to allow them to resume /maintain filter-feeding activity. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. requirements if there are many batches of live bivalve molluscs.Hygiene requirements for purification and dispatch centres A. 210 . and there are requirements for tanks and water storage containers. (iv) yessotoxins. dinophysistoxins and pectenotoxins together.3 Chapter III . for: (i) paralytic shellfish poison (PSP).3. B. 160 micrograms of okadaic acid equivalents per kilogram. the levels of marine biotoxins should not exceed the following limits. Before dispatch. 800 micrograms per kilogram. containers and a label to identify purification.

by way of derogation from directive 2000/13/EC. Registration document requirements also apply. Requirements of Chapters III and IV are to be complied with.7 Chapter VII .4. they cannot be re-immersed in or sprayed with water.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards. 160 micrograms of azaspiracid equivalents per kilogram. 9. the label.4. dispatch centre or processing establishment. 211 .4.4. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre.2.6 Chapter VI .1). (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.3.4.8 Chapter VIII . the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.3.LEGISLATION (v) azaspiracids. must be waterproof. 9.3.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.9 Chapter IX . (ii) Individual consumer-size packages of live bivalve molluscs must be closed and remain closed after leaving the dispatch centre and until presented for sale to the final consumer. the species of bivalve mollusc and date of packaging (at least the date and month) are required. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction. 9. including the identification mark.3.Specific requirements for pectinidae harvested outside classified production areas Requirements for harvesting and handling following harvesting and health standards for live bivalve molluscs must be met. Finally.Identification marking and labelling In addition to general identification marking (contained in 9. Also.

The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. echinoderms. 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. 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.5. This refers to official controls or production and placing on the market of fishery products.5 Regulation (EC) No. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. or (iii) bivalve molluscs. chemical. 9. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation.FISH AND SEAFOOD 9. Fishery products in particular must comply with the requirements of Annex III of the regulation. Chapter V of Regulation 853/2004 apply. (ii) fishery products not complying with the requirement concerning biotoxins. 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. In this regulation. or - - 212 . Therefore the health standards for fishery products as previously stated in Annex III. they derive from: (i) poisonous fish.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. Section VIII.

chapter V is reinforced. that microbiological criteria and chemical contaminants amongst other requirements are met.5. in 90% of the samples. A summary is given as follows: 9.1 Chapter II . 000 E.000 E. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. 213 . 4. The reference method for this analysis is the five-tube. live bivalve molluscs must not exceed 46. three dilutions MPN test specified in ISO 16649-3. Live bivalve molluscs from Class C areas must not exceed 46. Sampling plans and frequency are required. live echinoderms. The reference method for this analysis is the five-tube. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. In the remaining 10 % of samples.2. Regulation (EC) 854/2004. coli per 100 g of flesh and intravalvular liquid. section VII. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. Also. as previously stated. live tunicates and live marine gastropods.2 This states official controls for the production and placing on the market of live bivalve molluscs.5.Official controls concerning live bivalve molluscs from classified production areas A. Annex II 9. Classification of production and relaying areas: This section reinforces that specified in Annex III. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. coli per 100 g of flesh and intravalvular liquid. coli per 100 g of flesh and intravalvular liquid.600 E.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. - - - - B. Live bivalve molluscs from Class B areas must not exceed. Monitoring of classified relaying and production areas This is to ensure there is no malpractice.

boundaries or class of a production area or its closure. E. 9.2.FISH AND SEAFOOD C.2 Chapter III . 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No.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. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. 9. 9.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. which has applied since 1 January 2006. 2073/2005.5. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. the Member States may permit the import of bivalve molluscs and fishery products from the countries listed respectively in Annex I and Annex II to regulation (EC) 2076/2005. F. Procedures for re-opening a closed production area are stated D. 214 . establishes microbiological criteria for a range of foods. 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. 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.6 Regulation (EC) No. opening or closing a production area.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions.

9. presence or number of microorganisms. with chapter 2. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. volume. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. 215 .I.6. based on the absence. and/or on the quantity of their toxins or metabolites.LEGISLATION The aim of this legislation is to complement food hygiene requirements. per unit(s) of mass.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.II. The criteria are outlined in Table 9. Two kinds of criteria have been established: food safety criteria. area or batch. The relevant criteria are outlined in Table 9. and the legislation applies to all businesses involved in food production and handling. 9. ensuring that foods being placed on the market do not pose a risk to human health. a batch of foodstuffs or a process.6.4 referring to fishery products.

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 1.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life . monocytogenes 1.TABLE 9. other than those intended for infants and for special medical purposes(4) (8) L. monocytogenes. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1. 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.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 1.2 Ready-to-eat foods able to support the growth of L. monocytogenes.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L. monocytogenes L.

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 1.25 Live bivalve molluscs and live echinoderms.17 Live bivalve molluscs and live echinoderms. manufactured from fish species associated with a high amount of histidine (17) Histamine .26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1.27 Fishery products which have undergone enzyme maturation treatment in brine. coli (15) 1. tunicates and gastropods 1(16) 0 230 MPN/100 g of flesh and intravalvular liquid 100 mg/kg 200 mg/kg HPLC (19) ISO TS 16649-3 Salmonella 1. tunicates and gastropods 9 (18) 2 E.

. Bouquelet S. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. J. c = number of sample units giving values between m and M For point 1. This criterion shall apply to products before they have left the immediate control of the producing food business operator. live bivalve molluscs.92. — fresh. — — bottled or packed waters. when he is not able to demonstrate. shall not apply. 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. Dervin C. Scombresosidae. Malle P. according to which the whole batch is to be deemed unsafe. — bread. biscuits and similar products. Clupeidae. 1996. 43-9.. products heat treated in their final package). Single samples may be taken at retail level.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. E. to the satisfaction of the competent authority. 2. wine. to the satisfaction of the competent authority. coli is used here as an indicator of faecal contamination.25. that the product will not exceed the limit 100 cfu/g throughout the shelf life.1 . that the product will not exceed the limit of 100 cfu/g throughout the shelf life. m = M. The most recent edition of the standard shall be used. uncut and unprocessed vegetables and fruits. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . 5 This criterion shall apply if the manufacturer is able to demonstrate. A pooled sample comprising a minimum of 10 individual animals. 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. Valle M. Malle P. Assay of biogenic amines involved in fish decomposition..94.. monocytogenes. when recontamination is not possible after this treatment (for example. beer. Engraulidae. honey and confectionery. Coryfenidae. cider. References: 1.0 and aw ≤ 0. products with pH ≤ 5. 79. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. AOAC Internat.4 or aw ≤ 0. subject to scientific justification. Duflos G. sugar. spirits and similar products. Bouquelet S.. Pomatomidae. Products with pH ≤ 4. Particularly fish species of the families: Scombridae.1. excluding sprouted seeds. Other categories of products can also belong to this category. products with a shelf-life of less than five days shall be automatically considered to belong to this category. including cocoa and chocolate products. soft drinks.

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

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 2.1 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 .4.TABLE 9.

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

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

2006. The revised text of Regulation (EC) No. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. Commission Regulation (EC) No. Regulation (EC) No. 852/2004 is now set out in a Corrigendum (OJ No.6.10.2004. 1019/2008 and as read with Regulation 2073/2005. 25.4. 2075/2005 and Regulation (EC) No. 2076/2005.4. Regulation (EC) No. Regulation (EC) No. these are not be discussed here. The revised text of Regulation (EC) No. 1441/2007.6. 3) as amended by Regulation (EC) No. 25. 1. 30. L283.2004. 1). L226.LEGISLATION 9.9 Other Relevant Legislation The following regulations about fish designations need to be considered: • Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products Regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products • Note: Requirements for preserved fish are laid down elsewhere. L226. 206). 22) as amended by Regulation (EC) Nos. 9. L338. as amended. 2074/2005. 853/2004 is now set out in a Corrigendum (OJ No. 25. as read with the corrigenda at OJ No. L139.2005. L139. 1662/2006. 2073/2005 on microbiological criteria for foodstuffs (OJ No. 1688/2005. 30. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 14. Regulation (EEC) 2136/89. Regulation (EC) No. L139.2004. 83) as amended by Regulation Nos 882/2004.2004.2004. 223 . 55). 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. Regulation(EC) No. 62) as amended by Regulation (EC) No. laying down common marketing standards for preserved sardines. 22. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. The revised text of Regulation (EC) No.10 Further Reading Regulation (EC) No.12.4. 2076/2005. Regulation (EC) No. 1663/2006. namely. 2076/2005. L226. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. 2076/2005.6. 854/2004 is now set out in a Corrigendum (OJ No. 2074/2005.2004. 2074/2005 and Regulation (EC) No. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. 2074/2005. 30.

21. 22. 22.2005. (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004 ( OJ L 338.2001.10.2005. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. 23. 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 854/2004.12.1.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. 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. 6) 224 .12. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.2000. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.

0 .5 µm 10.5 minutes.1 10. 225 .10.10 (3).5 pH Under otherwise optimal conditions.1.0 x 1. Aeromonas has the potential to grow over a pH range of ca 4 .3 Temperature Aeromonas grows at temperatures of 2 . The minimum pH value for growth is thought to be pH 5. under optimal incubation temperatures. At pH 4.000-fold increase in number is observed after one week in food systems (1). 10. no growth is observed (1). At 5 °C. 10.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.45 °C (1).3 .1.1. a 10 to 1.1. 0. Some death will occur at this pH if the temperature is 4 °C.1. respectively (1). Morphology Gram-negative rods. The optimum growth temperature is 28 °C (2).3 minutes. 10.5. PATHOGEN PROFILES 10.2 Oxygen requirements Facultative anaerobe.3. and D-values at 45 and 51 °C have been reported as 29.5.1 Aeromonas spp. and 2.1.

6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. 10. botulinum are known. to a lesser extent. 226 .1. Thus an aerobically packed product may not support the growth of the organism on the surface. types B and F can be either.2 Clostridium botulinum Seven different types of C.2% sodium chloride (NaCl) (1). botulinum are not particularly heat resistant. botulinum types produce heat labile toxins. B.5 . 10. A to G. There are four main groupings of the organism. at 85 ºC for 5 minutes. botulinum is an obligate anaerobe.2 Oxygen requirements Although C. which may be inactivated by heating at 80 ºC for 20 .0 µm.2. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth.FISH AND SEAFOOD 10. and groups I and II are those responsible for the botulism cases. E and.22.5% NaCl (4).3 Temperature All strains of C.1 Morphology Gram-positive. spore-forming rods.4 Heat resistance The vegetative cells of C.2.4 x 1. All C. or at 90 ºC for a few seconds. but the spores of this organism are more so. 10. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C. All type A strains are proteolytic and type E strains are usually non-proteolytic. botulinum grow reasonably well in the temperature range of 20 45 ºC. 10. 0.even as low as 3 ºC (7). and is sensitive to > 4. botulinum is prevented. but Group II strains are psychrotrophic. F are the types that are responsible for most cases of human botulism (5.2.30 minutes. 10.2.2.7 . but the interior of the food may do so. forming at least seven different toxins. Types A. 6). Group I will not grow at temperatures of 10 ºC or less. being capable of slow growth and toxin production at low temperatures .

and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.9 x 2.3 . However.0 and aw > 0. respectively (8.is an obligate anaerobe. 9). 227 .6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. therefore.94 for Group I and 0. where oxygen levels are reduced. safety margins must be introduced to allow for process variability.0 µm.3 10. These values have been established under carefully controlled laboratory conditions.97) (10. vacuum packed. The organism will grow well in the centre of meat or poultry dishes.97 for Group II (5).2 Oxygen requirements C.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. Note also that if humectants other than salt are used. pH and temperature. In commercial situations.0 . but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. These concentrations correspond to limiting aw of 0.1 Clostridium perfringens Morphology Gram-positive spore-forming rods.2.g. parcooked meals with pH value > 5. the minimum aw for growth may be lower. they may survive mild heat treatments (70 . It will not.2.PATHOGEN PROFILES 10.10. 10.7 Characteristics of C. 10. grow on the surface of foods unless they are vacuum or gas-packed.like other clostridia .3. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. 10. 11)).6 and pH 5.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4.0. particularly by cooking. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. 10. botulinum spores The most heat resistant spores of Group I C. botulinum depends on solute.1. 0.3.2. perfringens .21 minutes) (9).

0 .3.3. perfringens in roast beef at 60 ºC is 14. 10.45 ºC. The D-value for C. the enterotoxin is not heat resistant .4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C.FISH AND SEAFOOD 10. will survive greater extremes of pH (and aw) (12. pH. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8.5 pH C. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. perfringens is not a tolerant organism with respect to pH. The lowest aw recorded to support the growth of C.it is destroyed by heating at 60 ºC for 10 minutes (14.93 and 0. Under otherwise ideal conditions. 10. It grows best at pH values between 6 and 7 (the same pH as most meats). Its optimum temperature for growth is 43 .3. lower concentrations may be effective in combination with other factors.50 ºC. 13).3. In addition. perfringens. although C.10 ºC. 10. perfringens has the potential ability to grow within the temperature range 15 .7 Characteristics of C. perfringens strains. Salt concentrations of 6 . Some studies indicate that the presence of 3% NaCl delays growth of C. type of solute. While some growth can occur at 50 ºC. At cold temperatures. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. 10. perfringens spores The spores of C. As in the case of other factors limiting the growth/survival of this organism. perfringens appears to be 0. 15. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. Recorded heat 228 . Spores. perfringens in vacuum packed beef (18). perfringens is not tolerant of low water activities. vegetative cells die rapidly (12). depending on strain and other conditions. 13). perfringens can vary quite considerably in their heat resistance.6 Aw/Sodium chloride C.3 Temperature The most significant characteristic of C.8% inhibit growth of most C.3. etc. and their heat resistance is also affected by the heating substrate. however. 16). the limits for aw are affected by temperature.3. 18).5 minutes (17). (It is unusual for spores to be formed in foods after the growth of this organism (14)).

0. growth at low temperatures can be very slow.3 .4.4 10. 10.4.98 and 0.1 Listeria spp. 229 .PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. and temperature. Morphology Gram-positive short rods. 1.5 .4. being potentially capable of growing . using hydrochloric acid as an acidulant).2 Oxygen requirements Aerobe or microaerophilic. In foods.0 µm.5 x 0.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. the organism is able to grow at pH values well below pH 5 (pH 4. and where acetic acid is used as an acidulant. -0.4 ºC is probably a more likely minimum for growth in foods (19). 10.2 has been suggested as the lowest working limit (22). However.albeit slowly .4. respectively (21).23. 60 and 65 ºC were reported as 10. D-values for L. is between 30 and 37 ºC.3 is the lowest value where growth has been recorded. monocytogenes is reported to be 45 ºC (20).19 minutes.2. the lowest limit for growth is likely to be considerably higher . It has been reported to have slightly greater heat resistance than certain other foodborne pathogens. requiring days or weeks to reach maximum numbers. The upper temperature limit for the growth of L.8 minutes for heat sensitive spores (12).0. it is not a sporeformer. 10. 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. monocytogenes is not a particularly heat resistant organism.4 Heat resistance L. or even below 0 ºC.especially at refrigeration temperatures.64.at refrigeration temperatures down to.0 minutes for heat resistant spores to 1. Under ideal conditions.4 .6 . monocytogenes in crawfish tail meat at 55. so can be destroyed by pasteurisation. however.4. 10. however. pH < 5.2. Its optimum growth temperature.

5. 25).3.5 (24.6 10. It grows best at aw of ≥ 0.5. Morphology Gram-negative short rods. 10. It is likely to survive.97.2 Oxygen requirements Facultative anaerobe. the optimum being 30 ºC (23). 10. at salt levels found in foods (10 .0.6.8. 10.0 µm. 0.5.3 µm.5. The bacterium may survive for long periods at aw values as low as 0.7 x 1.8 .3 Temperature Plesiomonas can grow in the temperature range of 8 .6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.0 .5.1 Plesiomonas Morphology Gram-negative.1. 10. 25).5.0 x 1 .5 pH Plesiomonas has a pH range for growth of pH 4.5 .4.12% NaCl or more).83 (20). monocytogenes is quite tolerant of high NaCl/low water activities.5 10. 10. 0. peritrichous flagella. 10. 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 0.45 ºC. or even grow.6 Aw/Sodium chloride L.5 . 230 .90.1 Salmonella spp.FISH AND SEAFOOD 10.

4% NaCl. It is not. depending on serotype. 10. some strains are able to grow at temperatures as low as 4 °C (26). and appropriate heat treatments must be determined experimentally for low-aw foods. The optimal pH for Salmonella growth is between 6.5 . under otherwise ideal conditions.20 times more heat resistant than the average strain of Salmonella at high aw (27). D60 ºC values normally range from about 1 to 10 minutes. and with an appropriate acid.7.6 Aw/Sodium chloride Where all other conditions are favourable. Some death will occur at pH values of less than about 4. 10.48 ºC. the optimum being 35 . Salmonella senftenberg 775W is about 10 . a heat resistant organism. Furthermore.37 ºC. Salmonellae are quite resistant to drying.6.6.2 Oxygen requirements Facultative anaerobe. Growth is slow at temperatures below about 10 ºC. with a z-value of 4 . substrate.3 Temperature Salmonellae can grow in the temperature range of 7 . temperature and pH. although salt tolerance increases with increasing temperature (28).6.5 pH Salmonella has a pH range for growth of pH 3.8 .5. The growth of Salmonella is generally inhibited by the presence of 3 . or possibly 0. However. Salmonellae are quite resistant to freezing. Salmonella has the potential to grow at aw levels as low as 0. 231 . 10. However. 10.9.4 Heat resistance Salmonella is not a spore-forming organism. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments.0. pasteurisation and equivalent treatments will destroy the organism under normal circumstances. therefore.6. depending on the type of acid and temperature.93 (as reported in dried meat and dehydrated soup).5 ºC.PATHOGEN PROFILES 10.5.945. strains vary in their ability to withstand heating.6.

but may cause some cell damage (30).7. the D60 ºC value is 2 . depending on aw.). with an optimum of 35 . The limits for toxin production are also narrower than for growth.2 to 9.7.0. Most strains of staphylococci can grow within the pH range 4. Staph.3 (optimum 7.20 minutes.0 . 232 .5 ºC. etc. In meat macerate. aw. The following relate to limits for growth only.1 Staphylococcus aureus Morphology Gram-positive cocci. 10.9 µm diameter.2 Oxygen requirements Facultative anaerobe. Under most circumstances.FISH AND SEAFOOD 10. 0.5). stressed cells can also be less tolerant of heating.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. 10.5 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. aw and temperature (sensitivity to acid increases with temperature).7 .37 ºC (29). It can survive well at low temperatures. NB: The growth of Staph. under otherwise ideal conditions (29. 10. fat content. the organism is heat sensitive and will be destroyed by pasteurisation. aureus than citric acid). As is the case with other bacteria.7. 10. aureus viability. Freezing and thawing have little effect on Staph.3 Temperature Under otherwise ideal conditions. aureus is more limited under anaerobic than aerobic conditions. however. aureus can grow within the temperature range 7 – 48.7.7. 31).7.7 10.

46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. In general. 0.99 little or no toxin production in anaerobically packed foods.8.10% NaCl. atmosphere.0 (optimum 7. aureus can grow in 7 .9.6 µm 10.4 .5 . Staph.0 . an aw of 0.7. with a temperature range for growth of 10 .86 is the generally recognised minimum in foods (32).5) (29.8 10. Staphylococci are more resistant to salt present in foods than other organisms. 34).0. and solute.2 Oxygen requirements Facultative anaerobe.PATHOGEN PROFILES 10. * dependent on temperature.7.2. pH.1 Vibrio cholerae Morphology Gram-negative curved rods. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31.3 Temperature The optimum temperature for growth has been reported as 30 ºC.8 x 1. 10.87 and > 0.>0.2 .99 aerobically under otherwise optimal conditions.6 Aw/Sodium chloride Staph.7. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities. It can grow over the aw range 0.8. 31) between 0. In general.7 Limits permitting toxin production 10 . strain.83 . 10. but certain strains can grow in 20% NaCl.8. 233 . An effect of increasing salt concentration is to raise the minimum pH of growth. 10.43 ºC (35). However. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.

984 (35). cholerae may persist on frozen fish for >180 days (36). but only moderately sensitive to freezing (40). 234 .9. cholerae is sensitive to heat at temperatures of >45 ºC. under otherwise favourable conditions.6.0% NaCl (35).1 Vibrio parahaemolyticus Morphology Gram-negative curved rods.9 10.2 Oxygen requirements Facultative anaerobe.8. and it is sensitive to refrigeration temperatures (0 . 10. are 5 and 43 ºC.5 ºC).998. 37.8.0. V.6 (35. 10.6 Aw/Sodium chloride V. with optimum at 7. 10.FISH AND SEAFOOD Also. with an optimum of 37 ºC (39). cholerae can grow over the aw range 0.9.8.9.4. cholerae in crab meat homogenate at 60 ºC is 2. parahaemolyticus is easily destroyed by drying. 10. It can grow in the presence of 0.65 minutes (35). there is evidence that V. 10.1 . but not acid conditions.9. with an optimum of 0. respectively. it has a pH range for growth of pH 5 . 38).97 .3 Temperature The minimum and maximum temperatures reported for growth in laboratory media. declining in number during storage.4 Heat resistance V.5 pH This organism is tolerant of high pH values. 10. The D-value for V.

8 .4 Heat resistance V. depending on solute.6 (41).43 ºC (42).10. respectively. vulnificus is heat sensitive.10 Vibrio vulnificus 10. and are readily destroyed by cooking.PATHOGEN PROFILES 10.2 Oxygen requirements Facultative anaerobe.6 Aw/Sodium chloride V.9. 10. 10. The organism can survive for long periods at freezer temperatures (42).10.5 minutes.5 pH The pH range for growth is 4.4 Heat resistance V. with a temperature range for growth of 8 .4% (39.11.0.10. 235 .29 minutes.6 .9. 10. parahaemolyticus will grow at aw levels as low as 0. respectively (39).3 Temperature The optimum temperature for growth has been reported at 37 ºC. The D-values at 55 ºC for clam and crab homogenate are 0.98 (35. 10. and 2. but the optimum concentration is 2 . The optimum concentration of NaCl equates to aw of approximately 0.02 . 10.5% NaCl. 41).922.9. parahaemolyticus is unable to grow in the absence of salt. 10. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.8. 41). with an optimum pH of 7.1 Morphology Gram-negative curved rods. V. parahaemolyticus are not heat resistant.10. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).

5-19. 1-27.W. Doyle M. Ed. in Foodborne Disease Handbook.P.10.W. Hui Y. Doyle M. Doyle M.8 (42).. 383-408. Foegeding P.O. Hauschild A. Clostridium botulinum. 6. Palumbo S. Betts G.H. 329-49. Marcel Dekker Inc. Marcel Dekker. Washington D.D.R. Dodds K. Marcel Dekker.A.K. Cliver D. 57-63. 236 . Food Control. in Clostridium botulinum: Ecology and Control in Foods... 97-131. 6 (1). Clostridium botulinum. London.. Montville T.O. in International Handbook of Foodborne Pathogens.... 357-67.H. New York. 2..5 .. Diseases Caused by Bacteria. Eds. Gorham J.W. Miliotis M. Beuchat L. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’. Austin J. ASM Press. Hui Y. 1995. 1993. Potential hazards associated with REPFEDS. 10. Kovacek K. in Clostridium botulinum: Ecology and Control in Foods. Clostridium botulinum and Clostridium perfringens. Stelma G. 7. Eds. Volume 1: Diseases Caused by Bacteria.. Microbiology and Molecular Biology.6 Aw/Sodium chloride V.10. Novak J. 1996.5% (42).H. Juneja V.. 2005.. 1-19. 1989.. in Foodborne Disease Handbook.L.L. New York. in Microorganisms in Foods. Volume 5: Microbiological Specifications of Food Pathogens. Kim J. 9. Eds.N.D. Smith J. in Food Microbiology: Fundamentals and Frontiers. 10. International Commission on Microbiological Specifications for Foods. New York.C. in Foodborne Pathogens.W. Gorman J. 8. New York. Volume 1.M. Lund B. 2001.. Stelma G.L.E. in Foodborne Bacterial Pathogens. 111-89. Aeromonas hydrophila group. vulnificus grows over the range 0.H. Gaze J.. Eds. Fratamico P..H.P.D.R. 4. 1989. Principles of control. 121-76. Marcel Dekker. References International Commission on Microbiological Specifications for Foods. Ed. in Foodborne Bacterial Pathogens. 5. Marcel Dekker. Bier J. Murrell K.P.. with an optimum pH of 7.M.. Bhunia A. Ed..FISH AND SEAFOOD 10.J. Dodds K. Caister Academic Press. Blackie. Johnson E.R.11 1.0% NaCl. Great Britain. Murrell K. Cliver D. New York.5. 2003. 279-303. with an optimum of 2. 3.5 pH The pH range for growth is 5 . New York.L. Notermans S. 1994.N. New York. Aeromonas species. Hauschild A. 11.W. Dodds K.. 1993. Eds. 10.D...M. Eds.10. Marcel Dekker. Eds.H. Peck M.. Hauschild A. Abeyta C. Faris A. 1994. Clostridium botulinum. Aeromonas hydrophila. Aeromonas. Marcel Dekker.

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

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

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

FISH AND SEAFOOD 240 .

org.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.holt@ifm.seafish.uk Institute of Fisheries Management (IFM) 22 Rushworth Avenue West Bridgford Nottingham NG2 7LF United Kingdom Tel: + 44 (0)115 9822317 Fax: + 44 (0)115 9826150 Email: info@ifm.uk/Topics/Fisheries Seafish Industry Authority 18 Logie Mill Logie Green Road Edinburgh EH7 4HS United Kingdom Tel: + 44 (0)131 5583331 Fax: + 44 (0)131 5581442 Web site: www.co.uk Web site: www.org.gov.gsi.uk Web site: www.ifm.defra.gov.gsi.org.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.scotland.gov.uk or v.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.uk Web site: www.

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

org Web site: www.cmar.gsi.uk Web site: www.ifr.uk Web site: www.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.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.com/ 243 .hpa.au Web site: www.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.dh.org.ac.org.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.gov.org.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.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.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.nz Web site: www.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: reception-cmar-hobart@csiro.gov.ifst.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.uk/en/index.fdf.seafood.com Web site: www.co.

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

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

FISH AND SEAFOOD 246 .

69-70 Aeromonas hydrophila. growth and survival in crustacean shellfish 86 Aeromonas sobria. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes.INDEX Achromobacter spp. planktonic source of spirolides in shellfish 163 Allergens. as autochthonous bacterial flora of fish 3 Aeromonas spp. pathogenic microorganisms in molluscan shellfish 59-60 as contaminant in enrobing process 37 as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45 growth in vegetable and rice 247 . 111 Astroviruses. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. 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. 102 Bacillus spp. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. in crustacean shellfish 87 Alteromonas spp. 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. 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. preserved by dry/wet salting 99 Anisakis spp. in crustacean shellfish 86-7 ATP. causing spoilage of dried fish 110. 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. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp.

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

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

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

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

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

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

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

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

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. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. 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 crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. 152-3 256 . 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. 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. of shellstock 61. processing for seafood recipe meal centres 39 Vibrio spp. 68 Vibrio cholerae non-01. 62 Yersinia spp. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139.

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