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

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6 Pathogens: Growth and Survival 1. 4.3 Initial Microflora 2.2 Initial Microflora 1.6 Pathogens: Growth and Survival 2.2 Definitions 2.6 Published Microbiological Criteria 4.7 References 3.5 Pathogens: Growth and Survival 3.7 References 4.5 Spoilage 2.3 Processing and its Effects on the Microflora 1. vii .1 Definitions 4.3 Processing and its Effects on the Microflora 4.1 Definitions 1.1 Introduction 2.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.8 References MOLLUSCAN SHELLFISH 3.4 Spoilage 4.7 Published Microbiological Criteria 1. CHILLED AND FROZEN RAW FISH 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 2.4 Spoilage 1.1 Definitions 3.5 Pathogens: Growth and Survival 4.2 Initial Microflora 3.8 References 1.5 Factors Affecting Fresh Fish Spoilage 1.3 Processing and its Effects on the Microflora 3.7 Published Microbiological Criteria 2.6 Published Microbiological Criteria 3.4 Spoilage 3.4 Processing and its Effects on the Microflora 2.2 Initial Microflora 4. 3.8 Further Reading CRUSTACEAN SHELLFISH 4.

8 Ciguatoxins 7.2 Legislative Structure 190 9.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.5. EU FOOD HYGIENE LEGISLATION 189 9. CURED. 8.5 Pathogens: Growth and Survival 6.3 Processing and its Effects on the Microflora 6.3 Stages of a HACCP Study 8.6 Published Microbiological Criteria 5.1 Definitions 5.9 Azaspiracids 7.7 Food Hygiene (England) Regulations 2006.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.I.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.11 Conclusion 7. SMOKED AND DRIED FISH 5.3 Tetrodotoxin 7.6 Lipophilic Shellfish Toxins (LST) 7.5 Pathogens: Growth and Survival 5. 9.7 Further Reading FISH AND SHELLFISH TOXINS 7.8 Further Reading FERMENTED FISH 6.4 Implementation and Review of the HACCP Plan 8.4 Regulation (EC) No. 14 (Hygiene requirements specific to the UK) 221 9.1 Introduction 189 9.7 References 5.3 Regulation (EC) No.2 Initial Microflora 5.8 Guidance 222 9.2 Definitions 8.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. S.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 6.4 Spoilage 5.1 Definitions 6.5 Regulation (EC) No. 7.4 Spoilage 6.6 References 6.2 Initial Microflora 6.6 Regulation (EC) No.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.3 Processing and its Effects on the Microflora 5. 852/2004 on the General Hygiene of Foodstuffs 191 9.1 Introduction 7. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9. 2006 No. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption 212 9.10 Further Reading 223 viii .9 Other Relevant Legislation 223 9.10 Cyclic Imines 7. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.1 Introduction 8.

3 Clostridium perfringens 10.1 Aeromonas spp.5 Plesiomonas 10.7 Staphylococcus aureus 10. 10.8 Vibrio cholerae 10. 10.10 Vibrio vulnificus 10. 10.4 Listeria spp.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .10 PATHOGEN PROFILES 10.6 Salmonella spp.2 Clostridium botulinum 10.9 Vibrio parahaemolyticus 10.

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

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

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

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

The lipid content. The connective tissue content of fish is also lower than that found in meat. However. Fish can be divided into fatty and lean fish. lean fish are those fish that store most of their fat in the liver. it shows a typical season pattern especially in migratory species such as herring or mackerel.21%. Protein content in fish fillet varies typically from 16 .20%. most fish contain omega 3-fatty acids and other essential nutrients. “fish” will be used for fresh and seawater finfish. and spawning season. fins and a streamline body. Fish are an important part of a healthy diet since they contain high quality protein. migratory patterns. which can be up to 67%.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. CHILLED AND FROZEN RAW FISH Associate Prof. typically fluctuates between 0. and is mostly interspersed between the muscle fibres. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. Fish fillets are a poor source of carbohydrates. crustaceans. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. This super class of vertebrates includes all the bony and cartilaginous finfish. In this chapter. and other forms of aquatic animal as part of their ‘fish’ definition. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. In addition. Fish flesh is dominated by the abundance of white muscle in relatively short segments. 1 .2 . In addition. offering less than 0.1. Fish fillet composition can vary significantly within the same species due to feed intake. while fatty fish have fat cells distributed along their bodies. respectively (1). The lipid fraction is the component showing the greatest variation. giving it its characteristically flaky structure. and excludes molluscs and Crustacea. Muscle composition and structure of fish also differ from those found in other meat. ‘fish’ also refers to the flesh of such animals used as food.5% (1). but typically present a low fat percent when compared to other meats. typically 3 and 15% of total weight.

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

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

Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.35 °C 8 .103 High (107 .14 . and risk associated with the consumption of seafood is low (2.II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp.106 cells/g.102 101 . and mud Ubiquitous. Plesiomonas shigelloides 3 .0.37 °C High for most species. estuarine environments. In addition. Toxin levels 0. 4 .FISH AND SEAFOOD TABLE 1. birds. vulnificus) 5 .1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.109 cells/g) * Some sources considered L. Escherichia coli Yersinia enterocolitica 101 .106 cells/g (exception: V. However. 105 . sewage.1 .26 °C 10 .I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. monocytogenes as part of the natural aquatic flora It is apparent from the above that there is potentially a very diverse range of organisms present on fish.37 °C Unknown Unknown TABLE 1. 4).19 µg/kg bodyweight from < 102 . stream water. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . numbers of pathogenic bacteria in raw fish tend to be low.>106 Salmonella spp.

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

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

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

It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). The problem of water loss in MAP fish is common. Although all fish species need to be treated separately. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. adequate product labelling in respect of storage temperature. TMAO) that can be reduced to produce 8 . Only the highest-quality fish. owing to the dissolution of cellular structures. should be used. 5. 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). The replacement gas should contain at least 40% carbon dioxide. Storage studies have shown that the extension in shelf life of MAP cod fillets was proportional to carbon dioxide concentration up to around 50% (V/V). may result in unacceptable colour and textural change and hence little shelf life extension (11). use of high-quality raw fish. Subsequent storage must be at or below 3 °C. In the United Kingdom. It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. 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. For fatty fish. at all times. grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. For other less fatty marine fish. but is recognised as a drawback of carbon dioxide application. however. and the use of a HACCP plan. however. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). The addition of high levels of carbon dioxide to these fish. botulinum. from both a microbiological and a chemical standpoint. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. trimethylamine and sulfides). adequate shelf life and cooking requirements.g. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. 3.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). this problem occurs to a lesser extent with meat. greater advantages are seen. These conditions include storage of the product at ≤ 3. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1.3 °C at all points from packaging onwards. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. as already stated. 4. The growth of these bacteria is reduced by high carbon dioxide concentrations but. 2.g.

loss of the fresh characteristics may be simply measured by comparative visual and smell analysis.4 1. reactions caused by activities of the fish’s own enzymes. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids). or for that matter evaluation of the taste of the fish. and this pattern is the basis of freshness grading schemes. as traditional spoilage patterns of the external organs are typically very distinct. A. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. the skin turns from iridescent to dull and bleached with bacterial slime. with no smell.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). 1. C (C=reject) Significantly. Degradation of whole fresh fish stored in ice generally follows a set pattern. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. B.III Fish-quality scales Name of scale Torry Scale Quality Index Method European Grading Scheme Scoring system 10 to 0 (≤ 4 reject) 0 to 24 (arbitrary reject) Extra. Taste panel evaluation of cooked fish may be used to score or grade colour. and metabolic activities of microorganisms present in the fish. so lower carbon dioxide concentrations may have better preservative effects. to brown and slimy with an intense off-odour. texture.4. Over time. When working with fillets. these are summarised in Table 1. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). The eyes turn from convex and clear to concave and opaque. 18) and are probably the most widely used. the gills from pink and shiny. smell and 9 . 17. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . TABLE 1.III. Loss of freshness and spoilage cannot be separated as processes. and the flesh turns from bright and elastic to dull and soft (1).CHILLED AND FROZEN RAW FISH off-odours. Various methods for whole fish freshness evaluation by a trained panel have been proposed.

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

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

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 example. by altering the gas atmosphere (extrinsic). This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. Fish spoil as a result of the chemical. Also. biochemical and microbiological reactions taking place within and on the fish. a minority are often associated with the majority of the spoilage. the slower the spoilage processes proceed (within limits).5.5. but also crucial for the assurance of consumer safety. the lower the temperature of storage. 34). The reasons for the differences between tropical. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks.5.and temperate-water fish spoilage rates may be many. The sum of these changes may result in a different spoilage profile. the results of altering one parameter may have far-reaching consequences on others. 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.3 Recognised specific spoilage organisms (SSOs) The degree of spoilage leading to sensory rejection of fish is partly dependent on the perception of the consumer. 5). Not all the bacteria growing on a food will lead to the production of objectionable characteristics. The factors are interactive and cannot be completely isolated. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent.5 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 . For fresh fish. 1.1 Factors Affecting Fresh Fish Spoilage Temperature By far the most effective method of reducing the rate of whole fresh fish spoilage is temperature control (1. The concept of specific spoilage organisms (SSOs) is not new. 1. careful temperature control during storage is not only important in terms of quality loss. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH.FISH AND SEAFOOD 1. For fresh fish iced immediately after capture and continually stored in ice.

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

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

where a particular 15 . they do not produce significant product spoilage).3 days after having reached 108 cfu/g (42). a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. botulinum recorded up to 1963. 1. these risks are substantially reduced (1. D. Huss (1. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). C1. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. thermosphacta did not produce off-flavours until 2 . 1. C2. 5).3.CHILLED AND FROZEN RAW FISH present in fish into histamine. algal toxins. E. types B. Acetate production has been reported as a good indicator of spoilage by this organism. However. but with proper cooking. botulinum. age. 1.5. bacteria and higher parasites all present some risk associated with fish consumption (1. Of these. B.6 Pathogens: Growth and Survival Endogenous chemicals.3 °C (Group II) and these are non-proteolytic (i.e.6. E.4 Brochothrix thermosphacta and lactic acid bacteria B. B. B. previous exposure and access to supportive treatment. human viruses. Of 404 intoxication outbreaks of type E C. presents a potential risk. a convenient but diverse species of bacterium with a number of different types. The heterogeneity of the types (A. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). Recent studies have investigated the dominance of B. including antisera. 75% (with a 35% mortality rate) occurred in Japan. it is bacterial risks that increase after capture of the fish. (and very rarely F and G) have been reported to cause human disease. lightly preserved fish products are associated with botulism. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. Some Ph. When inoculated into VP corned beef and sliced ham. especially type E botulinum intoxication. 6). Adult disease is caused by the production of the botulinum toxin. Review of outbreak data suggests fresh fish to be safe. this is probably due to spoilage occurring before toxin elaboration. Fewer still are capable of growing on fresh fish.1 Clostridium botulinum C. 5) reports that fresh fish consumption has never been shown to cause human botulism. E and F include bacteria with minimum growth temperatures of approximately 3. 5. F and G) is well documented (44). thermosphacta is a well-characterised psychrophilic spoilage organism of meat. Growing evidence suggests a role for B. Types A. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. thermosphacta in the spoilage of some MAP fish.

I-sushi. Other lightly or semi-preserved (e. parahaemolyticus outbreaks. which is believed to be required for virulence. is small. Despite the ubiquitous nature of type E C. storage and preparation (11). or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk.2005). V. France. In Japan. This strain has been responsible for 50 to 80% of all V. were associated with finfish between 1978 and 1998 (46). The first report of the organism being a foodborne agent was with shirasu. a Japanese boiled and semi-dried sardine dish. V. The bacterium causes primary septicaemia. 1. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). A unique clone of V. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. 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%) (2.2 Vibrio parahaemolyticus and other vibrios V. which was probably contaminated from an uncooked food source or an excreting foodhandler. USA. 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. the risk posed to the consumer of fresh fish. and North America. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . Russia. parahaemolyticus cases have increased across the world. smoked. due to the cultural preference for raw fish dishes. and proper cooking (boiling for 1 minute. 1978 . the majority of outbreaks are caused by consumption of raw fish products (e. parahaemolyticus O3:K6 is responsible for many of the recent V.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. sushi) (5). there is no specific guideline that describes a minimum level of V. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. parahaemolyticus is a marine organism.6. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. 11).3 °C) will eliminate all risk. Proper chilling.g. 5). and when in contact with contaminated water (no CDC reported cases from fish.g. including epidemics in India. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. whether stored aerobically or in modified-atmospheres. Japan. which can cause human gastroenteritis. accounted for the majority of cases (1. Since 1996. Proper temperature control (<3. salted or pickled) fish have also been associated with botulism. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. Generally. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). use of post harvest treatments and/or cooking of fresh seafood will reduce risks. Southeast Asia. Currently. vulnificus is associated with warm marine and estuarine waters. the majority are associated with products not cooked immediately before consumption. Only about 1% of marine isolates produce a thermostable haemolysin. again.

L.CHILLED AND FROZEN RAW FISH disorders). others are psychrotrophic mesophiles (49). which is often fatal (>50%). in assessing the risk of fishborne infection by P. are associated with human diarrhoeal illnesses. Again.6.4 Listeria monocytogenes L.20 °C.3 Aeromonas Aeromonas spp. sodium channel blocking and haemolytic activity (48). are present on fish and can grow to significant numbers during storage (1. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. but proper cooking should substantially reduce risk of infection. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. is historically associated with faecally contaminated water. Chill storage may not eliminate growth. in its habitat. with minimum growth temperatures around 0 °C and optima of 15 . Despite this. however. 17). It is known that Aeromonas spp. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. P. Although the psychrophiles are unable to grow at body temperature. V. the causative agent of cholera. 1. There is no risk associated with the consumption of properly chilled and cooked fresh fish. and especially Aeromonas hydrophila. and proper cooking will substantially reduce risk. Most cases have been sporadic rather than associated to large outbreaks. Since 2000. 5. coli on isolation media (50).6. 1. The major virulence factor appears to be the production of toxins with enterotoxic.. their potential to produce disease-causing exotoxin in food has not been fully examined (48).000 17 . They are aquatic organisms with an epidemiology that is yet to be fully understood. Some isolates are true psychrophiles. possibly due to under-reporting because of their likeness to E.26 to 0. and fish and shellfish are probably the major reservoirs for the organism (50). cytotoxic. shigelloides is similar to Aeromonas spp. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. proper chill storage will eliminate any risk associated with this organism. infection.55 cases per 100. shigelloides are encountered. Similar problems as those encountered for Aeromonas spp. cholerae O1. monocytogenes incidence has been between 0. monocytogenes has been well documented as a foodborne human pathogen. there is no risk from the consumption of properly handled and cooked fresh fish. In the US. and especially estuarine environment. but the bacterium is known to survive and grow in the shallow marine. although lack of data concerning infectious doses and the possibility that foodborne virulence is linked to the immune state of the host mean that full risk assessment is impossible. listeriosis cases have been reportable to the CDC.

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

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

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

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

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

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

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

2197-203.H. Trolle G. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. in Fish processing technology. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. 25 . Gram L.J. Hall G. London.9. Sala F. Journal of Applied Bacteriology.H.L. International Journal of Food Microbiology... Blackie Academic & Professional. 59. 1. Garcia M. 322-6. Eds. 20023.3 Pathogens Condon S. Modified atmosphere packaging of fish and fish products.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. 1997. 479-88. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila..CHILLED AND FROZEN RAW FISH Einarsson H...M. Jacobsen M.R. 1992. 5. Huss H. 1987. 1. Huss H. Liston J.. 1992. A rapid method for detection of histamine producing bacteria.9. Applied and Environmental Microbiology.. Huss H. Klausen N. Effect of culture age. 1987. Elsevier Science.europa. International Journal of Food Microbiology. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. in Quality Assurance in the Fish Industry. Amsterdam. 137-46.. Gram L.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. 65-72. http://eur-lex.H. Ed.eu/LexUriServ/LexUriServ. Otero A. 1993. 4.K. 72.

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

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

Again. in addition. tuna steaks) or band-sawn from a frozen block (e. Added colours can be included with the brine solution to meet specific requirements for products e. Fish steaks can be cut from a natural fillet or loin (e. haddock. and salmon. with a minimum residual salt content of 3.2. a protective glaze may be added to the frozen steak. Hot-smoked fish are prepared from brined.5 .g. sawdust with other woods adjusted for flavour profile and burn duration). swordfish steaks). mackerel. mackerel.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. This process in effect cooks the product and reduces or eliminates microbial contaminants. salmon.g. VP products) because of the high salt content acting as a barrier to spoilage.g. but may be increased (up to 1 month) by packaging methods like MAP or VP. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. Hot-smoked fillets are prepared by brining fish fillets in a 26% salt solution with added natural colours to provide a range of product types.5% in the aqueous phase. Pâtés are prepared with raw fish. etc. 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. natural fillets of fish such as herring. The cold smoking process does not cook the fish or destroy microorganisms so the expected shelf life is not significantly increased from that expected for wet fish fillets. and the product then shaped and heat-processed to cook the product before being chilled. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. be flavoured with herb dips or peppercorns.g. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. cod). the ingredients premixed. The shelf life for MAP fillets is typically up to 5 days. in VP or through mail order. The expected shelf life of these products is up to 8 days (or longer for some chilled. 29 . They are sold from wet fish counters. The result is fish with a typical residual salt content of 1. Fresh fillets are sold from wet fish counters. resulting in a product that is ready-to-eat without any further cooking.2 days) longer than that of non-prepacked fish.5%. or pre-packed frozen with added glaze. Cold smoking is essentially a flavouring process. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. which is approximately 30% (or 1 .. Some products may be salted for flavour only (e. annatto with smoked haddock. which may. or pre-packed in MAP or vacuum packaging (VP). They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes.

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

which may become a focus of contamination deep within the muscle. fat content.e. it may be used for other products or markets depending on the product specification.3. Dalgaard et al.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. and ingredients added to the final product. is not used for sushi or other high risk (and high value) products. whilst gaping is also dependent on a range of other factors that effect the muscle composition. including seasonal variation. or visible damage from gaff marks (4). species (Bluefin>Bigeye>Yellowfin>Albacore). After capture and during any subsequent processing. 31 . Contamination will. on the whole. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. and pathogenic flora associated with seafood. The edible tissue of some species of fish are more susceptible to gaping than others. The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. i. and the potentially toxic by-products of histamine-producing bacteria (5. For a summary of background information on the hazards associated with fisheries products and ingredients. as a result there is an increase in spoilage rate thereby reducing the product shelf life. searing or bar marking). specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). In summary. 6). Only after death. product handlers. handling or preparation stage. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). A primary role in fish spoilage has been assigned to this organism.I. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna.and turtlefriendly long lining to the less discriminatory netting. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. and have emphasised the origin and sustainability of the fish as key marketing points. skin. Tuna that does not meet the highest of grades based on colour. remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. see Table 2. and processing conditions such as dehydration or poor filleting practices. Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. and intestines of the fish. Grading is based on size of fish (>25 kg). Recent public opinion has favoured the use of dolphin. 2).3 2. will the saprophytic flora start to invade the surface flesh (1.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. sensory assessment of freshness etc. the microbial flora of seafood is associated with the gills. storage.

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

CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. Effects on the microflora are minimal during tempering (7). but will also need to ensure that the optimum shelf life can be safely obtained. It is critical that the time and product temperature during tempering be controlled to avoid elevated temperatures (>5 °C) on the product surface allowing microbial growth or the onset of spoilage in raw material.2. For some components. up to -5 °C so as to improve the cutting and handling properties during processing. Generic examples have been used to illustrate only the major points. Rapid controlled cooling of heat-processed components will help to minimise spoilage. Where the raw materials are frozen. Examples of tempering techniques include the use of heated air. but it is assumed that any band sawing or dice process will introduce heat from blade friction. and contamination from equipment to clean fish muscle.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. to evenly raise the temperature of the entire frozen product from -20 °C or lower.4. the core temperature.1 Coated products Seafood products are coated with a cereal-based batter or crumb with up to 30 50 % of the final product weight accounted for by the batter or crumb. line extensions and process reviews.1 . but it also poses a risk of cross-contaminating other products undergoing tempering. defined holding time at a chill temperature (0 – 3 °C). 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. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. there are a number of distinct methods used. Band-sawn materials or fish dice are normally promptly further processed. as a heat process is often required for functional or organoleptic reasons. It is recommended that the principles of HACCP are applied to novel processes.3.4.3 Preparation of fish component of chilled and frozen recipe dishes 2. 2. 2. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. depending on the size of the fillet 33 . as the full complexity of the processes used for specific products cannot be covered in a general text. and potential opportunities for growth of foodpoisoning bacteria. this may be an incidental effect. or the use of microwave technology. Figures 2.4. All have the same intention.

4. It is then size-graded. 2.1.1C . Raw material preparation stages for chilled or frozen coated fish products 34 .Shaped Fillets. frozen in a nitrogen blast freezer and weighed into sacks for later processing.Fish Cakes).3. the scampi is washed in potable water to remove foreign bodies.2. The processing of three examples types are described in Figure 2. Frozen cores are usually batter-enrobed.Scampi.1.7 days. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. with the coating heat-fixed. The “blown” meat is graded. after landing. may not rise above 0 ºC during processing.1 (2.3. 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. Fig. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.1A .1. 2.1B .4.2). by mincing and mixing with a binding agent (as described in section 2. quickfrozen. drained and then refrozen for storage until required for further processing. are placed into plastic boxes and covered in ice to preserve their condition. On arrival at the processing plant.FISH AND SEAFOOD processed. 2. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. chilled or sold chilled on defrost with an expected shelf life of 6 .1 Whole-fillet / scampi Scampi (Nephrops norvegicus).

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

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

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

1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes. 38 .4. Figure 2. The protein source can be fish fillet or block. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. and must not show evidence of disease.3. rice. The vegetables must withstand processing. A temperature no greater than 10 ºC for 12 hours cumulatively or 21. 2. to produce a specific product. but they are supplied without a source of carbohydrate e. For example.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe. Fig. canned fish may be used in their preparation owing to it having wider availability and low wastage. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7).2. as this could introduce off-flavours and reduce shelf life.5. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products.g. pasta or potato. There are two main categories.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. Flow diagram outline for recipe dish ingredients preparation 2.4.1 ºC for no more than 3 hours is recommended (1).

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

where composite meals are heat-treated. Placing such products in a standard chill store to cool down could result in condensation. If the sauce contains modified starches for freeze/chill stability.4. cereus are well known and are still a common issue in the UK (9). a process equivalent to at least a 5D Listeria kill will be necessary. 11). the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. B. 13). if subjected to temperatures between 20 and 40°C. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. Alternatively. and an increased risk of contamination from L. Typically. An incident involving Staph. On hydration. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. but temperature-abused pasta can also spoil rapidly by mould growth. rely on the use of microbiologically sound ingredients. For this reason all products. Sauces can be multi-component preparations or more simple preparations. Sauces or marinades are added to fish to broaden appeal or to accompany components.) to the product (12. such as butter garnish with black pepper. can germinate and rapidly multiply.g. a heat process in excess of 73 ºC will be required to rehydrate this component fully.4. 2. 40 . aureus toxin is well documented (10.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. Products or dried ingredients contaminated with either spores of aerobic bacteria (e. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. Potato toppings made from flake or powder will normally have little associated spoilage flora. cereus). Appropriate handling facilities or equipment will be required to protect the product. spore-formers can proliferate if adequate control of temperature is not maintained. cereus. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings.6. This is a particular issue especially where products are heat treated to eliminate vegetative cells. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. especially in those products that have a vegetable component or rice accompaniments. and then cooled slowly allowing germination and re-growth.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. with the range always expanding as new tastes and cuisines are developed.

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

Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. 42 . Flow diagram outline assembly and packaging of recipe dish 2. as it dissolves in the surface moisture to form a weak acid.4.7. generally carbon dioxide and nitrogen. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms.FISH AND SEAFOOD Fig. the method replaces air in the sealed packaging with a defined mixture of gases.2.7.4. which may restrict either microbial growth or enzyme activity.2 Modified-atmosphere packaging Originally developed for the red meat industry. and tuna loins are packed by placing the product in a plastic pouch and removing all the air. botulinum to grow. The oxygen-free conditions will however allow other anaerobic bacteria such as C.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. unless the temperature is strictly controlled to below 5 °C throughout the supply chain.5. 2.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions). The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow.4. These include: 2.

but they will also be readily adapted to the nutrient source and conditions. Beyond this. in numerical terms. It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage..5 Spoilage During processing. except for some crustacea. The literature has good information on spoilage of individual components of recipe meals. initially. as this organism is a common contaminant of freshwater fish species (19). the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). and finally flavour loss.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. Vibrionaceae will succeed (17 ). At refrigeration temperatures (5 . This is also true of chill storage and the distribution chain. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. but is not limited to. but little on in-pack spoilage. followed by moisture loss. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15). both organisms are capable of growth at chill temperatures. by a loss of texture. and Yersinia spp. phosphoreum. (20). is typically uncooked or at best lightly processed to retain eating characteristics. Not only will these organisms have a numerical advantage over other proposed spoilage organisms. Shewanella putrefaciens and Ph. However.1 Chilled storage For chilled products or chilled components of frozen products. 43 . a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood.10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. at slightly higher temperatures (10 ºC). Aeromonas spp. where the protein component is nearly always heat-processed. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18). the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. and in influencing the final eating quality of the product. The fish component of recipe dishes. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’. 2.5. Spoilage of this material is not expected to differ greatly from the processes described elsewhere. Spoilage will be most readily detected in proteinaceous components of ready meals. Nonetheless. temperature control is important in preserving chilled integrity of ingredients. This is considered to be the one major difference between fish and meat products. The case for growth and contamination of products with Aeromonas is stronger.

and Aerobacter spp.FISH AND SEAFOOD 2. 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. By retaining the intrinsic spoilage flora of seafood. within dairy products are well known. However. Bacillus spp. eliminating nematodes in products that do not have a heat treatment step.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. Pseudomonas spp. Still.g. where the variety of components and ingredients used in manufacture is varied and complex. however. These have been proved to impart off-flavours.. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. The ACMSF (22) issued clear advice on this topic and has recommended that the shelf life of products without barriers (23) is restricted to less than 10 days. auto-oxidation of fats/oils). spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e. The conditions and freezing process most favourable for good-quality products will. which would warn the consumer of the potentially unsafe nature of the product. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. producing characteristic off-flavours. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. 44 .. Products that are developed with an extended shelf life beyond the 6 . The type and nature of the enzymes and their activity will depend on the bacterial species.5.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). this should also take into account shelf life of product components used in the assembly of the final product. also be those that have the least effect on the viability of microorganisms (21). This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. Serratia spp. As a note of interest. botulinum. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. for example. For example. 2.5. This reinforces the need for prompt processing and temperature control of ingredients and finished products. freezing also has a protective role in improving the safety of raw materials by.. the shelf life of products is limited by the natural spoilage process described earlier.

Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs.3 Clostridium perfringens A study examining C. may be of benefit within any risk assessment for chilled and frozen prepared fish products. Incidents in such diverse materials as cream. low oxygen or anaerobic conditions. or a heat shock followed by holding at a temperature suitable for growth. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). and this issue may be of greater consequence for warmwater.104/ g should be treated with some concern (26). especially molluscan filter feeders.6.6. In addition. are dealt with elsewhere in this book. air. botulinum type E and Vibrio parahaemolyticus (25). This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. perfringens contaminating seafood will depend on the source of raw material.. etc. have been linked with food poisoning and should also be considered. cereus.1 Bacillus cereus This is a common contaminant of dust. spices and herbs and vegetable shoots have been reported (12. agricultural run-off or sewage can contaminate in-shore species. farmed or free-living crustacea (30). soil. Additional hazards from marine bio-toxins. Cases quoting contamination of seafood with organisms such as S. typhi. 2. 2. any risk of C. and Campylobacter are recorded in filter feeding molluscs such as mussels. Counts of between 103 . Inappropriate holding times and temperatures will. botulinum when held in favourable conditions for growth (28) i. Germination of spores requires favourable conditions. permit rapid growth of B.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. and many raw or processed foods (26). potato. Vibrio spp. A number of other Bacillus spp. 45 .6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. 27).CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. with pathogenic microorganisms or viruses. 2. and some effects related to product type and time to toxin production have been noted. along with previous discussion. Hazards attributable to the non-seafood components are listed below and this information.e. histamine intoxication.6. These are C. 13. Therefore. water. in some cases.

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

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

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

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

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

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

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Pacific Regional Laboratory Northwest U. Scaphopoda. oysters and clams can process about 3. clams. viruses. The class Pelecypoda includes all of the bivalves such as clams.E. mussels. where it is eventually swallowed and digested. cockles and scallops.1 Definitions General features of the mollusc The phylum Mollusca. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food.3. Food and Drug Administration 22201 23rd Dr. Pelecypods have little need for locomotion. For example. Gastropoda. chemical contaminants and other impurities on the mucus of the gills. respectively. cockles. P. Other species of shellfish such as crabs. can hold the shells tightly shut against enemies (1). The digestion process requires less than 2 hours in actively feeding adult bivalves (2). whelks. and Cephalopoda. Jr. scallops. Special tracts of cilia move this mucus toward the mouth. octopi. and shrimp. 4). lobster. the adductors. abalones. chitons. In these forms. the shell.7 litres of water per hour. The class having species of economic and public health importance is the Pelecypoda. In most species. while 53 . squid. Plankton and algae form the bulk of their diet from seawater. completely encloses the body. lying well within the mantle cavity. mussels. O. Box 3012 Bothell WA 98041-4421 United States of America 3.S.1. Strong muscles. MOLLUSCAN SHELLFISH Carlos Abeyta. cockles.1 3. Pelecypoda. The term shellfish in this context is limited to oysters. and scallops. which includes oysters. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. In the process of filtering water. composed of two valves hinged together dorsally. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. snails. mussels. and 2. clams. S. Many microorganisms ingested by shellfish survive the digestive process (3.6 litres. The phylum consists of five classes: Amphineura. they also trap bacteria. slugs. oysters.

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

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.

or siphons.1. extended above the surface to maintain water currents needed for respiration and feeding. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint. but 56 . A mantle tissue encloses the body of the clam. littleneck. mostly marine bivalve molluscs. Most clams are only a few centimetres in their maximum dimensions. hard-shell or soft-shell clam.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. cherrystone. geoduck.2.1 Clam Clam is the common name of a large group of often edible. The muscular foot is used to burrow in mud or sand. 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. Different species are variously called quahog. Buried clams leave tubes. laterally compressed.

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

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

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

Bacillus spp. Pseudomonas spp. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. Other microorganisms found in lesser numbers are the Achromobacter spp. dredges. Vibrios. In summary..3 3. shellstock (live) which is shellfish in the shell. Campylobacter coli Bacillus cereus Hepatitis non-A..1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. Flavobacterium spp.. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. 15). Micrococcus spp. responding to the environment in which they are harvested..FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. mussels. and shucked shellfish that is shellfish. and yeast cells of Rhodotorula rubra and Trichosporon spp. that have never been cultured on laboratory media. from which one or both shells have been removed. Alcaligenes spp. rakes. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp.1 Processing and its Effects on the Microflora Oysters. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100.. The largest groups found are the Pseudomonads. mechanical harvesters. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. 13). but have never been cultured (14. 11. At harvest. 12.1 and 3.1 Shellstock 3.1.. Temperature. observed microscopically.1. the initial flora composition and concentration of bivalves are highly variable.. They are termed viable non-culturable microorganisms. or by 60 . 3. Moraxella spp. and cockles The processing of bivalves takes two forms.2. Enterococci spp.3. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. and Aeromonads. salinity.000 /g. 3.3.. whole or in part. 10.1. Acinetobacter spp. clams. For example.3. spirochetes of the genus Cristispira and Saprospira. (9.. Water temperatures affect the concentrations of microflora in the water. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. nutrients. There are microorganisms.

Generic flow diagram of live shellfish from harvester to point of sale 61 . or in bay-approved shellfish-growing waters prior to sale or processing. for transportation to the point of sale. If by boat. 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. Secondly. after harvesting. or immediately placed in refrigerated trucks. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock. Fig.1. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. in containers from which they can be retrieved easily. Thirdly. There are three primary reasons for wet storage: first. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters.1). by truck or boat. The shellstock is transported dry.MOLLUSCAN SHELLFISH hand (Figure 3.3. or from other sources approved by the State Shellfish Control Agency (SSCA). Shellstock are placed in drycooler storage containers in the facility. Shellstock may be wet-stored at the processing facility in tanks of clean seawater. usually in open-mesh containers and/or burlap sacks. Water used for shellstock washing is from approved growing areas. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale.

handling. Water used for shellstock washing should be of good sanitary quality. 16. 21. and temperature storage of shellstock to point of sale. Klebsiella. 25). Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). 17). Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. there is evidence that in the absence of adequate refrigeration. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. shellstock are placed in refrigerated trucks for transport or placed in wet storage.1. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. An outbreak of infectious hepatitis in Sweden in 1956. when it reaches the processing facility. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. coli. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. Over a period.2 ºC (45 ºF) or below (19). Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. 23. Harvest conditions are another factor that influences the growth of bacteria in shellstock. The most important factor in controlling bacterial growth in shellstock is temperature. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). and thus contaminating the shucked meat at the point of sale. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. Furthermore. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. At harvest.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. vulnificus. the bacterial counts in the meats will increase. 22). Enterobacter. synthetic seawater for the purpose of de-sanding and salt uptake. some pathogenic organisms will survive and multiply in shellfish (20.3. including V. for a considerable length of time. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. 26). If long delays occur before reaching the processing facility and/or refrigeration. are prevented. without adequate refrigeration. and the sanitary practices applied during harvesting and shipping. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. At this temperature the multiplication of faecal coliforms (E. hydrophila (18). to avoid possible contamination of the shellstock. with prolonged storage the microbiological quality of the bivalve decreases. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . and Citrobacter species) and Vibrios. however. shellstock are not refrigerated on the harvest boat. bivalves tightly close their shells thus trapping water and microflora. Generally. with the exception of A. 24.1.FISH AND SEAFOOD 3. The washing of sediment and detritus soon after harvesting will prevent further contamination. Generally.

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

3.2. Generic flow diagram of shucked shellfish from harvester to point of sale Shellfish are subject to contamination during the shucking and packing process when the following are not adhered to (19): 1. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). There is a tendency to store such articles on the shucking benches or in packing rooms. Separate rooms or lockers for storage of clothing. which may lead to contamination of product. within one hour. Shucking containers are rinsed clean with running water. 4.2 ºC or less. and gloves help prevent the contamination of shellfish. 2. Fig 3. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. 64 . and sanitised before each filling. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. Shucked meats are thoroughly drained. aprons.

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

diarrhoea. ranging from epidemic cholera to sporadic cases of diarrhoea. parahaemolyticus. parahaemolyticus possibly producing illnesses (40). coastal areas). from harvesting to finished product (i. lot and retail outlet.48 hours with abdominal pain. and to the point of sale. It is a common marine bacterium. 3.5. V. They can be introduced to the shellfish from the environment or added during processing. Fortunately. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). shellstock and packing of shellfish in jars) (29). Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. where outbreaks occur regularly. headache and fever. however. primarily in brackish or saltwater. 44).e. approximately 23 cfu/g. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. during processing. widespread in both polluted and unpolluted waters. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. 66 . Major outbreaks are associated with the warmer months.000 population/year (46).5. This bacterium is present in all the major processing steps of a shellfish plant. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. prevention of cross-contamination. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). parahaemolyticus multiplies rapidly at temperatures above 20 ºC. and harvesting of shellfish in the cooler months. although non-hemolytic strains have been associated with illness (43.FISH AND SEAFOOD 3.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. and estimates based from a hospital study show less than 0. The incidence of V. of V. Sporadic cases are frequent along the coast of the United States. there is a recent report of low levels of V. Between May and September 1997.1 Vibrio parahaemolyticus V. 3. and very common in Japan. These organisms can be present in the shellfish from the point of harvesting. V. Products involved were from the same processor. only a small percentage of strains in the marine environment are potentially pathogenic (45). Diseases caused by Vibrios are enteric in nature.1. nausea. parahaemolyticus causes gastroenteritis lasting 24 .5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. 41). parahaemolyticus from shellfish is low.5 cases/100. One of 250 cases reported involved shucked products with low levels.

cirrhosis. cholerae. fever.8 case/100. Oysters are the most hazardous food associated with V. Control of oyster-associated 67 . 48).1. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. which have been implicated in shellfish-related disease outbreaks (31). nausea. 57. chills.48 hours of onset. Hazards from this bacterium can be controlled by thorough cooking of shellfish. gastroenteritis.7 days. It is one of the most severe foodborne infectious diseases. vulnificus.2 Vibrio vulnificus The symptoms of V. Healthy individuals are susceptible to gastroenteritis. vulnificus are fever. like V. and nausea within 24 . Onset of symptoms occurs in 48 hours and can be severe. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. are particularly susceptible to primary septicaemia. 56). lasting 6 . and vomiting.II (8. 3. V. 46. 53. 3. The organism attaches to the intestine and release of a toxin is suspected. 23 of 24 major USA-West Coast estuaries were positive for V. 55. and preventing cross contamination once the product is cooked. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). They are sensitive to cold.1. vulnificus is by rapid refrigeration during warm weather months. The occurrence of V. these individuals are advised not to eat raw shellfish. V.5.5. abdominal cramps. Inadequate refrigeration will accelerate the growth of V. High-risk individuals. or immunosuppression. and rapidly become inactive at low storage temperatures. Again. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness.0.4 . parahaemolyticus the estimated annual incidence rate is very low.000 population/year (8. Death has been reported to occur within 36 hours from onset. cholerae non-01 is ubiquitous in the estuarine environment. It causes diarrhoea. All isolates are considered pathogenic. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. cholerae non-01 (51). however. cholerae non-01 has been associated with septicaemia. V. In one study. 54. or primary septicaemia. Primarily. the organism has been found in the Gulf of Mexico. 46. in the range of 0. parahaemolyticus growth in shellfish meats is temperature dependent. leukaemia. Control of V. 49). The bacterium is an estuarine species that causes wound infections. 58).3 Vibrio cholerae non-01 Illness caused by V. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. diabetes. with a fatality rate of 50% for individuals with septicaemia.MOLLUSCAN SHELLFISH Control of V. those who have liver disease. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. parahaemolyticus. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47).

S. cholerae non-01 illnesses is by cooking seafood thoroughly. 3. undercooked. The frequency of infection of these organisms is no different from other Vibrios.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools.2 Campylobacter jejuni C. vulnificus. nausea. (8. and the transmission of the disease by consumption of raw. or cross-contaminated shellfish (57. e. It causes diarrhoea.FISH AND SEAFOOD V.5. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. imported product from a country experiencing an epidemic. and southern Asia including India and Bangladesh. cholerae 01 is transmitted by faecal contamination of food or water. 68 . 62. cholerae 01. headache and fever. There is evidence of strains of V. fever. cholerae 01 in molluscan shellfish.5 Other Vibrios spp. V. In addition. V. widespread in both polluted and unpolluted waters. 60). associated with shellfish-borne illness outbreaks. Death may occur after severe fluid and electrolyte loss.1.1. and shock. The control of V. the pathogenic severity is lower than V. fluvialis. 3. and illness lasts 7 . and nausea. 3.5. nausea.g. processors should know the product source. is the same as previously discussed for the non-01 strains. and V. cholerae 01 establishing in the U.10 days. vomiting. acidity and heat. They are common marine bacteria. The presence of these strains appears to be in low numbers and non-virulent (59). particularly in oysters. including V. 61.5%) and 2 . these organisms cause gastroenteritis lasting 24 . respectively. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. jejuni requires reduced levels of oxygen (3 . low pH. dehydration. Onset of disease is 6 hours to 5 days. diarrhoea. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American.5. hollisae. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). 63). Illnesses are associated with the warmer months. There are several other Vibrio spp. drying. V. Gulf coast environment. In general. preventing crosscontamination of cooked seafood. Onset of illness is from 3 to 5 days after ingestion of contaminated food. however. salinity. abdominal pain. In developing countries.48 hours with abdominal pain. parahaemolyticus and V. Control of infection from these organisms is no different from the other Vibrios. mimicus. Illness can include abdominal cramps. Poor sanitation and contaminated water supplies will spread the disease.10% carbon dioxide for optimal growth conditions.

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

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

2.R. Journal of the Mississippi Academy of Sciences. The American Oyster. Journal of Milk and Food Technology. W. 544-8. Cook D. 30.R. in Multilingual Dictionary of Fish and Fish Products. 29.A. in Fishery Bulletin 1964 No. 447-52. in Microbiology of Marine Food Products.N. 13.A. Applied and Environmental Microbiology.S. 3.C. Galtsoff P.MOLLUSCAN SHELLFISH 3. Washington D.C. 1936.E. Applied and Environmental Microbiology. Fate of enteric bacteria in estuarine sediments and oyster faeces. 1926. 1963. 12. Effects of harvesting waters and storage conditions on yeast population in shellfish. 1940.. 30. 105-8. Ward D. 72-8. 104-9. 1994. Liston J. 6. 1983..R. Eds. New York.W. American Journal of Public Health. Lee J. 1984. Microbiological characteristics of Dungeness crab (Cancer magister).L. Walker W.F. 42.K.. Applied Microbiology. 19-34. 1968. Journal of Food Protection. Vanderzant C.. 36. USA. Smith F. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves.. 33-6.W. Organisation for Economic Co-operation and Development. 71 . Villee C. 14.. 5. in General Zoology. 7.A. Ed. 633-40. Hackney C. 44. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. 71-6. Lovelace T. Government Printing Office.. 1960.D. Applied Microbiology. water and sediment from Galveston Bay. Colwell R. Rowse A. Eliot C. Eds. Tall B. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). 1970.J. Infectious diseases associated with molluscan shellfish consumption. Bacterial flora of the market oyster.. Philadelphia. 1981.S. Thompson C. Biology of the oyster in relation to sanitation. American Journal of Public Health. Jr. 46. 7. 1973. Fleet G.H. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. 4. Clinical Microbiology Review. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). 245-7. 9. U.. 17. 1982.R. 58. 10.7 1. Wiley-Blackwell.R. Galtsoff P. 755-76.R. 16. Bernard F. Microbiology of bivalve molluscan shellfish. Fish and Wildlife Service. 8. 231-51. Van Nostrand Reinhold. Viability and release Salmonella charity and Escherichia coli from oyster faeces. Pfeifer D. American Journal of Hygiene.E. Ray S.M. Nauman R.B. Rippey S. References The Phylum Mollusca. 82-7..S. Old H. 64. Veliger. 15.K.S. Hood M.. A typhoid fever epidemic caused by carrier bootlegging oysters. 8. Gill S. 1975. Tubiash H.. Proceedings of the National Shellfisheries Association. 1991. Saunders Company. Jr. 26. 1995. 33643. 419-25. 1964. 6. Cook D. 11. Colwell R. 13.. Schmidt C.

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

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

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

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

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

Pathogen monitoring of selected high risk foods (FY 88/89). Fish Inspection.E.. Lancaster. Washington D.W. Food and Drug Administration. Part I Sanitation of Shellfish Growing Areas. Inc. Food and Drug Administration. Washington D. Seafood Safety.. Food and Drug Administration Compliance Policy Guides. in National Shellfish Sanitation Program Manual of Operations. The Microbiological Safety and Quality of Food.S.H. Burdge D..1987. fresh or frozenadulteration by bacteriological contamination Sec. Ed. in U. oysters.L. U. Government Printing Office .. Food and Drug Administration.M. Aspen Publishers Inc..MOLLUSCAN SHELLFISH 88. 77 . in U.600 (CPG 7119. Martin R. 7.W. Quality Control. 1995. U.C.C. 3.12). U. Food and Drug Administration.. Chapter 5: Foods. Washington D. Technomic Publishing Co. 91.S. Food and Drug Administration. 90. Ed..C.S. Ed. Baird-Parker T. 2000. and HACCP: A Global Focus.A. Sherlock C.8 Further Reading Ahmed F.S. Food and Drug Administration. Food and Drug Administration Compliance Program Guidance Manual. Gaithersburg. Food and Drug Administration. U. National Academy Press.. 1991. Smith J. Food and Drug Administration. Slavin J.S.. 1987. 560. 1995. 1997. Gould G. mussels. 89.C.R. 63-8.030. Collette R. U. Clams. Lund B.S.S. U. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. Program 7303. Pittsurgh.S.E.

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0 kg polythene bags. Parapeneopsis spp. with additional water to form a glaze. lobsters. crayfish and shrimp (1). FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. Shrimps.. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi).5 and 1..1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. and frozen in 1 and 2. 79 . with shells and heads on or off are usually packed. and are usually aquatic. (deep-water shrimp). lobsters and crabs have ten legs.5 and 1. the front pair ends in claws.g. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. jointed limbs and a chitinous exoskeleton.Ph. and available in 0. Crayfish. CRUSTACEAN SHELLFISH Linda Nicolaides. M. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. e. Species eaten from tropical waters include Penaeus spp. Pandalus borealis (deep-water shrimp).4. Pandalus montagui and Crangon crangon (inshore shrimp).0 kg polythene bags. crabs. They possess a segmented body. Such products are usually only partially cooked before freezing. Peeled ready-to-cook shrimp are raw. peeled product. packed and frozen in 0.5 kilo pack (boxes or bags). Peeled and cooked shrimp are ready-to-eat after controlled thawing.. and Trachypenaeus spp.

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

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

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

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

later surveys in 1990 recovered the pathogen from retail raw.cooked prior to consumption. Aquatic birds and the hands of workers have also been identified as sources of the pathogen.5. Further surveys identified Listeria spp. However. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 .1 Pathogens: Growth and Survival Salmonella spp. shrimp and shellfish (11). 16). shrimps and cockles sold (17). in domestic/imported shrimps (15). Following this. in 24. cooked and smoked fish. 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 genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. Hence. even if GMP is followed. The presence of L. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) .FISH AND SEAFOOD 4.2 Listeria spp. 7). 5.5 4. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. although efficient cooking will quickly eliminate it (6). An import alert was put in place in June 1987. Although foodborne illness had not been associated with the product. and usually for both. nearly 30 years elapsed before the potential hazard of L. as well as for Salmonella. fish fingers. 4. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L.5% of frozen raw shrimps from different countries (13). a common pond fertiliser used in aquaculture. 6. animal manure. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. Moreover. Salmonella species are more commonly found in crustaceans reared by aquaculture or growing in faecally contaminated brackish water compared with species of marine origin (4. monocytogenes in cooked ready-to-eat foods. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). Salmonella cannot be completely removed from raw shrimp during processing.5. The above results prompted the FDA to develop a compliance programme for the pathogen. monocytogenes was absent from 40 samples of cooked prawns.

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

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. or freezing at a temperature of -23 °C for 7 days (30. to date. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). However. non-B Hepatitis viruses (31). 32). It is more commonly implicated in countries where crustacean shellfish are eaten raw. 4.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. Norovirus.5. South America. jejuni has not.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. hydrophila is able to grow well at refrigeration temperatures (0 . although A. hydrophila and A. Quantitative levels were below limits of detection in all cases (<0. vulnificus is an invasive and lethal pathogen associated with wound infections. Of these. a marine halophile. A. in a survey carried out in the United States between 1977 and 86 . coli. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. V. but has the potential to cause fatal foodborne illness (26). Hepatitis A virus presents the most serious hazard. V.g. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. 4. hydrophila has been isolated from prawns (29). occurs in crustaceans from warm environments. whilst 5.30 MPN/g) (27). the potential hazard exists as C.5.2 °C) so that GHP coupled with controlled low temperature storage is essential to prevent the pathogen from reaching levels that may present a hazard.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. However. parahaemolyticus.5. 4. Parasitic infections are controlled by either cooking infected material prior to consumption. however.5. been implicated in a case of foodborne infection associated with crustaceans. Caliciviruses. Astroviruses and non-A. 4.8% of samples contained C.6 Campylobacter jejuni and Campylobacter coli C. 31. Japan. e.7 Aeromonas hydrophila and Aeromonas sobria A. particularly trematodes (6).

2073/2005. The risk of such hazards is far higher with fin fish and molluscan shellfish. risk-based system based upon the seven principles of Hazard Analysis and Critical Control Point (HACCP) supported by pre-requisite or good practices programmes at all stages of the food chain. 87 . which results in a persistent and apparently permanent loss of short-term memory (36). 38. No foodborne incidents have been attributed to this source to date.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish.6. 4. produced or imported into the European Union (EU). Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. Norwalk viruses can also withstand heating to 60 °C. from farm to fork (37. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). although it is one of the European Union (EU) legislated allergens. They can survive low pH levels. 4. or chlorine concentrations similar to those found in drinking water. A summary of microbiological criteria for crustacean shellfish is listed below.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. are included in the Commission Regulation (EC) No.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. Selected parts from this regulation is presented below.5.5.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). 39). consistent with that in the stomach. which could be transferred to the meat during unskilled removal from the shell (35). 4. The toxin causes amnesic shellfish poisoning (ASP) when consumed. Provisions laid down concerning the use of additives would also apply to these types of products. only 0.

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

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

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

This should be distinguished from the salt content.6 132.2 105. Thus. it is important to state the temperature.80º brine (184. it is impossible to establish standardised processing stages in the curing of fish. processes and producers’ private recipes.8 211. or salt in the water phase. % 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. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. sometimes the salt crystallises onto the surface of the fish.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. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat.0 158.6 g of salt or more per litre of brine) may cause salt burn in this way.I). For smoking salmon. The dissolution of salt in water is dependent on the temperature. smoking or marinating processes. Salt concentration means the percentage of salt in the water contained in the fish. When a 100º brine is used. When a milder brine.8 211. TABLE 5.4 52. Different brine strengths are required according to the curing processes and/or the type of product. all using salting steps.4% salt (Table 5. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. The strength of the solution is described as degrees of saturation (brineometer degrees).8 79.2 g of salt per litre of brine). At 18 °C a 100º brine is produced when the solution contains 26. Therefore. it causes swelling because the fish absorbs water from the salt solution (1). about 50º (132.4 184.2 237. only the most common processes can be described here.0 g of salt per litre of brine).FISH AND SEAFOOD 5. the fish is usually salted with a 70 .3 Processing and its Effects on the Microflora Owing to the wide variety of products. a 90º or above brine (237. it is important to understand the concept of salt concentration.0 . a 0º brine saturation means pure water. When working with salting. etc. Salt in the water phase links the two conditions of moisture and saltiness.6 264. and salt content. is employed.

The most common fish used in the northern hemisphere is cod (Gadus morhua). producing characteristic pink patches. respectively. 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. caught in North Atlantic waters. and for a heavy salting a ratio of 1:3 salt:fish is used (1). having a short section at the end of the tail. lean white fish is used for this purpose. SMOKED AND DRIED FISH 5. On the other hand. outgrowth of halophilic bacteria may occur. When pure salt is used. which include calcium and magnesium. and the salt has penetrated throughout the fish (3). low in calcium and magnesium. can be dried further to produce ‘klipfish’ or ‘bacalão’. Salting can be done in a number of different ways. are likely to cause hydroscopicity of the product and.15 . respectively (1). mainly in tropical conditions. Also.15% calcium and magnesium. which consequently inhibits the diffusion of the water from the bulk of the product to the surface.3. if very pure sodium chloride is used for curing purposes.3.1. 97 .1. the fish is restacked with fresh salt after about a week. however.CURED. The salt used should be of food-grade quality. For a light salting. the fish is mixed with one part of salt per eight parts fish.35% and 0. However. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. and salt can speed this oxidation process. ‘Salt burn’ might occur when the surface of the fish dries out too quickly as a result of the small grain size of the salt. Usually it is followed by other preservation techniques such as drying and/or smoking. ‘Green cured’ split cod with about 60% of the backbone removed. and a ‘putty’ fish product is also obtained (1).1 ppm and 10 ppm. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). are recommended to help make the product whiter. This is because fatty fish turn rancid quickly. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. and such delays. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. when the air humidity is high. 5.1 Dry salting / Kench curing Usually. See Figure 5. for which there is a high demand in Mediterranean countries. it tends to lead to a slight yellowing and softening of the product. addition of levels of 0. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. This condition is known as ‘putty fish’ (4).0. and essentially free from iron and copper (3).1 Salting Salting of fish for preservation purposes has been used since pre-history. but the concentration of copper and iron must be kept under 0. The calcium and magnesium impurities.

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

76 4.41 3. SMOKED AND DRIED FISH smoking.23 8.41 5. This is why it is the preferred method for fatty fish. but as it is placed in containers.5 4. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.3 days.0 3.76 99 .66 5. and can be collected and replaced again over the fish.80 5.17 5. 5.85 3. the brine is then replaced over the fish.47 6.26 3. Histamineforming bacteria do not generally grow at water activities below 0. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.46 4.10 1.76 4.86 2.88 5.26 7.5 3. but mackerel.51 3.64 4.95 3.60 2.85 3.33 5. Here. The main objective of the brining method of salting fish is to limit air penetration during processing.17 4.78 2.61 3.23 2. the product obtained is called gaspé-cured (5).44 2.26 1. can be cured in a saturated brine solution for 6 days and.II.35 4. thus. Fresh sardines.41 6.70 3.74 9.1. and a red burgundy colour will develop in the product. herrings and other fatty fish can be used.56 6. See Table 5. the liquid that comes out of the fish forms a brine. Histamine formation is of importance when using scombroid and some clupeidean species. after being drained and pressed at 465 kg/m2 for 15 hours.84 1.90 (1).51 5.19 3.5 3. the fish is dry-salted.41 4. The fish are kept immersed in their own brine near saturation point for several days.99 2.26 2.0 4.44 2.3.98 6.26 4. headed and gutted.70 6. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 . TABLE 5. a saturated brine is used.25 5.54 5.23 3.76 4.05 7.76 4. There are several variations of this process.11 3. limiting the amount of rancidity that can develop.65 2.98 5. For preservation purposes.09 8.91 2.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) 2.25 5.03 2.76 4.30 2. For example.00 3.09 8.0 2. sardines.11 4. if split fish is dry-salted.73 1.45 3. the fish enzymes play an important role by maturing or ripening the product.49 4.03 4.78 6. Anchovies are the main fish used for these purposes.22 6.11 9.66 5.78) (1).96 1.04 5.3 Dry / wet salting In this process.98 6.CURED. with pressure being applied to the top of the container to help the brine to drain away. they can be stored at 28 °C.5 10.16 7.23 2.06 4. with a shelf life of about 4 weeks (water activity (aw) about 0.61 3.

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

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

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

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

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

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

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

Aspergillus ostianus. They are generally not cooked prior to consumption. The classical liquid smoke is 107 .3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output.3½ hours using the typical temperature set for hot-smoking referred to above (12). similar to masmeen. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. moulding it into long thick sections. 5. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). They will keep well frozen.3.5. break down of fats and the moderation of smoke flavour. to have no further advantage over other packing methods for other fish (12). When refrigerated at 3 °C. rewetted and remoulded to give the desired shape and consistency. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. The product is removed from the drying cycle. but mechanical drying is used instead of sun drying. Katsuobushi. and substances unconnected with smoke but with a smoky flavour or smell. fatty fish will remain in good condition for 6 days. 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). 23). resulting in a characteristic flavour (20. Natural smoke extracts. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. 17. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. oysters and mussels are usually wet-salted in 50º brine for 5 minutes and hot-smoked at 80 °C for less than 1 hour (12). Aspergillus oryzae. In terms of safety. cooking in brine. 21. Boiled or steamed clams. such as yeast derivatives. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). but seems. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). 22. then hot smoking and finally sun drying it. a very expensive and fashionable smoked product in the Netherlands. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. Vacuum packaging retards the onset of rancidity when fatty fish is used. apart from the improved appearance. Hot-smoked products made from white fish will usually keep better than those from fatty fish. are usually 80º brined for 5 . (Penicillum cyclopium and Penicillium puterellii) (19). are used. synthetic smoke flavours. and many closely related Aspergilli (Aspergillus ochraceus. Growth of these moulds on the tuna results in the extraction of moisture. Aspergillus tamarii) and several Penicillium spp. The production method is similar to masmeen. including ‘umami’ (16. including Eurotium repens and Eurotium rubrum. white fish about 8 days.6 hours and smoked for 3 . 18). chilled or freeze-thawed.CURED. Mackerel. Eels. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24).

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.03 μg/kg of benzopyrene. 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.3. When liquid smoke was used.6 to 2. 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. 5.8% and still provide protection for 7 days (25). The European Scientific Commitee for Food. nutritional neutrality and antioxidant activity (27). or applying it as an atomised spray within a modern automatic smoke kiln. 5. or smoke extracts in organic solvents. so as to impart an unnatural red colouration to the flesh. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). injecting it into the fish flesh. drying. Fresh fish spoilage may be measured by 108 . In experiments comparing traditionally smoked fish with liquid smoke. consistency of flavour and colour. smoking.06 μg/kg of benzoanthracene in the final foodstuff (28). There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). the salt level could effectively be reduced from 4. both of which are considered potential carcinogens.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. It has been shown that liquid smoke can reduce the formation of PAHs (3). applying it as a dip after brining. The synergistic effect of liquid smoke with other preservatives could reduce the amount of salt in the product when light diets are necessary whilst still meeting the necessary safety criteria (26). the result of this process is seen as a manipulation of the natural degradation of tuna colour that hides the true nature of the product quality and therefore has been made illegal in the EU. Liquid smoke was considered to provide advantages of safety.4 Spoilage Brining. spraying is preferable because it is easiest to control flavour and acceptable with this method (3).5. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. and less than 0. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. According to some recent research.

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

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

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

. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. sebi and B. + to +++ strength of preservative effect 4 Post-process contaminants 112 . Dun. LAB.not used/unchanged. boiled.W. 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. 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.variable.FISH AND SEAFOOD TABLE 5. 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. halophila Halophilic spoilage organisms Halophilic spoilage organisms.IV Spoilage of salted. and Halobacterium spp.

0-9. Before.5 5.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). In the United States. Examination of 165 outbreaks of botulism caused by fish products showed that smoked.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). sodium 113 . the fermented products being the most dangerous group (7). with this change. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13).86 0.61 Organisms Most Pseudomonadaceae C. However. but if the fish products are stored at temperatures <10 °C. the non-proteolytic type E strains can grow in up to 5% sodium chloride. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. salted and pickled products accounted for 10.5-8. The concentration required to prevent growth at room temperature can vary from as low as 3.5% to 5%. 9 and 8 of the outbreaks.97 0. types A and B are not affected by 3 . smoked and dried. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. botulinum type E and production of toxin. botulinum. could not grow and produce toxin. aureus Yeasts Moulds Lactic acid bacteria pH 5.5-10. SMOKED AND DRIED FISH TABLE 5.5 0-11 3. when present in the raw material.9 0. popular demand for healthy foods has led to a change in the tastes preferred by the public and. under optimal conditions.5-8. The presence of salt in cured products has a great effect on the growth of the bacteria. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14).5 5. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. and C.5 4. the importance of cold-chain integrity must be emphasised. respectively. and as botulism is a serious hazard in foods. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C.97 0.8 1. botulinum Staph. botulinum type E Staph.8 0.5. pressure on the traditional methods of curing.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. It has been shown that.5-10 4. the products were heavily salted. botulinum type E. Since neither the smoking nor the drying parts of this curing process are particularly severe. although the combination of the two factors.75 0.CURED.

the organism can survive most of the light curing processes.86 and 10% sodium chloride. are particularly susceptible to contamination and. but. they do not grow well in the raw material. it should contain not less than 3. if the person handling the product is a carrier of Staph.5% salt in the water phase.94 (7).5.3 Enterobacteriaceae The pathogens Salmonella. but. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). related to the consumption of a raw smoked fish meal. Shigella and E. As they are poor competitors. if it contains sodium nitrite ≥100 ppm. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. aureus growth. Once the product is contaminated. 5. these parameters change as a result of the interference of food components and other limiting factors that can act in an additive or synergistic way to prevent Staph. the organism may contaminate the product.5. Usually. if storage temperatures are over 10 °C. has been reported (42). An outbreak of Shigella in an Italian cruise ship.30% of the population positive for enterotoxin-producing strains (41). Sliced smoked fish products. more importantly. 5.4 Listeria monocytogenes The psychrotrophic L. as they require more handling. gills and guts as well as in several lightly preserved fish 114 . growth and toxin formation may occur.5. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . The FDA established that hot.0% salt in the water phase (30). but requires higher temperatures for toxin production (>10 °C). after processing. 5. In foodstuffs.4.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. and the minimum pH for growth is 4.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 risk of infection can be eliminated by proper cooking. if recontamination occurs and storage conditions are favourable. aureus. the human nose. when the bacterial load of the product is lower. It is a halotolerant bacterium able to grow at water activities as low as 0. coli all occur on fish products as a result of contamination from animal/human reservoirs.5. However. they will grow rapidly (7). throat and skin. The organism is mesophilic with a minimum growth temperature of 7 °C.or cold-smoked fish should have at least 3. Infection by Shigella sonnei has increased in the UK.0 . they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. As with other organisms. Thus.

the round worm Anisakis spp. and in CT95-1207FAIR). Some LAB. monocytogenes’ in 25 g of the finished product. and other local traditional products (7).9. monocytogenes in levels at or above 102 per gram (46). the CT 3162-FAIR program. do not survive 1 min at 60 °C. most of them are of little concern to public health.12%.6 and water activities below 0. Unlike some European countries. the only concern to public health is when eating raw or uncooked fish products. As they are heat-sensitive.514. The use of LAB in fish preservation is a topic that has been studied in projects (UP. a short period of freezing either of the raw material or of 115 . 5. Two outbreaks in the United States have been reported to be linked to salmon (10). favours growth of psychrotrophic LAB. which means that the process must conform to a level of ‘no detectable L. which include intermediate hosts. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod.95 (45). 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. or nematodes. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes.45 °C. there were no reported outbreaks related to cured fish. are considered unsafe. both being detected in cold-smoked salmon.5 Parasites Despite the presence of parasites in fish being very common. Thus. This includes matjes herring. salt levels of 8 . As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. pH 4. such as ‘lightly preserved’ fish products. from freshwater fish. marinades (ceviche) (43) and gravad fish (44). Pseudoterranova dicipiens. gravad fish. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48).0.5. which. The round worms. Until 1997. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). in combination with curing agents and refrigerated storage. lightly salted caviar.CURED. monocytogenes pathogenic for only specific segments at risk. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. the USA established ‘zero tolerance’ for the organism. has also been detected in cod (7).2. widely used as starter cultures in food processes. have been shown to have an anti-listerial activity in many food products. and the tape worm Diphyllobothrium spp. They have complicated life cycles. a round worm. from seawater fish.8 .92 . a number of fish products. Its limiting conditions for growth are 0 . cold-smoked fish. SMOKED AND DRIED FISH products such as cold-smoked fish. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. which considered L. Of microbiological concern in the northern hemisphere are two parasites . in the EU Fisheries R&D-FLAIR program.

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

1990. Fernandes R. SMOKED AND DRIED FISH TABLE 5. Wareing P. 7. 82 (revised).H.F. 9. Flick G. Marinades. 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. Eds. USA. Wareing P. 56. except in the viscera of dungeness crab. Smoked. or presence of toxin 0.. Handling and Processing Eels. parahaemolyticus Vibrio vulnificus C. Birnie K.CURED. in The Seafood Industry. 1993. New York. 3. 120-38. 6. coli (ETEC) L. Moody M. London..W. Assurance of Seafood Quality. Torry Advisory Note No. Hall G. 4. Hot Smoking of Fish. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5.A. in Micro-Facts: The Working Companion for Food Microbiologists. 381-406.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. Introduction to Fish Technology. 1984. 1982. 254-82 5. Eds. 3273..F. salting and smoking). Fernandes R. References Regenstein J. Van Nostrand Reinhold. monocytogenes Salmonella spp. FAO Fisheries Technical Paper. Regenstein C. 8. 334. Van Nostrand Reinhold. Huss H. Urner Barry Publications Inc.7 1. Doré I. cured and dried fish. Flick G. Food spoilage fungi. McLay R. 117 .. Preservation of fish by curing (drying. 1979. 2. Torry Advisory Note No. Torry Advisory Note No. in Fish Processing Technology. 37(revised).. Leatherhead Publishing. * Staph. 1994. botulinum * Limit of tolerance 1x 103 ETEC/g.M. Chapman & Hall. 1991. Leatherhead. Ed.. 2007. Martin R.E. Bannerman A. Catching. Horne J. New York. aureus * Vibrio cholerae V.VII US Food and Drug Administration (FDA) guidelines related to ready-to-eat fishery products (45) Product Ready-to-eat fishery products (minimal cooking by consumer) Organisms and/or toxins Enterotoxigenic E. Horner W.M. The Smoked and Cured Seafood Guide. 1997.

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

1991.1988. Huss H. New York. Institute of Medicine. 1996. Pszczola D. 30. 1996..F. 74. CRC. 37. 27. 3 (4). Ed. Interactions among xerophilic fungi associated with dried salted fish. 1971. 43. Pitt J. Incidence of Listeria monocytogenes in an acidified product.Y. Journal of the Fisheries Research Board of Canada. Journal of Applied Bacteriology. Maga J. 1994. Hocking A. 22. in Fish and Fishery Products Hazards and Controls Guide. Oxford. Cox J. Commission of the European Communities. 12. Røntved S. Borgstrøm G.. Hocking A.D. 49 (1). Tour highlights production and uses of smoke-based flavours. 1985. in Smoke in Food Processing. 36.R. 38. Studies on Sporendonema epizoum from ‘dun’ salt fish.I. 351-7. 29 (2). Pitt J. Loessner M. 70-2.M.J. in Fish as Food. 40.M.. Boca Raton. 33. Diyaolu S.A. Shewan J.D. US Food and Drug Administration. Springer Science. Sprouse R. 27686.H. Journal of Tropical Medicine and Hygiene. Ed. Fuchs R. Mycotaxon. 39.. 38 (3). 32. Hess E. Hansen L.CURED. 107-11. Smoked and smoke-flavoured fishery products. 311-7. ceviche. Committee on Evaluation of the Safety of Fishery Products. CEC.. Wheeler K. 74 (2). V. 88-90. 218-25. Simini B. Ahmed F.K. 14. Antimicrobial properties of smoke. Adebajo L.C. 35. Studies on salt fish. 1986. 1979. An outbreak of Shigella. Journal of Food Technology. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. 1993.. Press Inc.M. SMOKED AND DRIED FISH 26. Golden D. 2005. Academic Press.A. The microbiology of sea-water fish.. Fungi and Food Spoilage. Wheeler K.. 1994.A. Nahrung. Jay J. 31. Hocking A. Lucas F.A.D. Ed..F. 41. 1961. Lancet. The control of botulism in hot-smoked trout and mackerel. Cann D. 42..E. London.T. in Reports of the Scientific Committee for Food (34th Series). 742. Food Research International.. 5. Food Technology. 1941... 1995. 1991. Food and Drug Administration. Frank M. 28. 74..B. National Academic Press. Scientific Committee for Food. Letters in Applied Microbiology.E. 181-8. 34. Anggawati A. 1997. 164-9. Smoke flavourings.. Washington DC. Food Microbiology. Importance of autolysis and microbiological activity on quality of cold-smoked salmon.I. Luxembourg. Ed. 197-208. Fungi associated with Indonesian dried fish. 29. Modern Food Microbiology. 98100.I. Food and Drug Administration. 1995. 348. Townsend J.S. New species of fungi from Indonesian dried fish.. Gill T..O. Seafood Safety.D. Rockville. Taylor L. 123-9.V. 119 .. Hocking A. Fungal flora of South Vietnamese fish and rice. Sirvas S.. Pitt J. Blackie.

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

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

.

FERMENTED FISH Prof. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. The relative importance of these two activities depends on the formulation of the product. though some are produced elsewhere. Those to which only salt is added tend to be mainly autolytic. whereas in those where a carbohydrate source such as rice or sugar is also added. they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. lactic fermentation can play a significant role. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation.I. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. North American and other markets and are. As with most traditional products that are still produced principally on a cottage industry or domestic scale.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. in economic terms. the most important of all fermented fish products. they are exported to European. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein.1 Definitions As applied to fish products. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu.6.25% and are used mainly as a condiment. koji.1. added during preparation. producing the fish sauces and pastes. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6. 6. 123 . Carbohydrates are added to some fish sauces during their preparation. Fermented fish products are largely confined to east and south-east Asia. For example. typically in the range 15 . there are numerous variants of some common themes and a host of local names used to describe them. Essentially. The fish sauces are produced on the largest scale.

a variety of products can be produced within a single country. This is well exemplified by some of the fish/salt products of Thailand described in Table 6. made from whole fish in France. 6. cheese-like aroma Burma Indonesia Ngapi ketjap-ikan Paste Red/brown salty paste nga-ngapi trassi-ikan trassi-udang (shrimps) prahoc mam-ruoc (shrimps) Kampuchea nuoc-mam nuoc-mam-gau-ca (livers only) Korea myulchljeot jeots. to products analogous to other lactic fermented foods such as salami. who had adopted them from the Greeks.1. These have long since fallen from use. although garos. salty taste.II. made from fish livers in Greece. may be direct descendants. In Europe. 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. Fermented fish products are of very minor importance outside Asia. and pissala. where the bacterial production of lactic acid is a major feature.FISH AND SEAFOOD TABLE 6. A selection of Asian products is presented in Table 6.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. and the more widely known Worcestershire sauce a very distant relative. 124 . fish sauces known as liquamen or garum were ubiquitous condiments for the Romans.

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

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

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

Belacan (Condiment) Fig.2.12). 6. Patis (Condiment) 128 .1. 6. Fig.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 .

Ika-Shiokara (side dish) Fig. the 129 .FERMENTED FISH Fig. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are. In fish sauce production. thought to be largely the result of autolytic breakdown of the fish tissues.3.6. eviscerated fish are sometimes used to slow the autolytic process. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. To produce semi-solid fish pastes as opposed to fish sauces.4. however. 14). 6. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13.

volatile fatty acids and nucleotides. snack) 130 .FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. which is eventually separated from the residual fish tissues.5.8.3. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. In contrast to free-swimming fish.5). Fig. Hoi-Malangpu-Dong (side dish.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. Some examples of products and their formulation are presented in Table 6. particularly glutamate.V and Figures 6. shellfish such as mussels often have appreciable levels of carbohydrate in their tissues and in these cases addition of carbohydrate is not necessary for a lactic fermentation to occur (see. Figure 6. for example. filtered and bottled. 6. 6.6 – 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.

NE Formulation Fish (whole or pieces): salt: roasted rice.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.12 months Main dish 2 Pla-jao C. C NE. NE 10 . Kung-Chom (staple food. N. In these cases.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .5-1. N.6. side dish) 131 .2-4 Fish (pieces): salt:khaomark (fermented rice).6.FERMENTED FISH TABLE 6. south of Thailand.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. C. C NE. central. N. the prefix 'pla' is replaced by 'kung'. S.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE.12 days 3 . 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . N S 5 . NE.03 Preparation time 6 .4 are also produced using shrimps. north. north-east. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. 3:1:0.12 months Shelf life Use 1 .7 days 5 . Fig.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0.

(from Arroyo et al. The LAB isolated from these products are those with more marked salt tolerance but. 2. but there is considerable variation in this. their growth and metabolism are nonetheless slowed.. 106) Fig.3.salt and lactic acid.10%. typically in the range 2 .1 The use of higher salt levels results in a longer production phase but a better keeping quality product.6.2. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6.7. giving a reduced rate of acid generation. This is readily explained by the susceptibility of LAB to elevated salt concentrations. If acidity is a desired sensory property of the product. these products are salted at lower levels than the fish sauces and pastes. Phillipines Journal of Food Science and Technology. while they are able to grow at high salt levels.VI). Red burong isda 132 . then this will take longer to develop than if lower salt levels are used. the salt is of far greater importance in determining the product’s long-term stability.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. of the two factors contributing to shelf life . Generally. 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. It also indicates that.

3 .2 7. Philippines Journal of Food Science and Technology.6.17.1 4.4 . 133 . White balao-balao TABLE 6.5 .8 .4.5. 106) Fig.0 Shelf life 3 weeks 6 .FERMENTED FISH (from Arroyo et al.5. 2.9 .3 5.5.0 .5.6.8 pH 4..6 3.2 4.8 2.9 4.5 .VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.0 .7 .0 .4.8.1 .9.12 months 1 .2 4.5 3.3 years 1 .9.5.8 .6.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).9 4.

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

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

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. outbreaks of 136 . Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. fish products are the most common source of botulism. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. 6. although these limits apply when all other factors are optimal (26). The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and.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. V. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. typically 30 °C. viable numbers will probably decline quite rapidly. There is little in the way of documented information associating fermented fish products with foodborne illness. Because of the severity of the illness it causes. In the fish sauces and pastes. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. which accounted for 88% of outbreaks in the period 1952 .5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. During fermentation. After capture. The bacterial pathogens most commonly associated with the living environment of fish. under conditions inimical to growth and high ambient storage temperatures. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. parahaemolyticus has been reported growing down to a pH of 4.8 and at salt levels as high 10%. are Vibrio parahaemolyticus and Clostridium botulinum. As in Norway. 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 epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). but we are unaware of any reports of this happening.87. Several measures have been suggested for reducing the risk of botulism. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. 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. however. and consumed without cooking. In Japan.

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

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

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

J. 23. Progress in Industrial Microbiology. 854-60. Miwa M. 661-70... Todoroki T. Hanai S.. 2005. 15998. Fermented flesh foods. Gaithersburg.... 6.. 110-1... Kikukawa H. 2005.7 Further Reading Adams M.R. Southeast Asian Journal of Tropical Medicine and Public Health. Honjo S. Uchida K.. 32. Survival of Haplorchis taichui metacercariae in lab-pla. Deerasamee S.. 1995. International Journal of Cancer. Aspen Publishers Inc.. Survival of Anisakis larvae in marinated herring fillets. Adams M. Nithikathkul C. Sriplung H. Roepstorff A.R. 33.FISH AND SEAFOOD 31.H.. northeast Thailand. Sripa B. 29. International Journal of Food Science & Technology. 1986. Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom. Huss H. Chuboon S. Ruamsuk A.R. Bloemsma B. 140 . Horst K. Nout M.. Wongsawad C. 2001. 117. Jedpiyawongse A.... Srivatanakul P.. Fermentation and Food Safety. 36. Thai traditional food preparation. Kittiwatanachot P.

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

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

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

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. a pre-column method has received Association of Analytical Communities (AOAC) approval (18). and islands of the South Pacific. as well as other parts. The latter test is accepted by the United States Food and Drug Administration (US FDA) for use in the United States National Shellfish Sanitation Program. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). Alternative HPLC methods have subsequently reduced this to a single HPLC run.2). Thailand. a criticism has been that such methods are time consuming. TTX most commonly occurs in many species of puffer fish. 15. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. Two approaches have been taken here.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. and while simple and relatively inexpensive to operate. China. other derivatives have been reported. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). 24) (Figure 7.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. and over ten analogs are known (22. The mouse bioassay has been in place for decades and the method has been standardised. Most intoxication events have occurred in South East Asia. including Japan.3. However it remains as the monitoring method of choice in many countries. 23.2.FISH AND SEAFOOD 7.8 µg/g) of shellfish or seafood tissue. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. specifically 144 . While the chemical detection methods are most promising for the future. 7. 17). Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). Taiwan. 7. After some development and various trials. Since its discovery. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). and has also been evaluated by a number of EU laboratories.

Severe poisoning causes paralysis and respiratory failure. Fig. and once again the mouse bioassay has been used to detect the presence of the toxin. it is served in speciality restaurants with qualified chefs to properly prepare the fish. Structure of tetrodotoxin 7. respiratory failure and coma. All of the observed toxicity is secondary to the action potential blockade. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24.2 mg of purified toxin can be lethal. 27). 25. preventing depolarisation and propagation of action potentials in nerve cells. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. and in mild cases results in distal muscle weakness. 7.3. and more recently by an actinomycete Norcardiopsis dassonvillei (26). Typically this has been accomplished using HPLC followed by post 145 . molluscs. and even newts.FISH AND SHELLFISH TOXINS in the liver. Since then. nausea and vomiting. gonads and roe (24). 26). A review of TTX detection methods has been published (12. where puffer fish of the genus Fugu are a delicacy. 23. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. The onset of symptoms usually occurs rapidly. 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. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. An oral dose of 1 .3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. and blocks diffusion of sodium through the sodium channel. 7. the blue-ringed octopus. raising speculation that TTX is produced by a bacterial source.2. hypotension. In Japan.2 Toxicology Like the PSP toxins.3. horseshoe crabs. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera.

7. Fig. but it is known that such congeners are less toxic than DA itself (30). 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). both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28). However this method has been criticised for being complex.4.FISH AND SEAFOOD column oxidation to the fluorescent derivatives. Currently 12 isomers of DA have been reported (see Figure 7.3.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). Immunoassay methods have also been investigated.4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). a polar glutamate agonist. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. but all of them are considerably less toxic than DA itself (29). These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule.3 for examples). need further development (24). Structure of domoic acid and three naturally occurring photoisomers 7. 7. but 146 . and while promising.

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

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

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

nausea. Due to the lipophilic nature of the toxins. 7. A regulatory level of 80 . Since the late 1970s. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. and abdominal pain. 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. and while this provides critical seafood safety information. 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. even with the minimal of clean up steps (48). vomiting. New Zealand and Chile. Human symptoms include diarrhoea. 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. In recent years. DSP-Check is a commercially 150 .2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. various European countries.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48).5. 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).FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. it does not provide information on the DSP toxin profile. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity.3 days. although a comparative study of in vitro phosphatase inhibition with mouse bioassay data has shown that OA is about twice as potent as DTX-2 (51).3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. DSP episodes have been reported worldwide and are included in many monitoring programs (47).5. DSP episodes have been reported from Japan. which can lead to an underestimation of the total DSP content in a sample. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. 7. and a detection limit of 100 µg/kg has been reported (12). and DTX-2. 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.

58). Some representative examples of both groups of toxins are shown in Figures 7. for many years these compounds were included in the DSP toxin complex. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. with good agreement with the mouse bioassay and LC–based methods (54. with a claimed detection limit of 20 µg/kg. 7.6. 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. However. Within this group. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49).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). Indeed.5. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. 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. and another rapid antibodybased test kit for DSP toxins has also been reported (53). 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. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . 55). and the latter group are usually found to be responsible for any reported shellfish toxicity (59).FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. Fig. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. 7. In yet another approach.5 and 7.

7. New Zealand and South America (49. eastern Canada. YTX and homo-YTX 7.6. 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). 57. many parts of Europe and the Mediterranean. Japan. As is typical for many algal species. Structure of the two parent yessotoxins. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59).6.FISH AND SEAFOOD Fig.6. While 152 . which may co-occur in blooms with other organisms. 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. 58).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. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. the toxin content and profile may vary among different strains.

but various congeners are acutely toxic in the mouse bioassay following IP injection. 64). but less so following oral dosing (61).6.FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. The crude extract may be used directly in the mouse bioassay. chemical analysis is best carried out by LC/MS and several approaches have been described (57.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. a regulatory level of 1 mg/kg shellfish tissue has been set. the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present. Some of the PTXs are potent cytotoxins in vitro (60). 7. Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. though its cell effects may be linked to modulation of calcium channels (62). The YTX group has not been reported to cause human illness. which is further compounded by differences in the experimental design. Their toxicology and mechanism of action remain uncertain. the PTXs show considerably less toxicity when administered orally. it may be compromised by other biological materials in the matrix. and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7. The structures and chemical data including the molecular weight of each member have been collated (56). Because of the number and complexity of the YTX group. For many years it was believed that 153 . The regulatory level of 160 µg/kg shellfish tissue has been established and the mouse bioassay is often the primary detection method although LC/MS methods are preferred and this requires some simple clean-up of the crude extract prior to chemical analysis. Estimates of acute toxicity of the parent compound following IP injection vary considerably. 7. The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. some of which are found as glycosides and sulfates.7). However. 58). Many YTX toxins contain a conjugated diene system. but a fluorescent derivative has been prepared for others. The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds. While LC/UV detection is possible for PTXs. Although there is no evidence that yessotoxins are responsible for human illness. which aids detection by LC/UV. There are no clear cases of human illness resulting from ingestion of PTXs (59).3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue.

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

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

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

7. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. pruritus. which in turn are consumed by carnivores such as snappers. groupers. arthralgia. there are neither standards nor an official testing program. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. although not all CTXs are equipotent. and there are suggestions that such incidents are increasing and spreading worldwide (74). Other symptoms include a metallic taste. all of which can be linked to the effect of CTX on excitable membranes. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. Even when suspect samples are available.3 Detection methods As a consequence of their potency. and although the gastronomical problems may dissipate within a day or so. These include paresthesias (numbness and tingling of the extremities). 75. 7. where cold objects feel hot and vice versa. Another symptom that can develop over time is extreme fatigue. and moray eels. Globally. Local island populations attempt to avoid poisoning by catching only smaller fish. 75). The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. 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.8. only trace amounts of CTXs can cause illness following consumption of contaminated fish. myalgia. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. weeks or even months (74 . and it is estimated that ingestion of as little as 0.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. and in fact CTX poisoning is often only identified after consumption of contaminated fish.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. which can return following the consumption of alcohol and certain foods such as oily fish (74. the neurological symptoms can persist for days. sharks.1 µg of ciguatoxin can result in human illness (74. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . which can be confused with chronic fatigue syndrome. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. and temperature sensation reversal. Like the brevetoxins. and the sensation of loose teeth (77). barracudas.8. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area.76). 78). The onset of symptoms can occur rapidly within 24 hours. Patients have reported symptoms lasting weeks and even months.

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

Mytilus galloprovincialis). oysters (Crassostrea gigas. The bivalves affected include mussels (Mytilus edulis. it has been impossible to establish that P.9. and to date this has been the only source from which these toxins have been found at toxic levels. 7. 90). but current data indicates that unlike the DSP compounds. cockles (Cerastoderma edule).9. Donax spp. diarrhoea and stomach cramps. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87).). clams (Tapes philipinarum. Portugal. AZA toxins do 159 . Ostrea edulis). Structure of the three principal azaspiracids found in shellfish 7. and most recently in a crustacean (Cancer pagurus). Scandinavia.9. these molluscs were never considered to be the biogenetic source. cassipes produces AZAs in culture. and some time after the original episode in Ireland. Ensis siliqua. Following the original episode. and to date no long-term effects have been recorded (83. AZA occurrence has been reported by several European countries including France.3 days. which can persist for 2 . These symptoms are not unlike those experienced following the consumption of DSP toxins. vomiting. and off the coast of Morocco (83. While this result is not necessarily in debate.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. 7. 89). which are ingested by P. 88.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. 84. However. scallops (Pecten maximus).FISH AND SHELLFISH TOXINS Fig. cassipes (83). In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. as in other cases of shellfish toxins. There has been considerable debate over the source of azaspiracids.

Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. though the possibility exists that they may inhibit other phosphatases.12). the pinnatoxins. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. 85).3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. but all contain a spirocyclic or cyclic imine function within their structure. 7. it is recommended that the entire shellfish be used for AZA analysis since it has been shown that these toxins can migrate to other tissues. Based on IP minimum lethal dose data. 92).10 Cyclic imines These toxins are represented by several distinct structural types. Once again. which are particularly amenable to detection by positive ion electrospray mass spectrometry. and closely related ptierotoxins (Figures 7. 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).FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. and ASP. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). The mouse bioassay has been used to monitor shellfish extracts.10 – 7. and erosion and bleeding in the stomach. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. and necrosis of epithelial cells on the microvilli (91). Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes.9. the spirolides. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. parenchymal cell pyknosis in the pancreas. abnormalities in the thymus and spleen. Importantly these pathological outcomes were reported as being different from those caused by DSP. Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. However. Using the same approach. PSP. 91. 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. AZAs also display cytotoxicity towards various mammalian cell lines (83. 7. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . 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.

7. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure.. Subsequent taxonomic studies revised this again to a new dinoflagellate K.520 Da (94. Fig. cockles. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis).10. At the time of the initial observation of gymnodimine.10). two additional congeners. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94.10. 95). it was proposed that a dinoflagellate identified as Gymnodinium spp. In only two cases (gymnodimines and spirolides) has a planktonic source been identified. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . but they are included here for completeness. selliformis (97). a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin.1 Gymnodimines The observation of neurotoxic symptoms in the mouse bioassay of lipophilic extracts of dredge oysters (Tiostrea chilensis) collected from around the South Island of New Zealand. 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. scallops. 95). Structure of the cyclic imine gymnodimine 7. surf clam and New Zealand abalone. Some time later.10.1. they are sometimes referred to as emerging toxins of uncertain human health concern. Pacific oyster.FISH AND SHELLFISH TOXINS health. 7.1 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.

11). and there was a further ten-fold reduction in toxicity when the toxin was supplied with food.2 Toxicology A review of the toxicology of all cyclic imines has been published (98).to10-fold reduction of toxicity (LD50 600 . Since their original discovery which resulted in the characterisation of spirolides A-F (99. A later study using purified toxin established an LD50 dose (Lethal Dose. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. 95). This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. abdominal breathing. 50%) of 96 µg/kg.3 Detection methods The characteristic “fast-acting. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay.10. Oral administration of the toxin to mice by gavage resulted in about an 8. Based on this diminished oral toxicity. 95) (Figure 7. However. paralysis of the hind quarters and eventually complete immobility.FISH AND SEAFOOD 7. There is a rapid onset of symptoms including respiratory distress.10. Most cyclic imines lack an extended chromophore.900 µg/kg).100). The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . 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. and so spectroscopic detection by UV methods is not favoured. Whether isolated from dinoflagellate cells directly or from shellfish tissue.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. almost a dozen derivatives in total have been identified to date (94.1. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. 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. 7. together with a lack of evidence of human intoxication. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. The time lapse between ingestion and death is between 5 and 15 minutes. and this is the detection method of choice.1. and lack of responsiveness to stimuli. finishing with 100% methanol. They were originally discovered in extracts of mussels (M. it has been stated that regulation against gymnodimine is not required (98). In particular. 7. Reduction of the imine group eradicated the toxicity.10.

2. Interestingly. by gavage (87 .707 µg/kg. those spirolides possessing the vicinal dimethyl grouping (see Figure 7. in fjords in Denmark and Norway. While oral toxicity is considerably less than by IP injection. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. 7. Fig.8 µg/kg). fasted mice). R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). 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. the gulf of Maine. or by oral ingestion (381 . the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). All data reported here are for desmethyl spirolide C. southern Ireland.2 Toxicology The spirolides are the most studied group of the cyclic imines.10. fasted mice). 7.11.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia.166 µg/kg. Structure of four spirolide toxins A-D 7. 95). 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).2. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). the western coast of Scotland. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 .11. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 .10.

10. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . and in fact these compounds share a number of other structural features. consistent with modulation of acetyl cholinergic receptors (103). four members of the group have been identified – two of them being the C-34 diastereomers. A number of methods following this procedure have been described (94. Three members of this group have been reported. However.FISH AND SEAFOOD (100). the mouse bioassay can be used to detect these fast acting toxins. more useful information on the amount and distribution of toxins present in a shellfish extract is obtained by LC-MS analysis usually following a simple clean-up step.12). To date.3 Pinnatoxins and pteriatoxins Pinnatoxins A . The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. 7. Fig.12.2. 102). The compounds are zwitterionic and therefore somewhat more polar than the spirolides.B contain a spiro imine moiety similar to that found in the spirolides.3 Detection methods As discussed for gymnodimine. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. 7. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C. which includes two diastereomers. 95. While the mechanism of action remains to be clearly defined. 7.10.

7. like the gymnodimines and spirolides. The LD99 dose for the pteriatoxins is reported to be in the range (8 . has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. Despite this progress.3. research and certified standards of the key toxins are often unavailable or in short supply and in the future considerable logistical effort will be required to address this issue. 7. which in many cases are now found to be more widespread than previously believed.3.3.10. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). but this remains to be conclusively established (106). once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. 99%) values for pinnatoxins A-C are in the range (22 .2 Toxicology There is little information describing the toxicology of these compounds (98). The LD99 (Lethal Dose. There is strong circumstantial evidence to suspect that these toxins. However. which occupy tropical waters around Okinawa and the south east coast of China.1 Origins and distribution Shellfish of the genus Pinna.most likely a dinoflagellate. With increasing knowledge of the chemistry of toxins. while pinnatoxin D was essentially inactive. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104).10.180 µg/kg).11 Conclusion There has been considerable progress in the detection of known marine toxins. 7. arise from an algal source . to date action or 165 . While this is recognised as a real possibility for a wide range of toxins. there is no compelling evidence to suggest that they have resulted in human toxicity (98). and more advanced means of detection.100 µg/kg).3 Detection methods No formal detection methods for these compounds have been reported. have long been suspected of causing human illness after ingestion.FISH AND SHELLFISH TOXINS 7. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other.10. Only the acute toxicity in mice following IP injection has been reported. Coupled with this. This is generally a result of increased monitoring and continually improving detection technologies and methods. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. new toxin groups such as the azaspiracids and cyclic imines have also been reported.

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

Stobo L. Roberts R... Rapid detection methods for marine toxins: needs and limits. 283-317. Christian B. 61. Eds. CRC Press. Lacaze J-P.. in Manual on Harmful Marine Microalgae. 2008. 505-8. 1989. 15.. 2002. 6. in Methods of Analysis of the Association of Official Analytical Chemists. 1995. in parallel with the regulatory AOAC mousse bioassay.. 22. 19. 3-20. Analytical and Bioanalytical Chemistry. in FAO Food and Nutrition Paper 80.06. 219-27. Wageningen Academic Publishers. 16. 20.. Paralytic shellfish poisoning toxins in shellfish. Larsen J. Petrie J. in Rapid Methods for Biological and Chemical Contaminants in Food and Feed. Fukuyo Y. 12. 2002. Yotsu-Yamashita M..M. Toxicon.E. Anderson D.FISH AND SHELLFISH TOXINS 11. Lauwaars A. Journal of Shellfish Research.R. Risk assessment of marine toxins.C. method 2005.V. Analytical Biochemistry.L.. Pure and Applied Chemistry. Aune T.. Mackintosh F.. 10-7.M. Shoji Y. Tetrodotoxin – distribution and accumulation in aquatic organisms and cases of human intoxication. Food and Agriculture Organisation.. Eds. 5-52. Interspecies distribution and biogenetic origin of tetrodotoxin and its derivatives. Determination of marine biotoxins for regulations: from the mouse bioassay to coupled LC-MS methods. a new rapid test. Yasumoto T. Gaithersburg. Chemical analysis. AOAC International. 17.L. 1407-25. 40. Laycock M. Cembella C. Jellet J..M. Evaluation of MIST Alert rapid test kits for the detection of paralytic shellfish poisoning toxins in shellfish.F.. 13.A. 167 . Barug D. Yasumoto T. Ed. Taxonomy of harmful dinoflagellates. Van Amerongen A. 2004. 177-96. 2006. 2008. Neurotoxins from marine dinoflagellates: A brief review. Smith E. 230-42.. in Seafood and Freshwater Toxins.. Wageningen. Marine Biotoxins. Villar-Gonzalez A. 349-71. Boca Raton. Naoki H. Association of Official Analytical Chemists. Miyazawa T. Yotsu M. Scott A. and liquid chromatography/tandem mass spectrometry.. Electrospray ionisation mass spectrometry of tetrodotoxin and its analogs: Liquid chromatography/mass spectrometry.. 6.. 635-48. Taylor F. Wang D-Z. Rome. UNESCO. Krys S. Noguchi T.M. 21.C. Marine Drugs. Luckas B. Paralytic Shellfish Poisoning. 21.A. 18..H.. Arakawa O. 117-34.. 2008. Barrett R. Toxicon.J.. Endo A. 51.. Surveillance of algal toxins in shellfish from Scottish waters. Ben-Gigirey B. in Seafood and Freshwater Toxins.. Food and Agriculture Organisation of the United Nations. Boca Raton. Paris. 2008. Turrell E. 290.A. prechromatographic oxidation and liquid chromatography with fluorescence detection. Botana L. Quilliam M. Botana L. 391 (1). 23. 2008. 2005.M. Marine Drugs. 24. 14. Ed. tandem mass spectrometry.. CRC Press. Detection of paralytic shellfish poisoning (PSP) toxins in shellfish tissue using MIST Alert. Murata M. Hallegraeff G. Ed. 2001. Food and Agriculture Organisation of the United Nations.A. 455-60. 2008.

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

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

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

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

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

80. 81.

82.

83.

84.

85.

86.

87.

88.

89. 90.

91.

172

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

93.

94. 95.

96.

97.

98.

99.

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

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

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

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

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

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

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

7. but for which no control exists at that step or any other. It is prudent to set stricter limits (often-called target or process limits/levels) to ensure that any trend towards a loss of control is noted before the critical limit is exceeded. structured approach to decision making. If a significant hazard has been identified at a step where control is required for safety.2) to provide a logical.3.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. biological or sensory parameters. However. For example. application of the decision tree should be flexible. Where possible. and the reader is referred to the appropriate product chapter. given the enormous variety of fish products and processes in use. it is necessary to specify both time and temperature for a thermal process. it is not possible to generalise on likely CCPs. it can be said that effective control measures are likely to include the following: • • • • • 8. critical limits should be specified and validated for each CCP.3.3. However.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8.FISH AND SEAFOOD 8. Criteria used to set critical limits must be measurable and may include physical.3. 8. then the process must be modified to include a control measure. More than one critical limit may be defined for a single step. 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 .8.8 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. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively.1 Fish and seafood Again. and its use may not always be appropriate. 8. chemical.

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

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

and sets out what those enforcing the provisions have to do.9. European Communities L 268. 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. 9. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. 7. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption Regulation (EC) No. 190 • • . as last amended by Regulation (EC) No 806/2003.15). handling and offering for sale or supply to the consumer. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. packaging.1991.B. processing. distribution. 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.1992. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No.2 Legislative Structure From 1 January 2006. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. p. p. J.7.1991. 24. Subsequently.1). N. Council Directive 92/48/EEC of 16 June 1992 laying down the minimum hygiene rules applicable to fishery products caught on board certain vessels in accordance with Article 3(1)(a)(i) of Directive 91/493/EEC (OJ L 187. storing. transportation. manufacturing. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation.9. A number of more detailed implementing and transitional measures have been adopted at EC level. 41). p. 24.

and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. Primary products are defined as products of primary production including products of the soil. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. domestic preparation or handling. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland.3 Regulation (EC) No. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. The regulation lays down general hygiene provisions for which food business operators carrying out primary production must comply with as laid down in Part A of Annex 1. national legislation in the form of a Statutory Instrument (S.I. for example.11) The EU hygiene regulations apply to all stages of food production including primary production. The Food Hygiene (England) Regulations 2006 (S. and equivalent legislation in Scotland. of stock farming. etc. right through to the implementation of procedures based on HACCP principles. 19. with primary production. revocation of existing implementing legislation. As regulations. There are some exemptions. 9. Additionally the requirements of EC Regulation 853/2004 must be complied with. 2073/2005.1993. 1 . 2006 No. and binding in its entirety on all member states from the date of entry into force. Although the regulations have the force of law.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. setting offences. is required to give effect to the EU regulations.I. and of hunting and fishing. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. the legislation is directly applicable law. food storage that is for private or domestic consumption. penalties and powers of entry.g. which will be covered later in this chapter. Although EU food hygiene regulations are directly applicable in the individual Member States there are some aspects where Member States are required or allowed to adopt certain provisions into their national laws. p. 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. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. Wales and Northern Ireland).7. 191 . e.14. 852/2004.) in England. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. Wales and Northern Ireland.

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

please refer to Regulation (EC) 853/2004. a new mark must be applied to the product. Section I of Regulation (EC) 853/2004. EY. In this case.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. (iv) If the identification mark is applied in an establishment within the Community. EG. EÜ.2.2 Fishery product requirements 9.4. 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. (ii) The mark must indicate the name of the country in which the establishment is located and written out in full or shown as a two-letter code in accordance with the relevant ISO standard. EC. EO. and the characters easily decipherable and be clearly displayed. EK. (ii) However. Section I of Regulation (EC) 853/2004 are given as follows: 9.2.4.1. when a product's packaging and/or wrapping is removed or is further processed in another establishment.4. (iii) Food business operators must have in place systems and procedures to identify food business operators from whom they have received and to whom they have delivered products of animal origin. ES. the new mark must show the approval number of the establishment where this occurred.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. Extracts of the requirements of Annex II. EB or WE. for other codes. EK.2 Form of the identification mark (i) The mark must be legible and indelible. the mark must be oval shaped and include the abbreviation CE. the food business operator may apply the same identification mark to both types of food.4. (iii) Additionally.FISH AND SEAFOOD 9.1. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. 9. For the UK. 198 . the mark must show the approval number of the establishment.2. EF. the code would be UK.

With the exception of Chapters I and II. specifically relating to fishery products are given below. the approval number of the establishment and the abbreviation for the community where appropriate. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. the colourings used must be EC authorised colours. (ii) For products of animal origin that are placed in transport containers or large packages. including vessels. The requirements of this section accompany those in Regulation (EC) 852/2004. they would need to be obtained following Section VII on live bivalve molluscs. i.e.LEGISLATION 9. wrapping or packaging in another establishment. the mark may be applied to the outer surface of the container or packaging. reference should be made to the actual Regulation. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . as amended. C and D. tunicates and marine gastropods when placed on the market live. Part A and Chapter V apply to retail. Chapter IV.2. processing. granulate and powdered products of animal origin carried in bulk. (iii) In the case of liquid. it is sufficient to apply the mark to the exterior of that package only. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed.1.4.4. 9.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. (v) When the mark is applied directly to products of animal origin. Extracts of the requirements of Annex III. for full requirements. its wrapping or the packaging. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. echinoderms. it applies to those animals when not placed on the market live. and fishery products carried in bulk. This section does not apply to bivalve molluscs. and are intended for further handling. or be printed on a label affixed to the product. a) requirements for establishments.2. Therefore. Section VIII of Regulation 853/2004. the wrapping or the packaging. Chapter III.3 Method of marking (i) The mark is to be applied directly to the product. Parts A.

clean seawater may be used for the handling and washing of fishery products.4. i.. or to handle or process them after harvesting. bleeding. The definition of fishery products covers primary production as well as its associated operations. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking.2. if a vessel has water coming in that is used with fishery products. A. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material.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. removing fins. gutting. 2. refrigeration and wrapping. Chapter VII of Regulation 852/2004. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. fishing and collection of live fishery products with a view to their being placed on the market. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. There is a derogation from point a.FISH AND SEAFOOD c) for the water supply.) 9.1 Chapter I . it must be placed somewhere that will not contaminate the water supply. Structural and equipment requirements refer to the following points 1. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products.2. (a) primary production covers the farming. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 . and (b) related operations include any of the following operations if done on board fishing vessels: slaughter.e. heading. this supplements the requirements of Annex II. 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. from the place of production to the first establishment of destination.

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

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

packaged and chilled as quickly as possible after preparation. Requirements for frozen products An establishment on land that freezes fishery products needs to have equipment that meets that for freezer vessels as stated in Section VIII. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice.LEGISLATION consumption.3 Chapter III .2. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. 9. They may also continue to be transported in cooled water after landing and be transported from aquaculture establishments until they reach the first establishment on land that carries out an activity other than transport or sorting.Requirements for establishments. They should be wrapped. they must be stored under ice in appropriate facilities. Additionally. C. prepared or processed immediately after arrival at an establishment on land. Food business operators need to co-operate with the relevant competent authorities so as to allow them to carry out official controls as laid down in Regulation (EC) No. and where necessary.2. Fillets and slices must then be removed from work tables as soon as possible after preparation. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. including vessels. (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. C. part I. (ii) Heading and gutting must be carried out hygienically. points 1 and 2. dispatched. handling fishery products A. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . B. 854/2004.4. Chapter I. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed.

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

2. pressing.LEGISLATION approaching that of melting ice is reached. refining and purification steps before being placed on the market for the final consumer. B. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. 2073/2005.2. 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. and after this. separation. 2074/2005. there is an exception: food business operator may refrain from chilling the fishery products when whole fishery products are used directly in the preparation of fish oil for human consumption. Shelling/shucking is to be done hygienically. C. centrifugation.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No.4. 205 . processing. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. the following conditions need to be met: A. (ii) The production process for fish oil must ensure that all raw material intended for the production of crude fish oil is subject to a treatment including (where relevant) heating. However.5 Chapter V . 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. Organoleptic properties of fishery products An organoleptic examination of fishery products must be carried out to ensure that fishery products comply with any freshness criteria (such criteria have yet to be set). 2074/2005). 2073/2005). B. raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. and the raw material is processed within 36 hours after loading. 9.

prepared and processed fishery products belonging to the family Gempylidae. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. In the case of whole frozen fish in brine intended for the manufacture of canned food. 9.4. food business operators must ensure that wrapping material is not contaminated and is stored in such a way that it is not exposed to contamination and is easy to clean where necessary. The scientific name must accompany the common name on the label.FISH AND SEAFOOD D. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. Diodontidae and Canthigasteridae. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. thawed unprocessed fishery products. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption. Molidae.2.4. and when fishery products are wrapped on board fishing vessels. may only be placed on the market in wrapped/packaged form and must be appropriately labeled to provide information to the consumer on preparation/cooking methods and on the risk related to the presence of substances with adverse gastrointestinal effects. 9.2. it should be kept at no more than -9 °C.Wrapping and packaging of fishery products (i) Containers in which fresh fishery products are kept under ice must be waterresistant and ensure that melt water does not remain in contact with the products.2.6 Chapter VI .Storage of fishery products (i) Fresh fishery products.7 Chapter VII . 206 . Fishery products contaminated with parasites may not be placed on the market for human consumption. E. Fresh.2.

4. Sebastichthys capensis. The temperature of frozen fishery products must not fluctuate above -15°C. Helicolenus dactylopterus. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability.Transport of fishery products The above points for Chapter VII apply in relation to transport.8 Chapter VIII . The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. provided that the journey is short and the competent authority allows this.2. melt water must not remain in contact with the products. 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.2. (ii) If fishery products are kept under ice. species belonging to the Merlucciidae family.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. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. 9. (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 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.). (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar.2. species belonging to the Gadidae family. 2074/2005. 9.

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

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

3 Chapter III .3. Before dispatch. requirements if there are many batches of live bivalve molluscs. for: (i) paralytic shellfish poison (PSP). (iv) yessotoxins. the levels of marine biotoxins should not exceed the following limits. and there are requirements for tanks and water storage containers.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. 800 micrograms per kilogram. 20 milligrams of domoic acid per kilogram (iii) okadaic acid.4. shells of live bivalve molluscs are to be washed throughout with clean water and the molluscs are to come from one of the following: (i) Class A production area (ii) Relaying area (iii) Purification centre (iv) Another dispatch centre 9.FISH AND SEAFOOD 9.4.3.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. containers and a label to identify purification. 210 . the purification system to allow them to resume /maintain filter-feeding activity. 1 milligram of yessotoxin equivalent per kilogram.5 Chapter V . 9.4 Chapter IV . Requirements for dispatch centres Handling should not result in contamination to any degree. (ii) amnesic shellfish poison (ASP).Hygiene requirements for purification and dispatch centres A.3. Requirements for purification centres There are requirements for live bivalve molluscs to be washed. 160 micrograms of okadaic acid equivalents per kilogram. dinophysistoxins and pectenotoxins together. B.4.

9. the label.8 Chapter VIII .Identification marking and labelling In addition to general identification marking (contained in 9.3.3.Specific requirements for pectinidae harvested outside classified production areas Requirements for harvesting and handling following harvesting and health standards for live bivalve molluscs must be met.6 Chapter VI .4. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.3. 9. the species of bivalve mollusc and date of packaging (at least the date and month) are required.4.3.2. they cannot be re-immersed in or sprayed with water. must be waterproof.4. Also.7 Chapter VII . by way of derogation from directive 2000/13/EC. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. 9. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.4. Requirements of Chapters III and IV are to be complied with. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction. Finally.1).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. including the identification mark. 160 micrograms of azaspiracid equivalents per kilogram. dispatch centre or processing establishment.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.LEGISLATION (v) azaspiracids.4. (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. Registration document requirements also apply.9 Chapter IX . 211 .

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

Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. that microbiological criteria and chemical contaminants amongst other requirements are met. in 90% of the samples. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. live bivalve molluscs must not exceed 46. 000 E. live tunicates and live marine gastropods.600 E. three dilutions MPN test specified in ISO 16649-3. 213 . Also. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. - - - - B.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. as previously stated. Live bivalve molluscs from Class C areas must not exceed 46. coli per 100 g of flesh and intravalvular liquid. Regulation (EC) 854/2004.2. In the remaining 10 % of samples. Live bivalve molluscs from Class B areas must not exceed. The reference method for this analysis is the five-tube. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3.000 E. A summary is given as follows: 9. Classification of production and relaying areas: This section reinforces that specified in Annex III.2 This states official controls for the production and placing on the market of live bivalve molluscs. 4. section VII. coli per 100 g of flesh and intravalvular liquid. The reference method for this analysis is the five-tube. chapter V is reinforced.5.1 Chapter II .5. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. Annex II 9. live echinoderms. Sampling plans and frequency are required. coli per 100 g of flesh and intravalvular liquid.Official controls concerning live bivalve molluscs from classified production areas A.

opening or closing a production area. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No.5. Additional monitoring requirements This concerns the competent authority in relation to monitoring of classified production areas where bivalve mollusc harvesting is forbidden and of relaying and production zones to ensure there is no malpractice etc. the competent authority is to close the production area. 214 .Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. 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) 2076/2005.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. Procedures for re-opening a closed production area are stated D. 9. E. which has applied since 1 January 2006. 2073/2005.6 Regulation (EC) No. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying.5.2 Chapter III .2. F. boundaries or class of a production area or its closure. Recording and exchange of information Requirements are stated for competent authorities in relation to establishing and maintaining a current list of approved production and relaying areas and their role to inform interested parties of any changes of location.FISH AND SEAFOOD C. 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. 9. 9.

per unit(s) of mass. and/or on the quantity of their toxins or metabolites. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. and the legislation applies to all businesses involved in food production and handling. ensuring that foods being placed on the market do not pose a risk to human health. a batch of foodstuffs or a process. Two kinds of criteria have been established: food safety criteria. 9.6. based on the absence. 9. volume.4 referring to fishery products.6. area or batch.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. The relevant criteria are outlined in Table 9.I. with chapter 2.II.1 Food safety criteria Chapter 1 of the regulation focuses on food safety criteria that cover foods such as ready to eat foods and fishery products and live bivalve molluscs. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product.LEGISLATION The aim of this legislation is to complement food hygiene requirements. The criteria are outlined in Table 9. presence or number of microorganisms. 215 .

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.2 Ready-to-eat foods able to support the growth of L.TABLE 9.3 Ready-to-eat foods unable to support the growth of L. monocytogenes. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life . other than those intended for infants and for special medical purposes EN/ISO 11290-1 Products placed on the market during their shelf life Before the food has left the immediate control of the food business operator. monocytogenes 1. monocytogenes L.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. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1. monocytogenes. other than those intended for infants and for special medical purposes(4) (8) L.

coli (15) 1.17 Live bivalve molluscs and live echinoderms.27 Fishery products which have undergone enzyme maturation treatment in brine. tunicates and gastropods 9 (18) 2 E.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. 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. 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.

. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. AOAC Internat. beer. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . J. shall not apply.. Malle P. soft drinks. — — bottled or packed waters. 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. live bivalve molluscs. wine. Scombresosidae. Valle M. 79. when he is not able to demonstrate. according to which the whole batch is to be deemed unsafe. — bread.94.. Products with pH ≤ 4.1. — fresh. to the satisfaction of the competent authority. products with a shelf-life of less than five days shall be automatically considered to belong to this category.0 and aw ≤ 0. products with pH ≤ 5.. to the satisfaction of the competent authority.1 . spirits and similar products. m = M. 43-9. cider.4 or aw ≤ 0. subject to scientific justification.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. 1996. References: 1. coli is used here as an indicator of faecal contamination. sugar. excluding sprouted seeds. when recontamination is not possible after this treatment (for example. This criterion shall apply to products before they have left the immediate control of the producing food business operator. Bouquelet S. The most recent edition of the standard shall be used. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. Coryfenidae. Dervin C. Bouquelet S.. Clupeidae. honey and confectionery. including cocoa and chocolate products.25.92. Other categories of products can also belong to this category. c = number of sample units giving values between m and M For point 1. Pomatomidae. 2. Particularly fish species of the families: Scombridae. 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. Single samples may be taken at retail level. Assay of biogenic amines involved in fish decomposition. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. Duflos G. E. uncut and unprocessed vegetables and fruits. A pooled sample comprising a minimum of 10 individual animals. monocytogenes. that the product will not exceed the limit 100 cfu/g throughout the shelf life. Malle P. 5 This criterion shall apply if the manufacturer is able to demonstrate. Engraulidae. biscuits and similar products. products heat treated in their final package).

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

TABLE 9.1 Shelled and E.4. coli 5 shucked products of cooked crustaceans and molluscan shellfish 2 100 cfu/g 1000 cfu/g EN/ISO 6888-1 or 2 FISH AND SEAFOOD Coagulasepositive staphylococci 5 Improvements in production hygiene 220 .II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2.

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

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

Regulation (EC) No.2004. 1019/2008 and as read with Regulation 2073/2005. 83) as amended by Regulation Nos 882/2004. L139. 9. 1662/2006. 25. Commission Regulation (EC) No. 62) as amended by Regulation (EC) No.4. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. The revised text of Regulation (EC) No. 2076/2005. 22. as amended.6. Regulation (EC) No. 2074/2005.2004.LEGISLATION 9. 854/2004 is now set out in a Corrigendum (OJ No. Regulation (EC) No. 2074/2005. L226.12.4.2005.6. 1441/2007. 22) as amended by Regulation (EC) Nos. 2076/2005. 206). 25.9 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. 25.2006. Regulation (EC) No. 30.2004. The revised text of Regulation (EC) No. Regulation (EC) No. 1663/2006. Regulation(EC) No. 2076/2005. laying down common marketing standards for preserved sardines. 55).2004.4. 2075/2005 and Regulation (EC) No. 1). 1791/2006 and 1020/2008 and as read with EC Directive 2004/41.2004. L226. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No.2004. 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. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. L283. L139. Regulation (EEC) 2136/89. 2074/2005. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 223 . L139. 853/2004 is now set out in a Corrigendum (OJ No.10 Further Reading Regulation (EC) No.6. 2076/2005. 14. 1. The revised text of Regulation (EC) No. 1688/2005. 30. L226. 2073/2005 on microbiological criteria for foodstuffs (OJ No. namely. 852/2004 is now set out in a Corrigendum (OJ No. as read with the corrigenda at OJ No. 30. 2074/2005 and Regulation (EC) No.10. 3) as amended by Regulation (EC) No. L338. these are not be discussed here.

22.2005. 22. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. 23.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.10. 21.12. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.1.2000. 6) 224 .2005. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.2001. 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 854/2004.12. 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. (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.

0 . 0.3 .5 pH Under otherwise optimal conditions.5. At pH 4. a 10 to 1.1.1 10.5 minutes.1. no growth is observed (1).1. PATHOGEN PROFILES 10.1. Morphology Gram-negative rods. 225 .4 Heat resistance Aeromonads are sensitive to moderately high temperatures.10. The optimum growth temperature is 28 °C (2).5.000-fold increase in number is observed after one week in food systems (1).3. respectively (1). under optimal incubation temperatures. and 2. At 5 °C.5 µm 10.1 Aeromonas spp. 10. The minimum pH value for growth is thought to be pH 5.3 minutes. 10.1.1. Aeromonas has the potential to grow over a pH range of ca 4 . 10.2 Oxygen requirements Facultative anaerobe. Some death will occur at this pH if the temperature is 4 °C.3 Temperature Aeromonas grows at temperatures of 2 .10 (3). and D-values at 45 and 51 °C have been reported as 29.0 x 1.45 °C (1).

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

10. the minimum aw for growth may be lower. The organism will grow well in the centre of meat or poultry dishes.21 minutes) (9).97 for Group II (5).94 for Group I and 0. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains.3.2 Oxygen requirements C. 11)).3 . 9).6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. 10. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. It will not.g. they may survive mild heat treatments (70 .is an obligate anaerobe.97) (10.PATHOGEN PROFILES 10. particularly by cooking. safety margins must be introduced to allow for process variability. grow on the surface of foods unless they are vacuum or gas-packed. These concentrations correspond to limiting aw of 0.2.0 .0. 227 . These values have been established under carefully controlled laboratory conditions.1 Clostridium perfringens Morphology Gram-positive spore-forming rods.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e.7 Characteristics of C.0 µm.9 x 2. 0. therefore. 10. In commercial situations. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. parcooked meals with pH value > 5. Note also that if humectants other than salt are used. where oxygen levels are reduced.1. vacuum packed. botulinum spores The most heat resistant spores of Group I C. botulinum depends on solute.6 and pH 5.10. respectively (8.3. 10.2.0 and aw > 0. However.like other clostridia . pH and temperature.3 10. perfringens .5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4.2.

however. Some studies indicate that the presence of 3% NaCl delays growth of C. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. perfringens in roast beef at 60 ºC is 14. While some growth can occur at 50 ºC. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. although C.8% inhibit growth of most C. 16).97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. perfringens spores The spores of C.3. Its optimum temperature for growth is 43 . although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. perfringens is not tolerant of low water activities. The D-value for C. will survive greater extremes of pH (and aw) (12. 13).3.3. Recorded heat 228 . pH. etc.50 ºC. It grows best at pH values between 6 and 7 (the same pH as most meats).10 ºC.3. Under otherwise ideal conditions. As in the case of other factors limiting the growth/survival of this organism. perfringens is not a tolerant organism with respect to pH. and their heat resistance is also affected by the heating substrate. In addition. The lowest aw recorded to support the growth of C.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C.3. perfringens in vacuum packed beef (18).FISH AND SEAFOOD 10. 0 . Spores. 10.5 minutes (17). (It is unusual for spores to be formed in foods after the growth of this organism (14)). perfringens. depending on strain and other conditions.45 ºC.93 and 0. the limits for aw are affected by temperature. perfringens has the potential ability to grow within the temperature range 15 .3 Temperature The most significant characteristic of C. perfringens strains.7 Characteristics of C.3.it is destroyed by heating at 60 ºC for 10 minutes (14. perfringens appears to be 0.6 Aw/Sodium chloride C. 15. 10. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. 18). perfringens can vary quite considerably in their heat resistance. 10. type of solute. lower concentrations may be effective in combination with other factors. 13).5 pH C. Salt concentrations of 6 . At cold temperatures. the enterotoxin is not heat resistant . vegetative cells die rapidly (12). 10.

is between 30 and 37 ºC. growth at low temperatures can be very slow. Under ideal conditions. 10. monocytogenes is not a particularly heat resistant organism.4 .4.0 µm. pH < 5. 10. 0. 229 . 60 and 65 ºC were reported as 10.4. monocytogenes in crawfish tail meat at 55. 10. the lowest limit for growth is likely to be considerably higher .3 . using hydrochloric acid as an acidulant). D-values for L.0.2 has been suggested as the lowest working limit (22). 10. Morphology Gram-positive short rods.0 minutes for heat resistant spores to 1. requiring days or weeks to reach maximum numbers. However.2.at refrigeration temperatures down to. so can be destroyed by pasteurisation.4.3 is the lowest value where growth has been recorded. monocytogenes is reported to be 45 ºC (20).4 ºC is probably a more likely minimum for growth in foods (19). being potentially capable of growing . In foods. 10. however.5 x 0. the organism is able to grow at pH values well below pH 5 (pH 4.19 minutes.2 Oxygen requirements Aerobe or microaerophilic.1 Listeria spp. -0. or even below 0 ºC.4.5 .3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. it is not a sporeformer. and where acetic acid is used as an acidulant. respectively (21).4.4 Heat resistance L.albeit slowly .8 minutes for heat sensitive spores (12).23.64.6 . Its optimum growth temperature.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. however. and temperature.4 10.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17.especially at refrigeration temperatures. The upper temperature limit for the growth of L. 1.2. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.98 and 0.

Morphology Gram-negative short rods.5. monocytogenes is quite tolerant of high NaCl/low water activities.5.83 (20).3 Temperature Plesiomonas can grow in the temperature range of 8 .5.5 pH Plesiomonas has a pH range for growth of pH 4. 230 . 0.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.5 .FISH AND SEAFOOD 10. 0.12% NaCl or more).4. 10. 10. or even grow. The bacterium may survive for long periods at aw values as low as 0.5.3.90.6 10.45 ºC.0 .0 µm. but has been shown to be able to grow at an aw level of 0. 10.8. the optimum being 30 ºC (23).5 .5.5.1 Plesiomonas Morphology Gram-negative. peritrichous flagella.97.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).6 Aw/Sodium chloride L.5 10. 25).0. 10. 10.0 x 1 .7 x 1. 10.6.2 Oxygen requirements Facultative anaerobe. It grows best at aw of ≥ 0.8 . 25). at salt levels found in foods (10 .5 (24. It is likely to survive. 10.1.3 µm.1 Salmonella spp.

although salt tolerance increases with increasing temperature (28). depending on serotype. therefore. D60 ºC values normally range from about 1 to 10 minutes. Salmonellae are quite resistant to drying.PATHOGEN PROFILES 10. 10. Salmonellae are quite resistant to freezing.4 Heat resistance Salmonella is not a spore-forming organism. and with an appropriate acid.8 . high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. pasteurisation and equivalent treatments will destroy the organism under normal circumstances.48 ºC. substrate. Salmonella has the potential to grow at aw levels as low as 0. However.0.5 ºC. The growth of Salmonella is generally inhibited by the presence of 3 . However. or possibly 0.9. and appropriate heat treatments must be determined experimentally for low-aw foods.2 Oxygen requirements Facultative anaerobe. 10. The optimal pH for Salmonella growth is between 6.3 Temperature Salmonellae can grow in the temperature range of 7 .7.6 Aw/Sodium chloride Where all other conditions are favourable. under otherwise ideal conditions.5.6.93 (as reported in dried meat and dehydrated soup). some strains are able to grow at temperatures as low as 4 °C (26). with a z-value of 4 . Furthermore. It is not. a heat resistant organism. 10. strains vary in their ability to withstand heating. the optimum being 35 .4% NaCl. depending on the type of acid and temperature.945. Some death will occur at pH values of less than about 4. 231 . temperature and pH. Salmonella senftenberg 775W is about 10 . 10.6.5 pH Salmonella has a pH range for growth of pH 3.20 times more heat resistant than the average strain of Salmonella at high aw (27).5 .6.6.37 ºC.5.6. Growth is slow at temperatures below about 10 ºC.

5 ºC. etc.20 minutes. Most strains of staphylococci can grow within the pH range 4.7. Freezing and thawing have little effect on Staph.1 Staphylococcus aureus Morphology Gram-positive cocci.3 Temperature Under otherwise ideal conditions. Staph. aureus can grow within the temperature range 7 – 48.9 µm diameter. The limits for toxin production are also narrower than for growth. In meat macerate. aureus is more limited under anaerobic than aerobic conditions.).4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH.7. aureus than citric acid). Under most circumstances. aw. the D60 ºC value is 2 .7 .7. under otherwise ideal conditions (29. As is the case with other bacteria.7. 31).2 Oxygen requirements Facultative anaerobe.37 ºC (29). 10. It can survive well at low temperatures. the organism is heat sensitive and will be destroyed by pasteurisation.0. 232 . with an optimum of 35 .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. 10. NB: The growth of Staph.7 10.2 to 9. 0.0 . aw and temperature (sensitivity to acid increases with temperature). but may cause some cell damage (30). 10.7. fat content. however.7.5).FISH AND SEAFOOD 10.3 (optimum 7. 10. The following relate to limits for growth only. aureus viability. stressed cells can also be less tolerant of heating. depending on aw.

9.99 aerobically under otherwise optimal conditions. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.2.2 .7. An effect of increasing salt concentration is to raise the minimum pH of growth.8 x 1.7 Limits permitting toxin production 10 .0 .46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5.8. 0.8 10.86 is the generally recognised minimum in foods (32).5 .6 µm 10.5) (29.0 (optimum 7.87 and > 0.0. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant. Staphylococci are more resistant to salt present in foods than other organisms. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. an aw of 0. 34).7. However.3 Temperature The optimum temperature for growth has been reported as 30 ºC. It can grow over the aw range 0.8.>0. atmosphere.10% NaCl. 31) between 0. 233 .4 . and solute.1 Vibrio cholerae Morphology Gram-negative curved rods. pH.7. 10. 10.43 ºC (35). aureus can grow in 7 . but certain strains can grow in 20% NaCl. 10. with a temperature range for growth of 10 . In general.2 Oxygen requirements Facultative anaerobe. In general. Staph.83 . strain.PATHOGEN PROFILES 10.8. * dependent on temperature.6 Aw/Sodium chloride Staph.99 little or no toxin production in anaerobically packed foods.

cholerae in crab meat homogenate at 60 ºC is 2. there is evidence that V.4. 10. respectively.9. with an optimum of 0.9. 37. declining in number during storage.65 minutes (35). 10.97 . parahaemolyticus is easily destroyed by drying.5 pH This organism is tolerant of high pH values.0% NaCl (35). V.9.1 .6 Aw/Sodium chloride V. with optimum at 7.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.8. it has a pH range for growth of pH 5 .998.FISH AND SEAFOOD Also. It can grow in the presence of 0. cholerae may persist on frozen fish for >180 days (36). but only moderately sensitive to freezing (40).1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. The D-value for V.5 ºC).9.6.2 Oxygen requirements Facultative anaerobe.9 10. 10. under otherwise favourable conditions. 234 .6 (35. cholerae is sensitive to heat at temperatures of >45 ºC. 10. cholerae can grow over the aw range 0. are 5 and 43 ºC.984 (35).4 Heat resistance V.0. and it is sensitive to refrigeration temperatures (0 .8. 38). 10. with an optimum of 37 ºC (39). 10. but not acid conditions.8.

10 Vibrio vulnificus 10. depending on solute.10.11. and are readily destroyed by cooking.98 (35.PATHOGEN PROFILES 10. but the optimum concentration is 2 .3 Temperature The optimum temperature for growth has been reported at 37 ºC. 235 . vulnificus is heat sensitive.10. 10. 10. 41).10.10. The optimum concentration of NaCl equates to aw of approximately 0. with an optimum pH of 7. parahaemolyticus are not heat resistant. 10. 41).8 .5% NaCl. and 2.5 pH The pH range for growth is 4.8. parahaemolyticus will grow at aw levels as low as 0. respectively. respectively (39). it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).4 Heat resistance V. V. The organism can survive for long periods at freezer temperatures (42). 10.4% (39.5 minutes.0.9.1 Morphology Gram-negative curved rods.922.9. 10. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.6 Aw/Sodium chloride V. The D-values at 55 ºC for clam and crab homogenate are 0.29 minutes.9. parahaemolyticus is unable to grow in the absence of salt.4 Heat resistance V. 10.02 .43 ºC (42). with a temperature range for growth of 8 .6 .2 Oxygen requirements Facultative anaerobe.6 (41).

8 (42)..W. Aeromonas hydrophila. Marcel Dekker.. Hui Y. in Foodborne Bacterial Pathogens. Montville T.R. in Foodborne Disease Handbook.H. Doyle M. in Food Microbiology: Fundamentals and Frontiers. 236 . Marcel Dekker. Marcel Dekker. 4. Eds.L. Microbiology and Molecular Biology.P. 6 (1). Clostridium botulinum. with an optimum pH of 7. Clostridium botulinum. Foegeding P.. 11.W. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.10.L. Austin J. ASM Press. 6. Palumbo S. Ed.H. Hauschild A. Eds.R. Aeromonas hydrophila group. 1993. in International Handbook of Foodborne Pathogens.W.FISH AND SEAFOOD 10.H. 2001. 5. 1994. Doyle M.H.. International Commission on Microbiological Specifications for Foods.. References International Commission on Microbiological Specifications for Foods. 97-131. Blackie. Gorham J. Clostridium botulinum and Clostridium perfringens. Juneja V... New York. Diseases Caused by Bacteria.5.5 . 57-63.. Eds. Hauschild A. Ed.. Bhunia A.P.5 pH The pH range for growth is 5 . in Clostridium botulinum: Ecology and Control in Foods. Ed. 5-19. 1-19. Principles of control.N.. 9. Murrell K.N. Stelma G.. Kovacek K. Notermans S. with an optimum of 2. 1-27.5% (42).M..D. Hui Y. 383-408. 2005. Fratamico P. Novak J.E. 10. 1996..O. in Foodborne Pathogens. Eds.. Doyle M.. Marcel Dekker.J. Cliver D. vulnificus grows over the range 0. Volume 1: Diseases Caused by Bacteria. Potential hazards associated with REPFEDS.. 329-49.D. Gorman J. Marcel Dekker. Peck M. 111-89.0% NaCl. 1989.10. Lund B. Kim J..D. 10. 1994. 1995. Washington D. London. Dodds K. Volume 1.W..10. Aeromonas.M. Johnson E..M. 1993. New York. 10. Stelma G. in Foodborne Disease Handbook. Cliver D. New York. Eds.C. Miliotis M.D. Beuchat L.A. New York.11 1. Marcel Dekker Inc. New York.W. Great Britain. 279-303. Caister Academic Press. 2003. 3. 7. Abeyta C. 8.O. 357-67.L. in Microorganisms in Foods. Bier J. Food Control.L. Eds. Clostridium botulinum. Dodds K.. New York.H.6 Aw/Sodium chloride V. Faris A.. Eds. Aeromonas species. Hauschild A. Dodds K. in Clostridium botulinum: Ecology and Control in Foods.K. New York. Gaze J.. 121-76.P. Murrell K.. 1989. 2.H. Smith J. in Foodborne Bacterial Pathogens.R. Volume 5: Microbiological Specifications of Food Pathogens. Marcel Dekker. Betts G.

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

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

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

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gov.gsi.org.uk Web site: www.uk or v.org.org.org/ .gov.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.ifm.uk Web site: www.gov.scotland.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.co.defra.gsi.uk Web site: www.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: cawoods@denholmseafoods.uk/Topics/Fisheries Seafish Industry Authority 18 Logie Mill Logie Green Road Edinburgh EH7 4HS United Kingdom Tel: + 44 (0)131 5583331 Fax: + 44 (0)131 5581442 Web site: www.uk Prepared Fish Products Association (PFPA) Federation House 6 Catherine Street London WC2B 5JJ United Kingdom Tel: + 44 (0) 208 78362460 The Scottish Government Fisheries Group Environment and Rural Affairs Department Pentland House 47 Robb's Loan Edinburgh EH14 1TY United Kingdom Tel: + 44 (0) 8457 741741 or + 44 (0)131 5568400 Fax: + 44 (0)1397 795001 Email: ceu@scotland.seafish.gov.holt@ifm.

Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo.dk Web site: www.bim.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.gov. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.matis. Northern Ireland Fisheries and Rural Policy Division 4th Floor North Dundonald House Stormont Estate Belfast BT4 3SB United Kingdom Tel: + 44 (0) 28 90524999/ 4991 Fax: + 44 (0) 28 90378323 Email: dardhelpline@dardni.uk/index/fisheriesfarming-and-food.gov.suite.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: matis@matis.dardni.dk Web site: www.uk Web site: www.ie/templates/homepage.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.is/ 242 .fiskeriforskning.is Web site: www.C.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: post@fiskeriforskning.dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .eapo.dk Danmarks Fiskehandlere H.FISH AND SEAFOOD Department of Agriculture and Rural Development.fiskehandlerne.no Web site: www.org/fishery/ European Association of Fish Producers’ Organisations H. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.difta.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.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.fao.ifpea@eircom. Andersens Boulevard 37.org Web site: www.com Web site: www.

fdf.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.nz Web site: www.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: reception-cmar-hobart@csiro.co.ifst.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: generalenquiries@fdf.org.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.cmar.uk 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.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.ifr.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.com/ 243 .gsi.seafood.hpa.org Web site: www.co.dh.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.uk Web site: www.gov.bsi-global.org.gov.org.au Web site: www.uk/en/index.com Web site: www.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.ac.

int Web site: www.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.org.who.sofht.co.scot.uk Web site: www.nhs.nhs.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.hps.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: hpsenquiries@hps.int/foodsafety World Health Organization (WHO) Headquarters CH .se Web site: www.uk/index.The Swedish Institute for Food and Biotechnology Box 5401 SE-402 29 Göteborg Sweden Tel: + 46 31 3355600 Fax: + 46 31 833782 Email: info@sik.asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.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.who.who.sik.codexalimentarius.org Web site: www.euro.sfam.uk Web site: www.php Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: admin@sofht. Web site: www.who.net/web/indexen.int/en/ WHO Regional Office for Europe Scherfigsvej 8 DK-2100 Copenhagen Ø Denmark Tel: + 45 39 171717 Fax: + 45 39 171818 Email: postmaster@euro.de 244 .int.scot.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: foodsafety@euro.co.who. jsp Food Safety 10 .int Web site: http://www.uk/scieh.euro.int/ SIK .

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

FISH AND SEAFOOD 246 .

growth and survival in crustacean shellfish 86 Aeromonas sobria. 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. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. 111 Astroviruses. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. to test microbiological criteria 19 Aeromonadaceae. preserved by dry/wet salting 99 Anisakis spp. 102 Bacillus spp.INDEX Achromobacter spp. 69-70 Aeromonas hydrophila. source of PSP toxins 142 Alexandrium ostenfeldii. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. causing spoilage of dried fish 110. 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 . 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. as autochthonous bacterial flora of fish 3 Aeromonas spp. in crustacean shellfish 87 Alteromonas spp. planktonic source of spirolides in shellfish 163 Allergens.

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

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

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

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

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

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

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

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

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

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