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

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7 Published Microbiological Criteria 1.4 Spoilage 4.1 Introduction 2.2 Initial Microflora 1.7 References 3.8 References 1.8 References MOLLUSCAN SHELLFISH 3.3 Processing and its Effects on the Microflora 1.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.5 Factors Affecting Fresh Fish Spoilage 1.4 Processing and its Effects on the Microflora 2.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. CHILLED AND FROZEN RAW FISH 1.4 Spoilage 1. vii .6 Pathogens: Growth and Survival 2.2 Definitions 2.4 Spoilage 3.8 Further Reading CRUSTACEAN SHELLFISH 4.3 Processing and its Effects on the Microflora 3. 3.6 Pathogens: Growth and Survival 1.3 Processing and its Effects on the Microflora 4.2 Initial Microflora 3.6 Published Microbiological Criteria 3.5 Spoilage 2.3 Initial Microflora 2.5 Pathogens: Growth and Survival 3.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.2 Initial Microflora 4. 4.6 Published Microbiological Criteria 4.1 Definitions 3.1 Definitions 1.1 Definitions 4.7 Published Microbiological Criteria 2.5 Pathogens: Growth and Survival 4.7 References 4.

I.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.1 Introduction 189 9.5 References 93 93 95 96 108 113 116 117 120 123 123 126 127 135 136 137 140 141 141 142 144 146 148 151 153 155 158 160 165 166 175 175 176 177 184 186 6.6 Published Microbiological Criteria 5.2 Legislative Structure 190 9. S. 8.9 Other Relevant Legislation 223 9.6 Regulation (EC) No.7 References 5.10 Cyclic Imines 7.2 Initial Microflora 6. 9.10 Further Reading 223 viii . 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.8 Guidance 222 9.9 Azaspiracids 7.8 Ciguatoxins 7. 14 (Hygiene requirements specific to the UK) 221 9.11 Conclusion 7.8 Further Reading FERMENTED FISH 6. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9. CURED.1 Definitions 5.3 Regulation (EC) No.5. 852/2004 on the General Hygiene of Foodstuffs 191 9. SMOKED AND DRIED FISH 5.3 Processing and its Effects on the Microflora 6.4 Spoilage 6.3 Processing and its Effects on the Microflora 5. 7.6 Lipophilic Shellfish Toxins (LST) 7.1 Introduction 8.4 Regulation (EC) No. EU FOOD HYGIENE LEGISLATION 189 9. 2006 No.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.3 Stages of a HACCP Study 8.5 Regulation (EC) No.5 Pathogens: Growth and Survival 5.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption 212 9.4 Spoilage 5.6 References 6.7 Further Reading FISH AND SHELLFISH TOXINS 7.1 Definitions 6.3 Tetrodotoxin 7.1 Introduction 7.2 Initial Microflora 5.7 Food Hygiene (England) Regulations 2006.4 Implementation and Review of the HACCP Plan 8.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.2 Definitions 8.5 Pathogens: Growth and Survival 6.

10.6 Salmonella spp.10 PATHOGEN PROFILES 10.4 Listeria spp. 10.9 Vibrio parahaemolyticus 10.3 Clostridium perfringens 10.7 Staphylococcus aureus 10.1 Aeromonas spp.2 Clostridium botulinum 10.10 Vibrio vulnificus 10.5 Plesiomonas 10. 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .8 Vibrio cholerae 10.

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

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

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

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

Fish flesh is dominated by the abundance of white muscle in relatively short segments. CHILLED AND FROZEN RAW FISH Associate Prof. offering less than 0. lean fish are those fish that store most of their fat in the liver. Fish fillet composition can vary significantly within the same species due to feed intake. Protein content in fish fillet varies typically from 16 . fins and a streamline body. The lipid content. which can be up to 67%.20%.5% (1). This super class of vertebrates includes all the bony and cartilaginous finfish. In addition. The connective tissue content of fish is also lower than that found in meat. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. giving it its characteristically flaky structure. while fatty fish have fat cells distributed along their bodies. migratory patterns. and other forms of aquatic animal as part of their ‘fish’ definition. 1 . In this chapter. Fish fillets are a poor source of carbohydrates. crustaceans. and excludes molluscs and Crustacea. but typically present a low fat percent when compared to other meats. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. and spawning season. Muscle composition and structure of fish also differ from those found in other meat. most fish contain omega 3-fatty acids and other essential nutrients. However.2 . “fish” will be used for fresh and seawater finfish. typically 3 and 15% of total weight.1. it shows a typical season pattern especially in migratory species such as herring or mackerel. typically fluctuates between 0. Fish are an important part of a healthy diet since they contain high quality protein. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. In addition.21%. respectively (1). and is mostly interspersed between the muscle fibres. Fish can be divided into fatty and lean fish. The lipid fraction is the component showing the greatest variation. ‘fish’ also refers to the flesh of such animals used as food.

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

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

sewage.>106 Salmonella spp. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. 4 . However. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp.102 101 .109 cells/g) * Some sources considered L.14 .0. and risk associated with the consumption of seafood is low (2. numbers of pathogenic bacteria in raw fish tend to be low. Escherichia coli Yersinia enterocolitica 101 .37 °C Unknown Unknown TABLE 1.103 High (107 .19 µg/kg bodyweight from < 102 . stream water. 105 . during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.26 °C 10 .37 °C High for most species. Toxin levels 0.FISH AND SEAFOOD TABLE 1.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00.1 .106 cells/g (exception: V. vulnificus) 5 . Plesiomonas shigelloides 3 . 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. and mud Ubiquitous.II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil. In addition.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp. estuarine environments. 4).106 cells/g.35 °C 8 . human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . birds.

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

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

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

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

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

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

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

and temperate-water fish spoilage rates may be many.5. Fish spoil as a result of the chemical. For fresh fish. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33.5. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. the results of altering one parameter may have far-reaching consequences on others. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. 1. The sum of these changes may result in a different spoilage profile. 34).1 Factors Affecting Fresh Fish Spoilage Temperature By far the most effective method of reducing the rate of whole fresh fish spoilage is temperature control (1.FISH AND SEAFOOD 1. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. The factors are interactive and cannot be completely isolated.3 Recognised specific spoilage organisms (SSOs) The degree of spoilage leading to sensory rejection of fish is partly dependent on the perception of the consumer. The concept of specific spoilage organisms (SSOs) is not new. 5).5 1. the slower the spoilage processes proceed (within limits). 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. but also crucial for the assurance of consumer safety. the lower the temperature of storage. 1. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 .5. Also. a minority are often associated with the majority of the spoilage. biochemical and microbiological reactions taking place within and on the fish. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. For example. by altering the gas atmosphere (extrinsic). The reasons for the differences between tropical. careful temperature control during storage is not only important in terms of quality loss.2 Fish-spoilage bacteria The parameters affecting the multiplication of microorganisms in foods have been categorised into two general groups: intrinsic (inherent qualities of the food) and extrinsic (qualities of the food environment) (35). For fresh fish iced immediately after capture and continually stored in ice.

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

but Sh.how to reduce a molecule that they are not able to take into their cells efficiently. and recent taxonomic description using modern molecular methods bifurcated this species. Sh. and Shewanella profunda were described. Another distinct feature of the physiology of Sh. Shewanella livingstonensis.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. putrefaciens growth has been collected (12. owing to its cell size and activity. This type of intoxication occurs when bacteria convert the histidine 14 . phosphoreum has also been identified as responsible for histamine fish poisoning. 1. It is also able to produce hydrogen sulphide and a range of other off-odour compounds. putrefaciens has been studied. phosphoreum were required for 50% of taste panellists to reject a sample. putrefaciens was not present in such numbers (23.5. putrefaciens lies in its chemotaxis: the bacterium. The ferric reductase activity of Sh. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). 38). and Shewanella aquimarina. Later. whereas >108 cfu/g Sh. the bacteria have a significant problem to overcome . and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. as the ferric iron is insoluble. Research has shown that approximately 107 cfu/g of Ph. 41). phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). it was proposed that this organism. Shewanella oneidensis. The prevalence of the bacterium in so many environments. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39).3. Ph. additional psychrotrophic species such as Shewanalla baltica. does not show carbon-source chemotaxis. a population of Ph. Shewanella frigidimarina. In 50% N2/50% CO2 MAP cod. This wide-ranging respiratory capability is thought to be unique (39). Sh. it is strictly respiratory. Shewanella denitrificans. putrefaciens were required. unlike many other motile bacteria. with several new species being described. putrefaciens strains are characterised as microaerophiles or anaerobes. 41). In terms of its taxonomy. Shewanella affinis. In fact. Shewanella waksmanii. Ph. mesophilic strains of Sh. Shewanella olleyana. Sh. was capable of being in a significant numerical minority on the MAP fish but still able to yield the majority of the TMA thought to be responsible for the rejection (23. they are heterogeneous. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). Shewanella gelidimarina. although. phosphoreum sufficient to cause spoilage was found. Initially. at the time of rejection. putrefaciens were identified as Shewanella algae. baltica has been identified as the main H2S producer in cod during cold storing (37). like all members of the Pseudomonadaceae.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh.

previous exposure and access to supportive treatment.CHILLED AND FROZEN RAW FISH present in fish into histamine. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). Growing evidence suggests a role for B. Fewer still are capable of growing on fresh fish. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. lightly preserved fish products are associated with botulism. bacteria and higher parasites all present some risk associated with fish consumption (1. Huss (1. Types A. 1. Acetate production has been reported as a good indicator of spoilage by this organism. where a particular 15 . 1. thermosphacta in the spoilage of some MAP fish. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. algal toxins.3 days after having reached 108 cfu/g (42). a convenient but diverse species of bacterium with a number of different types. Recent studies have investigated the dominance of B. 1. When inoculated into VP corned beef and sliced ham. human viruses.4 Brochothrix thermosphacta and lactic acid bacteria B. this is probably due to spoilage occurring before toxin elaboration. 5. but with proper cooking. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. E. (and very rarely F and G) have been reported to cause human disease. C2. 75% (with a 35% mortality rate) occurred in Japan.5. B. 5) reports that fresh fish consumption has never been shown to cause human botulism. especially type E botulinum intoxication.6 Pathogens: Growth and Survival Endogenous chemicals. E.3 °C (Group II) and these are non-proteolytic (i. these risks are substantially reduced (1.3. they do not produce significant product spoilage). B. Review of outbreak data suggests fresh fish to be safe. Of these. 5). it is bacterial risks that increase after capture of the fish. 6). botulinum recorded up to 1963. D. The heterogeneity of the types (A.e. Some Ph. B. However. E and F include bacteria with minimum growth temperatures of approximately 3. age.6. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). Of 404 intoxication outbreaks of type E C. botulinum. thermosphacta did not produce off-flavours until 2 . including antisera. F and G) is well documented (44). presents a potential risk. types B. Adult disease is caused by the production of the botulinum toxin. C1.1 Clostridium botulinum C.

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

the causative agent of cholera. but the bacterium is known to survive and grow in the shallow marine. Since 2000. but proper cooking should substantially reduce risk of infection. in its habitat. possibly due to under-reporting because of their likeness to E. shigelloides are encountered. with minimum growth temperatures around 0 °C and optima of 15 . is historically associated with faecally contaminated water. Again. In the US. It is known that Aeromonas spp. which is often fatal (>50%). They are aquatic organisms with an epidemiology that is yet to be fully understood. There is no risk associated with the consumption of properly chilled and cooked fresh fish.000 17 . infection. Similar problems as those encountered for Aeromonas spp. 5. V. Chill storage may not eliminate growth.4 Listeria monocytogenes L. coli on isolation media (50). 1. monocytogenes incidence has been between 0. and especially estuarine environment. however. Most cases have been sporadic rather than associated to large outbreaks..6. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. there is no risk from the consumption of properly handled and cooked fresh fish.55 cases per 100. are associated with human diarrhoeal illnesses. are present on fish and can grow to significant numbers during storage (1. and proper cooking will substantially reduce risk. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. listeriosis cases have been reportable to the CDC. L. others are psychrotrophic mesophiles (49). in assessing the risk of fishborne infection by P. proper chill storage will eliminate any risk associated with this organism. and fish and shellfish are probably the major reservoirs for the organism (50). cholerae O1. Despite this. P.6.20 °C.3 Aeromonas Aeromonas spp.26 to 0. Although the psychrophiles are unable to grow at body temperature. 17). and especially Aeromonas hydrophila. sodium channel blocking and haemolytic activity (48). monocytogenes has been well documented as a foodborne human pathogen. although lack of data concerning infectious doses and the possibility that foodborne virulence is linked to the immune state of the host mean that full risk assessment is impossible. 1. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. The major virulence factor appears to be the production of toxins with enterotoxic.CHILLED AND FROZEN RAW FISH disorders). their potential to produce disease-causing exotoxin in food has not been fully examined (48). cytotoxic. shigelloides is similar to Aeromonas spp. Some isolates are true psychrophiles.

alcoholics. at risk groups . 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. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. full risk analysis is not possible (52). Klebsiella pneumoniae and Hafnia alvei. D60 of 1. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. especially pickled fish.. 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. and Lactobacillus spp. Infections are mostly associated with the consumption of raw or 18 . It is regularly isolated from seafoods (51). leads to the production of histamine. The organism poses a serious risk in chilled products not cooked before consumption. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. neonates. are exceedingly difficult to produce within the legal limits of histamine as the temperatures used during production can lead to rapid growth of histidine decarboxylating bacteria.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. Clostridium spp. some Vibrio spp. it is not apparent to the consumer and it cannot be destroyed by cooking. Lightly preserved products. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. but proper cooking will reduce any risk. foetuses. but more especially by Morganella morganii. there is a possibility for growth of the organism on chill-stored fish..FISH AND SEAFOOD persons. Like other toxins.. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . 1.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. 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. If fish are not properly refrigerated.48 minutes in salmon have been reported (53). AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses.pregnant women. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6).6.6. Fortunately. Photobacterium spp. but cooking will significantly reduce the risk of infection. Being a psychrotroph. 1.2002 (55)) and is also common worldwide.98 minutes in cod and D60 of 4. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54).

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

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

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

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

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

International Journal of Food Microbiology.. Journal of Food Science. International Journal of Food Microbiology. in Fish as Food. Bulletin of the Faculty of Fisheries (Hokkaido University).FISH AND SEAFOOD 1...9. 1997. 1. Torry Research Station. Jones Y. Huss H. 1985.). 15. 4.M. Taoukis P. 1989. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Labuza T. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. 65-72. 1992. Akiba M. 271-85. Ed. Estimation of the bacteriological quality of fish by automated conductance measurements. Application of predictive microbiology to assure the quality and safety of fish and fish products.. Houston N.. 1984.A. Handling and processing of scad.S.. 741-50..H. Food and Agriculture Organisation of the United Nations. in Seafood Science and Technology.. Fisheries and Food.. Ogden I. in FAO Fisheries Report No. Ross T.L.. Eds. Cho Y. Trolle G. 295-307. 51.2 Spoilage Shewan J. Sumner J.1 Further Reading Processing Anon.9. McMeekin T. 62-70. 6. 127-34. Olley J. Technomic Publishers. 1. 317 (Suppl.. 1984.. The Microbiology of Seawater Fish.. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches. Kitts D. Torry Research Station of the Ministry of Agriculture. Aberdeen.P. Smith G. Changes in microflora and chemical compounds in mackerel stored by partial freezing. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. Huss H. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. Florida.D. Shahidi F. 1989. 24 . Lancaster.. Picard G.. 55.. Journal of Food Protection. Gram L. Aberdeen. Food and Agriculture Organisation. Dalgaard P. International Journal of Food Microbiology.D. 1986. 1988.H. Ravn Jørgensen B. Magno-Orejana F. 1992. Hobbs G. 317-21. Ed. Shinano H. Huss H.M. in Torry Advisory Note No 93. Cann D.G. Borgstrom G.M.. Fisheries and Food. 13-32.H.E. 35.9 1.. Gibson D. Gibson D. Academic Press. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. 1961. Torry Research Station of the Ministry of Agriculture. 1.C.1983. 73-89.. Villemure G. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. International Journal of Food Microbiology. Microbiological quality and shelf life prediction of chilled fish.. Rome. Simard R.

Sala F. Journal of Applied Bacteriology. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. Eds. 1992.. 1987. Garcia M. 5. 1992. 25 . 322-6. A rapid method for detection of histamine producing bacteria. 72. Modified atmosphere packaging of fish and fish products. in Quality Assurance in the Fish Industry. 1987.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products... London. 1. in Fish processing technology. Huss H. 1997. Huss H. Amsterdam. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C.9. 1.M. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish.. Hall G. 479-88. 2197-203.. Klausen N.. Blackie Academic & Professional. Gram L.CHILLED AND FROZEN RAW FISH Einarsson H. Trolle G. 59. http://eur-lex..L.H. Ed.R. Applied and Environmental Microbiology. 4.. International Journal of Food Microbiology. Gram L.K. Huss H. 20023. 65-72. International Journal of Food Microbiology.europa.H.eu/LexUriServ/LexUriServ. Otero A. Liston J.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A.J. 1993. 137-46. Jacobsen M. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures.3 Pathogens Condon S.9. Effect of culture age.H. Elsevier Science.

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

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

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

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

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

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

3 Preparation of fish component of chilled and frozen recipe dishes 2. to evenly raise the temperature of the entire frozen product from -20 °C or lower. but it also poses a risk of cross-contaminating other products undergoing tempering. the core temperature. Figures 2. and contamination from equipment to clean fish muscle.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing.3. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. 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. Examples of tempering techniques include the use of heated air. and potential opportunities for growth of foodpoisoning bacteria. or the use of microwave technology. but will also need to ensure that the optimum shelf life can be safely obtained.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. Generic examples have been used to illustrate only the major points. this may be an incidental effect. Rapid controlled cooling of heat-processed components will help to minimise spoilage. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. It is recommended that the principles of HACCP are applied to novel processes. 2. depending on the size of the fillet 33 . All have the same intention.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.2.4. 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.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. line extensions and process reviews.4. defined holding time at a chill temperature (0 – 3 °C). 2. up to -5 °C so as to improve the cutting and handling properties during processing. Band-sawn materials or fish dice are normally promptly further processed. there are a number of distinct methods used.4. Effects on the microflora are minimal during tempering (7). as a heat process is often required for functional or organoleptic reasons. but it is assumed that any band sawing or dice process will introduce heat from blade friction. as the full complexity of the processes used for specific products cannot be covered in a general text.1 . For some components. Where the raw materials are frozen.

1A .3.Fish Cakes). Fig.2. with the coating heat-fixed. may not rise above 0 ºC during processing.3. 2.1 (2. are placed into plastic boxes and covered in ice to preserve their condition. by mincing and mixing with a binding agent (as described in section 2.7 days.1. chilled or sold chilled on defrost with an expected shelf life of 6 . the scampi is washed in potable water to remove foreign bodies. Frozen cores are usually batter-enrobed. frozen in a nitrogen blast freezer and weighed into sacks for later processing.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.1B .4.Scampi.1.1. The “blown” meat is graded. The processing of three examples types are described in Figure 2.4.Shaped Fillets. 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. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process.2). Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product.1C . drained and then refrozen for storage until required for further processing. quickfrozen. 2. after landing. 2. Raw material preparation stages for chilled or frozen coated fish products 34 . On arrival at the processing plant.FISH AND SEAFOOD processed. It is then size-graded.

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. Therefore. It is for this reason that all by-products from primary processing. and water is added to rehydrate the components. Fish cake cores may be batter-enrobed.1.3. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. but.block-sawn dust. iii) Fresh/fillet/minced fish block. This is especially the case with block-sawn dust. as the physical structure of the material has been destroyed. chilled.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.4. with an expected shelf life of 6 . The fish component of the cake will.g. or those where the trimmings. additional spoilage organisms may be introduced.1. spoil more rapidly than the dry ingredients. ii) Fish-filleting waste .7 days. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. material will need to be transferred swiftly to a chiller. those that use frozen blocks of whole fillets.2 Shaped products This type of product falls into two categories. are handled and stored as any other food product. that are to be reused for other foods. Waste material produced from this type of processing is often reused in the third category (fish cakes). Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. once the dry components have been rehydrated. 35 . from potato flake. pathogens such as Salmonella and Listeria. The remaining dried ingredients are mixed with the fish.4. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing 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 promptly processed to avoid further quality loss. such as aerobic spore formers and Staphylococcus spp.3.. 2. more seriously. fish fingers). or sold as a chilled product after thawing. with the coating heat-fixed. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e.off-cuts or belly flaps. unless it is poorly handled. quickfrozen. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. because of its nature.

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

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

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

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

1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. B. a heat process in excess of 73 ºC will be required to rehydrate this component fully. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. with the range always expanding as new tastes and cuisines are developed. Sauces or marinades are added to fish to broaden appeal or to accompany components. cereus. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. Alternatively. such as butter garnish with black pepper.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B.6 Preparation of non-fish components of chilled and frozen recipe dishes 2.4. especially in those products that have a vegetable component or rice accompaniments. a process equivalent to at least a 5D Listeria kill will be necessary. For this reason all products. Appropriate handling facilities or equipment will be required to protect the product. This is a particular issue especially where products are heat treated to eliminate vegetative cells. spore-formers can proliferate if adequate control of temperature is not maintained. and then cooled slowly allowing germination and re-growth. 11). rely on the use of microbiologically sound ingredients. Placing such products in a standard chill store to cool down could result in condensation. can germinate and rapidly multiply. Typically. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. if subjected to temperatures between 20 and 40°C. If the sauce contains modified starches for freeze/chill stability. Sauces can be multi-component preparations or more simple preparations. aureus toxin is well documented (10. cereus). and an increased risk of contamination from L. Potato toppings made from flake or powder will normally have little associated spoilage flora.) to the product (12.6. 2. 40 . Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. cereus are well known and are still a common issue in the UK (9).4. Products or dried ingredients contaminated with either spores of aerobic bacteria (e. On hydration. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. but temperature-abused pasta can also spoil rapidly by mould growth. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. An incident involving Staph. 13). the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. where composite meals are heat-treated.g.

41 . Materials prepared with regard to good hygienic practice need only be assembled in a tray or pouch before chilling or freezing in readiness for distribution.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig.5 illustrate the recipe dish ingredients preparation and assembly and packaging. suspended in an acidified sauce with an oil component. flour products. etc. and handling of processed sauce can be less rigorous than. 13). that of a dairy-based sauce. Without barriers. Marinade-type products are becoming more popular. spices. It is expected that this product format would have a restricted shelf life to limit potential for growth of psychrotrophic bacilli or any survival of enteric pathogens (12.herbs.2.4 and 2. The format uses flavour compounds .4. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. for instance. respectively. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. 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. Figures 2.

and tuna loins are packed by placing the product in a plastic pouch and removing all the air. 2.4.5.FISH AND SEAFOOD Fig. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow.4.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. botulinum to grow. 42 . The oxygen-free conditions will however allow other anaerobic bacteria such as C. as it dissolves in the surface moisture to form a weak acid.2. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).4. These include: 2. Flow diagram outline assembly and packaging of recipe dish 2. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. which may restrict either microbial growth or enzyme activity. the method replaces air in the sealed packaging with a defined mixture of gases.2 Modified-atmosphere packaging Originally developed for the red meat industry.7.7. unless the temperature is strictly controlled to below 5 °C throughout the supply chain. generally carbon dioxide and nitrogen.

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

5. This reinforces the need for prompt processing and temperature control of ingredients and finished products. and Aerobacter spp. By retaining the intrinsic spoilage flora of seafood. However. 44 .5. producing characteristic off-flavours. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. Products that are developed with an extended shelf life beyond the 6 . Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. For example. within dairy products are well known. this should also take into account shelf life of product components used in the assembly of the final product. The conditions and freezing process most favourable for good-quality products will.. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. Still. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e.. As a note of interest. freezing also has a protective role in improving the safety of raw materials by.g. 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. also be those that have the least effect on the viability of microorganisms (21). eliminating nematodes in products that do not have a heat treatment step. These have been proved to impart off-flavours. which would warn the consumer of the potentially unsafe nature of the product. the shelf life of products is limited by the natural spoilage process described earlier. Pseudomonas spp.FISH AND SEAFOOD 2. botulinum. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. The type and nature of the enzymes and their activity will depend on the bacterial species. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). auto-oxidation of fats/oils).3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10).. however.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. 2.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24). enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage. Serratia spp. where the variety of components and ingredients used in manufacture is varied and complex. for example. Bacillus spp.

Counts of between 103 . in some cases. air.e. Hazards attributable to the non-seafood components are listed below and this information. Vibrio spp. Incidents in such diverse materials as cream.1 Bacillus cereus This is a common contaminant of dust. perfringens contaminating seafood will depend on the source of raw material. etc. 2. have been linked with food poisoning and should also be considered. are dealt with elsewhere in this book. agricultural run-off or sewage can contaminate in-shore species. any risk of C. and this issue may be of greater consequence for warmwater.. low oxygen or anaerobic conditions.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. especially molluscan filter feeders. botulinum type E and Vibrio parahaemolyticus (25). and many raw or processed foods (26). Inappropriate holding times and temperatures will. histamine intoxication.6.3 Clostridium perfringens A study examining C.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. soil. Germination of spores requires favourable conditions. 13. 45 . This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. These are C. In addition. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). and Campylobacter are recorded in filter feeding molluscs such as mussels. permit rapid growth of B. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. A number of other Bacillus spp. along with previous discussion.6. Cases quoting contamination of seafood with organisms such as S. 2.6. and some effects related to product type and time to toxin production have been noted. with pathogenic microorganisms or viruses. 2. or a heat shock followed by holding at a temperature suitable for growth. 27). potato. botulinum when held in favourable conditions for growth (28) i. cereus. typhi. spices and herbs and vegetable shoots have been reported (12. Additional hazards from marine bio-toxins. may be of benefit within any risk assessment for chilled and frozen prepared fish products. farmed or free-living crustacea (30). Therefore. water.104/ g should be treated with some concern (26).2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C.

and will therefore cause “food poisoning” even in a product that is cooked before consumption. will colonise factory environments in a similar fashion except for possibly Vibrio spp. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). and inadequate cooking or holding within the temperature growth range (27). All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. but under the appropriate conditions. However.6. if any other potentially pathogenic organisms. It is unclear. growth may occur in foods (26). These include inadequate refrigeration and preparing foods far in advance of planned service. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32).6. the restrictions on growth rate seen with seafood will not apply. L.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. which is not affected by any subsequent cooking process. An acceptance has now developed in the UK of the presence at low levels of L.6. L. 2. at the present time. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. These conditions will allow the production of the toxin. when introduced or inoculated into sterile components of chilled prepared recipe dishes. 2. Chilling may help Shigella to survive. Shigella spp. but.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). 46 . Rates of contamination with smoked seafood are much higher than those with unsmoked products.5 Shigella spp. and this is believed to be due to a tolerance to brine used in the smoking process (32). aureus (35). monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. aureus enterotoxins in foods. indigenous or other. Sources of contamination are many because of the widespread distribution of this organism in the environment. infected food handlers. monocytogenes as a contaminant in foods (31). are considered primarily to be waterborne organisms but epidemiology indicates that contamination of vegetables and possibly in-shore caught seafood could act as vehicles for infection by Shigella (34).6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. 2.

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

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

8. Food and Drug Administration. Microbiology of processed spices. 663-87. Kumeta H. 14.H. 1991.C.. London. 38 (11). Outbreaks of foodborne illness in humans. 13. Wienke A. Huss H. Martin R. . 1311-7. Pathogenic microorganisms associated with fresh produce. 59 (2). 1976.8 1. Hoshino T. 1997. Incidence and levels of Bacillus cereus in processed spices. The bacteriology of fresh and spoiling fish and some related chemical changes. 204-16.. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. 2001.T. 167-93. Journal of Food Protection. Ohgiya S.M.J. 49 4. Powers E. IBH Publishing Co.M. 25 (1). Lawyer R. Kluwer Academic. 9. 47. Importance of Photobacterium phosphoreum in relation to spoilage. Journal of Milk Food Technology.E. Tu A. 11. Pvt. Journal of Hygiene. England and Wales Quarterly Report.L. Oxford. Gilbert R.G. Yumoto I. Masuoka Y. Ward D.. 15. Communicable Disease Report. 233-7. 373-8.. Animals and Spoilage.. 1986. 3. International Commission on Microbiological Specifications for Foods.. 2005. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. 7 (24).. 1976. Eds.. Hobbs G. Murrel H. 668 -70. New York. Recent Advances in Food Science. 2. Proper processing and handling of raw tuna. Mejholm O.K.A. Aureli P. Fish: microbiological spoilage and safety. Brown T.. Toxin Related Diseases: Poisons Originating from Plants. 1. 1962. 6.. Bartram P. 2. 96. Cambridge. 1991. 215. Bacterial histamine production as a function of temperature and time of incubation... 70-2...R. Ishizaki K.R. Dalgaard P. Shewan J. Sakamoto T. Journal of Food Science.J. 24 (5). 5. 1997. Taylor S. 39 (10). 67-73. Latt T.. Hackney C.R. 1997. 1993. Taylor and Francis.. Communicable Disease Report. 12. in Fish Inspection Quality Control and HACCP: A Global Focus.W. Slavin J. Ltd. 10. Letters in Applied Microbiology.M.L.. Journal of Milk Food Technology.. Microbiology of Marine Food Products.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). Matsuyama H...R.. Letters in Applied Microbiology. Bartlett C.L.C. Christiansen T. Fish and Fisheries Products: Hazards and Controls Guidance.C. 166-73. Washington D.R. 1997. Beuchat L. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. 7. Food Science and Technology Today. London. Collette R. 5 (3). Woolaway M. 1982. Powers E. Behling A.. References US Food and Drug Administration. Van Nostrand Reinhold.

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

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

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

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

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

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

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

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

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

13).2. Flavobacterium spp.. Other microorganisms found in lesser numbers are the Achromobacter spp. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. Micrococcus spp.1 and 3. Pseudomonas spp. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. Water temperatures affect the concentrations of microflora in the water.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs.1 Shellstock 3.. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. that have never been cultured on laboratory media.3. 10. Enterococci spp. 3. the initial flora composition and concentration of bivalves are highly variable. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. whole or in part.1. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. Campylobacter coli Bacillus cereus Hepatitis non-A. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. mechanical harvesters. In summary.1 Processing and its Effects on the Microflora Oysters.. mussels. and cockles The processing of bivalves takes two forms. or by 60 . 15). At harvest.. and shucked shellfish that is shellfish.1.3. Temperature. nutrients. They are termed viable non-culturable microorganisms. (9. The largest groups found are the Pseudomonads.. dredges.... For example. There are microorganisms. and Aeromonads. 11. Alcaligenes spp. Bacillus spp. 12. observed microscopically. but have never been cultured (14. spirochetes of the genus Cristispira and Saprospira.. and yeast cells of Rhodotorula rubra and Trichosporon spp. responding to the environment in which they are harvested. from which one or both shells have been removed.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. clams.3 3. shellstock (live) which is shellfish in the shell.3. Moraxella spp. rakes.000 /g. Acinetobacter spp.1. 3. salinity. Vibrios. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves.

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

1. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. and the sanitary practices applied during harvesting and shipping. 21. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). with the exception of A. however. An outbreak of infectious hepatitis in Sweden in 1956.1. hydrophila (18). 22). Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. Harvest conditions are another factor that influences the growth of bacteria in shellstock. to avoid possible contamination of the shellstock. At this temperature the multiplication of faecal coliforms (E. and thus contaminating the shucked meat at the point of sale. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. without adequate refrigeration. metabolic waste accumulates in the shell serving as nutrients for bacterial growth.3.FISH AND SEAFOOD 3. shellstock are not refrigerated on the harvest boat. If long delays occur before reaching the processing facility and/or refrigeration. 23.2 ºC (45 ºF) or below (19). and temperature storage of shellstock to point of sale. 26). Water used for shellstock washing should be of good sanitary quality. some pathogenic organisms will survive and multiply in shellfish (20. for a considerable length of time. synthetic seawater for the purpose of de-sanding and salt uptake. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . At harvest. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. there is evidence that in the absence of adequate refrigeration. 25). Enterobacter. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. with prolonged storage the microbiological quality of the bivalve decreases. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. Generally. shellstock are placed in refrigerated trucks for transport or placed in wet storage. handling. Klebsiella. Over a period. Furthermore. 17). Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. 16. and Citrobacter species) and Vibrios. Generally. The most important factor in controlling bacterial growth in shellstock is temperature. coli. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. when it reaches the processing facility. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. including V. bivalves tightly close their shells thus trapping water and microflora. are prevented. the bacterial counts in the meats will increase. vulnificus. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. The washing of sediment and detritus soon after harvesting will prevent further contamination. 24.

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

2. and sanitised before each filling. which may lead to contamination of product. Shucked meats are thoroughly drained. There is a tendency to store such articles on the shucking benches or in packing rooms. 2.2 ºC or less. 4.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). Separate rooms or lockers for storage of clothing. 3. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. 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 . Shucking containers are rinsed clean with running water. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. within one hour. and gloves help prevent the contamination of shellfish. Fig 3. aprons. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash.

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

and harvesting of shellfish in the cooler months. parahaemolyticus from shellfish is low. approximately 23 cfu/g. and estimates based from a hospital study show less than 0. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams.e. This bacterium is present in all the major processing steps of a shellfish plant. Fortunately. ranging from epidemic cholera to sporadic cases of diarrhoea. Products involved were from the same processor. nausea. shellstock and packing of shellfish in jars) (29). parahaemolyticus possibly producing illnesses (40). and to the point of sale. 41). primarily in brackish or saltwater.1. Major outbreaks are associated with the warmer months. 3. during processing. Between May and September 1997.FISH AND SEAFOOD 3.5. where outbreaks occur regularly. V. prevention of cross-contamination. there is a recent report of low levels of V.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. parahaemolyticus. 3. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). These organisms can be present in the shellfish from the point of harvesting.5 cases/100. One of 250 cases reported involved shucked products with low levels. lot and retail outlet.1 Vibrio parahaemolyticus V. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. The incidence of V. widespread in both polluted and unpolluted waters.5. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. Diseases caused by Vibrios are enteric in nature.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. 66 . although non-hemolytic strains have been associated with illness (43. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. only a small percentage of strains in the marine environment are potentially pathogenic (45). however.000 population/year (46). diarrhoea. headache and fever. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). and very common in Japan. from harvesting to finished product (i.48 hours with abdominal pain. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. Sporadic cases are frequent along the coast of the United States. 44). It is a common marine bacterium. parahaemolyticus causes gastroenteritis lasting 24 . of V. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. V. They can be introduced to the shellfish from the environment or added during processing. coastal areas).

or primary septicaemia. like V. vulnificus are fever. 54. however.4 . Onset of symptoms occurs in 48 hours and can be severe. 46. cholerae. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). Inadequate refrigeration will accelerate the growth of V. Control of oyster-associated 67 . with a fatality rate of 50% for individuals with septicaemia. 56). Primarily. abdominal cramps. cholerae non-01 (51). vulnificus. lasting 6 . The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. 3. High-risk individuals. which have been implicated in shellfish-related disease outbreaks (31).3 Vibrio cholerae non-01 Illness caused by V. leukaemia.II (8. The bacterium is an estuarine species that causes wound infections. diabetes. Healthy individuals are susceptible to gastroenteritis. in the range of 0. V.1. and preventing cross contamination once the product is cooked.5. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. parahaemolyticus. 46. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. 58). parahaemolyticus growth in shellfish meats is temperature dependent. parahaemolyticus the estimated annual incidence rate is very low.0.48 hours of onset. nausea. cholerae non-01 has been associated with septicaemia. In one study. V. or immunosuppression. 3. these individuals are advised not to eat raw shellfish. All isolates are considered pathogenic.8 case/100. 57. those who have liver disease. V.7 days. the organism has been found in the Gulf of Mexico.5. and vomiting.1. vulnificus is by rapid refrigeration during warm weather months. 55. are particularly susceptible to primary septicaemia. The occurrence of V. The organism attaches to the intestine and release of a toxin is suspected. cholerae non-01 is ubiquitous in the estuarine environment. cirrhosis. 23 of 24 major USA-West Coast estuaries were positive for V. and rapidly become inactive at low storage temperatures. 49). chills.2 Vibrio vulnificus The symptoms of V. 53. gastroenteritis. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. fever. It causes diarrhoea. Hazards from this bacterium can be controlled by thorough cooking of shellfish. Death has been reported to occur within 36 hours from onset. 48). Control of V.MOLLUSCAN SHELLFISH Control of V. and nausea within 24 .000 population/year (8. They are sensitive to cold. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. It is one of the most severe foodborne infectious diseases. Oysters are the most hazardous food associated with V. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). Again.

There is evidence of strains of V.5.1. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera.5. imported product from a country experiencing an epidemic. and nausea. 3.10% carbon dioxide for optimal growth conditions. Gulf coast environment. V.FISH AND SEAFOOD V. drying. nausea. is the same as previously discussed for the non-01 strains. however. and V. (8. Illness can include abdominal cramps. fluvialis.2 Campylobacter jejuni C. mimicus. widespread in both polluted and unpolluted waters. cholerae non-01 illnesses is by cooking seafood thoroughly. respectively.S. In general. cholerae 01 establishing in the U. They are common marine bacteria. cholerae 01 in molluscan shellfish.5 Other Vibrios spp. processors should know the product source. 3. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). There are several other Vibrio spp. particularly in oysters. It causes diarrhoea. acidity and heat. 60). salinity. vomiting. Death may occur after severe fluid and electrolyte loss. vulnificus. including V. 3.5%) and 2 . Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. V. dehydration. jejuni requires reduced levels of oxygen (3 . In addition. cholerae 01 is transmitted by faecal contamination of food or water. e.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. fever. Control of infection from these organisms is no different from the other Vibrios. parahaemolyticus and V.48 hours with abdominal pain. Poor sanitation and contaminated water supplies will spread the disease. or cross-contaminated shellfish (57. associated with shellfish-borne illness outbreaks. headache and fever. Onset of disease is 6 hours to 5 days. 62. The control of V. The presence of these strains appears to be in low numbers and non-virulent (59). nausea.5. preventing crosscontamination of cooked seafood. cholerae 01. 63). and the transmission of the disease by consumption of raw.g. 61. 68 . the pathogenic severity is lower than V. hollisae. and shock. diarrhoea. and illness lasts 7 . low pH. V. and southern Asia including India and Bangladesh. Illnesses are associated with the warmer months. abdominal pain. undercooked. Onset of illness is from 3 to 5 days after ingestion of contaminated food.1. these organisms cause gastroenteritis lasting 24 . It is considered a fragile and sensitive organism to environmental stresses such as oxygen.10 days. In developing countries. The frequency of infection of these organisms is no different from other Vibrios.

80) are ubiquitous in the marine environment. 69. 65. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors.II Molluscan shellfish associated outbreaks caused by Vibrio spp. 77. jejuni were recovered. V. although their role as a pathogen is highly controversial and circumstantial. they survive well at refrigerated temperatures. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. V. jejuni begins in the harvest of shellstock from approved waters. 3. 68. The seasonality of 69 . This finding is significant in that C. Campylobacter appear to survive well in oysters during storage. Recently.3. Campylobacter spp. from Pacific shellstock oysters purchased from a retail grocery store (74). vulnificus.3 Other organisms of concern 3. V. 73). of Cases 4 362 0 1 0 a Includes V. of Incidents 4 279 0 1 0 No. Although. parahaemolyticus. cholerae 01 and non-01. 71).4 to 114 cells. fluviallis. C. C. hydrophila group (75. mimicus. 66). They have been implicated in limited shellfishassociated gastroenteritis cases (8.500 cells/gram of Campylobacter may cause illness (64. V. and V. jejuni does not multiply in shellstock at refrigerated temperatures. however. In one case. 84. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. control of C. shigelloides (79. 85).a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. To reduce the risk of shellfish-borne campylobacteriosis.5. 72.5. Human feeding studies suggest that about 400 . 83. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). hollisae Infective dose varies with host susceptibility and virulence of the strain. however.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. 76. and their immediate proper refrigeration. 70. 81. whereas at 10 ºC Campylobacter survive only for 5 days (71). jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. 82. have been isolated from shellfish beds throughout the world (67. 78) and P.MOLLUSCAN SHELLFISH TABLE 3. infection occurred among participants of a firemen's banquet. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. ranging from 0.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Control should be achieved by promoting GHP and staff training programmes. pasteurisation and cooking.5. was associated with seafood consumption. cholerae is divided into two groups. the causative agent of cholera. 4. is widely distributed in aquatic environments. aureus on their hands. 19). which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. especially in products that have been vacuum packaged. Researchers have demonstrated that inoculated shrimp supported toxin production by C. aureus is not a member of the intrinsic flora associated with crustacean shellfish.3 Staphylococcus aureus Staph. particularly crabmeat (18. pregnant women and the immuno-compromised. The former. 25). botulinum type E is commonly associated with fish and fishery products. therefore.70% of healthy individuals carry toxin-producing strains of Staph. cholerae. so is able to survive heat treatments that eliminate vegetative cells. about 50 . whilst boils and cuts are also common sources of the pathogen. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21.4 Clostridium botulinum C. no incidence of foodborne illness caused by L. the pathogen can grow at temperatures as low as 3 °C. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. V. 85 . The main source of contamination is the hands of processing personnel. 4. An epidemic that began in South America in 1991. 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.5. combined with the degree of handling of these types of product. 24.5. Crabs are covered in a chitin exoskeleton. e. To date. as part of the natural bacterial flora.g. Crustaceans have been linked with food intoxications attributed to Staph.CRUSTACEAN SHELLFISH old. The pathogen has resistant spores. Once the competing microflora has been removed by a heat treatment.5 Vibrio spp. aureus because of the ubiquity of this organism in the processing environment. However. V. cholerae non-O1. cholerae O1 and V. Vibrio cholerae. 4. monocytogenes has been attributed to crustaceans. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. although contamination by types A and B may occur from the environment. botulinum type E at 10 °C but not at 4 °C (20). which can become colonised by V. 22) as well as from undercooked crustacea when harvested from contaminated waters (23.

hydrophila is able to grow well at refrigeration temperatures (0 . Of these. in a survey carried out in the United States between 1977 and 86 . e. a marine halophile. South America.2 °C) so that GHP coupled with controlled low temperature storage is essential to prevent the pathogen from reaching levels that may present a hazard.8% of samples contained C.5. However. 4. but has the potential to cause fatal foodborne illness (26). Japan. 4. Caliciviruses.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. 32).7 Aeromonas hydrophila and Aeromonas sobria A. coli. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. vulnificus is an invasive and lethal pathogen associated with wound infections. or freezing at a temperature of -23 °C for 7 days (30. Astroviruses and non-A. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. occurs in crustaceans from warm environments. hydrophila and A. Parasitic infections are controlled by either cooking infected material prior to consumption. A. hydrophila has been isolated from prawns (29). However. been implicated in a case of foodborne infection associated with crustaceans. whilst 5. Norovirus. 31.5. particularly trematodes (6). to date. Quantitative levels were below limits of detection in all cases (<0. although A. the potential hazard exists as C.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. Contamination by this organism can be controlled by rapid chilling of crustaceans after harvesting and avoidance of time/temperature abuse during distribution and processing chains. Hepatitis A virus presents the most serious hazard.6 Campylobacter jejuni and Campylobacter coli C. 4.30 MPN/g) (27). V.g. V. 4. It is more commonly implicated in countries where crustacean shellfish are eaten raw.5. non-B Hepatitis viruses (31). jejuni has not. however. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). parahaemolyticus.5.

although it is one of the European Union (EU) legislated allergens.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control.5. 4. Selected parts from this regulation is presented below. 38. They can survive low pH levels.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. only 0. The toxin causes amnesic shellfish poisoning (ASP) when consumed.6. are included in the Commission Regulation (EC) No. which results in a persistent and apparently permanent loss of short-term memory (36). Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). 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. consistent with that in the stomach. The risk of such hazards is far higher with fin fish and molluscan shellfish.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). produced or imported into the European Union (EU).11 Chemical pollutants and biotoxins Researchers have demonstrated that crabs feeding in waters contaminated with algae do have potentially high levels of algal toxin domoic acid (DA) in their viscera.CRUSTACEAN SHELLFISH 1981. 4. A summary of microbiological criteria for crustacean shellfish is listed below. Provisions laid down concerning the use of additives would also apply to these types of products. No foodborne incidents have been attributed to this source to date. Norwalk viruses can also withstand heating to 60 °C.5. or chlorine concentrations similar to those found in drinking water. from farm to fork (37. 87 .10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. 4. 2073/2005. 39). which could be transferred to the meat during unskilled removal from the shell (35). 4.

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

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

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

8 211.3 Processing and its Effects on the Microflora Owing to the wide variety of products.80º brine (184. is employed. % 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. For smoking salmon. When a milder brine. When a 100º brine is used.4 52. or salt in the water phase. it causes swelling because the fish absorbs water from the salt solution (1). At 18 °C a 100º brine is produced when the solution contains 26. all using salting steps. etc.8 211. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.6 264. it is important to state the temperature. it is important to understand the concept of salt concentration. Different brine strengths are required according to the curing processes and/or the type of product.0 . The strength of the solution is described as degrees of saturation (brineometer degrees). Therefore.0 g of salt per litre of brine).2 g of salt per litre of brine).4 184. smoking or marinating processes.FISH AND SEAFOOD 5. a 90º or above brine (237. Salt in the water phase links the two conditions of moisture and saltiness. When working with salting. This should be distinguished from the salt content. Salt concentration means the percentage of salt in the water contained in the fish.0 158.6 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. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat.6 132.4% salt (Table 5. about 50º (132.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) 26.2 105.I).2 237. TABLE 5. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. only the most common processes can be described here. processes and producers’ private recipes. Thus. and salt content. a 0º brine saturation means pure water. sometimes the salt crystallises onto the surface of the fish.8 79. the fish is usually salted with a 70 . it is impossible to establish standardised processing stages in the curing of fish.

1. 5. 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. and essentially free from iron and copper (3). respectively.0. Salting can be done in a number of different ways. respectively (1). The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. For a light salting. are likely to cause hydroscopicity of the product and. low in calcium and magnesium. are recommended to help make the product whiter.1 ppm and 10 ppm. 97 . A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). See Figure 5.CURED. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. outgrowth of halophilic bacteria may occur. and for a heavy salting a ratio of 1:3 salt:fish is used (1). however.15% calcium and magnesium. On the other hand. if very pure sodium chloride is used for curing purposes.1. the fish is restacked with fresh salt after about a week. the fish is mixed with one part of salt per eight parts fish. The calcium and magnesium impurities. for which there is a high demand in Mediterranean countries. Also.3. but the concentration of copper and iron must be kept under 0.15 . which include calcium and magnesium. and such delays. and salt can speed this oxidation process. This is because fatty fish turn rancid quickly. when the air humidity is high. However. SMOKED AND DRIED FISH 5. ‘Green cured’ split cod with about 60% of the backbone removed. lean white fish is used for this purpose.1 Salting Salting of fish for preservation purposes has been used since pre-history. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. It takes about 3 weeks until proper salt penetration and moisture loss are achieved.35% and 0. This condition is known as ‘putty fish’ (4).1 Dry salting / Kench curing Usually. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. having a short section at the end of the tail. and a ‘putty’ fish product is also obtained (1). addition of levels of 0.3. producing characteristic pink patches. Usually it is followed by other preservation techniques such as drying and/or smoking. it tends to lead to a slight yellowing and softening of the product. can be dried further to produce ‘klipfish’ or ‘bacalão’. mainly in tropical conditions. and the salt has penetrated throughout the fish (3). caught in North Atlantic waters. ‘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. When pure salt is used. The most common fish used in the northern hemisphere is cod (Gadus morhua). The salt used should be of food-grade quality.

when a light brine is used.1. the process is mainly for flavouring purposes prior to the use of other preservation processes. Processing of dried salted cod 5. it appears that water activities much less than 0.1.5.3. such as 98 . Food-poisoning organisms.2 Wet salting or brining As referred to above. The greatest barriers to salt penetration are the skin and scales. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. Where fish are salted in their whole form. so that the risk of spoilage is high. eviscerated and/or split). such as Staphylococcus aureus could survive and grow at such water activities in the product. Fig.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. 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.90 are rarely achieved.

limiting the amount of rancidity that can develop.90 (1).3. Here. headed and gutted.76 4.05 7.85 3.26 2.09 8.33 5.78) (1). It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.51 5.09 8.76 4.23 3.91 2. herrings and other fatty fish can be used. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 . 5.26 3.51 3.41 4.65 2.06 4.26 1.98 6.5 3.1.23 2.74 9. the brine is then replaced over the fish.96 1. SMOKED AND DRIED FISH smoking. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance. the liquid that comes out of the fish forms a brine.60 2.99 2.10 1. with pressure being applied to the top of the container to help the brine to drain away.00 3.11 3.23 8.54 5.76 4.26 4.5 3.3 days. Fresh sardines.30 2.70 3.64 4. Histamineforming bacteria do not generally grow at water activities below 0.80 5.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) 2.78 2.70 6. See Table 5.66 5.5 10.35 4.46 4.0 2.41 5. but as it is placed in containers. There are several variations of this process. and can be collected and replaced again over the fish.0 4.49 4.47 6.41 3.86 2. Histamine formation is of importance when using scombroid and some clupeidean species.85 3.61 3.23 2. TABLE 5.84 1.44 2. thus.11 9.11 4.26 7. The main objective of the brining method of salting fish is to limit air penetration during processing.25 5. with a shelf life of about 4 weeks (water activity (aw) about 0.76 4.22 6. can be cured in a saturated brine solution for 6 days and. a saturated brine is used.78 6.03 4. the product obtained is called gaspé-cured (5). the fish enzymes play an important role by maturing or ripening the product.56 6.61 3.76 4.25 5. the fish is dry-salted. after being drained and pressed at 465 kg/m2 for 15 hours.95 3. For example. For preservation purposes.3 Dry / wet salting In this process. they can be stored at 28 °C.88 5.03 2.44 2. and a red burgundy colour will develop in the product.16 7.73 1. The fish are kept immersed in their own brine near saturation point for several days.41 6.19 3.17 4.98 6.45 3. if split fish is dry-salted.76 99 .66 5. Anchovies are the main fish used for these purposes. but mackerel. This is why it is the preferred method for fatty fish.CURED.04 5. sardines.0 3.98 5.17 5.II.5 4.

placed in a marinade of lime juice. In a second step. 5.2 months.4% of salt) for distribution and sale (1). is made of different varieties of fish. they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. which is dipped into a boiling brine for 10 . As referred to above. or can be boiled by being placed in a basket.3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. containing 1 kg of salt for every 25 kg of fish. up to 1 . Another variation uses sour cream instead of the second brine step. Usually. because of the salt content of the product. Afterwards. Herring can also be cured for several days in an 80 to 90º brine with 2. The fish can be soaked in a high concentration of vinegar (about 4 . The lime juice plays the role of the vinegar. Ceviche. After this. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. and lasts for 3 .2% acetic acid and 2 . outgrowth of bacterial spores after processing may occur (4). underlines the need for careful control (4). even if the product is hermetically sealed.3. The first process can give a longer shelf life. or a pH <5 and they are stabilised for a limited period. has a shelf life of several years. and spices) is added to the brine. giving a cooked appearance to the product. the occasional implication of pindang in cases of foodborne illness. a Peruvian product. and 6% salt. A much firmer product is obtained when the fish is salted prior to acidification. Another salt-boiled Indonesian product. drying and salt-boiling steps until a hard translucent block is obtained (4). until the flesh is soft. herring or other fatty species are used.6 days without refrigeration (1).FISH AND SEAFOOD 5.5% 120grain distilled vinegar.30 minutes. and even death. They will have a salt content >6% (w/w) in the water phase. Traditional jellied eels are a product where pieces of fish are cooked in boiling water. flavouring (sugar. Other preservation techniques can be used for extending the shelf life of these products. then packed in barrels with a 70º brine.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. usually requiring refrigeration below 10 °C for longer-term storage. usually flavoured with onions and hot chillies. ‘ikan kaju’.3. Once cooled. There is epidemiological evidence that these products have been the cause of some 100 . only the nonspore-forming organisms are eliminated in the process and.7%) for as long as 3 weeks. it can be repacked in milder brine (1 . they are packed for consumption within 2 weeks at chill temperature (6). salt. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. but it has further smoking. However. and packed (3). All these products are ‘semi-preserves’. whereas the latter method preserves the fish for only a few days. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar.

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

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

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

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

Smoked salmon fillets are usually vacuum packed whole or after slicing. botulinum type E present an even longer time than before to grow and produce toxin (14). typically mesophilic.5. To avoid growth of non-proteolytic C. Neither the smoking temperature nor the salt content is enough to kill the organism. secondly. Length of the smoking process at each stage will depend on the species and the type of product. can grow with salt levels of 10% in the water phase. refrigerated storage and vacuum packaging still allow its growth (12). a drying period at 30 °C. in order to produce toxin. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1).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. stored at 4 °C and packed. which underlines the danger of storage at abuse temperatures of >10 °C (13). there is no point in the process that can fully assure the absence of L. SMOKED AND DRIED FISH During cold-smoking. A typical temperature set is as follows: firstly. The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. Furthermore. a partial cooking period at 50 °C. if they are taken from a chill room at 3 °C and packed. Smoked products can be also packed under controlled-atmosphere packaging. if present. See Figure 5.3. monocytogenes.CURED. Hot-smoked products require a longer cooling stage. This temperature should be maintained during storage and distribution (3). The exception is vacuum packed fish. On the other hand. botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. Pathogenic vibrios. but the proteolytic types A and B. During smoking. Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. as. a final cooking period at 80 °C (12). thirdly. Vacuum packing is recommended only for coldsmoked salmon. further cooled to 3 °C within 12 hours and then packed.2. If the refrigeration chain is maintained. thus allowing any psychrotrophic C. gravad salmon (also called gravad lax or gravlax). they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). and hotsmoked salmon (kippered) (3). Afterwards. 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. condensation will occur inside the pack. and their sensitivity to salt (13). Although other human pathogens that can grow at refrigeration temperature. and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). the products are cooled. such as Yersinia enterocolitica. These products should be vacuum packed at room temperature. can be isolated from seafood. but they should be cooled to a temperature of 10 °C within 3 hours. 5. if a 105 . are of concern when smoking fish from tropical waters (13).

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

Aspergillus tamarii) and several Penicillium spp. (Penicillum cyclopium and Penicillium puterellii) (19). Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. but mechanical drying is used instead of sun drying. break down of fats and the moderation of smoke flavour. The classical liquid smoke is 107 . Boiled or steamed clams. 22. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). white fish about 8 days. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). 18). Mackerel. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. resulting in a characteristic flavour (20. such as yeast derivatives. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. chilled or freeze-thawed. When refrigerated at 3 °C.3.3½ hours using the typical temperature set for hot-smoking referred to above (12). 5. Eels.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. moulding it into long thick sections. synthetic smoke flavours. The product is removed from the drying cycle. apart from the improved appearance. Aspergillus ostianus. In terms of safety. rewetted and remoulded to give the desired shape and consistency. including Eurotium repens and Eurotium rubrum. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. They are generally not cooked prior to consumption.6 hours and smoked for 3 . and many closely related Aspergilli (Aspergillus ochraceus. then hot smoking and finally sun drying it. Hot-smoked products made from white fish will usually keep better than those from fatty fish.5. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. fatty fish will remain in good condition for 6 days. including ‘umami’ (16. Aspergillus oryzae. Katsuobushi. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. 21. The production method is similar to masmeen. are usually 80º brined for 5 . 23). and substances unconnected with smoke but with a smoky flavour or smell. but seems. They will keep well frozen. cooking in brine. similar to masmeen. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). 17.CURED. are used. 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). a very expensive and fashionable smoked product in the Netherlands. to have no further advantage over other packing methods for other fish (12). Growth of these moulds on the tuna results in the extraction of moisture. Vacuum packaging retards the onset of rancidity when fatty fish is used. 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). Natural smoke extracts.

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

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

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

botulinum type E. so the bacteria spoiling these products will be psychrotrophic or psychrophilic. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. with a lower aw limit for growth of 0. where a high internal temperature is reached. Vacuum packing limits chemical oxidation. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). and microbial spoilage is more likely.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). Table 5. SMOKED AND DRIED FISH infrequently at low levels. The heat during hot-smoking will kill vegetative microorganisms but not all spores. which does not create the salty conditions required for spoilage by those halophilic microorganisms. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations. Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. it was. For more information on spoilage see Table 5.747.CURED. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. Pseudomonadaceae. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants.IV. flavus. LAB. Marine vibrios were the dominant flora in smoked salmon that was drysalted. will have a different residual post-smoking flora from 'hot-smoked' fish. Smoked fish products tend to have a light brine treatment. In Vietnam. able to grow on dried salted fish. 111 . Cold-smoked fish will contain microbial flora representative of that of the raw fish. but this will not prevent the growth of spoilage bacteria. including A. not the growth of spoilage organisms. but. Refrigeration is necessary. however. 33. Fish processed by cold-smoking. The preservative effect of the smoke will inhibit some spoilage microorganisms. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life. 36). around the water activity of saturated sodium chloride.5% sodium chloride in the water phase will inhibit some bacteria. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. The salt in the water phase must be high enough to inhibit the growth of C. It has been reported that the type of salting (dry. Aspergillus spp. as already stated. Aspergillus clavatus and Aspergillus niger. has been isolated from dried fish (35).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.. The brining to 3 .

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

although the combination of the two factors.0-9. botulinum type E. the non-proteolytic type E strains can grow in up to 5% sodium chloride.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades.97 0.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C.86 0. Since neither the smoking nor the drying parts of this curing process are particularly severe. the products were heavily salted.9 0. when present in the raw material. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. In the United States. The presence of salt in cured products has a great effect on the growth of the bacteria. types A and B are not affected by 3 . It has been shown that. The concentration required to prevent growth at room temperature can vary from as low as 3.5 5. but if the fish products are stored at temperatures <10 °C. sodium 113 .5-8.5. and C.97 0. under optimal conditions. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). Before.5-8.8 1. botulinum type E and production of toxin. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C.8 0.5 5.5-10. respectively. and as botulism is a serious hazard in foods.CURED. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E.61 Organisms Most Pseudomonadaceae C. with this change. pressure on the traditional methods of curing. aureus Yeasts Moulds Lactic acid bacteria pH 5. popular demand for healthy foods has led to a change in the tastes preferred by the public and. botulinum type E Staph. SMOKED AND DRIED FISH TABLE 5. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). the fermented products being the most dangerous group (7).5-10 4. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). could not grow and produce toxin. botulinum. botulinum Staph.75 0.5 4.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). Examination of 165 outbreaks of botulism caused by fish products showed that smoked.5 0-11 3. However. salted and pickled products accounted for 10. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. smoked and dried. 9 and 8 of the outbreaks. the importance of cold-chain integrity must be emphasised.5% to 5%.

but. as they require more handling. but requires higher temperatures for toxin production (>10 °C).30% of the population positive for enterotoxin-producing strains (41). if the person handling the product is a carrier of Staph. more importantly. The FDA established that hot.94 (7). after processing.5% salt in the water phase.5. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. As they are poor competitors. are particularly susceptible to contamination and. An outbreak of Shigella in an Italian cruise ship. coli all occur on fish products as a result of contamination from animal/human reservoirs. gills and guts as well as in several lightly preserved fish 114 . the organism may contaminate the product. However. 5. if storage temperatures are over 10 °C. growth and toxin formation may occur. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and.3 Enterobacteriaceae The pathogens Salmonella. if recontamination occurs and storage conditions are favourable. aureus growth.0% salt in the water phase (30). It is a halotolerant bacterium able to grow at water activities as low as 0.or cold-smoked fish should have at least 3. when the bacterial load of the product is lower. 5. they do not grow well in the raw material. In foodstuffs. Infection by Shigella sonnei has increased in the UK. 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. throat and skin. As with other organisms. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). Usually.86 and 10% sodium chloride.5.0 . Shigella and E. 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.FISH AND SEAFOOD nitrite can be added to the brine but this is not permitted in EU countries because nitrites might lead to formation of the carcinogenic nitrosamines. the organism can survive most of the light curing processes. The organism is mesophilic with a minimum growth temperature of 7 °C. has been reported (42). they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0.4. it should contain not less than 3.5. but.5. aureus. the human nose. the risk of infection can be eliminated by proper cooking. Thus. related to the consumption of a raw smoked fish meal. Sliced smoked fish products. they will grow rapidly (7). 5. Once the product is contaminated.4 Listeria monocytogenes The psychrotrophic L. if it contains sodium nitrite ≥100 ppm.

monocytogenes’ in 25 g of the finished product. a round worm. a number of fish products.95 (45). cold-smoked fish. in combination with curing agents and refrigerated storage. lightly salted caviar.8 . the USA established ‘zero tolerance’ for the organism. Some LAB. marinades (ceviche) (43) and gravad fish (44). SMOKED AND DRIED FISH products such as cold-smoked fish. both being detected in cold-smoked salmon. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. monocytogenes pathogenic for only specific segments at risk. As they are heat-sensitive. such as ‘lightly preserved’ fish products. has also been detected in cod (7). the CT 3162-FAIR program.2. which means that the process must conform to a level of ‘no detectable L.0.CURED.6 and water activities below 0. pH 4. from seawater fish. there were no reported outbreaks related to cured fish. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). or nematodes. widely used as starter cultures in food processes. monocytogenes in levels at or above 102 per gram (46).5. Pseudoterranova dicipiens.514. salt levels of 8 . They have complicated life cycles.9. most of them are of little concern to public health. from freshwater fish. Of microbiological concern in the northern hemisphere are two parasites . in the EU Fisheries R&D-FLAIR program. Unlike some European countries. The use of LAB in fish preservation is a topic that has been studied in projects (UP. do not survive 1 min at 60 °C. Two outbreaks in the United States have been reported to be linked to salmon (10). Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. Its limiting conditions for growth are 0 . Thus. the only concern to public health is when eating raw or uncooked fish products. The round worms. Until 1997. and other local traditional products (7).the round worm Anisakis spp. 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. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. have been shown to have an anti-listerial activity in many food products. 5.5 Parasites Despite the presence of parasites in fish being very common. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). and in CT95-1207FAIR). which considered L. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. This includes matjes herring. gravad fish. are considered unsafe.12%. favours growth of psychrotrophic LAB. and the tape worm Diphyllobothrium spp. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. which. which include intermediate hosts. a short period of freezing either of the raw material or of 115 .45 °C.92 .

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

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

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

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

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

R. 1987. Roberts T. Applied Microbiology. Beuchat L. Contamination pattern of Listeria monocytogenes and other Listeria spp. Eklund M. Interacting effects of pH.. 19.R. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. Davies A.A. New York. 94-8.A.. 1995.. 27 (6)..E.W. 1973. Histamine production by psychrophilic flora. 11.P. Journal of Food Safety. Rørvik L. Marine toxins of microbial origin. detection and growth of Listeria monocytogenes in seafoods: a review. Patel T.. Yndestad M. Food Technology. Fuchs R. 61 (12). Beuchat L.. 14. 1992.178-80. Competition of food bacteria with Listeria monocytogenes during enrichment culture. 1995. 1075-80.. in Seafood Quality Determination. 1994. 1991. and salt concentration on growth and survival of Vibrio parahaemolyticus. Slade A. Poindexter C. 1994. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout.. Dallas H. Dillon R.R. 121 .. 23 (1). 309-18. van Spreckens K. Ratnam S. Combined effects of water activity. International Journal of Food Microbiology.. 394-6.. 866-70. Prevalence of Listeria in smoked fish.. Eds. Pelroy G. 309-16. pH and sodium chloride. in a salmon slaughterhouse and smoked salmon processing plant. Applied Microbiology.A. Letters in Applied Microbiology.C. 1974. Elsevier. 42. Food Microbiology. Hitchins A. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. 55 (11).. 1992. Nicolaides L. SMOKED AND DRIED FISH Sutherland J. 217-36.. 58 (5). 21 (3). 4389-95. Hudson J. 502-8. Taylor S. International Journal of Food Microbiology.. 844-6. Kramer D. Paranjpye R. 19-27. Journal of Food Protection. Bayliss A.N. Caugant D.. and temperature on the growth of Vibrio parahaemolyticus. 1994. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. Letters in Applied Microbiology. Journal of Food Protection. solute. 1734. Jemmi T..T. Lashbrook L. 1994. Peterson M. temperature. Presence. 25 (5). 1988.J.. Tran T.CURED. International Journal of Food Microbiology. Poysky F. Letters in Applied Microbiology. 11. 1995. 25. Ben Embarek P.S. 293301. Incidence of Listeria in hot and cold-smoked fish.K. Liston J. Keusch A.

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

although garos.1. A selection of Asian products is presented in Table 6. These have long since fallen from use. This is well exemplified by some of the fish/salt products of Thailand described in Table 6. to products analogous to other lactic fermented foods such as salami.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. salty taste. where the bacterial production of lactic acid is a major feature. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. 6. and the more widely known Worcestershire sauce a very distant relative. a variety of products can be produced within a single country. 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. 124 .FISH AND SEAFOOD TABLE 6.II. In Europe. made from whole fish in France. who had adopted them from the Greeks. 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.III.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. and pissala. made from fish livers in Greece. may be direct descendants.

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

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

In some cases. they can contribute to the initial microflora. Generally. were found to be the predominant isolates.3. mean that microorganisms have been generally held to play little or no role in the process that follows. 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.FERMENTED FISH Where other ingredients are used. These preparations contain a mixed microflora of moulds and yeasts. 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. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. however. either directly or as the partially saccharified rice they produce. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7).3 6. in many cases. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). the Philippine fish sauce. Endomycopsis and Saccharomyces . the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp.1 – 6. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. Bacillus spp.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. principally species of the genera Rhizopus. and the anaerobic conditions that prevail in the bulk of the material. after one month. 4). Hansenula. traditional starter culture preparations such as koji (Japan). In another microbiological study of four fish sauces. and studies 127 . When this is a simple carbohydrate source such as sugar.4. however. sufficient to saturate the water phase present and produce an aw of 0. 6. (3.74 or below. It also demonstrated their significant proteolytic activity during the first month of fermentation. or boiled or roasted rice. angkak (Philippines and Thailand) and look-pang (Thailand) are used. Similar results were seen with patis. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. which included nam-pla and patis. Mucor. This. Other halophilic species have been identified from fish/salt products. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). More recently. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that.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.

2. 6.1. Patis (Condiment) 128 . Fig. 6.12). Belacan (Condiment) Fig.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 .

eviscerated fish are sometimes used to slow the autolytic process. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are. To produce semi-solid fish pastes as opposed to fish sauces. thought to be largely the result of autolytic breakdown of the fish tissues.3. 6. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13.4. 14). This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. In fish sauce production. Ika-Shiokara (side dish) Fig. the 129 .6. however.FERMENTED FISH Fig.

Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. particularly glutamate. 6. Some examples of products and their formulation are presented in Table 6.3.5).5. volatile fatty acids and nucleotides. for example. snack) 130 .2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids.8. which is eventually separated from the residual fish tissues.6 – 6. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. 6.V and Figures 6. In contrast to free-swimming fish. Hoi-Malangpu-Dong (side dish. 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. Figure 6. filtered and bottled. Fig.

12 months Shelf life Use 1 .6. N. NE 10 .5-1. S. N. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C. north-east. south of Thailand. the prefix 'pla' is replaced by 'kung'.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. C NE.FERMENTED FISH TABLE 6. C NE. NE. N S 5 . 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. 3:1:0.03 Preparation time 6 . north. N. NE Formulation Fish (whole or pieces): salt: roasted rice.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE.4 are also produced using shrimps.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0.7 days 5 .12 days 3 . Fig. In these cases. central.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . C. side dish) 131 .6. Kung-Chom (staple food.2-4 Fish (pieces): salt:khaomark (fermented rice).5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0.

VI). their growth and metabolism are nonetheless slowed.6. while they are able to grow at high salt levels.salt and lactic acid. The LAB isolated from these products are those with more marked salt tolerance but. these products are salted at lower levels than the fish sauces and pastes. giving a reduced rate of acid generation. (from Arroyo et al. It also indicates that. but there is considerable variation in this. Generally. the salt is of far greater importance in determining the product’s long-term stability. This is readily explained by the susceptibility of LAB to elevated salt concentrations.2.. Phillipines Journal of Food Science and Technology.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. 2.1 The use of higher salt levels results in a longer production phase but a better keeping quality product. Red burong isda 132 . typically in the range 2 . of the two factors contributing to shelf life .7. then this will take longer to develop than if lower salt levels are used. 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. 106) Fig. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6.10%.3. If acidity is a desired sensory property of the product.

2 4.8.5.5.8 2.6.4 .8 .9 4.6 3.3 .0 .2 4.5.8 .5 .2 7.8 pH 4.5 3..0 Shelf life 3 weeks 6 . Philippines Journal of Food Science and Technology.6. 133 .0 .2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).9 . 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 2.12 months 1 .1 4.5.3 5.5 .4.5.9 4.0 .17.FERMENTED FISH (from Arroyo et al. White balao-balao TABLE 6.3 years 1 .9.7 . 2.6.4.9.1 .

although the final population of LAB may be similar. 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).5) despite other factors. also lower the unit cost of the product and this is probably a significant factor in many cases. being similar (18.9) compared with the white product (pH 4. 21). a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. of course. a common ingredient in these products.1 . In several products.2. inulin. 19). also serves as a valuable source of the more readily fermented carbohydrate. In som-fak 9% of LAB isolated were able to ferment rice whereas 19% could ferment garlic and omission of garlic from the formulation resulted in lower acid production during fermentation (20. Where they are used. TABLE 6.VII). Garlic. Addition of carbohydrate will. When added as boiled or roasted rice alone.3. the rice may be added in a partially saccharified form such as kao-mark (Thailand). Rice is the most common carbohydrate source used.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 survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. For example.3. they increase the amount of soluble sugars produced and thus the range of LAB that can grow.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. This results in a more active lactic fermentation and a lower pH.0 .FISH AND SEAFOOD 6.4. Although the ability to ferment starch is not widespread in the LAB. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. alternatively. 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 . such as salt content.

such as wrapping in banana leaves.7 to below 4. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. lead to an increase in pH. 135 . which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. The growth of yeasts and moulds could.8 . the Korean fermented fish product. 6.5. Clearly. the level of salt employed is on the high side for fish/salt/carbohydrate products. Generally.9%.2 at the end of fermentation. There is little published information on the factors that limit the shelf life of the low-salt products. The mixture is allowed to ferment for 2 .17. In a successful fermentation.3 weeks at around 20 °C. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet.2. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. around 12%. 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. Traditional packing techniques used with lactic fermented fish products. allowing other spoilage organisms to grow. 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). the pH drops from 6. 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.7 up to 6.4 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).5 in the first 2 . markedly extending the shelf life (25). in turn.3 days.5:9:3. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). will not reliably exclude air. Excessive growth and acid production by the LAB could render products too sour for acceptability. and there is a marked regional variation in the preferred type.FERMENTED FISH Sikhae. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. but they are likely to behave similarly to lactic fermented products produced from other commodities. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. which ranged from 7. reporting the product’s pH to range from 4. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. presumably correlating with the salt content. thereafter declining to about 4. is something of an exception since millet is used along with garlic as the carbohydrate source.

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

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

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

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

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

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

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

143 .30 minutes of consumption. 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. Neurophysiological studies have shown that PSP toxins all possess the same mode of action though some derivatives such as STX and neoSTX are considerably more potent than the C-toxins (15).2.2 Toxicology PSP toxins affect the central nervous system of mammals. 7. and fingers within 5 . and humans seem to be particularly susceptible. Human symptoms begin with tingling and numbness of the lips. 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. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis. tongue.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.1. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. this is never sufficient to render the seafood safe to consume. However.

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. 15. 23. 7. Since its discovery. Most intoxication events have occurred in South East Asia. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). a criticism has been that such methods are time consuming.3. specifically 144 . as well as other parts. While the chemical detection methods are most promising for the future. However it remains as the monitoring method of choice in many countries. other derivatives have been reported. 7. Taiwan.2. Two approaches have been taken here.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. TTX most commonly occurs in many species of puffer fish. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. 17). One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. The mouse bioassay has been in place for decades and the method has been standardised.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. Thailand. 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. and has also been evaluated by a number of EU laboratories. and while simple and relatively inexpensive to operate. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®).2). Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. 24) (Figure 7. 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. China.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. 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. a pre-column method has received Association of Analytical Communities (AOAC) approval (18).FISH AND SEAFOOD 7. After some development and various trials. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). including Japan. and over ten analogs are known (22.8 µg/g) of shellfish or seafood tissue.

Structure of tetrodotoxin 7. raising speculation that TTX is produced by a bacterial source.2 mg of purified toxin can be lethal. and more recently by an actinomycete Norcardiopsis dassonvillei (26). Since then. 27). 7. and even newts. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. All of the observed toxicity is secondary to the action potential blockade. Severe poisoning causes paralysis and respiratory failure. In Japan. horseshoe crabs. nausea and vomiting. respiratory failure and coma. hypotension. the blue-ringed octopus. it is served in speciality restaurants with qualified chefs to properly prepare the fish. An oral dose of 1 . where puffer fish of the genus Fugu are a delicacy. A review of TTX detection methods has been published (12. and in mild cases results in distal muscle weakness.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions.2. and blocks diffusion of sodium through the sodium channel. 7. preventing depolarisation and propagation of action potentials in nerve cells. Fig. 26).FISH AND SHELLFISH TOXINS in the liver. 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. molluscs. and once again the mouse bioassay has been used to detect the presence of the toxin. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. 23.3. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods.3. gonads and roe (24). The onset of symptoms usually occurs rapidly. Typically this has been accomplished using HPLC followed by post 145 .2 Toxicology Like the PSP toxins. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. 25. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24.

Immunoassay methods have also been investigated. 7.FISH AND SEAFOOD column oxidation to the fluorescent derivatives. 7. Fig. and while promising.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31).3 for examples). Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28).3.4. Currently 12 isomers of DA have been reported (see Figure 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). a polar glutamate agonist. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. Structure of domoic acid and three naturally occurring photoisomers 7. However this method has been criticised for being complex.4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). but it is known that such congeners are less toxic than DA itself (30). but all of them are considerably less toxic than DA itself (29). need further development (24). but 146 . Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA.

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

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

Structure of okadaic acid and the two related DSP toxins 7. Lately these compounds.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. The parent acids are the only DSP congeners to be found in shellfish tissue. In comparison. While these esters are not toxic in vitro. are also recognised as a threat to human health. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. 149 . 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. Dinophysis acuta. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. This was probably not the first DSP episode. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. 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. In this case Dinophysis was suspected as the culprit organism. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. such as Prorocentrum lima. Dinophysis caudata.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. Today over ten Dinophysis spp.. and Dinophysis norvegica among others (47.4. Prorocentrum maculosum. 49). DTX-5. and include Dinophysis acuminata. and Prorocentrum belizeanum. Dinophysis fortii. are recognised as DSP toxin producers.5. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. 7. DSP toxins produced by benthic Prorocentrum spp. Fig. which include the pectenotoxins and the yessotoxins.

and while this provides critical seafood safety information. 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. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48).5. DSP episodes have been reported worldwide and are included in many monitoring programs (47). and DTX-2. The 7-O acyl derivatives (DTX-3) of OA that can form in shellfish also show poor cross reactivity and this further exacerbates the problem. most progress in DSP toxin detection has been using new or improved LC/MS methods and a detection limit of 10 µg/kg has been reported. 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). New Zealand and Chile. In recent years. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. and abdominal pain. Due to the lipophilic nature of the toxins. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. even with the minimal of clean up steps (48). Since the late 1970s. Human symptoms include diarrhoea. both critical enzymes involved in many key metabolic pathways in mammalian cells (50) and consequently affect a host of other biological processes in a cell including tumor promotion. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. DSP-Check is a commercially 150 .160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. which can lead to an underestimation of the total DSP content in a sample. vomiting. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. various European countries.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. it does not provide information on the DSP toxin profile. A regulatory level of 80 . nausea. 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).3 days. 7. DSP episodes have been reported from Japan. 7. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. and a detection limit of 100 µg/kg has been reported (12).5.

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

many parts of Europe and the Mediterranean. 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. Structure of the two parent yessotoxins.6. 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).2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). which may co-occur in blooms with other organisms.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). 7. the toxin content and profile may vary among different strains. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. 58). Japan. YTX and homo-YTX 7.FISH AND SEAFOOD Fig. As is typical for many algal species. New Zealand and South America (49. 57. eastern Canada. While 152 . 7. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia.6.6.

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

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

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. malaise.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). In this application. 69). In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay.7.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . The mouse bioassay is used as the primary detection method. 7. shellfish homogenates. and diarrhoea. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). Ingestion of brevetoxins results in a variety of symptoms including parathesia. 7. with a mean duration of symptoms of around 1 day. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. While the mouse bioassay continues to be used. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68).7. The mean time to onset of symptoms is around 3 hours. 66. the competitive ELISA method correlated most closely with the mouse bioassay results. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. and in severe cases seizure. and mammalian body fluids without pretreatment or dilution (72). Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. with hot spots in the coral reef areas of the Caribbean and French Polynesia. nausea. 7.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). which can result in a cascade of associated events in a cell (66). The human health implications following exposure to NSPs have been reviewed (69). vertigo. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). this family of toxins has proven to be a challenge both to identify and monitor.

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

weeks or even months (74 . and moray eels. pruritus. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. all of which can be linked to the effect of CTX on excitable membranes. Even when suspect samples are available. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. and the sensation of loose teeth (77). Local island populations attempt to avoid poisoning by catching only smaller fish. 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. the neurological symptoms can persist for days. 7. there are neither standards nor an official testing program.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish.3 Detection methods As a consequence of their potency. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . Like the brevetoxins. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. barracudas. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. sharks.1 µg of ciguatoxin can result in human illness (74. 78). groupers. which can be confused with chronic fatigue syndrome. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. which can return following the consumption of alcohol and certain foods such as oily fish (74. and there are suggestions that such incidents are increasing and spreading worldwide (74). and it is estimated that ingestion of as little as 0. 75. Other symptoms include a metallic taste. Another symptom that can develop over time is extreme fatigue. arthralgia. These include paresthesias (numbness and tingling of the extremities). and although the gastronomical problems may dissipate within a day or so. Patients have reported symptoms lasting weeks and even months. only trace amounts of CTXs can cause illness following consumption of contaminated fish. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. and in fact CTX poisoning is often only identified after consumption of contaminated fish. Globally.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. myalgia. 7. where cold objects feel hot and vice versa.8. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue.76). The onset of symptoms can occur rapidly within 24 hours. 75).8. which in turn are consumed by carnivores such as snappers. although not all CTXs are equipotent. and temperature sensation reversal.

158 . and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). 84). but this only has application in a clinical setting (81).g. This is then subjected to LCMS/MS analysis. and following synthesis of the molecule some time later. only a few laboratories are able to undertake such analyses. Recently. and the corrected structure was published in 2004 (86) (Figure 7. Immunoassays may be an alternative detection method (79). Acetone extracts of fish tissue are subjected to solvent partitioning. another in vitro sodium channel-based assay. at C-3 and C-23).FISH AND SEAFOOD Lethal Dose (MLD) 0.25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. Since the original discovery and identification of the toxin. although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). following consumption of contaminated mussels Mytelius edulis (82).9). has been developed.85). 7.20%) present in lesser amounts (Figure 7. The original structure proposed revealed an entirely new class of shellfish toxin. Consequently. almost 20 additional derivatives have been reported (83 . Recent reviews of this new group of toxins have appeared (83. and only a few structures (AZA 1 . The majority of these have been proposed based on LC/MS analytical data.80%). Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. with AZA2 (10 . which features characteristic fragment ions corresponding to sequential losses of three molecules of water. which are based on an LC-MS/MS approach. specific for CTX activity. and another dealing with the chemistry and detection methods has been published (85).9). high-throughput screen (74). and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. minor revisions of a few stereocenters were made. Chemical detection methods are severely compromised by the lack of standards.30%) and AZA3 (5 . 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. Usually the major AZA derivatives found in shellfish are AZA1 (50 .9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland.

2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. AZA occurrence has been reported by several European countries including France. which are ingested by P. which can persist for 2 . and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp.9. and some time after the original episode in Ireland. While this result is not necessarily in debate.FISH AND SHELLFISH TOXINS Fig. 89). 7. but current data indicates that unlike the DSP compounds. 88.9. AZA toxins do 159 . clams (Tapes philipinarum. The bivalves affected include mussels (Mytilus edulis. and to date no long-term effects have been recorded (83.9. and to date this has been the only source from which these toxins have been found at toxic levels. these molluscs were never considered to be the biogenetic source. 84. vomiting. There has been considerable debate over the source of azaspiracids. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). 7. Following the original episode. Mytilus galloprovincialis). scallops (Pecten maximus). as in other cases of shellfish toxins. Scandinavia. 90). cockles (Cerastoderma edule). diarrhoea and stomach cramps. Ostrea edulis). Structure of the three principal azaspiracids found in shellfish 7. Donax spp. and off the coast of Morocco (83. However. it has been impossible to establish that P.). cassipes produces AZAs in culture. and most recently in a crustacean (Cancer pagurus). Portugal. oysters (Crassostrea gigas. These symptoms are not unlike those experienced following the consumption of DSP toxins. In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. Ensis siliqua. cassipes (83).3 days.

Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. AZAs also display cytotoxicity towards various mammalian cell lines (83. 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. 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). PSP. abnormalities in the thymus and spleen. However. Based on IP minimum lethal dose data. 7. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . 91.12). the spirolides. 92). Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. the lack of sufficient material and other logistical problems have prevented complete studies of any chronic effects arising from long term exposure to low levels of the AZA toxins. Using the same approach. but all contain a spirocyclic or cyclic imine function within their structure. and closely related ptierotoxins (Figures 7.9. The mouse bioassay has been used to monitor shellfish extracts. and ASP. 7. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. and necrosis of epithelial cells on the microvilli (91). Importantly these pathological outcomes were reported as being different from those caused by DSP.10 Cyclic imines These toxins are represented by several distinct structural types. Once again. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue.10 – 7. 85). it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. the pinnatoxins. 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. and erosion and bleeding in the stomach. though the possibility exists that they may inhibit other phosphatases. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. parenchymal cell pyknosis in the pancreas. which are particularly amenable to detection by positive ion electrospray mass spectrometry.

161 . Cyclic imines are often referred to as fast acting toxins due to their “all or nothing” effect in the mouse bioassay over a very narrow dose range. selliformis (97). it was proposed that a dinoflagellate identified as Gymnodinium spp. but they are included here for completeness. surf clam and New Zealand abalone. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin. two additional congeners.FISH AND SHELLFISH TOXINS health.10). Pacific oyster.1. In only two cases (gymnodimines and spirolides) has a planktonic source been identified. 95). cockles.. Some time later. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94. 7. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . scallops. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure.520 Da (94.10. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). At the time of the initial observation of gymnodimine. Structure of the cyclic imine gymnodimine 7. Subsequent taxonomic studies revised this again to a new dinoflagellate K. 7. 95). led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7.10.1 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. Fig.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. they are sometimes referred to as emerging toxins of uncertain human health concern.

They were originally discovered in extracts of mussels (M. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. together with a lack of evidence of human intoxication. A later study using purified toxin established an LD50 dose (Lethal Dose.10.11). almost a dozen derivatives in total have been identified to date (94. Reduction of the imine group eradicated the toxicity.FISH AND SEAFOOD 7. Whether isolated from dinoflagellate cells directly or from shellfish tissue. 50%) of 96 µg/kg.100). paralysis of the hind quarters and eventually complete immobility.10.10. There is a rapid onset of symptoms including respiratory distress. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. abdominal breathing.to10-fold reduction of toxicity (LD50 600 . and there was a further ten-fold reduction in toxicity when the toxin was supplied with food.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. finishing with 100% methanol. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. 7. Most cyclic imines lack an extended chromophore. However. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. and lack of responsiveness to stimuli. 95) (Figure 7. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode.900 µg/kg). The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . Since their original discovery which resulted in the characterisation of spirolides A-F (99.3 Detection methods The characteristic “fast-acting.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). and this is the detection method of choice. 7. Based on this diminished oral toxicity. In particular. 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. 95). it has been stated that regulation against gymnodimine is not required (98). and so spectroscopic detection by UV methods is not favoured. Oral administration of the toxin to mice by gavage resulted in about an 8.1.1. The time lapse between ingestion and death is between 5 and 15 minutes.

Fig. those spirolides possessing the vicinal dimethyl grouping (see Figure 7.2.707 µg/kg. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). While oral toxicity is considerably less than by IP injection. 7. in fjords in Denmark and Norway. fasted mice). Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . Interestingly.166 µg/kg.11.10. fasted mice). or by oral ingestion (381 . the gulf of Maine. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). southern Ireland. 95).11. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . the western coast of Scotland.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. by gavage (87 .2 Toxicology The spirolides are the most studied group of the cyclic imines. All data reported here are for desmethyl spirolide C. 7.2. 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. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra).10. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). Structure of four spirolide toxins A-D 7.8 µg/kg).

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

which occupy tropical waters around Okinawa and the south east coast of China. Coupled with this. With increasing knowledge of the chemistry of toxins. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. 7. there is no compelling evidence to suggest that they have resulted in human toxicity (98). to date action or 165 . has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. and more advanced means of detection.10. The LD99 dose for the pteriatoxins is reported to be in the range (8 .180 µg/kg).100 µg/kg). There is strong circumstantial evidence to suspect that these toxins. arise from an algal source . have long been suspected of causing human illness after ingestion.10.3. 7.11 Conclusion There has been considerable progress in the detection of known marine toxins. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105).10. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds.most likely a dinoflagellate. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. 7.3. while pinnatoxin D was essentially inactive. 99%) values for pinnatoxins A-C are in the range (22 .FISH AND SHELLFISH TOXINS 7. However. new toxin groups such as the azaspiracids and cyclic imines have also been reported.3. The LD99 (Lethal Dose. Despite this progress. but this remains to be conclusively established (106). Only the acute toxicity in mice following IP injection has been reported.2 Toxicology There is little information describing the toxicology of these compounds (98). like the gymnodimines and spirolides. 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.1 Origins and distribution Shellfish of the genus Pinna. While this is recognised as a real possibility for a wide range of toxins. This is generally a result of increased monitoring and continually improving detection technologies and methods. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104).3 Detection methods No formal detection methods for these compounds have been reported. which in many cases are now found to be more widespread than previously believed.

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

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

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

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

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8. HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
Rhea Fernandes and Dr. Peter Wareing Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 8.1 Introduction

The Hazard Analysis Critical Control Point (HACCP) system is a structured, preventative approach to ensuring food safety. HACCP provides a means to identify and assess potential hazards in food production and establish preventive control procedures for those hazards. A critical control point (CCP) is identified for each significant hazard, where effective control measures can be defined, applied and monitored. The emphasis on prevention of hazards reduces reliance on traditional inspection and quality control procedures and end-product testing. A properly applied HACCP system is now internationally recognised as an effective means of ensuring food safety. The HACCP concept can be applied to new or existing products and processes, and throughout the food chain from primary production to consumption. It is compatible with existing standards for quality management systems such as the ISO 9000-2000 series, and HACCP procedures can be fully integrated into such systems. The new ISO 22000 food safety standard formally integrates HACCP within the structure of a quality management system. HACCP is fully integrated into the British Retail Consortium (BRC) Global Standard for Food Safety, and is one of the ‘fundamental’ requirements of that system. The application of HACCP at all stages of the food supply chain is actively encouraged, and increasingly required, worldwide. For example, the Codex Alimentarius advises that "the application of HACCP systems can aid inspection by regulatory authorities and promote international trade by increasing confidence in food safety". In many countries, there is a legal requirement for all food business operators to have some form of hazard analysis based on HACCP as a means of ensuring food safety. For example, within the European Union, Regulations 852/2004 and
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853/2004 require a fully operational, and maintained HACCP system, according to Codex, to be in place. 8.2 Definitions

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

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HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . procedures. 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. 6.A point.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. food microbiology. The team should also include 177 . in addition to monitoring to determine compliance with the HACCP plan. 2. hygiene managers. The team should include members familiar with all aspects of the production process as well as specialists with expertise in particular areas such as production. 8. Validation . Verification . quality assurance or control. procedure. ingredient and packaging buyers. 5. Establish procedures for verification to confirm that the HACCP system is working effectively.The application of methods. Conduct a hazard analysis.1 Assemble HACCP team HACCP requires management commitment of resources to the process. operation or stage in the food chain including raw materials. Establish documentation concerning all procedures and records appropriate to these principles and their application. 7. Establish a system to monitor control of the CCP. 3.3. Determine the CCPs. Establish the critical limit(s). 8. food chemistry or engineering. from primary production to final consumption. Figure 8.1 is a flow diagram illustrating this 12-stage logic sequence.Obtaining evidence that the elements of the HACCP plan are effective. tests and other evaluations. 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. 4. Establish the corrective action to be taken when monitoring shows that a CCP is not under control.

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

2 Describe the product It is important to have a complete understanding of the product.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. shelf life. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation.3. pH. etc. contaminated fish and shellfish should not be used. or in aquaculture operations. there may be species-specific hazards related to certain fish and shellfish.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.3. 8.). Expertise on such inherent hazards.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora. The description should include information such as the product name. 8. marine toxins associated with particular fish and shellfish species. It is also important to identify the consumer target groups. These factors are likely to have a significant effect on some potential hazards.g. storage and distribution conditions and instructions for use. Vulnerable groups of the population. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments.3. etc. However. 8.1. aquaculture drug residues and potentially pathogenic Vibrio spp. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . freezing.3.3. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. 8. is a cooking process required?). packaging. fermentation.3. In addition.). composition. naturally present on fish from warm waters (> 15 °C). for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. such as enteric viruses in shellfish. including practices onboard fishing vessels.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. such as children or the elderly. may need to be considered specifically. processing conditions (e. which should be described in detail. physical and chemical structure (including water activity (aw). heat treatment.g.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. If the processing does not include a kill step.

conduct a hazard analysis. 8.3. The operation should be observed at all stages. because the hazard analysis and decisions regarding CCPs are based on these data. Many fish-processing operations have relatively few steps and the flow diagrams appear simple.FISH AND SEAFOOD product.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters.3. If HACCP studies are applied to proposed new process lines or products. It is essential that the diagram is accurate. and therefore a detailed description of the procedure applied must be available. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. 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. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised.4. the qualitative and/or quantitative evaluation of the presence of hazards. Ideally. survival or multiplication of pathogenic microorganisms.3. it should also include details of CCP steps. For example. It may include details of the factory and equipment layout. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. once determined. features of equipment design. 8.6 List all potential hazards associated with each step. it is essential that the details of each step are fully appreciated and recorded. and identify any measures to control identified hazards The HACCP team should list all hazards that may reasonably be expected to occur at each step in the production process. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. time/temperature data. ingredient specifications. 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. However. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. 180 ● ● . cleaning and hygiene procedures and storage conditions. or circumstances of any reclaim or rework activity.

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

it is necessary to specify both time and temperature for a thermal process. application of the decision tree should be flexible.FISH AND SEAFOOD 8. 8. structured approach to decision making. Where possible.8. but for which no control exists at that step or any other.7.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. it can be said that effective control measures are likely to include the following: • • • • • 8. However. and the reader is referred to the appropriate product chapter. For example. Criteria used to set critical limits must be measurable and may include physical. and its use may not always be appropriate. critical limits should be specified and validated for each CCP. If a significant hazard has been identified at a step where control is required for safety. More than one critical limit may be defined for a single step. it is not possible to generalise on likely CCPs.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. chemical.3.2) to provide a logical.3.3. biological or sensory parameters. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively.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.1 Fish and seafood Again.3. However. then the process must be modified to include a control measure. 8. given the enormous variety of fish products and processes in use. 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 . 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.

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

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

1991. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. 24. 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.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. 7. and sets out what those enforcing the provisions have to do. manufacturing. Subsequently.9. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. processing.7. N.15).2 Legislative Structure From 1 January 2006. storing. 9. 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. transportation.1991.1992. A number of more detailed implementing and transitional measures have been adopted at EC level. 24.B. p. distribution.9. p. 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. 41). J. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. 190 • • . 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. 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. as last amended by Regulation (EC) No 806/2003. p.1). as last amended by Regulation (EC) No 806/2003. packaging. European Communities L 268. handling and offering for sale or supply to the consumer.

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

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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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and chopping. Fresh fishery products means unprocessed fishery products. live tunicates and live marine gastropods. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. mincing or processing. gutting and removal of fins and. 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. 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. parts and products of such animals. slicing. shelling. if necessary: chilling or freezing: filleting. 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. and all mammals. Conditioning means the storage of live bivalve molluscs coming from class A production areas. to remove mud. purification centres or dispatch centres in tanks or any other installation containing clean seawater. or in natural sites. where appropriate. skinning. Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. shucking. live echinoderms. after preparatory work such as bleeding. such as gutting. slicing. in particular as a result of feeding on plankton containing toxins. heading. to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness. reptiles and frogs) whether wild or farmed and including all edible forms. 197 . whether whole or prepared. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. filleting. slime or sand. heading. where necessary.

9. In this case.4. For the UK. (iv) If the identification mark is applied in an establishment within the Community. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not.2.1. EK.2. the mark must be oval shaped and include the abbreviation CE.2 Fishery product requirements 9. (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. EB or WE. the code would be UK. and the characters easily decipherable and be clearly displayed. EO.2 Form of the identification mark (i) The mark must be legible and indelible. (iii) Additionally. EF. EY. 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. for other codes.4. 198 .1.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. please refer to Regulation (EC) 853/2004. ES. EG. EC. (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.FISH AND SEAFOOD 9. Section I of Regulation (EC) 853/2004. when a product's packaging and/or wrapping is removed or is further processed in another establishment.4. the mark must show the approval number of the establishment.4. Section I of Regulation (EC) 853/2004 are given as follows: 9. (ii) However. EÜ. a new mark must be applied to the product. the food business operator may apply the same identification mark to both types of food.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production.2. Extracts of the requirements of Annex II. the new mark must show the approval number of the establishment where this occurred. EK.

tunicates and marine gastropods when 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. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. Part A and Chapter V apply to retail. the wrapping or the packaging.1. a) requirements for establishments. With the exception of Chapters I and II. processing. C and D. they would need to be obtained following Section VII on live bivalve molluscs. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. 9. the colourings used must be EC authorised colours.2. the approval number of the establishment and the abbreviation for the community where appropriate. (ii) For products of animal origin that are placed in transport containers or large packages. i. and fishery products carried in bulk. The requirements of this section accompany those in Regulation (EC) 852/2004.LEGISLATION 9. specifically relating to fishery products are given below. Chapter III. it applies to those animals when not placed on the market live. (iii) In the case of liquid.4.2. Section VIII of Regulation 853/2004. its wrapping or the packaging. wrapping or packaging in another establishment. as amended. reference should be made to the actual Regulation. the mark may be applied to the outer surface of the container or packaging. echinoderms.e. Extracts of the requirements of Annex III. Parts A. Chapter IV.4.3 Method of marking (i) The mark is to be applied directly to the product. it is sufficient to apply the mark to the exterior of that package only. and are intended for further handling. This section does not apply to bivalve molluscs. including vessels. Therefore. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. or be printed on a label affixed to the product. granulate and powdered products of animal origin carried in bulk. (v) When the mark is applied directly to products of animal origin.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . for full requirements.

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

(iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. and allow the monitoring and. to ensure that melt water does not stay in contact with the products. tanks must incorporate devices that allow a uniform temperature throughout the tanks. 3. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas. recording of temperatures. any waste processing unit needs to have a separate hold to store the waste. (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. 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.LEGISLATION (i) Such vessels need to have holds. Requirements for factory vessels a) Factory vessels (i) These must have at a receiving area reserved for taking fishery products on board so each catch can be separated (ii) There must be a hygienic system for conveying fishery products from the receiving area to the work area (iii) Large work areas are needed to allow fishery products to be prepared and processed hygienically. 4. where necessary. Storage areas for the finished products also need to be sufficiently large and cleanable. where necessary. 201 . (iii) In vessels equipped for chilling fishery products in cooled clean seawater. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. Also. A temperature recording device must be present and placed where the temperature in the hold is the highest. 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.

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

C.LEGISLATION consumption. 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. Additionally. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. and where necessary. They should be wrapped. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . (ii) Heading and gutting must be carried out hygienically. 854/2004. packaged and chilled as quickly as possible after preparation. B. dispatched. 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. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. they must be stored under ice in appropriate facilities. Fillets and slices must then be removed from work tables as soon as possible after preparation. part I. (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. Chapter I.Requirements for establishments. 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.3 Chapter III . (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels.2.4. points 1 and 2. prepared or processed immediately after arrival at an establishment on land. handling fishery products A. 9. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. including vessels. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices.2. C.

raw materials must have no guts. Once produced. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . and whole fish must be gutted and washed beforehand.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.4. except when supplied to the final consumer. (ii) Compliance with the above is not necessary if epidemiological data indicates that the fishing grounds of origin do not present a health hazard in relation to the presence of parasites and this has been authorised by the competent authority. (iii) A document from the manufacturer. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. must accompany fishery products referred to above when placed on the market. cooling must continue until a temperature 204 .FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. (iv) (wild) Atlantic and Pacific salmon. or a stabilising treatment.Requirements for certain processed fishery products A.2. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked.4 Chapter IV . stating the type of process they have undergone. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. they must be cooled rapidly. 9. If no other method of preservation is used. if the processing is insufficient to destroy nematode larvae. D. and (c) marinated and/or salted fishery products. (b) fishery products from the following species. (iii) sprat. (ii) mackerel. 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.

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. B. B.2. 2073/2005. 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). 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. However.LEGISLATION approaching that of melting ice is reached. separation. 2074/2005. pressing. and the raw material is processed within 36 hours after loading. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. 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. centrifugation. 2073/2005). Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. and after this. 9. refining and purification steps before being placed on the market for the final consumer. (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.5 Chapter V .Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No.4.2. 2074/2005). 205 . Shelling/shucking is to be done hygienically. the following conditions need to be met: A. C. processing.

prepared and processed fishery products belonging to the family Gempylidae. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption.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.7 Chapter VII . thawed unprocessed fishery products.2.2. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. 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. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. 9. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. Fishery products contaminated with parasites may not be placed on the market for human consumption. Molidae. Diodontidae and Canthigasteridae.2. Fresh. In the case of whole frozen fish in brine intended for the manufacture of canned food.6 Chapter VI . and when fishery products are wrapped on board fishing vessels.Storage of fishery products (i) Fresh fishery products. 206 .FISH AND SEAFOOD D. it should be kept at no more than -9 °C.2. 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.4. 9. The scientific name must accompany the common name on the label.4. E. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice.

(d) 60 mg of nitrogen/100 g of whole fishery products used directly for the preparation of fish oil for human consumption as referred to in the second sub-paragraph of Part B(1) of Chapter IV of Section VIII of Annex III to 207 . The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. The following in relation to TVB-N limits is extracted from the regulation: Unprocessed fishery products shall be regarded as unfit for human consumption where organoleptic assessment has raised doubts as to their freshness and chemical checks reveal that the following TVB-N limits are exceeded: (a) 25 mg of nitrogen/100 g of flesh for the species: Sebastes spp.2.2. melt water must not remain in contact with the products. species belonging to the Gadidae family. provided that the journey is short and the competent authority allows this. (ii) If fishery products are kept under ice. 9.2.8 Chapter VIII .LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability.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.4.4. 9. The temperature of frozen fishery products must not fluctuate above -15°C. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. species belonging to the Merlucciidae family. Sebastichthys capensis. Helicolenus dactylopterus.). 2074/2005. 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.Transport of fishery products The above points for Chapter VII apply in relation to transport.

3. The information required in the registration document is given in point 4 and includes the health status of the production area. The sample must consist of about 100 g of flesh. (b) In the case of other operations. shellfish species etc. The copy of 208 . 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.4.FISH AND SEAFOOD Regulation (EC) No 853/2004. 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. this Section also applies to live echinoderms. Extracts of the requirements of Annex III. 9.4. and point 3 of Chapter VII applies to retail. Apart from the provisions on purification. (b) and (c) of Part B(1) of that Chapter. reference should be made to the actual regulation. where the raw material complies with points (a).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. Live bivalve mollusc requirements - 9. Section VII of Regulation 853/2004. as amended. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. they supplement the requirements of Annex II to that Regulation. taken from at least three different points and mixed together by grinding. specifically relating to bivalve molluscs are given below. for full requirements. they supplement the requirements of Annex I to that Regulation. VIII and IX. VI. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. please refer directly to the regulation. Chapters V.1 Chapter I . tunicates and marine gastropods.3 The regulation details specific hygiene requirements for bivalve molluscs. however. Chapter IX applies to pectinidae harvested outside those areas. 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.

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

dinophysistoxins and pectenotoxins together.4. the purification system to allow them to resume /maintain filter-feeding activity. shells of live bivalve molluscs are to be washed throughout with clean water and the molluscs are to come from one of the following: (i) Class A production area (ii) Relaying area (iii) Purification centre (iv) Another dispatch centre 9.3. Requirements for purification centres There are requirements for live bivalve molluscs to be washed.3 Chapter III .4.4 Chapter IV . 210 .Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. 800 micrograms per kilogram. containers and a label to identify purification. requirements if there are many batches of live bivalve molluscs.5 Chapter V . 20 milligrams of domoic acid per kilogram (iii) okadaic acid. 160 micrograms of okadaic acid equivalents per kilogram. and there are requirements for tanks and water storage containers.3. Requirements for dispatch centres Handling should not result in contamination to any degree. for: (i) paralytic shellfish poison (PSP).Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements.FISH AND SEAFOOD 9. 9. B.4. (iv) yessotoxins.3. 1 milligram of yessotoxin equivalent per kilogram. Before dispatch. (ii) amnesic shellfish poison (ASP). the levels of marine biotoxins should not exceed the following limits.Hygiene requirements for purification and dispatch centres A.

7 Chapter VII .3.6 Chapter VI .4. 9.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.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.3. 211 . the label.Identification marking and labelling In addition to general identification marking (contained in 9.4. 9.4. must be waterproof.8 Chapter VIII . the species of bivalve mollusc and date of packaging (at least the date and month) are required. Also. Finally.4. dispatch centre or processing establishment.2.9 Chapter IX .3.LEGISLATION (v) azaspiracids. Registration document requirements also apply. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.3. they cannot be re-immersed in or sprayed with water. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.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. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.4. 9. 160 micrograms of azaspiracid equivalents per kilogram. including the identification mark. by way of derogation from directive 2000/13/EC. (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. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. Requirements of Chapters III and IV are to be complied with.1).

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

Classification of production and relaying areas: This section reinforces that specified in Annex III. The reference method for this analysis is the five-tube. 4. Also.000 E. live tunicates and live marine gastropods. - - - - B. coli per 100 g of flesh and intravalvular liquid.5.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. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. chapter V is reinforced.Official controls concerning live bivalve molluscs from classified production areas A. live bivalve molluscs must not exceed 46. In the remaining 10 % of samples. as previously stated. that microbiological criteria and chemical contaminants amongst other requirements are met. three dilutions MPN test specified in ISO 16649-3. The reference method for this analysis is the five-tube. coli per 100 g of flesh and intravalvular liquid. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. live echinoderms. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. A summary is given as follows: 9.5. Live bivalve molluscs from Class B areas must not exceed. Sampling plans and frequency are required. Annex II 9. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3.2. in 90% of the samples. coli per 100 g of flesh and intravalvular liquid.1 Chapter II .2 This states official controls for the production and placing on the market of live bivalve molluscs. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. 000 E. Regulation (EC) 854/2004. 213 .600 E. Live bivalve molluscs from Class C areas must not exceed 46. section VII.

Procedures for re-opening a closed production area are stated D. boundaries or class of a production area or its closure. the competent authority is to close the production area.6 Regulation (EC) No. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met.2 Chapter III .3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. 214 .5. 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. 2073/2005. F.2.FISH AND SEAFOOD C. 9. opening or closing a production area. 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. E. which has applied since 1 January 2006. 9. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. establishes microbiological criteria for a range of foods. the Member States may permit the import of bivalve molluscs and fishery products from the countries listed respectively in Annex I and Annex II to regulation (EC) 2076/2005. 9.5.

LEGISLATION The aim of this legislation is to complement food hygiene requirements. a batch of foodstuffs or a process.6. volume. and/or on the quantity of their toxins or metabolites. presence or number of microorganisms.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria.I. 9. The criteria are outlined in Table 9.II. per unit(s) of mass. 215 . applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. 9. Two kinds of criteria have been established: food safety criteria. area or batch.4 referring to fishery products. with chapter 2. ensuring that foods being placed on the market do not pose a risk to human health. and the legislation applies to all businesses involved in food production and handling. based on the absence. The relevant criteria are outlined in Table 9.6.1 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.

monocytogenes 1. monocytogenes. 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. other than those intended for infants and for special medical purposes(4) (8) L. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1. monocytogenes. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.1 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.2 Ready-to-eat foods able to support the growth of L.3 Ready-to-eat foods unable to support the growth of L.TABLE 9.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 .

27 Fishery products which have undergone enzyme maturation treatment in brine. tunicates and gastropods 9 (18) 2 E. 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. 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 1.17 Live bivalve molluscs and live echinoderms. coli (15) 1.25 Live bivalve molluscs and live echinoderms.Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.

0 and aw ≤ 0. 79. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. Bouquelet S. Coryfenidae. — fresh. Dervin C. 2. — bread. that the product will not exceed the limit 100 cfu/g throughout the shelf life. 5 This criterion shall apply if the manufacturer is able to demonstrate. Malle P. References: 1.1 . Single samples may be taken at retail level. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. subject to scientific justification. 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.4 or aw ≤ 0. 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.92. Particularly fish species of the families: Scombridae.25. products with pH ≤ 5. Pomatomidae. to the satisfaction of the competent authority. 43-9. AOAC Internat. when recontamination is not possible after this treatment (for example. — — bottled or packed waters.. biscuits and similar products. honey and confectionery. spirits and similar products. Assay of biogenic amines involved in fish decomposition. Valle M. This criterion shall apply to products before they have left the immediate control of the producing food business operator. wine. cider. Malle P. The most recent edition of the standard shall be used. Clupeidae. J. m = M. beer. 1996. Bouquelet S.. Duflos G.. Products with pH ≤ 4. products with a shelf-life of less than five days shall be automatically considered to belong to this category. A pooled sample comprising a minimum of 10 individual animals.. products heat treated in their final package). Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . according to which the whole batch is to be deemed unsafe.. uncut and unprocessed vegetables and fruits. coli is used here as an indicator of faecal contamination.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. sugar. Scombresosidae. Other categories of products can also belong to this category. monocytogenes. to the satisfaction of the competent authority. when he is not able to demonstrate.1. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002.94. c = number of sample units giving values between m and M For point 1. including cocoa and chocolate products. Engraulidae. soft drinks. shall not apply. excluding sprouted seeds. live bivalve molluscs. E.

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

1 Shelled and E.TABLE 9.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. 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.

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

such as that below. Alternatively. was kept at above 8 °C or the recommended temperature for an unavoidable reason. S. or. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions.gov. for a limited period consistent with food safety. 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. 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. 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. 2006 No. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S.K.I.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. 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. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment. and was kept at above 8 °C or the recommended temperature. or temporary breakdown of equipment For Scotland there are separate provisions to include requirements to hold food under refrigeration or in a cool ventilated place. 2006 No. For other food. 3).uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . 9.I. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. or at a temperature above 63 °C and to reheat food to a temperature of at least 82 °C (The Food Hygiene (Scotland) Regulations 2006. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures.8 Guidance In the U.S.food. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing.

2004. L283. 2076/2005. 62) as amended by Regulation (EC) No. 223 . namely. 2075/2005 and Regulation (EC) No. as read with the corrigenda at OJ No. 9.2004. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito.6. 55). 2076/2005. 206). 30. Regulation(EC) No. these are not be discussed here. Regulation (EC) No. 853/2004 is now set out in a Corrigendum (OJ No. 854/2004 is now set out in a Corrigendum (OJ No. 1). Regulation (EC) No. Commission Regulation (EC) No. L226. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. Regulation (EC) No. Regulation (EEC) 2136/89. 25. 2074/2005.2004. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 25. L226.12. 2074/2005 and Regulation (EC) No. Regulation (EC) No. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No.LEGISLATION 9. 2074/2005.4.2004. 2076/2005.10. laying down common marketing standards for preserved sardines. 30. 22) as amended by Regulation (EC) Nos. L139.4. 1.4. L139. Regulation (EC) No.2004. The revised text of Regulation (EC) No.2006.6.2005. The revised text of Regulation (EC) No.2004. 2073/2005 on microbiological criteria for foodstuffs (OJ No. 852/2004 is now set out in a Corrigendum (OJ No. 2076/2005. 2074/2005. 30. 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. The revised text of Regulation (EC) No. as amended. 25.6.9 Other Relevant Legislation The following regulations about fish designations need to be considered: • Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products Regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products • Note: Requirements for preserved fish are laid down elsewhere. L139. 1441/2007. 1688/2005. L338. 83) as amended by Regulation Nos 882/2004. 1019/2008 and as read with Regulation 2073/2005. 1662/2006.10 Further Reading Regulation (EC) No. 1663/2006. 14. L226. 3) as amended by Regulation (EC) No. 22.

22.2001. derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17. 21.10.2000. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004.12. 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. 23.1. (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004 ( OJ L 338.2005.12. 6) 224 . as amended by Commission Regulation (EC) No 1022/2008 OJ L 338. 22.FISH AND SEAFOOD Commission Regulation (EC) No 2074/2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC)No 882/2004 of the European Parliament and of the Council.2005.

1.1 10.3 Temperature Aeromonas grows at temperatures of 2 .1 Aeromonas spp. 10.1. no growth is observed (1).10. The optimum growth temperature is 28 °C (2).5 µm 10.0 .3 minutes. 10.1.5 minutes.2 Oxygen requirements Facultative anaerobe.1. and 2. Morphology Gram-negative rods. Aeromonas has the potential to grow over a pH range of ca 4 . At pH 4. 0.000-fold increase in number is observed after one week in food systems (1).10 (3).45 °C (1).3 . under optimal incubation temperatures.5 pH Under otherwise optimal conditions. a 10 to 1.4 Heat resistance Aeromonads are sensitive to moderately high temperatures. PATHOGEN PROFILES 10. Some death will occur at this pH if the temperature is 4 °C.1. 225 .5. The minimum pH value for growth is thought to be pH 5.0 x 1. At 5 °C.5.3. respectively (1). 10.1. and D-values at 45 and 51 °C have been reported as 29.

There are four main groupings of the organism. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C. or at 90 ºC for a few seconds.30 minutes. All type A strains are proteolytic and type E strains are usually non-proteolytic. Thus an aerobically packed product may not support the growth of the organism on the surface. botulinum are not particularly heat resistant. B. but the spores of this organism are more so.1 Morphology Gram-positive. types B and F can be either.even as low as 3 ºC (7).0 µm.4 Heat resistance The vegetative cells of C. botulinum is an obligate anaerobe. botulinum is prevented. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. spore-forming rods.7 . Types A. 226 .2. F are the types that are responsible for most cases of human botulism (5. forming at least seven different toxins. 0. being capable of slow growth and toxin production at low temperatures . 6). at 85 ºC for 5 minutes. 10.1.22. but Group II strains are psychrotrophic. All C.2 Clostridium botulinum Seven different types of C. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. 10. to a lesser extent.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 .2.2. but the interior of the food may do so.2% sodium chloride (NaCl) (1).4 x 1.2. Group I will not grow at temperatures of 10 ºC or less.2 Oxygen requirements Although C. E and. which may be inactivated by heating at 80 ºC for 20 . 10. A to G. and is sensitive to > 4.2. 10. botulinum are known.5 . 10.3 Temperature All strains of C.FISH AND SEAFOOD 10.5% NaCl (4). botulinum types produce heat labile toxins. and groups I and II are those responsible for the botulism cases. botulinum grow reasonably well in the temperature range of 20 45 ºC.

PATHOGEN PROFILES 10.0 . botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. 0.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I.97) (10.2.6 and pH 5. 10.97 for Group II (5). they may survive mild heat treatments (70 . 9). These concentrations correspond to limiting aw of 0. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. 11)).g.3 10. pH and temperature.1 Clostridium perfringens Morphology Gram-positive spore-forming rods.0 and aw > 0. botulinum depends on solute.10. 10. parcooked meals with pH value > 5. However. The organism will grow well in the centre of meat or poultry dishes.21 minutes) (9).2.2 Oxygen requirements C. particularly by cooking.0.like other clostridia . perfringens .94 for Group I and 0. botulinum spores The most heat resistant spores of Group I C.3. grow on the surface of foods unless they are vacuum or gas-packed.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. In commercial situations.0 µm. 10. It will not. 10.9 x 2.3.is an obligate anaerobe.7 Characteristics of C.1. where oxygen levels are reduced. therefore.2. the minimum aw for growth may be lower.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. vacuum packed. These values have been established under carefully controlled laboratory conditions. respectively (8. safety margins must be introduced to allow for process variability.3 . 227 . Note also that if humectants other than salt are used.

Its optimum temperature for growth is 43 .45 ºC. however. etc. Recorded heat 228 . perfringens can vary quite considerably in their heat resistance. (It is unusual for spores to be formed in foods after the growth of this organism (14)). and their heat resistance is also affected by the heating substrate.3. 18).FISH AND SEAFOOD 10. Spores. perfringens is not tolerant of low water activities. In addition. the limits for aw are affected by temperature.50 ºC. pH.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. Salt concentrations of 6 . perfringens has the potential ability to grow within the temperature range 15 . perfringens in vacuum packed beef (18). Under otherwise ideal conditions.93 and 0. vegetative cells die rapidly (12).97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. depending on strain and other conditions. 13). 10.10 ºC. 16).3 Temperature The most significant characteristic of C. 10. perfringens appears to be 0. 15.3. perfringens is not a tolerant organism with respect to pH.3. lower concentrations may be effective in combination with other factors. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. type of solute. perfringens strains. the enterotoxin is not heat resistant . will survive greater extremes of pH (and aw) (12. although C. perfringens in roast beef at 60 ºC is 14.it is destroyed by heating at 60 ºC for 10 minutes (14.5 pH C.8% inhibit growth of most C. At cold temperatures.3. 13). perfringens. The lowest aw recorded to support the growth of C. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. perfringens spores The spores of C. As in the case of other factors limiting the growth/survival of this organism.5 minutes (17). death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. 10.3. It grows best at pH values between 6 and 7 (the same pH as most meats). perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures.3. Some studies indicate that the presence of 3% NaCl delays growth of C.7 Characteristics of C. 0 . The D-value for C. 10.6 Aw/Sodium chloride C. While some growth can occur at 50 ºC.

it is not a sporeformer.4. However.4 Heat resistance L.64.98 and 0. monocytogenes is reported to be 45 ºC (20). growth at low temperatures can be very slow.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.2. -0. In foods. pH < 5.0 minutes for heat resistant spores to 1. however. 10. requiring days or weeks to reach maximum numbers. using hydrochloric acid as an acidulant). is between 30 and 37 ºC.2 Oxygen requirements Aerobe or microaerophilic.4.albeit slowly .2. being potentially capable of growing . Morphology Gram-positive short rods. monocytogenes in crawfish tail meat at 55. 10.2 has been suggested as the lowest working limit (22). or even below 0 ºC. the lowest limit for growth is likely to be considerably higher .3 .PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. D-values for L.6 .1 Listeria spp. 10. 0. and temperature. 60 and 65 ºC were reported as 10. monocytogenes is not a particularly heat resistant organism. the organism is able to grow at pH values well below pH 5 (pH 4.4 .8 minutes for heat sensitive spores (12).4 10. The upper temperature limit for the growth of L. 229 .3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. 10.0 µm.19 minutes. 10.especially at refrigeration temperatures. and where acetic acid is used as an acidulant.3 is the lowest value where growth has been recorded. Under ideal conditions. Its optimum growth temperature.5 . 1. so can be destroyed by pasteurisation. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.4. however.0.4 ºC is probably a more likely minimum for growth in foods (19).5 x 0.at refrigeration temperatures down to.23. respectively (21).4.4.

5 pH Plesiomonas has a pH range for growth of pH 4. 10.5. 10. 0.1 Salmonella spp.6 Aw/Sodium chloride L.12% NaCl or more).0 x 1 . 10.7 x 1. the optimum being 30 ºC (23). 10.8.1. 0.45 ºC.5.5 .0 µm.4. The bacterium may survive for long periods at aw values as low as 0. It is likely to survive.5 .5.FISH AND SEAFOOD 10. 10.5 (24. peritrichous flagella.3 Temperature Plesiomonas can grow in the temperature range of 8 . monocytogenes is quite tolerant of high NaCl/low water activities. 230 .3 µm. 10.97. at salt levels found in foods (10 . but has been shown to be able to grow at an aw level of 0.5.1 Plesiomonas Morphology Gram-negative.6.83 (20).6 10.0. It grows best at aw of ≥ 0. 10. Morphology Gram-negative short rods. 25).90.5 10.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.5.2 Oxygen requirements Facultative anaerobe.0 .4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).8 . or even grow.3. 25).5.

2 Oxygen requirements Facultative anaerobe.6 Aw/Sodium chloride Where all other conditions are favourable. The optimal pH for Salmonella growth is between 6. However. a heat resistant organism. pasteurisation and equivalent treatments will destroy the organism under normal circumstances.4 Heat resistance Salmonella is not a spore-forming organism.5. and appropriate heat treatments must be determined experimentally for low-aw foods. depending on serotype.5. Some death will occur at pH values of less than about 4.93 (as reported in dried meat and dehydrated soup). Salmonella has the potential to grow at aw levels as low as 0.7. with a z-value of 4 . some strains are able to grow at temperatures as low as 4 °C (26). Salmonellae are quite resistant to drying. substrate.5 . therefore. 10. It is not. and with an appropriate acid. or possibly 0. Salmonella senftenberg 775W is about 10 . although salt tolerance increases with increasing temperature (28).3 Temperature Salmonellae can grow in the temperature range of 7 .0. under otherwise ideal conditions.48 ºC. depending on the type of acid and temperature.6.4% NaCl. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments.6.5 pH Salmonella has a pH range for growth of pH 3.8 . 231 . 10. Furthermore.5 ºC. Growth is slow at temperatures below about 10 ºC.6.PATHOGEN PROFILES 10.9. However. 10.20 times more heat resistant than the average strain of Salmonella at high aw (27). The growth of Salmonella is generally inhibited by the presence of 3 . D60 ºC values normally range from about 1 to 10 minutes. temperature and pH.6.37 ºC.6. 10. the optimum being 35 .945. strains vary in their ability to withstand heating. Salmonellae are quite resistant to freezing.

aureus can grow within the temperature range 7 – 48. with an optimum of 35 .3 Temperature Under otherwise ideal conditions.7. 0. under otherwise ideal conditions (29. 31).0 . 10. In meat macerate.37 ºC (29). Most strains of staphylococci can grow within the pH range 4. As is the case with other bacteria.7 10.0. 10.FISH AND SEAFOOD 10.7 .5). It can survive well at low temperatures. however. Freezing and thawing have little effect on Staph. aureus viability. The limits for toxin production are also narrower than for growth. depending on aw.7. etc.).5 ºC. but may cause some cell damage (30).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.20 minutes. Staph.7.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. Under most circumstances. stressed cells can also be less tolerant of heating. NB: The growth of Staph.3 (optimum 7.7.7.9 µm diameter.7. the organism is heat sensitive and will be destroyed by pasteurisation. aureus than citric acid). aw and temperature (sensitivity to acid increases with temperature).2 Oxygen requirements Facultative anaerobe.2 to 9. 232 . aureus is more limited under anaerobic than aerobic conditions. The following relate to limits for growth only. 10. fat content. aw. the D60 ºC value is 2 .1 Staphylococcus aureus Morphology Gram-positive cocci. 10.

99 aerobically under otherwise optimal conditions.7. 0.5) (29. with a temperature range for growth of 10 . especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.1 Vibrio cholerae Morphology Gram-negative curved rods. Staph.8.83 .0 . 10.6 Aw/Sodium chloride Staph. and solute.8 x 1. * dependent on temperature. It can grow over the aw range 0.3 Temperature The optimum temperature for growth has been reported as 30 ºC.5 . 233 .43 ºC (35). strain.10% NaCl. 31) between 0.0 (optimum 7. However. but certain strains can grow in 20% NaCl.87 and > 0.6 µm 10.8 10.0.7.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5.PATHOGEN PROFILES 10. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31.>0. 10.86 is the generally recognised minimum in foods (32). atmosphere.8. In general.7 Limits permitting toxin production 10 .7. Staphylococci are more resistant to salt present in foods than other organisms. 34).4 .8.2 . 10. aureus can grow in 7 . an aw of 0. An effect of increasing salt concentration is to raise the minimum pH of growth.2 Oxygen requirements Facultative anaerobe. pH.2. In general.9.99 little or no toxin production in anaerobically packed foods.

9.9.5 ºC).97 . but only moderately sensitive to freezing (40). with an optimum of 0. 10.5 pH This organism is tolerant of high pH values.65 minutes (35). under otherwise favourable conditions. The D-value for V. 10. and it is sensitive to refrigeration temperatures (0 . 37. cholerae can grow over the aw range 0. with an optimum of 37 ºC (39). parahaemolyticus is easily destroyed by drying. declining in number during storage. cholerae in crab meat homogenate at 60 ºC is 2.984 (35). It can grow in the presence of 0.9 10.8. 234 .FISH AND SEAFOOD Also. with optimum at 7.4 Heat resistance V.6 Aw/Sodium chloride V.2 Oxygen requirements Facultative anaerobe. are 5 and 43 ºC.0% NaCl (35). respectively.0. 38).1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. cholerae is sensitive to heat at temperatures of >45 ºC. 10. there is evidence that V.6 (35.998.8. 10.1 . but not acid conditions. 10.6.9. V. cholerae may persist on frozen fish for >180 days (36).9.8.4. 10.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media. it has a pH range for growth of pH 5 .

with a temperature range for growth of 8 . with an optimum pH of 7. 10.9.8.6 (41).43 ºC (42). parahaemolyticus will grow at aw levels as low as 0. and are readily destroyed by cooking. depending on solute. parahaemolyticus is unable to grow in the absence of salt.10. 235 . 10. but the optimum concentration is 2 . respectively (39). it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43). The optimum concentration of NaCl equates to aw of approximately 0. parahaemolyticus are not heat resistant.2 Oxygen requirements Facultative anaerobe. respectively.10 Vibrio vulnificus 10.9. 10.1 Morphology Gram-negative curved rods.29 minutes.4 Heat resistance V. The organism can survive for long periods at freezer temperatures (42).9. The D-values at 55 ºC for clam and crab homogenate are 0.8 .5 minutes.922.98 (35.10.0.6 .02 .6 Aw/Sodium chloride V.PATHOGEN PROFILES 10. 10. 41). 41).4 Heat resistance V.10. vulnificus is heat sensitive.4% (39.10. 10. 10. and 2.11.5 pH The pH range for growth is 4. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0. V.3 Temperature The optimum temperature for growth has been reported at 37 ºC.5% NaCl.

O. Smith J.. Clostridium botulinum. 1993. Food Control. Kovacek K. Murrell K. Stelma G. in Foodborne Pathogens. 11. Eds.R. Abeyta C.. Principles of control.L. Ed. Hui Y.. 357-67.H.J. Eds. Clostridium botulinum and Clostridium perfringens. Foegeding P. Eds. 7. Peck M. Aeromonas.L.D. Great Britain. 1995. International Commission on Microbiological Specifications for Foods. 5-19. Kim J. Faris A. 383-408. Eds.C. Austin J. 1-27. 10.5% (42).H..M.11 1. Dodds K. 1996. in Clostridium botulinum: Ecology and Control in Foods. Ed. Marcel Dekker.L. Doyle M. Marcel Dekker Inc. 5. New York. Clostridium botulinum. Gaze J. 97-131. Aeromonas hydrophila.. Hui Y.W.N.. Ed. Blackie. 2003. 121-76. in Foodborne Bacterial Pathogens. 57-63.. 2. Aeromonas hydrophila group. New York. Betts G. Bier J. Eds. Stelma G. in Clostridium botulinum: Ecology and Control in Foods. Gorham J. New York.D. Hauschild A. London. New York. 10. 2005..M. in Foodborne Bacterial Pathogens.P.10. Hauschild A..8 (42)... 6.K.E.10.W.. 4. Clostridium botulinum. Bhunia A. in Foodborne Disease Handbook. 8.H.. vulnificus grows over the range 0.O. 1-19. New York. Gorman J. Dodds K.P. with an optimum of 2.W. Palumbo S.L..P. 279-303. 236 . 329-49. Beuchat L.R.10. 10. 2001.H.. Marcel Dekker. New York.0% NaCl. Dodds K. in Food Microbiology: Fundamentals and Frontiers. New York.6 Aw/Sodium chloride V. Eds. Volume 1. 1994.H. Marcel Dekker. Microbiology and Molecular Biology. Diseases Caused by Bacteria.5 pH The pH range for growth is 5 .. Marcel Dekker. Cliver D. with an optimum pH of 7.R. Novak J.5 . Potential hazards associated with REPFEDS.D. Montville T. Eds.. in Foodborne Disease Handbook. Doyle M.. 111-89. Doyle M. 9.H. Aeromonas species. 1994. in International Handbook of Foodborne Pathogens.5.W. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’... 1989. Johnson E. Notermans S. 1989. 1993. Volume 5: Microbiological Specifications of Food Pathogens. Murrell K.. in Microorganisms in Foods.M. Fratamico P. References International Commission on Microbiological Specifications for Foods. Hauschild A.A. Cliver D. Marcel Dekker. Volume 1: Diseases Caused by Bacteria.D. Miliotis M. ASM Press. 6 (1). Washington D. Marcel Dekker..W. Lund B.N. 3.FISH AND SEAFOOD 10. Caister Academic Press.. Juneja V.

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

1996. 327-52. Loessner M. Japanese Journal of Experimental Medicine.S. in Foodborne Disease. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems. Jay J. Vibrio parahaemolyticus. 38. CDR Review. 36.62. 543-68.M. Cambridge. Cliver D. 1983. 238 . Marcel Dekker. in Foodborne Bacterial Pathogens. International Commission on Microbiological Specifications for Foods. Volume 5: Microbiological Specifications of Food Pathogens.C.S. Stewart G. New York. International Commission on Microbiological Specifications for Foods. Vibrio parahaemolyticus. Staphylococcal Food Poisoning. 2005. 8. London. Salmonellae... 35. 327-445. New York. 1996.R. Cholera: current epidemiology.. 32. 33. Gustafson J. Tohyama Y. 2005. 273-84. 85-106. Bergdoll M. in Microorganisms in Foods.. Bergdoll M. Crowcroft N. 1990. Microbiological Specifications of Food Pathogens. 2005.P. Volume 5. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report).O. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures.S. Dairy. 414-25. Academic Press.L. Microbiological Specifications of Food Pathogens. 4 (13). Marcel Dekker. Woodhead Publishing Ltd. Bhunia A.A. in Foodborne Pathogens: Microbiology and Molecular Biology. Reed G. in Foodborne Infections and Intoxications. Academic Press. Staphylococcus aureus. International Commission on Microbiological Specifications for Foods. 545-66. 31. 30. Wymondham.FISH AND SEAFOOD 27.M. Applied and Environmental Microbiology. Loessner M. 39. Ed. Doyle M. 1989. 34. Blackie.A. in Microorganisms in Foods.J. 1993. Blackie.C. 217-64. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. 2005. New York. in Foodborne Pathogens.H. Eds. Volume 5. 1996.. Lee-Wong A. Ed. London. Salmonella. Vibrio cholerae.O. 45 (3). Cleland D. Fratamico P.. International Commission on Microbiological Specifications for Foods.. 40.A. Wilkinson B. Doyle M. Eds. 426-35. Staphylococcal gastroenteritis. Ed. Eds. Ed. Warner R. International Commission on Microbiological Specifications for Foods. Cliver D. Stress Response and Resistance. 13 (11). Ed.J. Ed. 29. 1989. Oliver J. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. Ed. Staphylococcal intoxications.K. Food and Environmental Sanitation. 28. 523.P.M. Riemann H.. in Microorganisms in Foods. R15764. Caister Academic Press. Twedt R.M. in Modern Food Microbiology. 1930. International Commission on Microbiological Specifications for Foods. 985-98. Blackie. 331-57. D’Aoust J-Y. 37.. London. Griffiths M..D. Golden D. 1994. in Understanding Pathogen Behaviour Virulence. 642. Golden D.P. Smith J. Jay J. London. Springer Science. London.

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

FISH AND SEAFOOD 240 .

gov.defra.holt@ifm.uk Web site: www.seafish.gsi.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.gsi.uk Institute of Fisheries Management (IFM) 22 Rushworth Avenue West Bridgford Nottingham NG2 7LF United Kingdom Tel: + 44 (0)115 9822317 Fax: + 44 (0)115 9826150 Email: info@ifm.scotland.gov.uk Web site: www.gov.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.ifm.uk/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: helpline@defra.org.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.co.uk or v.org.uk Web site: www.org.

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

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.seafood.fdf.dh.org.csiro.uk Web site: www.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.ac.uk/en/index.bsi-global.au Web site: www.co.hpa.ifr.uk Web site: www.com/ 243 .uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.co.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.org.org.cmar.gov.com Web site: www.gov.ifst.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: reception-cmar-hobart@csiro.uk Institute of Food Science and Technology (IFST) 5 Cambridge Court 210 Shepherds Bush Road London W6 7NL United Kingdom TeI: + 44 (0) 207 6036316 Fax: + 44 (0) 207 6029936 Email: info@ifst.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.org Other Sources of Information British Standards Institution 389 Chiswick High Road London W4 4AL United Kingdom Tel: + 44 (0) 208 9969001 Fax: + 44 (0) 208 9967001 Email: cservices@bsigroup.org Web site: www.gsi.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.nz Web site: www.

hps.se Web site: www.int/ SIK .codexalimentarius.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.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: hpsenquiries@hps.who.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.euro.nhs.uk/index.org.sfam.nhs.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.uk Web site: www.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: foodsafety@euro.de 244 .who.uk Web site: www.co.int Web site: http://www.int/foodsafety World Health Organization (WHO) Headquarters CH .sik.who.euro.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. jsp Food Safety 10 . Web site: www.who.int Web site: www.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.who.co.int.net/web/indexen.scot.uk/scieh.scot.asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.org Web site: www.sofht.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.

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

FISH AND SEAFOOD 246 .

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 . as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. planktonic source of spirolides in shellfish 163 Allergens. growth and survival in crustacean shellfish 86 Aeromonas sobria.INDEX Achromobacter spp. preserved by dry/wet salting 99 Anisakis spp. 102 Bacillus spp. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. to test microbiological criteria 19 Aeromonadaceae. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. source of PSP toxins 142 Alexandrium ostenfeldii. as autochthonous bacterial flora of fish 3 as spoilage organisms in oysters 65 as spoilage organisms in temperate crustacean shellfish 80 as spoilage organisms in tropical crustacean shellfish 81 in molluscan shellfish 60 Aerobic plate counts. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. as autochthonous bacterial flora of fish 3 Aeromonas spp. 111 Astroviruses. in crustacean shellfish 86-7 ATP. 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. 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. causing spoilage of dried fish 110. in crustacean shellfish 87 Alteromonas spp. 69-70 Aeromonas hydrophila.

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

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

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

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

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

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

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

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

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

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