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

The contents of this publication are copyright and reproduction in whole, or in part, is not permitted without the written consent of the Chief Executive of Leatherhead International Limited. Leatherhead International Limited uses every possible care in compiling, preparing and issuing the information herein given but accepts no liability whatsoever in connection with it.

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

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4 Spoilage 1.6 Published Microbiological Criteria 4.2 Initial Microflora 4.8 References MOLLUSCAN SHELLFISH 3. vii .6 Published Microbiological Criteria 3.2 Initial Microflora 3. 4.7 References 3.6 Pathogens: Growth and Survival 1.3 Processing and its Effects on the Microflora 1.3 Processing and its Effects on the Microflora 4.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2. CHILLED AND FROZEN RAW FISH 1.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.8 Further Reading CRUSTACEAN SHELLFISH 4.5 Pathogens: Growth and Survival 3.7 References 4.2 Definitions 2.3 Processing and its Effects on the Microflora 3.5 Spoilage 2. 3.5 Pathogens: Growth and Survival 4.4 Spoilage 4.1 Definitions 1.8 References 1.7 Published Microbiological Criteria 2.4 Spoilage 3.6 Pathogens: Growth and Survival 2.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.2 Initial Microflora 1.5 Factors Affecting Fresh Fish Spoilage 1.4 Processing and its Effects on the Microflora 2.7 Published Microbiological Criteria 1.1 Definitions 3.1 Definitions 4.3 Initial Microflora 2.1 Introduction 2.

2 Definitions 8.3 Processing and its Effects on the Microflora 5.8 Guidance 222 9.8 Ciguatoxins 7.3 Regulation (EC) No.4 Implementation and Review of the HACCP Plan 8. EU FOOD HYGIENE LEGISLATION 189 9. 7.10 Cyclic Imines 7. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.10 Further Reading 223 viii . 2006 No. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.8 Further Reading FERMENTED FISH 6.3 Processing and its Effects on the Microflora 6.3 Stages of a HACCP Study 8.1 Introduction 189 9. 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. S.5 Pathogens: Growth and Survival 6.5.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.6 References 6.2 Initial Microflora 6.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.5 Regulation (EC) No.1 Introduction 7.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.9 Other Relevant Legislation 223 9.4 Amnesic Shellfish Poisoning (ASP) Toxins 7. SMOKED AND DRIED FISH 5.7 References 5.9 Azaspiracids 7.4 Spoilage 5.5 Pathogens: Growth and Survival 5.11 Conclusion 7.7 Food Hygiene (England) Regulations 2006.1 Definitions 5.6 Regulation (EC) No.7 Further Reading FISH AND SHELLFISH TOXINS 7.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.3 Tetrodotoxin 7. 14 (Hygiene requirements specific to the UK) 221 9. 8. 9.6 Published Microbiological Criteria 5.2 Initial Microflora 5.1 Introduction 8.2 Legislative Structure 190 9. 852/2004 on the General Hygiene of Foodstuffs 191 9. CURED.6 Lipophilic Shellfish Toxins (LST) 7.1 Definitions 6.4 Regulation (EC) No.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 6.I.

10 PATHOGEN PROFILES 10.3 Clostridium perfringens 10.8 Vibrio cholerae 10.4 Listeria spp.10 Vibrio vulnificus 10.9 Vibrio parahaemolyticus 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .7 Staphylococcus aureus 10. 10. 10.1 Aeromonas spp.2 Clostridium botulinum 10.6 Salmonella spp. 10.5 Plesiomonas 10.

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

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

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

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

5% (1). and other forms of aquatic animal as part of their ‘fish’ definition. Fish fillets are a poor source of carbohydrates. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. In this chapter. Fish can be divided into fatty and lean fish.21%. and excludes molluscs and Crustacea. most fish contain omega 3-fatty acids and other essential nutrients. typically 3 and 15% of total weight. Fish flesh is dominated by the abundance of white muscle in relatively short segments.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. offering less than 0.2 . migratory patterns. giving it its characteristically flaky structure. but typically present a low fat percent when compared to other meats. Fish fillet composition can vary significantly within the same species due to feed intake. ‘fish’ also refers to the flesh of such animals used as food. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. Protein content in fish fillet varies typically from 16 .20%. and is mostly interspersed between the muscle fibres.1. In addition. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. In addition. Fish are an important part of a healthy diet since they contain high quality protein. Muscle composition and structure of fish also differ from those found in other meat. which can be up to 67%. lean fish are those fish that store most of their fat in the liver. fins and a streamline body. crustaceans. The lipid content. “fish” will be used for fresh and seawater finfish. while fatty fish have fat cells distributed along their bodies. CHILLED AND FROZEN RAW FISH Associate Prof. The connective tissue content of fish is also lower than that found in meat. The lipid fraction is the component showing the greatest variation. and spawning season. This super class of vertebrates includes all the bony and cartilaginous finfish. typically fluctuates between 0. it shows a typical season pattern especially in migratory species such as herring or mackerel. However. 1 . respectively (1).

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

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

1 . sewage.37 °C Unknown Unknown TABLE 1.102 101 .14 .26 °C 10 . human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 .0. birds.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.FISH AND SEAFOOD TABLE 1.106 cells/g (exception: V. 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.>106 Salmonella spp.103 High (107 . estuarine environments.35 °C 8 . and risk associated with the consumption of seafood is low (2. Toxin levels 0. vulnificus) 5 .109 cells/g) * Some sources considered L.II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil. Plesiomonas shigelloides 3 . 4 .19 µg/kg bodyweight from < 102 .37 °C High for most species. In addition.106 cells/g. numbers of pathogenic bacteria in raw fish tend to be low. 4). 105 . Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. stream water. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. Escherichia coli Yersinia enterocolitica 101 . However. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria. and mud Ubiquitous.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00.

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

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

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

It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. TMAO) that can be reduced to produce 8 . the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). this problem occurs to a lesser extent with meat. from both a microbiological and a chemical standpoint. adequate product labelling in respect of storage temperature. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. The replacement gas should contain at least 40% carbon dioxide. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. use of high-quality raw fish. These conditions include storage of the product at ≤ 3. Only the highest-quality fish. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. For other less fatty marine fish. The addition of high levels of carbon dioxide to these fish. 2. botulinum. Another problem facing the application of MAP to marine fish is the existence of carbon dioxide-resistant bacteria that can rapidly produce off-odours (e. as already stated. Storage studies have shown that the extension in shelf life of MAP cod fillets was proportional to carbon dioxide concentration up to around 50% (V/V). 3. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. Although all fish species need to be treated separately. adequate shelf life and cooking requirements. may result in unacceptable colour and textural change and hence little shelf life extension (11). The growth of these bacteria is reduced by high carbon dioxide concentrations but. should be used. For fatty fish. however. greater advantages are seen.3 °C at all points from packaging onwards. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1.g.g. The problem of water loss in MAP fish is common. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13). Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. In the United Kingdom. 5. but is recognised as a drawback of carbon dioxide application.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). trimethylamine and sulfides). at all times. owing to the dissolution of cellular structures. Subsequent storage must be at or below 3 °C. 4. however. and the use of a HACCP plan.

Degradation of whole fresh fish stored in ice generally follows a set pattern. 1. so lower carbon dioxide concentrations may have better preservative effects. 17.4. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids).III. The eyes turn from convex and clear to concave and opaque. and metabolic activities of microorganisms present in the fish.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. C (C=reject) Significantly. A. B. reactions caused by activities of the fish’s own enzymes. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). texture. the skin turns from iridescent to dull and bleached with bacterial slime.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). loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. with no smell. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. When working with fillets. TABLE 1. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. Loss of freshness and spoilage cannot be separated as processes.CHILLED AND FROZEN RAW FISH off-odours. as traditional spoilage patterns of the external organs are typically very distinct. Over time. or for that matter evaluation of the taste of the fish.4 1. to brown and slimy with an intense off-odour. the gills from pink and shiny. smell and 9 . and the flesh turns from bright and elastic to dull and soft (1). Taste panel evaluation of cooked fish may be used to score or grade colour. Various methods for whole fish freshness evaluation by a trained panel have been proposed. and this pattern is the basis of freshness grading schemes. these are summarised in Table 1.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. 18) and are probably the most widely used.

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

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

2 Fish-spoilage bacteria The parameters affecting the multiplication of microorganisms in foods have been categorised into two general groups: intrinsic (inherent qualities of the food) and extrinsic (qualities of the food environment) (35). For fresh fish iced immediately after capture and continually stored in ice.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. Also.5 1. The factors are interactive and cannot be completely isolated. The sum of these changes may result in a different spoilage profile. 34). For fresh fish. 1. The concept of specific spoilage organisms (SSOs) is not new. 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. careful temperature control during storage is not only important in terms of quality loss. a minority are often associated with the majority of the spoilage. Fish spoil as a result of the chemical. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity.and temperate-water fish spoilage rates may be many. 5).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 application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 . the lower the temperature of storage. 1. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. but also crucial for the assurance of consumer safety. by altering the gas atmosphere (extrinsic). the slower the spoilage processes proceed (within limits).FISH AND SEAFOOD 1.5. the results of altering one parameter may have far-reaching consequences on others. The reasons for the differences between tropical.5.5. biochemical and microbiological reactions taking place within and on the fish. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. For example.

putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s. frozen) fish products. obligate respiratory bacteria. 36). The Pseudomonadaceae family represent a large and poorly defined group of microorganisms. putrefaciens growth may be used as indicators of spoilage (30). As previously noted. It is able to grow as fast as. ‘oniony’ and ‘faecal’ odours from the production of ketones. Sh. it is often isolated as about 1 .g.1 Pseudomonas spp. more especially. It is thus an important spoilage organism of gadoid fish such as cod. Pseudomonas spp.5. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. 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. putrefaciens spoilage of fish is due to its biochemical action on muscle. 35. therefore. motile with polar flagella. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. produce hydrogen sulphide (H2S) from cysteine. plus other characteristic compounds of the species responsible for spoilage.5. During the 1970s. the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. 35). They are generally characterised as Gram-negative rods. i.2 Shewanella putrefaciens Sh. oxidase-positive. (9) and they are considered SSO of iced freshwater fish (35).CHILLED AND FROZEN RAW FISH started in the 1940s. Members of the genus are able to produce pigments. 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. or faster than the rest of the flora of ice-stored fresh marine fish (9). processed (e. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. which produced the characteristic spoilage compounds of the fish (30.3. It is also present in fresh water. 13 . on fish are diverse and in many cases species-specific.e. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. mediated spoilage is characterised by ‘fruity’. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp. its ability to reduce TMAO to TMA. 37). The importance of Sh. aldehydes. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp.3. 1. These organisms are described as potential spoilers. the spoilage of a product may be strongly influenced by the conditions under which the product is held. although only since the late 1960s has it been realised that specific metabolites of Sh. catalase-positive.10% of the total flora of fresh fish from temperate marine waters.

owing to its cell size and activity. It is also able to produce hydrogen sulphide and a range of other off-odour compounds. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39).how to reduce a molecule that they are not able to take into their cells efficiently. Shewanella gelidimarina. The prevalence of the bacterium in so many environments. putrefaciens lies in its chemotaxis: the bacterium. Sh. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). Shewanella frigidimarina. and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. 38). like all members of the Pseudomonadaceae. as the ferric iron is insoluble. and Shewanella aquimarina. a population of Ph. The ferric reductase activity of Sh. phosphoreum sufficient to cause spoilage was found. at the time of rejection. Initially. putrefaciens growth has been collected (12. putrefaciens has been studied. Shewanella oneidensis. unlike many other motile bacteria. Later. 41). Ph. it is strictly respiratory. Shewanella waksmanii. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). phosphoreum were required for 50% of taste panellists to reject a sample. Shewanella affinis. Shewanella livingstonensis. Another distinct feature of the physiology of Sh. In fact. although. In 50% N2/50% CO2 MAP cod. Sh. putrefaciens were identified as Shewanella algae. This wide-ranging respiratory capability is thought to be unique (39). Research has shown that approximately 107 cfu/g of Ph. baltica has been identified as the main H2S producer in cod during cold storing (37). Shewanella denitrificans. 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 (23.5. Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). putrefaciens strains are characterised as microaerophiles or anaerobes. Shewanella olleyana. 1. 41). putrefaciens was not present in such numbers (23. phosphoreum has also been identified as responsible for histamine fish poisoning. does not show carbon-source chemotaxis.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. with several new species being described. putrefaciens were required. mesophilic strains of Sh. In terms of its taxonomy. and recent taxonomic description using modern molecular methods bifurcated this species. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). Ph. it was proposed that this organism. Sh. but Sh. additional psychrotrophic species such as Shewanalla baltica. the bacteria have a significant problem to overcome . they are heterogeneous. and Shewanella profunda were described. This type of intoxication occurs when bacteria convert the histidine 14 . whereas >108 cfu/g Sh.3.

B. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). especially type E botulinum intoxication. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. thermosphacta in the spoilage of some MAP fish. B. 5.4 Brochothrix thermosphacta and lactic acid bacteria B.CHILLED AND FROZEN RAW FISH present in fish into histamine. When inoculated into VP corned beef and sliced ham. Recent studies have investigated the dominance of B. 75% (with a 35% mortality rate) occurred in Japan.6 Pathogens: Growth and Survival Endogenous chemicals. 1. thermosphacta did not produce off-flavours until 2 . Review of outbreak data suggests fresh fish to be safe. The heterogeneity of the types (A. lightly preserved fish products are associated with botulism. this is probably due to spoilage occurring before toxin elaboration. including antisera. 1. algal toxins. 5) reports that fresh fish consumption has never been shown to cause human botulism. Growing evidence suggests a role for B. Of 404 intoxication outbreaks of type E C. Fewer still are capable of growing on fresh fish. botulinum. these risks are substantially reduced (1. Of these.1 Clostridium botulinum C. it is bacterial risks that increase after capture of the fish. human viruses. presents a potential risk.e. However. Types A. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. they do not produce significant product spoilage).6. Some Ph. where a particular 15 . a convenient but diverse species of bacterium with a number of different types. C1. C2. (and very rarely F and G) have been reported to cause human disease. Adult disease is caused by the production of the botulinum toxin. botulinum recorded up to 1963. E and F include bacteria with minimum growth temperatures of approximately 3. but with proper cooking. 6).3. E. bacteria and higher parasites all present some risk associated with fish consumption (1. F and G) is well documented (44). D.5. age. types B. Huss (1. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. E.3 °C (Group II) and these are non-proteolytic (i. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. B.3 days after having reached 108 cfu/g (42). 5). MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). previous exposure and access to supportive treatment. 1. Acetate production has been reported as a good indicator of spoilage by this organism.

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

Although the psychrophiles are unable to grow at body temperature.3 Aeromonas Aeromonas spp. is historically associated with faecally contaminated water. cytotoxic. The major virulence factor appears to be the production of toxins with enterotoxic. with minimum growth temperatures around 0 °C and optima of 15 . Since 2000. shigelloides is similar to Aeromonas spp. and proper cooking will substantially reduce risk. infection. P. and especially estuarine environment. but the bacterium is known to survive and grow in the shallow marine. their potential to produce disease-causing exotoxin in food has not been fully examined (48).4 Listeria monocytogenes L. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp.6. 5. in assessing the risk of fishborne infection by P. L. Similar problems as those encountered for Aeromonas spp. Again. but proper cooking should substantially reduce risk of infection. coli on isolation media (50). listeriosis cases have been reportable to the CDC. sodium channel blocking and haemolytic activity (48). others are psychrotrophic mesophiles (49). proper chill storage will eliminate any risk associated with this organism. shigelloides are encountered. Chill storage may not eliminate growth. and fish and shellfish are probably the major reservoirs for the organism (50). monocytogenes has been well documented as a foodborne human pathogen. Some isolates are true psychrophiles. There is no risk associated with the consumption of properly chilled and cooked fresh fish. in its habitat. They are aquatic organisms with an epidemiology that is yet to be fully understood.CHILLED AND FROZEN RAW FISH disorders). It is known that Aeromonas spp. 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. In the US. Despite this.55 cases per 100. are present on fish and can grow to significant numbers during storage (1. 1. monocytogenes incidence has been between 0. and especially Aeromonas hydrophila. however. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis.000 17 . though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. possibly due to under-reporting because of their likeness to E. cholerae O1. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. V. 17).26 to 0.20 °C. the causative agent of cholera. there is no risk from the consumption of properly handled and cooked fresh fish. are associated with human diarrhoeal illnesses.. 1.6. which is often fatal (>50%). Most cases have been sporadic rather than associated to large outbreaks.

Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . Klebsiella pneumoniae and Hafnia alvei. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). Clostridium spp. Like other toxins. The organism poses a serious risk in chilled products not cooked before consumption. there is a possibility for growth of the organism on chill-stored fish. it is not apparent to the consumer and it cannot be destroyed by cooking.. If fish are not properly refrigerated.48 minutes in salmon have been reported (53). It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. neonates. D60 of 1. and Lactobacillus spp. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae.. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. but cooking will significantly reduce the risk of infection. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). foetuses..6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans.98 minutes in cod and D60 of 4. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. Larvae of parasites such as Anisakis simplex are resistant to curing and marinating but can be easily destroyed by freezing at -17 to -20 °C for 2 hrs. but more especially by Morganella morganii.pregnant women. Fortunately. 1. 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. some Vibrio spp. but proper cooking will reduce any risk. 1. Being a psychrotroph.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. at risk groups . leads to the production of histamine. 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. alcoholics. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. full risk analysis is not possible (52). Photobacterium spp. especially pickled fish. 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. Infections are mostly associated with the consumption of raw or 18 .6. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. Lightly preserved products.FISH AND SEAFOOD persons. It is regularly isolated from seafoods (51).6.

Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers.IV) (57). coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF).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. No.. which in many cases will eliminate the risk.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. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it. 1997) Huss H. For fish from inshore or inland waters of doubtful microbiological quality. including parasites and/or quantity of their toxins / metabolites per unit of mass. especially in warm-water areas and where fish are to be consumed raw. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration. The most widely accepted microbiological criteria for chilled and frozen raw fish are those set for aerobic plate counts (APC) at 25 °C and E. it may be desirable to test for Salmonella and V. Faecal coliform counts may be used instead of E. volume. In summary. TABLE1.1997. Gram L.. EC. Ababouch L. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. Rome.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. area or lot (CAC.H. Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease. coli counts where this method is preferred. aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . 2003. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). parahaemolyticus (see Table 1. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. 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. FAO. parahaemolyticus Staph. 1. 444. long storage under refrigeration or one of the former prior to freezing.

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

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

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

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

Fisheries and Food.. Shahidi F.. 1985. 1992. Simard R. 1997. 1. 271-85. Akiba M. Gibson D. 1986. 73-89. McMeekin T.).G.. in Torry Advisory Note No 93. 15. Lancaster. Kitts D.9.P. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. 55.L. Trolle G. Huss H. Gram L.1 Further Reading Processing Anon. 1961. in Seafood Science and Technology. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. Estimation of the bacteriological quality of fish by automated conductance measurements.. Aberdeen. Ross T.. 24 .S. The Microbiology of Seawater Fish. Houston N.1983. Smith G. Ravn Jørgensen B. 1988. Gibson D. 1984. Labuza T. International Journal of Food Microbiology. 6. Microbiological quality and shelf life prediction of chilled fish. 62-70. Torry Research Station of the Ministry of Agriculture. 317 (Suppl. 1992. Food and Agriculture Organisation. Hobbs G.H.E. International Journal of Food Microbiology. 1984. Sumner J.A.M. Eds. International Journal of Food Microbiology. 13-32. 1989. Florida... Bulletin of the Faculty of Fisheries (Hokkaido University). 35. Dalgaard P..M. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. 1989. Villemure G. 127-34..H. Academic Press. Ogden I.9. Magno-Orejana F. International Journal of Food Microbiology. Cann D. 317-21. 4. Changes in microflora and chemical compounds in mackerel stored by partial freezing.C... Olley J. Aberdeen... Huss H. Food and Agriculture Organisation of the United Nations.. Taoukis P. Cho Y. Application of predictive microbiology to assure the quality and safety of fish and fish products. Torry Research Station of the Ministry of Agriculture.H. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. Huss H... Picard G.. Ed. 1.9 1.D. Jones Y. Handling and processing of scad..D. Fisheries and Food. 65-72. in Fish as Food. Journal of Food Science. 51. Rome. 741-50. Borgstrom G.FISH AND SEAFOOD 1. Shinano H. Technomic Publishers. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere.2 Spoilage Shewan J. Ed. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches.. Torry Research Station. 1. 295-307.M. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Journal of Food Protection. in FAO Fisheries Report No..

Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. 1993. 1992.9.. 1.J..H. 4.. 1987. http://eur-lex.eu/LexUriServ/LexUriServ. Ed. in Quality Assurance in the Fish Industry. Hall G. Liston J.. Garcia M. Jacobsen M. Amsterdam.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. Huss H. 59.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. Gram L.. A rapid method for detection of histamine producing bacteria. Applied and Environmental Microbiology..M. 72. Journal of Applied Bacteriology. International Journal of Food Microbiology. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila.. Gram L.europa. 1997. 1. 5. London. 20023.9.R.CHILLED AND FROZEN RAW FISH Einarsson H. 2197-203. Eds. 322-6. International Journal of Food Microbiology.H. 479-88. 1987. 137-46.H. Huss H.3 Pathogens Condon S. Trolle G. Sala F. 65-72. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. Effect of culture age. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Huss H. Klausen N. 25 . Blackie Academic & Professional.. 1992. Otero A.L. Modified atmosphere packaging of fish and fish products. in Fish processing technology. Elsevier Science.K.

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

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

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

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

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

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

3 Preparation of fish component of chilled and frozen recipe dishes 2. Rapid controlled cooling of heat-processed components will help to minimise spoilage. All have the same intention.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. depending on the size of the fillet 33 . as a heat process is often required for functional or organoleptic reasons. but it is assumed that any band sawing or dice process will introduce heat from blade friction. 2. up to -5 °C so as to improve the cutting and handling properties during processing. or the use of microwave technology. but will also need to ensure that the optimum shelf life can be safely obtained.4. For some components. and contamination from equipment to clean fish muscle.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes.3. Generic examples have been used to illustrate only the major points.1 . defined holding time at a chill temperature (0 – 3 °C).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.4.4. there are a number of distinct methods used. but it also poses a risk of cross-contaminating other products undergoing tempering. 2. and potential opportunities for growth of foodpoisoning bacteria. Effects on the microflora are minimal during tempering (7). this may be an incidental effect. Figures 2. Block-sawn dust that is required for use in fish cakes is swiftly transferred to a chiller to avoid water loss and spoilage from process contaminants. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. to evenly raise the temperature of the entire frozen product from -20 °C or lower. Band-sawn materials or fish dice are normally promptly further processed. Where the raw materials are frozen. as the full complexity of the processes used for specific products cannot be covered in a general text. Examples of tempering techniques include the use of heated air. line extensions and process reviews. the core temperature. It is critical that the time and product temperature during tempering be controlled to avoid elevated temperatures (>5 °C) on the product surface allowing microbial growth or the onset of spoilage in raw material. It is recommended that the principles of HACCP are applied to novel processes. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface.2.

1A . chilled or sold chilled on defrost with an expected shelf life of 6 . On arrival at the processing plant.3. by mincing and mixing with a binding agent (as described in section 2.FISH AND SEAFOOD processed. Fig. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process.3. drained and then refrozen for storage until required for further processing. with the coating heat-fixed.1. after landing.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). Frozen cores are usually batter-enrobed. frozen in a nitrogen blast freezer and weighed into sacks for later processing. The processing of three examples types are described in Figure 2. It is then size-graded. are placed into plastic boxes and covered in ice to preserve their condition. The “blown” meat is graded.1.4. 2.1B .1C .4. 2.Scampi.Shaped Fillets. quickfrozen. Raw material preparation stages for chilled or frozen coated fish products 34 . The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.7 days.1 (2. may not rise above 0 ºC during processing.1. 2. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. 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.2.Fish Cakes). the scampi is washed in potable water to remove foreign bodies.2).

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

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

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

3. to produce a specific product. The vegetables must withstand processing. The protein source can be fish fillet or block. rice. and must not show evidence of disease. as this could introduce off-flavours and reduce shelf life. but they are supplied without a source of carbohydrate e. A temperature no greater than 10 ºC for 12 hours cumulatively or 21.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal.4.5. Flow diagram outline for recipe dish ingredients preparation 2. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe.1 ºC for no more than 3 hours is recommended (1).2. For example. Fig. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing.g. 2. There are two main categories. pasta or potato. canned fish may be used in their preparation owing to it having wider availability and low wastage.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes.4. Figure 2. 38 .

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

B. where composite meals are heat-treated. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. such as butter garnish with black pepper.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. Potato toppings made from flake or powder will normally have little associated spoilage flora. and then cooled slowly allowing germination and re-growth. Appropriate handling facilities or equipment will be required to protect the product.) to the product (12. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. Sauces can be multi-component preparations or more simple preparations. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. If the sauce contains modified starches for freeze/chill stability. This is a particular issue especially where products are heat treated to eliminate vegetative cells. 40 . cereus). Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. rely on the use of microbiologically sound ingredients. Placing such products in a standard chill store to cool down could result in condensation. cereus.6. spore-formers can proliferate if adequate control of temperature is not maintained. if subjected to temperatures between 20 and 40°C. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. 2.4. can germinate and rapidly multiply. Products or dried ingredients contaminated with either spores of aerobic bacteria (e. a process equivalent to at least a 5D Listeria kill will be necessary. Sauces or marinades are added to fish to broaden appeal or to accompany components. aureus toxin is well documented (10.g. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. especially in those products that have a vegetable component or rice accompaniments. Typically. 11).FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. but temperature-abused pasta can also spoil rapidly by mould growth. Alternatively. and an increased risk of contamination from L. with the range always expanding as new tastes and cuisines are developed. cereus are well known and are still a common issue in the UK (9). An incident involving Staph. For this reason all products. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients.4. On hydration. 13). a heat process in excess of 73 ºC will be required to rehydrate this component fully.

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

5. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. 2. botulinum to grow.4. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. 42 . The oxygen-free conditions will however allow other anaerobic bacteria such as C. Flow diagram outline assembly and packaging of recipe dish 2.2.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).2 Modified-atmosphere packaging Originally developed for the red meat industry.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. generally carbon dioxide and nitrogen. unless the temperature is strictly controlled to below 5 °C throughout the supply chain.4. These include: 2.FISH AND SEAFOOD Fig. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. as it dissolves in the surface moisture to form a weak acid.7.4.7. and tuna loins are packed by placing the product in a plastic pouch and removing all the air. the method replaces air in the sealed packaging with a defined mixture of gases. which may restrict either microbial growth or enzyme activity.

and Yersinia spp. in numerical terms. phosphoreum. both organisms are capable of growth at chill temperatures. except for some crustacea. the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. Aeromonas spp. The fish component of recipe dishes. temperature control is important in preserving chilled integrity of ingredients. Shewanella putrefaciens and Ph. but they will also be readily adapted to the nutrient source and conditions. Spoilage will be most readily detected in proteinaceous components of ready meals. This is also true of chill storage and the distribution chain. Vibrionaceae will succeed (17 ). It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage. followed by moisture loss. Beyond this. Spoilage of this material is not expected to differ greatly from the processes described elsewhere.. at slightly higher temperatures (10 ºC). and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15). However. This is considered to be the one major difference between fish and meat products. Nonetheless. by a loss of texture. is typically uncooked or at best lightly processed to retain eating characteristics. where the protein component is nearly always heat-processed. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). The case for growth and contamination of products with Aeromonas is stronger. Not only will these organisms have a numerical advantage over other proposed spoilage organisms. 2.1 Chilled storage For chilled products or chilled components of frozen products. The literature has good information on spoilage of individual components of recipe meals. At refrigeration temperatures (5 . have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18). but is not limited to.5 Spoilage During processing. (20).10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.5. initially. 43 . and in influencing the final eating quality of the product. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. but little on in-pack spoilage. as this organism is a common contaminant of freshwater fish species (19). and finally flavour loss. a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood.

Still. for example. eliminating nematodes in products that do not have a heat treatment step. However. The ACMSF (22) issued clear advice on this topic and has recommended that the shelf life of products without barriers (23) is restricted to less than 10 days. 2. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e. also be those that have the least effect on the viability of microorganisms (21). Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24).5. and Aerobacter spp.. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. the shelf life of products is limited by the natural spoilage process described earlier. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution.. By retaining the intrinsic spoilage flora of seafood.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. Serratia spp. Bacillus spp. Products that are developed with an extended shelf life beyond the 6 . botulinum. this should also take into account shelf life of product components used in the assembly of the final product.g.FISH AND SEAFOOD 2. This reinforces the need for prompt processing and temperature control of ingredients and finished products. producing characteristic off-flavours. The conditions and freezing process most favourable for good-quality products will. auto-oxidation of fats/oils).7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage.. For example. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. 44 . which would warn the consumer of the potentially unsafe nature of the product. Pseudomonas spp. within dairy products are well known. freezing also has a protective role in improving the safety of raw materials by.5.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). These have been proved to impart off-flavours. As a note of interest. where the variety of components and ingredients used in manufacture is varied and complex. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). The type and nature of the enzymes and their activity will depend on the bacterial species. however. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp.

and some effects related to product type and time to toxin production have been noted. Cases quoting contamination of seafood with organisms such as S. any risk of C. perfringens contaminating seafood will depend on the source of raw material. and this issue may be of greater consequence for warmwater. farmed or free-living crustacea (30).6. water. etc. histamine intoxication.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. with pathogenic microorganisms or viruses. are dealt with elsewhere in this book. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. 2. Counts of between 103 . 45 . permit rapid growth of B.6. typhi.e. low oxygen or anaerobic conditions. agricultural run-off or sewage can contaminate in-shore species.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. air. and Campylobacter are recorded in filter feeding molluscs such as mussels. may be of benefit within any risk assessment for chilled and frozen prepared fish products. 13. Hazards attributable to the non-seafood components are listed below and this information. in some cases. These are C. In addition.3 Clostridium perfringens A study examining C.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. potato. cereus.104/ g should be treated with some concern (26). A number of other Bacillus spp. Germination of spores requires favourable conditions. along with previous discussion. or a heat shock followed by holding at a temperature suitable for growth. botulinum when held in favourable conditions for growth (28) i. Inappropriate holding times and temperatures will. have been linked with food poisoning and should also be considered. soil..6. 27). 2. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. 2. and many raw or processed foods (26). perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). especially molluscan filter feeders. Therefore. Additional hazards from marine bio-toxins. spices and herbs and vegetable shoots have been reported (12.1 Bacillus cereus This is a common contaminant of dust. Incidents in such diverse materials as cream. Vibrio spp. botulinum type E and Vibrio parahaemolyticus (25).

Shigella spp. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). infected food handlers. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. and inadequate cooking or holding within the temperature growth range (27).5 Shigella spp. if any other potentially pathogenic organisms. 46 . and this is believed to be due to a tolerance to brine used in the smoking process (32). will colonise factory environments in a similar fashion except for possibly Vibrio spp. the restrictions on growth rate seen with seafood will not apply. indigenous or other. monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. aureus enterotoxins in foods. aureus (35). 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. An acceptance has now developed in the UK of the presence at low levels of L. which is not affected by any subsequent cooking process. 2. but under the appropriate conditions. growth may occur in foods (26).6. at the present time. 2. These include inadequate refrigeration and preparing foods far in advance of planned service. L. Chilling may help Shigella to survive. 2. Rates of contamination with smoked seafood are much higher than those with unsmoked products.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). L.6.6. when introduced or inoculated into sterile components of chilled prepared recipe dishes. 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). monocytogenes as a contaminant in foods (31). but.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. and will therefore cause “food poisoning” even in a product that is cooked before consumption. However. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). It is unclear. Sources of contamination are many because of the widespread distribution of this organism in the environment. These conditions will allow the production of the toxin.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph.

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

pasta.g.FISH AND SEAFOOD the microbial criteria for other products (rice. aureus V. fish fingers. 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. cereus Staph. TABLE 2. coli GMP* <102 Maximum 104 TABLE 2. L. Maximum= Levels at end of stated shelf life 48 . batter etc) should also be consulted when establishing the specifications for raw materials and final product. monocytogenes B. coated fish.fish products.II IFST Guidelines for part-cooked foods – e. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp.III IFST Guidelines for processed foods . coli GMP* <104 <102 <10 Maximum Product-dependent 104 103 GMP* Not detected in 25 g Not detected in 25 g <102 <20 Not detected in 25 g <50 ppm (<5 mg/100 g) Maximum Not detected in 25 g 103 104 103 102 <50 ppm (<5 mg/100 g) *GMP = Values found immediately after production under good manufacturing conditions. parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E.

5. 373-8. Taylor and Francis. Bacterial histamine production as a function of temperature and time of incubation.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). 5 (3).. 1997. Gilbert R. London. 7. 14.W. Hackney C. 9. Journal of Milk Food Technology.. 1991. Behling A. Christiansen T. 1982.. Bartlett C. International Commission on Microbiological Specifications for Foods. Sakamoto T. London... 1311-7.8 1. Tu A. Ohgiya S. Woolaway M.J.M. Powers E. 1986. Yumoto I. The bacteriology of fresh and spoiling fish and some related chemical changes.K. Shewan J. Communicable Disease Report. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. Proper processing and handling of raw tuna..L. Food and Drug Administration. 204-16. Van Nostrand Reinhold. 96. 1997. Fish: microbiological spoilage and safety. Eds. Fish and Fisheries Products: Hazards and Controls Guidance.E.R. Matsuyama H. Toxin Related Diseases: Poisons Originating from Plants. 38 (11). Microorganisms in Foods 6: Microbial Ecology of Food Commodities.R..G.L..M. Journal of Food Protection. Oxford... . IBH Publishing Co. 67-73. Microbiology of processed spices. Kumeta H. 2001. 2.R. Cambridge. Incidence and levels of Bacillus cereus in processed spices.. 215. Slavin J. References US Food and Drug Administration. Ltd. Masuoka Y. 12. Martin R. Dalgaard P. 10. Communicable Disease Report. Hobbs G. Outbreaks of foodborne illness in humans. Recent Advances in Food Science. England and Wales Quarterly Report.. Ward D. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. 1976. 11.. 59 (2). 1. Journal of Hygiene. Huss H. Pvt. Taylor S. 3. 1962.. Mejholm O.C.. 1976. Lawyer R. 70-2. 167-93. Microbiology of Marine Food Products. 1997. Journal of Food Science. Pathogenic microorganisms associated with fresh produce. Hoshino T.H.. Kluwer Academic. 49 4. 39 (10). 166-73..R. 1997. Food Science and Technology Today. 15.R. Brown T. 668 -70. Latt T.. 47. 8. Wienke A. Beuchat L. Washington D. 2005. 663-87. 6. 7 (24). Letters in Applied Microbiology.L. Journal of Milk Food Technology. 13. Ishizaki K. Bartram P. 233-7...M. Murrel H. Animals and Spoilage. 1993. in Fish Inspection Quality Control and HACCP: A Global Focus. 24 (5).. Importance of Photobacterium phosphoreum in relation to spoilage. Letters in Applied Microbiology.A. Powers E. Aureli P.T. New York. 2.J. 25 (1).C.. 1991. Collette R.C.

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

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

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

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

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

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

either whole or minced. uncooked meats packed unprocessed. Frozen: in shell. or shelled meats. These molluscs feed on minute plants and animals carried to them by the current. oyster soup. tidal waters or in brackish water in the mouths of rivers. all in cans. the meats are smoked and packed in oil in cans or jars. 3.2 Oyster Oysters are marine bivalve molluscs. after precooking and brining. Dried: shelled meats. skewered with bamboo sticks. Canned: meats removed from the shell by steaming. Live: in shells.2. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. oyster bisque. Soups: oyster stew. clam madrilène and clam liquor. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. 57 . Fresh: in shell. sun-dried. Dried: Hoshigai (Japan) shelled meat. then heat-processed. Semi-preserved: meats cooked and packed in spiced vinegar.1. hermetically sealed. smoked meats are also canned. Live: in shell Fresh: in shell or shelled meats. trimmed and packed. Canned soups include clam chowder. uncooked. Smoked: meats. packed in weak brine. in cans with either brine or clam liquor.MOLLUSCAN SHELLFISH the giant clam of the Indian and Pacific oceans may be nearly 130 cm long and weigh 227 kg. Canned: clams are steamed open and the meats removed from the shells. having rough and irregularly shaped shells that are closed by a single adductor muscle. uncooked (shucked). washed. or shelled meats. and no siphon. Frozen: shelled meats. Oysters lack a foot or have a reduced foot.

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

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

. and cockles The processing of bivalves takes two forms. 11.. whole or in part. that have never been cultured on laboratory media. Alcaligenes spp..FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. and yeast cells of Rhodotorula rubra and Trichosporon spp. responding to the environment in which they are harvested. They are termed viable non-culturable microorganisms. Micrococcus spp. Vibrios. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. salinity. Pseudomonas spp. For example.000 /g. from which one or both shells have been removed. 10. dredges.3. Campylobacter coli Bacillus cereus Hepatitis non-A.1. Temperature. At harvest.3.. shellstock (live) which is shellfish in the shell. observed microscopically. (9. In summary.3. There are microorganisms..1. 3. Other microorganisms found in lesser numbers are the Achromobacter spp. Water temperatures affect the concentrations of microflora in the water. mussels. and Aeromonads. Acinetobacter spp. and shucked shellfish that is shellfish.3 3. rakes. mechanical harvesters.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs.1 and 3..1 Processing and its Effects on the Microflora Oysters. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. 15). pollution point sources and hydrological events will greatly influence the initial microflora in bivalves.. Flavobacterium spp.. nutrients. Moraxella spp. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. clams.1. Bacillus spp. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. 13). but have never been cultured (14. Enterococci spp.. The largest groups found are the Pseudomonads. or by 60 . the initial flora composition and concentration of bivalves are highly variable. 3. spirochetes of the genus Cristispira and Saprospira. 12.2.1 Shellstock 3.

by truck or boat. The shellstock is transported dry. wet storage also may be used to de-sand those shellfish species that tend to accumulate sand in their mantles and gills thus making them more palatable. Thirdly. in containers from which they can be retrieved easily. Shellstock may be wet-stored at the processing facility in tanks of clean seawater. Water used for shellstock washing is from approved growing areas.MOLLUSCAN SHELLFISH hand (Figure 3.1. or immediately placed in refrigerated trucks. If by boat.3.1). usually in open-mesh containers and/or burlap sacks. or from other sources approved by the State Shellfish Control Agency (SSCA). Shellstock are placed in drycooler storage containers in the facility. Fig. or in bay-approved shellfish-growing waters prior to sale or processing. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. Secondly. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. Generic flow diagram of live shellfish from harvester to point of sale 61 . There are three primary reasons for wet storage: first. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock. for transportation to the point of sale. after harvesting.

There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. Harvest conditions are another factor that influences the growth of bacteria in shellstock. Generally.1. with prolonged storage the microbiological quality of the bivalve decreases. 23. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21.FISH AND SEAFOOD 3. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. Furthermore. At this temperature the multiplication of faecal coliforms (E. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. are prevented. Enterobacter. when it reaches the processing facility. the bacterial counts in the meats will increase. Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. bivalves tightly close their shells thus trapping water and microflora. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. however. and thus contaminating the shucked meat at the point of sale. shellstock are placed in refrigerated trucks for transport or placed in wet storage. including V. there is evidence that in the absence of adequate refrigeration. 22). The washing of sediment and detritus soon after harvesting will prevent further contamination. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. shellstock are not refrigerated on the harvest boat. An outbreak of infectious hepatitis in Sweden in 1956.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. and the sanitary practices applied during harvesting and shipping. 16. with the exception of A. and temperature storage of shellstock to point of sale. Generally. without adequate refrigeration. Over a period.3.1. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. coli. to avoid possible contamination of the shellstock. some pathogenic organisms will survive and multiply in shellfish (20. If long delays occur before reaching the processing facility and/or refrigeration. 24. and Citrobacter species) and Vibrios. 17). hydrophila (18).2 ºC (45 ºF) or below (19). for a considerable length of time. Klebsiella. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. At harvest. 21. The most important factor in controlling bacterial growth in shellstock is temperature. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). vulnificus. handling. 26). 25). Water used for shellstock washing should be of good sanitary quality. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. synthetic seawater for the purpose of de-sanding and salt uptake.

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

Separate rooms or lockers for storage of clothing. which may lead to contamination of product. Shucked meats are thoroughly drained. Generic flow diagram of shucked shellfish from harvester to point of sale Shellfish are subject to contamination during the shucking and packing process when the following are not adhered to (19): 1. Shucking containers are rinsed clean with running water.2 ºC or less. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. 4.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). 3. within one hour.2. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. 64 . aprons. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. and sanitised before each filling. 2. and gloves help prevent the contamination of shellfish. Fig 3. There is a tendency to store such articles on the shucking benches or in packing rooms.

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

V. The incidence of V. shellstock and packing of shellfish in jars) (29). and to the point of sale. 3.FISH AND SEAFOOD 3. from harvesting to finished product (i. V. Major outbreaks are associated with the warmer months.5 cases/100. 3. although non-hemolytic strains have been associated with illness (43.48 hours with abdominal pain. there is a recent report of low levels of V. Sporadic cases are frequent along the coast of the United States. It is a common marine bacterium. of V. This bacterium is present in all the major processing steps of a shellfish plant. Products involved were from the same processor. They can be introduced to the shellfish from the environment or added during processing.5. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). These organisms can be present in the shellfish from the point of harvesting. Fortunately. parahaemolyticus causes gastroenteritis lasting 24 . during processing. and very common in Japan. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. diarrhoea. and harvesting of shellfish in the cooler months. 44). Diseases caused by Vibrios are enteric in nature. parahaemolyticus possibly producing illnesses (40). parahaemolyticus from shellfish is low. 41). widespread in both polluted and unpolluted waters.000 population/year (46). parahaemolyticus multiplies rapidly at temperatures above 20 ºC. Between May and September 1997.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. ranging from epidemic cholera to sporadic cases of diarrhoea. and estimates based from a hospital study show less than 0. headache and fever. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. lot and retail outlet. parahaemolyticus. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. approximately 23 cfu/g. primarily in brackish or saltwater. coastal areas). only a small percentage of strains in the marine environment are potentially pathogenic (45). 66 .1. One of 250 cases reported involved shucked products with low levels. nausea. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39).1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. where outbreaks occur regularly.1 Vibrio parahaemolyticus V.5. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. however.e. prevention of cross-contamination.

diabetes. cholerae non-01 (51). Control of V.MOLLUSCAN SHELLFISH Control of V. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. V. parahaemolyticus. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). Death has been reported to occur within 36 hours from onset. parahaemolyticus the estimated annual incidence rate is very low. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. They are sensitive to cold. Onset of symptoms occurs in 48 hours and can be severe. It causes diarrhoea. vulnificus are fever. 3. are particularly susceptible to primary septicaemia. Hazards from this bacterium can be controlled by thorough cooking of shellfish. Again. Inadequate refrigeration will accelerate the growth of V. like V. 55. lasting 6 .000 population/year (8. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. V. which have been implicated in shellfish-related disease outbreaks (31). with a fatality rate of 50% for individuals with septicaemia. fever. The bacterium is an estuarine species that causes wound infections.5.5. Illnesses are usually associated with the consumption of raw oysters as described in Table 3.1.0. vulnificus is by rapid refrigeration during warm weather months. In one study. gastroenteritis.II (8.1.7 days. or primary septicaemia. Oysters are the most hazardous food associated with V.48 hours of onset. 23 of 24 major USA-West Coast estuaries were positive for V. 46. 54. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. chills. in the range of 0.8 case/100. cholerae non-01 is ubiquitous in the estuarine environment.3 Vibrio cholerae non-01 Illness caused by V. The organism attaches to the intestine and release of a toxin is suspected. these individuals are advised not to eat raw shellfish. abdominal cramps. 53. 46.4 .2 Vibrio vulnificus The symptoms of V. parahaemolyticus growth in shellfish meats is temperature dependent. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. cirrhosis. V. or immunosuppression. 3. 57. however. 49). nausea. 48). All isolates are considered pathogenic. High-risk individuals. and vomiting. Primarily. and nausea within 24 . Healthy individuals are susceptible to gastroenteritis. and preventing cross contamination once the product is cooked. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). vulnificus. those who have liver disease. leukaemia. It is one of the most severe foodborne infectious diseases. The occurrence of V. cholerae non-01 has been associated with septicaemia. the organism has been found in the Gulf of Mexico. and rapidly become inactive at low storage temperatures. 58). Control of oyster-associated 67 . 56). cholerae.

Illness can include abdominal cramps. abdominal pain.48 hours with abdominal pain. In developing countries.5. and V. and shock. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). parahaemolyticus and V. V. e.1. 3. Gulf coast environment. fluvialis. these organisms cause gastroenteritis lasting 24 . undercooked. vulnificus.S. hollisae. diarrhoea.FISH AND SEAFOOD V. It is considered a fragile and sensitive organism to environmental stresses such as oxygen.10% carbon dioxide for optimal growth conditions. including V. Onset of disease is 6 hours to 5 days. Onset of illness is from 3 to 5 days after ingestion of contaminated food. dehydration. Control of infection from these organisms is no different from the other Vibrios. and illness lasts 7 . Poor sanitation and contaminated water supplies will spread the disease. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. cholerae 01 is transmitted by faecal contamination of food or water.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. 68 . The presence of these strains appears to be in low numbers and non-virulent (59). Death may occur after severe fluid and electrolyte loss.5. V. processors should know the product source. cholerae 01 in molluscan shellfish.1.10 days. vomiting. There is evidence of strains of V. The frequency of infection of these organisms is no different from other Vibrios. They are common marine bacteria.g. widespread in both polluted and unpolluted waters. low pH. salinity. and the transmission of the disease by consumption of raw. and southern Asia including India and Bangladesh. headache and fever. respectively. however. 61. mimicus. 60). V. acidity and heat. cholerae 01. preventing crosscontamination of cooked seafood. the pathogenic severity is lower than V. In general. associated with shellfish-borne illness outbreaks. 63). jejuni requires reduced levels of oxygen (3 . imported product from a country experiencing an epidemic. 3. nausea.2 Campylobacter jejuni C. nausea. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. Illnesses are associated with the warmer months. The control of V. cholerae non-01 illnesses is by cooking seafood thoroughly. cholerae 01 establishing in the U.5%) and 2 . fever. 62. particularly in oysters. or cross-contaminated shellfish (57. is the same as previously discussed for the non-01 strains. 3. and nausea. There are several other Vibrio spp. (8. drying.5.5 Other Vibrios spp. In addition. It causes diarrhoea.

and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. 70.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. they survive well at refrigerated temperatures. 82. infection occurred among participants of a firemen's banquet. Human feeding studies suggest that about 400 . 69. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. They have been implicated in limited shellfishassociated gastroenteritis cases (8. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. 71). 84. V. 72. 85). 83. Campylobacter spp. control of C. jejuni were recovered. 73).3. 77.500 cells/gram of Campylobacter may cause illness (64. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). and their immediate proper refrigeration. 78) and P.5. Recently. 81.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. of Cases 4 362 0 1 0 a Includes V. however. hollisae Infective dose varies with host susceptibility and virulence of the strain.3 Other organisms of concern 3. The seasonality of 69 . jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. shigelloides (79. however. V. V. 66). To reduce the risk of shellfish-borne campylobacteriosis. 80) are ubiquitous in the marine environment.II Molluscan shellfish associated outbreaks caused by Vibrio spp. although their role as a pathogen is highly controversial and circumstantial. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. of Incidents 4 279 0 1 0 No. cholerae 01 and non-01. 3. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. and V. 65. 76. fluviallis. mimicus.4 to 114 cells. vulnificus. from Pacific shellstock oysters purchased from a retail grocery store (74). jejuni begins in the harvest of shellstock from approved waters. Although. In one case. have been isolated from shellfish beds throughout the world (67. jejuni does not multiply in shellstock at refrigerated temperatures. C. This finding is significant in that C. hydrophila group (75. V. ranging from 0. whereas at 10 ºC Campylobacter survive only for 5 days (71). parahaemolyticus.5. Campylobacter appear to survive well in oysters during storage.MOLLUSCAN SHELLFISH TABLE 3. C. 68.

action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. cholerae V. parahaemolyticus Staph. and where indicated are intended to be used for official control purposes. jejuni V. vulnificus in molluscan shellfish. aureus C. Similar results were seen by other investigators (86. FDA/Administrative Guidelines 3 Source (91).000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. prepared sandwiches.000 cfu/g1 <230/100 g2 <1000 per gram. being isolated more often during the warmer month. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. Enforcement 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. 70 .FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. Test Standard Plate Counts E. 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. In one study. They are easily destroyed with mild cooking temperatures.000/gram (29).000 to 3. dried milk/whey drink powders.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. coli Salmonella V. Control of these organisms in shellfish products is limited. FDA/ Administrative Guidelines. 2 Source (90). non-dairy frozen desserts. hydrophila group stored at refrigeration temperatures. soy products. Note: Although molluscan shellfish are not included in the selected high risk food.0001 negative for the presence3 Under review4 Source (89). FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. coli Enterotoxigenic E. Caution should be exercised by individuals with haematological malignancies (88). 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. 3. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500.600.

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

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

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

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

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

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

63-8.S. Pittsurgh. Food and Drug Administration. Gaithersburg.S. Food and Drug Administration Compliance Policy Guides. 90. in National Shellfish Sanitation Program Manual of Operations. National Academy Press. Food and Drug Administration Compliance Program Guidance Manual.W.H. in U. U.R. in U. 1995. Food and Drug Administration. Chapter 5: Foods.. Food and Drug Administration. Food and Drug Administration.1987. 1987. Baird-Parker T. Program 7303.600 (CPG 7119. U. 3.M. 89. Food and Drug Administration..030.. Slavin J.8 Further Reading Ahmed F.MOLLUSCAN SHELLFISH 88.12)..S. Lund B. Sherlock C.S. Lancaster. Clams. 1997.E. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. 77 .. Government Printing Office . U.C. 1991.W. Fish Inspection. Quality Control. 2000. Part I Sanitation of Shellfish Growing Areas.C. Martin R. fresh or frozenadulteration by bacteriological contamination Sec. 1995. Gould G. The Microbiological Safety and Quality of Food. Burdge D. Ed. 560.L. mussels. 7. U. Inc. Ed.. and HACCP: A Global Focus. 91.S. Food and Drug Administration.C. Seafood Safety. Technomic Publishing Co. Collette R. Ed. Food and Drug Administration.S. Smith J.Pathogen monitoring of selected high risk foods (FY 88/89).E. Aspen Publishers Inc.. Washington D. Food and Drug Administration.A.S. U.. U. Washington D. Washington D.C. oysters.S..

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crayfish and shrimp (1).1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. and are usually aquatic. Pandalus borealis (deep-water shrimp). Peeled and cooked shrimp are ready-to-eat after controlled thawing. CRUSTACEAN SHELLFISH Linda Nicolaides. Parapeneopsis spp. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. They possess a segmented body. the front pair ends in claws. e. lobsters. peeled product. and Trachypenaeus spp..5 and 1. Such products are usually only partially cooked before freezing. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). (deep-water shrimp). and available in 0. Species eaten from tropical waters include Penaeus spp.0 kg polythene bags.4.5 and 1. with shells and heads on or off are usually packed. jointed limbs and a chitinous exoskeleton. packed and frozen in 0.. with additional water to form a glaze. Peeled ready-to-cook shrimp are raw.5 kilo pack (boxes or bags). lobsters and crabs have ten legs.g.0 kg polythene bags. Crayfish. Pandalus montagui and Crangon crangon (inshore shrimp). 79 . M. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption.. Shrimps.Ph. crabs. and frozen in 1 and 2.

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

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

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

CRUSTACEAN SHELLFISH 4. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. the meat is either repacked and sold in the shells.4.4. wet and ‘soapy’ or ‘jammy’ to the touch. However. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. If the tail drops. With spoiled crabs and lobsters. spoiling prawns and shrimps have shells that are soft. and a strong ammoniacal odour is produced. there is no associated spoilage problem. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. The shells have a crisp. An offensive odour is also produced. The shell will be soft. with the growing demand for ready-to-eat products from crab and lobster meat. they include Sh. the joint between the tail and carapace is open and the limbs hang loosely. putrefaciens and Pseudomonas spp. or packaged and processed. If boiled. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail.2 Cooked crabs and lobsters As indicated earlier.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. for crabs. since crabs and lobsters are traditionally consumed immediately after cooking. 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. Once removed and cleaned. dull and sticky. immediately prior to boiling. viruses or parasites. 4. contaminating microflora may include enteric bacteria. 4. or in hermatically sealed cans that have undergone an appropriate heat treatment. 83 .2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. there is no indication of life. whilst boiled crustaceans have a pleasant slightly sweet smell. bacteria of public health significance. slightly iodine smell. In comparison. and have a sweetish. putrefaciens.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. 4.3. If hygienic procedures are not followed. bright and dry appearance.

However. 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 first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). 16). in domestic/imported shrimps (15). 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. a common pond fertiliser used in aquaculture. in 24.5 4. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. fish fingers. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12).5. Salmonella cannot be completely removed from raw shrimp during processing. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. monocytogenes was absent from 40 samples of cooked prawns. 4. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. shrimps and cockles sold (17). Further surveys identified Listeria spp. as well as for Salmonella. even if GMP is followed.2 Listeria spp. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L.5. and usually for both.5% of frozen raw shrimps from different countries (13). monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. animal manure. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . Hence. An import alert was put in place in June 1987. cooked and smoked fish.1 Pathogens: Growth and Survival Salmonella spp. later surveys in 1990 recovered the pathogen from retail raw. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. shrimp and shellfish (11). Although foodborne illness had not been associated with the product. monocytogenes in cooked ready-to-eat foods. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9).FISH AND SEAFOOD 4. although efficient cooking will quickly eliminate it (6). 5. 6. nearly 30 years elapsed before the potential hazard of L.cooked prior to consumption. The above results prompted the FDA to develop a compliance programme for the pathogen. Moreover. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. Following this. The presence of L. 7).

19). the causative agent of cholera. combined with the degree of handling of these types of product. whilst boils and cuts are also common sources of the pathogen.3 Staphylococcus aureus Staph. although contamination by types A and B may occur from the environment.5 Vibrio spp. 25). 4. The former. 85 . it is important that the cooking stage is controlled to eliminate the pathogen from the product and that procedures are in place to prevent any cross-contamination of the cooked product. Researchers have demonstrated that inoculated shrimp supported toxin production by C. pregnant women and the immuno-compromised. cholerae non-O1. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. botulinum type E at 10 °C but not at 4 °C (20). botulinum type E is commonly associated with fish and fishery products. V. An epidemic that began in South America in 1991. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21.4 Clostridium botulinum C.g. 4. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. 22) as well as from undercooked crustacea when harvested from contaminated waters (23. However.5. monocytogenes has been attributed to crustaceans. e. is widely distributed in aquatic environments. particularly crabmeat (18. aureus because of the ubiquity of this organism in the processing environment. Crustaceans have been linked with food intoxications attributed to Staph. Once the competing microflora has been removed by a heat treatment. the pathogen can grow at temperatures as low as 3 °C. 24. especially in products that have been vacuum packaged. was associated with seafood consumption. therefore.70% of healthy individuals carry toxin-producing strains of Staph. aureus is not a member of the intrinsic flora associated with crustacean shellfish. V. The main source of contamination is the hands of processing personnel. cholerae is divided into two groups. Control should be achieved by promoting GHP and staff training programmes. Crabs are covered in a chitin exoskeleton. so is able to survive heat treatments that eliminate vegetative cells. as part of the natural bacterial flora. The pathogen has resistant spores. which can become colonised by V.CRUSTACEAN SHELLFISH old. cholerae O1 and V. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru.5. no incidence of foodborne illness caused by L. aureus on their hands. about 50 . 4. To date. pasteurisation and cooking.5. Vibrio cholerae. cholerae.

It is more commonly implicated in countries where crustacean shellfish are eaten raw. hydrophila is able to grow well at refrigeration temperatures (0 . jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat.7 Aeromonas hydrophila and Aeromonas sobria A. Astroviruses and non-A.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.6 Campylobacter jejuni and Campylobacter coli C.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. V.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. parahaemolyticus. Parasitic infections are controlled by either cooking infected material prior to consumption. Quantitative levels were below limits of detection in all cases (<0.5. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. hydrophila has been isolated from prawns (29).8% of samples contained C. However. 4. Of these. e. although A. Contamination by this organism can be controlled by rapid chilling of crustaceans after harvesting and avoidance of time/temperature abuse during distribution and processing chains. the potential hazard exists as C. non-B Hepatitis viruses (31).5. occurs in crustaceans from warm environments. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. 4.30 MPN/g) (27). or freezing at a temperature of -23 °C for 7 days (30. whilst 5.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. V. Norovirus. hydrophila and A.5. jejuni has not. However. however. particularly trematodes (6). 4. Japan. 31. in a survey carried out in the United States between 1977 and 86 . South America. 4. Caliciviruses. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28).5. a marine halophile. been implicated in a case of foodborne infection associated with crustaceans. coli. 32). but has the potential to cause fatal foodborne illness (26). A. to date.g. vulnificus is an invasive and lethal pathogen associated with wound infections. Hepatitis A virus presents the most serious hazard.

Norwalk viruses can also withstand heating to 60 °C. although it is one of the European Union (EU) legislated allergens. Selected parts from this regulation is presented below. produced or imported into the European Union (EU). 2073/2005. 4.6. only 0. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34).47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). 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. 4.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. are included in the Commission Regulation (EC) No.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. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control.5. 87 . 39). They can survive low pH levels. The risk of such hazards is far higher with fin fish and molluscan shellfish. 4. Provisions laid down concerning the use of additives would also apply to these types of products.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. 38. consistent with that in the stomach.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish.CRUSTACEAN SHELLFISH 1981. The toxin causes amnesic shellfish poisoning (ASP) when consumed. from farm to fork (37.5. 4. which could be transferred to the meat during unskilled removal from the shell (35). No foodborne incidents have been attributed to this source to date. or chlorine concentrations similar to those found in drinking water. which results in a persistent and apparently permanent loss of short-term memory (36). A summary of microbiological criteria for crustacean shellfish is listed below.

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

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

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

When working with salting.0 g of salt per litre of brine).6 g of salt or more per litre of brine) may cause salt burn in this way. The dissolution of salt in water is dependent on the temperature.2 237. Salt in the water phase links the two conditions of moisture and saltiness.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. and salt content.6 264.8 211. For smoking salmon. At 18 °C a 100º brine is produced when the solution contains 26. is employed. a 0º brine saturation means pure water. etc.2 g of salt per litre of brine).8 211.4% salt (Table 5. it is important to understand the concept of salt concentration. a 90º or above brine (237. Thus.0 158.I).4 184. When a milder brine.FISH AND SEAFOOD 5. This should be distinguished from the salt content. Different brine strengths are required according to the curing processes and/or the type of product. only the most common processes can be described here. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. When a 100º brine is used. the fish is usually salted with a 70 . sometimes the salt crystallises onto the surface of the fish. processes and producers’ private recipes.2 105. about 50º (132.6 132. Therefore. smoking or marinating processes.3 Processing and its Effects on the Microflora Owing to the wide variety of products. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. all using salting steps.0 . it causes swelling because the fish absorbs water from the salt solution (1). or salt in the water phase. it is impossible to establish standardised processing stages in the curing of fish.4 52.80º brine (184. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. Salt concentration means the percentage of salt in the water contained in the fish. % 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. The strength of the solution is described as degrees of saturation (brineometer degrees). it is important to state the temperature. TABLE 5.8 79.

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

For large fish it is advisable to split it before salting. such as 98 .1. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. such as Staphylococcus aureus could survive and grow at such water activities in the product.90 are rarely achieved. so that the risk of spoilage is high.5. Processing of dried salted cod 5. Where fish are salted in their whole form. Only small fish can be salted whole.1. when a light brine is used.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. the process is mainly for flavouring purposes prior to the use of other preservation processes.2 Wet salting or brining As referred to above. Food-poisoning organisms. eviscerated and/or split). although toxin production would be unlikely (4).3. it appears that water activities much less than 0. Fig. The greatest barriers to salt penetration are the skin and scales.

61 3.19 3. the liquid that comes out of the fish forms a brine.11 4.5 3.3 Dry / wet salting In this process.0 2.1.60 2. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained. thus.98 6.76 4. the brine is then replaced over the fish.03 2.23 3.II. the fish is dry-salted.98 5.99 2.26 1.23 2.3 days.78 2.66 5. This is why it is the preferred method for fatty fish.5 3.26 3. with a shelf life of about 4 weeks (water activity (aw) about 0.51 5.70 6.II Salt in the water phase (after Ian Doré (5)) Moisture (%) 40 45 50 55 60 65 70 75 80 85 90 Percentage salt in the flesh (by weight) 2.51 3. sardines.46 4.41 6.88 5. and can be collected and replaced again over the fish.03 4. but mackerel.CURED.96 1.66 5. Histamine formation is of importance when using scombroid and some clupeidean species.56 6.84 1.41 4.11 9.22 6. they can be stored at 28 °C.33 5.65 2.23 8.04 5.76 4.85 3.35 4.25 5.76 4. with pressure being applied to the top of the container to help the brine to drain away.30 2.26 4.98 6.91 2. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.41 5. but as it is placed in containers. The main objective of the brining method of salting fish is to limit air penetration during processing.74 9.64 4. a saturated brine is used.73 1. the fish enzymes play an important role by maturing or ripening the product.44 2.3.85 3.06 4.76 4.5 10.5 4. headed and gutted.90 (1).47 6. if split fish is dry-salted. Here.17 5.70 3. Histamineforming bacteria do not generally grow at water activities below 0.78) (1). For preservation purposes.86 2. Anchovies are the main fish used for these purposes.17 4.26 2.76 99 .05 7. 5. For example.10 1.23 2. limiting the amount of rancidity that can develop.54 5.78 6.09 8. and a red burgundy colour will develop in the product. Fresh sardines.44 2. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .26 7.11 3.25 5. herrings and other fatty fish can be used.09 8. can be cured in a saturated brine solution for 6 days and.80 5.76 4.0 3.49 4. The fish are kept immersed in their own brine near saturation point for several days. See Table 5.00 3. the product obtained is called gaspé-cured (5).16 7. after being drained and pressed at 465 kg/m2 for 15 hours.61 3. TABLE 5.41 3.95 3.45 3. There are several variations of this process. SMOKED AND DRIED FISH smoking.0 4.

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

are commonly dried in Asian countries. Some bacteria and enzymes are still active in the presence of acid and salt. In Scandinavian countries. a very fine delicacy for Oriental food lovers.4 Air drying This is probably the oldest method of fish preservation. such as squid. by using either solar heaters or mechanical dryers. Italy and Spain (5). sugar and spices. split and dried gadoid species) are traditionally made in cold climates. sea cucumbers and seaweeds. Matjes are made of fat young herring. In some coastal areas of Portugal. air flows naturally. bacterial toxins and parasites (7).There are several products named marinades (matjes. after frying. Fresh dill is placed between the layers of fish. Usually. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). but sometimes it is dried without prior salting. the fish is dried naturally in a relatively cold environment. Stockfish (dressed. 5. are likely to survive in these mild-processed products. and a fine product is obtained. jellyfish. are sundried and. They are an important dried product. gutted. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. 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). such as Listeria monocytogenes and Clostridium botulinum. gravad fish. etc. It is important to assure the quality of the raw material. used as an appetiser. and the flesh will break down. These can be considered as ‘lightly preserved’ products with a salt content <6% sodium chloride (w/w) in the water phase and low acid content (pH >5. marinated products will keep for several months at a temperature of 4 °C. the temperature and the amount of pressure applied (5). and the stacks are placed in the refrigerator. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). South America and the Caribbean. Dried shrimps are widely used in Asia. Stockfish products have a golden brown colour and are very hard. depending on the thickness. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. mixed with salt.0) (7). are generally air-dried at sea. under a light pressure. The most common method used for fish products is the air-drying process.) where no acid is added to the process. 101 . Gravad fish is a Scandinavian product made of salmon fillets. It takes about 2 days to marinate completely.5. Shark pectoral fins.CURED. it will last for many years (5). which are placed in a mixture of salt. Stockfish is extensively used in Portugal. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. Other products. Foodborne pathogens. some fish species such as moray eel and varieties of dog fish. Dried octopus is also a very common dish in the interior of the country. In the first case. the fish is dried after salting or brining. At pH levels below 4.3. Provided the product is kept in dry and fairly cool conditions. still made without prior salting on a large scale.

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

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

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

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

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

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

spraying is preferable because it is easiest to control flavour and acceptable with this method (3). 5. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0.8% and still provide protection for 7 days (25).3. Liquid smoke was considered to provide advantages of safety. drying.5. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine.03 μg/kg of benzopyrene. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. Fresh fish spoilage may be measured by 108 .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. so as to impart an unnatural red colouration to the flesh. applying it as a dip after brining. the salt level could effectively be reduced from 4. The European Scientific Commitee for Food. nutritional neutrality and antioxidant activity (27). and less than 0. 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).FISH AND SEAFOOD smoke condensate that is dissolved in water or oil.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. or smoke extracts in organic solvents. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. When liquid smoke was used. injecting it into the fish flesh. consistency of flavour and colour. 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. It has been shown that liquid smoke can reduce the formation of PAHs (3). both of which are considered potential carcinogens. In experiments comparing traditionally smoked fish with liquid smoke. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). or applying it as an atomised spray within a modern automatic smoke kiln. 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. smoking.4 Spoilage Brining.6 to 2. 5. According to some recent research. 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. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25).

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

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

The salt in the water phase must be high enough to inhibit the growth of C. so the bacteria spoiling these products will be psychrotrophic or psychrophilic. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. flavus. botulinum type E. Vacuum packing limits chemical oxidation. including A. has been isolated from dried fish (35). Aspergillus clavatus and Aspergillus niger. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. Marine vibrios were the dominant flora in smoked salmon that was drysalted. and microbial spoilage is more likely. Pseudomonadaceae.IV. will have a different residual post-smoking flora from 'hot-smoked' fish. LAB. 33.747. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). For more information on spoilage see Table 5. 36). however.5% sodium chloride in the water phase will inhibit some bacteria. The preservative effect of the smoke will inhibit some spoilage microorganisms. It has been reported that the type of salting (dry. Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation.. but.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. Table 5. where a high internal temperature is reached.CURED. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. Cold-smoked fish will contain microbial flora representative of that of the raw fish. which does not create the salty conditions required for spoilage by those halophilic microorganisms. but this will not prevent the growth of spoilage bacteria. The brining to 3 . SMOKED AND DRIED FISH infrequently at low levels. it was. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. around the water activity of saturated sodium chloride. Smoked fish products tend to have a light brine treatment. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations. not the growth of spoilage organisms. with a lower aw limit for growth of 0. able to grow on dried salted fish. Aspergillus spp.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). as already stated. Fish processed by cold-smoking. 111 . Refrigeration is necessary. The heat during hot-smoking will kill vegetative microorganisms but not all spores. In Vietnam.

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

although the combination of the two factors.5 5. the fermented products being the most dangerous group (7).75 0.9 0.5-8.5% to 5%. The presence of salt in cured products has a great effect on the growth of the bacteria. Before. smoked and dried. botulinum Staph. respectively. aureus Yeasts Moulds Lactic acid bacteria pH 5.5 4.5 0-11 3. the importance of cold-chain integrity must be emphasised.86 0. botulinum type E.CURED.8 1. 9 and 8 of the outbreaks.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13).5-10. but if the fish products are stored at temperatures <10 °C. However.0-9. and C.5-8. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. In the United States.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. botulinum type E and production of toxin. popular demand for healthy foods has led to a change in the tastes preferred by the public and. the products were heavily salted. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E.5-10 4.61 Organisms Most Pseudomonadaceae C. pressure on the traditional methods of curing.97 0. the non-proteolytic type E strains can grow in up to 5% sodium chloride. and as botulism is a serious hazard in foods.5.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. sodium 113 .8 0. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. SMOKED AND DRIED FISH TABLE 5.97 0. Since neither the smoking nor the drying parts of this curing process are particularly severe. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. under optimal conditions. with this change.5 5. It has been shown that. when present in the raw material. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). types A and B are not affected by 3 . botulinum type E Staph. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). botulinum. could not grow and produce toxin. The concentration required to prevent growth at room temperature can vary from as low as 3. salted and pickled products accounted for 10.

It is a halotolerant bacterium able to grow at water activities as low as 0.4. aureus. However.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 human carrier rate can be as high as 60% of healthy individuals with an average of 25 . but requires higher temperatures for toxin production (>10 °C). Once the product is contaminated.94 (7). growth and toxin formation may occur. are particularly susceptible to contamination and.5% salt in the water phase. if recontamination occurs and storage conditions are favourable. when the bacterial load of the product is lower. aureus growth. Infection by Shigella sonnei has increased in the UK. As they are poor competitors. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin.86 and 10% sodium chloride. gills and guts as well as in several lightly preserved fish 114 . As with other organisms. 5. the risk of infection can be eliminated by proper cooking. if storage temperatures are over 10 °C.4 Listeria monocytogenes The psychrotrophic L.5. but. Sliced smoked fish products. throat and skin. after processing. if it contains sodium nitrite ≥100 ppm.5. Thus. The organism is mesophilic with a minimum growth temperature of 7 °C. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. but.3 Enterobacteriaceae The pathogens Salmonella.0 .30% of the population positive for enterotoxin-producing strains (41).2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. 5. it should contain not less than 3. and the minimum pH for growth is 4. as they require more handling. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. if the person handling the product is a carrier of Staph. An outbreak of Shigella in an Italian cruise ship. coli all occur on fish products as a result of contamination from animal/human reservoirs. In foodstuffs. they do not grow well in the raw material. Shigella and E. they will grow rapidly (7). 5.5.or cold-smoked fish should have at least 3. has been reported (42). more importantly. the organism may contaminate the product. related to the consumption of a raw smoked fish meal. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). the organism can survive most of the light curing processes. The FDA established that hot. the human nose. 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.5. Usually.0% salt in the water phase (30).

5 Parasites Despite the presence of parasites in fish being very common. lightly salted caviar.92 . from seawater fish. most of them are of little concern to public health. pH 4. has also been detected in cod (7).12%. which means that the process must conform to a level of ‘no detectable L. Pseudoterranova dicipiens. 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.8 . a short period of freezing either of the raw material or of 115 . which include intermediate hosts. from freshwater fish. marinades (ceviche) (43) and gravad fish (44). there were no reported outbreaks related to cured fish. They have complicated life cycles. monocytogenes in levels at or above 102 per gram (46). and other local traditional products (7). such as ‘lightly preserved’ fish products. cold-smoked fish.6 and water activities below 0. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. both being detected in cold-smoked salmon. As they are heat-sensitive. have been shown to have an anti-listerial activity in many food products. do not survive 1 min at 60 °C. and in CT95-1207FAIR). Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod.5. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. Unlike some European countries. Two outbreaks in the United States have been reported to be linked to salmon (10). Some LAB. a round worm. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. a number of fish products. which. gravad fish. widely used as starter cultures in food processes. This includes matjes herring. in the EU Fisheries R&D-FLAIR program. monocytogenes pathogenic for only specific segments at risk. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. are considered unsafe.95 (45). when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). favours growth of psychrotrophic LAB. in combination with curing agents and refrigerated storage. The use of LAB in fish preservation is a topic that has been studied in projects (UP. Thus.the round worm Anisakis spp. Of microbiological concern in the northern hemisphere are two parasites . salt levels of 8 . which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). Its limiting conditions for growth are 0 . the CT 3162-FAIR program. The round worms.514. the USA established ‘zero tolerance’ for the organism. SMOKED AND DRIED FISH products such as cold-smoked fish. or nematodes.45 °C.2. which considered L. and the tape worm Diphyllobothrium spp. 5.CURED. the only concern to public health is when eating raw or uncooked fish products. monocytogenes’ in 25 g of the finished product.9.0. Until 1997.

<100 20 . coli O157 & other VTEC Vibrio.<100 >104 >100 N/A N/A <20 20 . E. 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. soft cheese. monocytogenes Staph. † Guidelines for aerobic colony counts may not apply to certain fermented foods. coli (total) Listeria spp. These foods fall into category 5. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E.<100 20 . for example. smell. 5. texture. aureus C.<100 200 .<100 10 .<107 ≥107 N/A <100 <20 100 . monocytogenes) (cfu/g unless stated) <106 106 . cholerae Vibrio parahaemolyticus L.<104 100 .<104 20 .<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 .100 >100 N/A Pathogens Salmonella spp. not detected in 25 g Campylobacter spp. and the levels or absence of indicator organisms or pathogens. the organism should be identified # N/A denotes not applicable 116 .<104 104 . perfringens B.VI Guidelines for the microbiological quality of ready-to-eat smoked fish. salami.<105 ≥105 * Prosecution based solely on high colony counts and/or indicator organisms in the absence of other criteria of unacceptability is unlikely to be successful. (not L. Acceptability is based on appearance.<103 N/A 100 .FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). cereus and Bacillus subtilis# <20 <20** <20 <10 20 . If the Bacillus counts exceed 104/g.6 Published Microbiological Criteria TABLE 5. ‡ On occasions some strains may be pathogenic. and unpasteurised yoghurt.

F. SMOKED AND DRIED FISH TABLE 5. Assurance of Seafood Quality. except in the viscera of dungeness crab. Torry Advisory Note No. Preservation of fish by curing (drying.7 1. monocytogenes Salmonella spp. Horner W. Marinades. References Regenstein J. Wareing P. 2007.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. * Staph. 120-38.. 1979. Leatherhead Publishing. in The Seafood Industry. 8.. 1994. Food spoilage fungi. Eds. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. 56. 334. Smoked. Ed. 9. 254-82 5. Leatherhead. 1990. Fernandes R. 3273. Horne J. Introduction to Fish Technology. in Fish Processing Technology.W. USA. Torry Advisory Note No.CURED. 1991. Catching. Fernandes R..M. Flick G. Chapman & Hall. Moody M. cured and dried fish. FAO Fisheries Technical Paper. Martin R. Bannerman A.. Regenstein C. 1984. New York. Flick G. 4. London. 381-406.H. 37(revised). Huss H. Urner Barry Publications Inc. or presence of toxin 0.F. Eds. 7. 1997. Handling and Processing Eels. Hot Smoking of Fish. 3. Hall G.E. Van Nostrand Reinhold. McLay R.A. 82 (revised). Van Nostrand Reinhold. aureus * Vibrio cholerae V. 1982. in Micro-Facts: The Working Companion for Food Microbiologists. Birnie K. 2.. parahaemolyticus Vibrio vulnificus C. coli (ETEC) L. 6. Doré I. 117 .. The Smoked and Cured Seafood Guide.M. Torry Advisory Note No.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. Wareing P. 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. New York. 1993. botulinum * Limit of tolerance 1x 103 ETEC/g. salting and smoking).

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

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

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

. Poindexter C.N.. 1988. International Journal of Food Microbiology.. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. Liston J. 1974. Presence. Prevalence of Listeria in smoked fish. Poysky F. Competition of food bacteria with Listeria monocytogenes during enrichment culture. 21 (3). Tran T. Beuchat L.CURED. 61 (12).. 25 (5). 866-70. International Journal of Food Microbiology. Food Technology. pH and sodium chloride. and temperature on the growth of Vibrio parahaemolyticus..A. Letters in Applied Microbiology. Histamine production by psychrophilic flora.J.. 844-6. 19. 217-36. Jemmi T. and salt concentration on growth and survival of Vibrio parahaemolyticus. 11. solute. Interacting effects of pH. Journal of Food Safety.A. Peterson M.R.K..P.. 1995. 42. 1734. Beuchat L. 502-8. 11. 1992. SMOKED AND DRIED FISH Sutherland J. Taylor S. 1075-80. Davies A. 1994. Lashbrook L. Journal of Food Protection. Dillon R. Hudson J.. International Journal of Food Microbiology. Applied Microbiology. 14. Incidence of Listeria in hot and cold-smoked fish. 1994. 1991. Letters in Applied Microbiology. Dallas H. Yndestad M. Journal of Food Protection.T. in a salmon slaughterhouse and smoked salmon processing plant. Kramer D. 55 (11). 1995.178-80.. 58 (5). Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. detection and growth of Listeria monocytogenes in seafoods: a review. Combined effects of water activity. 394-6.S.R. Roberts T. 121 . 23 (1).A. Applied Microbiology. Contamination pattern of Listeria monocytogenes and other Listeria spp. Ben Embarek P. Elsevier. 4389-95. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. Letters in Applied Microbiology. 1987. Keusch A.. Caugant D. 27 (6). 94-8.. 1994.W. 1992. Rørvik L. Nicolaides L. Fuchs R. 1995. Eklund M. 309-16. Bayliss A.. Pelroy G.. 25. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. Slade A. Food Microbiology. Hitchins A. in Seafood Quality Determination. 1994..C..R. 293301. Patel T. New York. 309-18. Marine toxins of microbial origin.. Ratnam S. temperature. 1973.E. Eds. Paranjpye R. 19-27.. van Spreckens K.

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

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

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

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

mean that microorganisms have been generally held to play little or no role in the process that follows.3.4. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. Mucor. Similar results were seen with patis. When this is a simple carbohydrate source such as sugar. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. they can contribute to the initial microflora. Hansenula. More recently. either directly or as the partially saccharified rice they produce. were found to be the predominant isolates. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. or boiled or roasted rice. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). These preparations contain a mixed microflora of moulds and yeasts. in many cases. which included nam-pla and patis. sufficient to saturate the water phase present and produce an aw of 0. In some cases. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. (3. angkak (Philippines and Thailand) and look-pang (Thailand) are used. however. and studies 127 . and the anaerobic conditions that prevail in the bulk of the material. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. 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. traditional starter culture preparations such as koji (Japan). however. after one month. Bacillus spp. 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. 4). 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. It also demonstrated their significant proteolytic activity during the first month of fermentation. Generally. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). This.FERMENTED FISH Where other ingredients are used. In another microbiological study of four fish sauces. Endomycopsis and Saccharomyces . principally species of the genera Rhizopus. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns.74 or below.1 – 6.3 6. the Philippine fish sauce. Other halophilic species have been identified from fish/salt products.

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

and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. In fish sauce production. 6.3. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are. thought to be largely the result of autolytic breakdown of the fish tissues. To produce semi-solid fish pastes as opposed to fish sauces.4. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. 14). eviscerated fish are sometimes used to slow the autolytic process. the 129 .6. however.FERMENTED FISH Fig. Ika-Shiokara (side dish) Fig.

5. volatile fatty acids and nucleotides. Some examples of products and their formulation are presented in Table 6.3. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. Hoi-Malangpu-Dong (side dish. In contrast to free-swimming fish. 6. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. 6. for example.6 – 6. snack) 130 .5). Figure 6. which is eventually separated from the residual fish tissues. 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. filtered and bottled.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. particularly glutamate.V and Figures 6.8. Fig.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.

12 days 3 .FERMENTED FISH TABLE 6. NE 10 . N.5-1. In these cases. the prefix 'pla' is replaced by 'kung'. C NE. C NE. Kung-Chom (staple food. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. Fig.4 are also produced using shrimps.12 months Shelf life Use 1 .6.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . S.6. NE. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. side dish) 131 . north-east.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C. N S 5 . 3:1:0. south of Thailand.2-4 Fish (pieces): salt:khaomark (fermented rice).7 days 5 . N. N. central. north. C.03 Preparation time 6 . NE Formulation Fish (whole or pieces): salt: roasted rice.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE.12 months Main dish 2 Pla-jao C.

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

VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.2 7.3 5.9 4.8 .0 Shelf life 3 weeks 6 . Philippines Journal of Food Science and Technology.0 .9.4.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).5.5.9 .8.8 .17. 133 .1 4.2 4.0 .FERMENTED FISH (from Arroyo et al.5..4 . White balao-balao TABLE 6.7 .3 .1 .8 2.8 pH 4.4.5 .6 3.6.5 3.6.9 4.0 .5 .5.2 4.9. 2.3 years 1 .5.12 months 1 .6. 106) Fig.

a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. In several products.2 Inclusion of more carbohydrate gives a faster fermentation and a stronger acid taste Increasing the proportion of carbohydrate in the formulation decreases the product’s buffering capacity and provides the LAB with more substrate on which to act. Where they are used. Garlic.9) compared with the white product (pH 4. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. This results in a more active lactic fermentation and a lower pH.1 . TABLE 6.0 . 19).FISH AND SEAFOOD 6. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6.3.VII). such as salt content. of course. For example. alternatively. Rice is the most common carbohydrate source used. also lower the unit cost of the product and this is probably a significant factor in many cases. When added as boiled or roasted rice alone.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi.5) despite other factors. although the final population of LAB may be similar. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. also serves as a valuable source of the more readily fermented carbohydrate. Addition of carbohydrate will. Although the ability to ferment starch is not widespread in the LAB.2. a common ingredient in these products. 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. being similar (18. 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). the rice may be added in a partially saccharified form such as kao-mark (Thailand). inulin. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. 21).3.4. 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 .

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). which ranged from 7.2 at the end of fermentation. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. There is little published information on the factors that limit the shelf life of the low-salt products.4 Spoilage The ASCA handbook on fermented foods in the region ascribes shelf lives to various fermented fish products ranging from 2 weeks in the case of some of the low-salt lactic fermented products to 5 years for some of the high salt sauces and pastes (16).9%. the pH drops from 6. the level of salt employed is on the high side for fish/salt/carbohydrate products. around 12%. the Korean fermented fish product. Results of a product survey were consistent with this.3 days.8 . In a successful fermentation. Traditional packing techniques used with lactic fermented fish products. The growth of yeasts and moulds could.3 weeks at around 20 °C. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products.5:9:3.FERMENTED FISH Sikhae.7 up to 6. The same product can range from a brown turbid liquid where the fish has been subject to extensive autolysis to brown partly dried pieces of fish. Excessive growth and acid production by the LAB could render products too sour for acceptability. lead to an increase in pH. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB.2. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23).17. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. The mixture is allowed to ferment for 2 . will not reliably exclude air. such as wrapping in banana leaves. markedly extending the shelf life (25). reporting the product’s pH to range from 4. is something of an exception since millet is used along with garlic as the carbohydrate source. in turn. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. Generally. thereafter declining to about 4.7 to below 4. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). Clearly.5. presumably correlating with the salt content.5 in the first 2 . allowing other spoilage organisms to grow. 6. 135 . It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. and there is a marked regional variation in the preferred type. but they are likely to behave similarly to lactic fermented products produced from other commodities. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases.

87. The bacterial pathogens most commonly associated with the living environment of fish. viable numbers will probably decline quite rapidly. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. There is little in the way of documented information associating fermented fish products with foodborne illness. In the fish sauces and pastes. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. As in Norway. which accounted for 88% of outbreaks in the period 1952 . In Japan. outbreaks of 136 . fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage.8 and at salt levels as high 10%. 6. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. During fermentation. are Vibrio parahaemolyticus and Clostridium botulinum. typically 30 °C. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. parahaemolyticus has been reported growing down to a pH of 4. but we are unaware of any reports of this happening. although these limits apply when all other factors are optimal (26). V. Several measures have been suggested for reducing the risk of botulism. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. Because of the severity of the illness it causes. under conditions inimical to growth and high ambient storage temperatures. and consumed without cooking.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. however. After capture. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. the high salt concentrations preclude the growth of all food-poisoning bacteria and those initially present would not survive well the prolonged exposure to high salt during production. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. such as soaking the fish at temperatures below 5 °C before fermentation and the use of starter cultures to ensure a rapid and reliable decrease in pH. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. fish products are the most common source of botulism. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27).

botulinum.. 1966. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). they could act as an important vehicle of transmission. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. Greenland and Alaska. walrus nose. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31). and salmon heads and eggs. Kasemsarn B. Lutefisk. Mahidol University.. Foodborne trematode infections are emerging as a major public health problem. Salting levels are often insufficiently low to prevent growth of C. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. Where these products are consumed without final cooking. Puwastien P. 3. particularly in south-east Asia and the western Pacific regions (30). Saisithi P. 1990. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). 137 . Raw materials subject to this process include seal flipper. giving an alkaline fermentation. 31.6 1. Novus Press. One study with the trematode Haplorchis taichui. 2. and warm weather can result in rapid growth of C. Thailand. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find.FERMENTED FISH botulism have become less frequent in recent years. although the absence of any source of fermentable carbohydrate means that the process is putrefactive. though 27% survival was still recorded (33). Institute of Nutrition. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. which will persist into the product.. whale meat.. Microbiology and chemistry of fermented fish. Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. botulinum and the production of toxin. With this particular organism. which is generally consumed raw (29). while increasing the acetic acid content of the formulation had little or no effect (32).M. and these are often used in the production of fermented fish products. Rakefisk and Herring in the Norwegian Tradition. salt content was found to have the greatest impact on the viability of larvae. Oslo. beaver tail and paw. affecting an estimated 40 million people. Baseline Study (1987-1988).O. 105-10. Journal of Food Science. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. Liston J. Dhanamitta S. 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).. Dollar A. 6. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada. 1989. Fermented Fish Products Study (Thailand) Phase 1. Many of these products are described as fermented. References Riddervold A. Yhoung-Aree J.

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

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

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

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

neoSTX and Gonyautoxins (GTX) 1-4. In addition. Not all the toxins found in shellfish extracts are microalgal products.1). even crustaceans such as lobsters. Consequently. some are the result of biotransformation processes by the filter-feeding host. Many of these species are extremely common and are found throughout the world in a variety of cold.2. all of which contain a tetrahydropurine skeleton (Figure 7. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). The Ncarbamoyl sulfates are readily hydrolysed to STX. 7.2 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. a number of useful and comprehensive reviews and book chapters have appeared dealing with various topics related to individual toxin groups and these are referred to in each appropriate section. and another. 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. with intoxications occurring in many countries even today (15). 7. PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. Gymnodinium catenatum. and a review on the risk assessment of marine toxins has also been produced (14). Saxitoxin Group instead of the Paralytic Shellfish Poisons).g. a great variety of shellfish from all latitudes. The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). and sub-tropical waters and habitats (15). dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. All PSPs degrade rapidly in alkaline conditions. temperate. Around twenty PSP toxins are known. can become contaminated with these toxins.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. 142 . and a tropical species Pyrodinium bahamense (16).FISH AND SEAFOOD has been produced (9). An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10).

the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins.1. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. 7.2. and humans seem to be particularly susceptible. Human symptoms begin with tingling and numbness of the lips. Neurophysiological studies have shown that PSP toxins all possess the same mode of action though some derivatives such as STX and neoSTX are considerably more potent than the C-toxins (15).2 Toxicology PSP toxins affect the central nervous system of mammals. this is never sufficient to render the seafood safe to consume.30 minutes of consumption. and fingers within 5 . 143 .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. However. tongue. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. PSPs bind to site 1 of sodium channels reversibly blocking sodium conductance in nerve and muscle membranes by binding to a specific receptor site located on the outside surface of the membrane close to the ion channel opening.

Taiwan. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). China. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19).3.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. The mouse bioassay has been in place for decades and the method has been standardised. specifically 144 . Alternative HPLC methods have subsequently reduced this to a single HPLC run. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. Two approaches have been taken here. However it remains as the monitoring method of choice in many countries. a pre-column method has received Association of Analytical Communities (AOAC) approval (18). After some development and various trials. and while simple and relatively inexpensive to operate. other derivatives have been reported. 17). and over ten analogs are known (22. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. a criticism has been that such methods are time consuming.8 µg/g) of shellfish or seafood tissue. including Japan. 7. 23.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0.FISH AND SEAFOOD 7. 7.2). Since its discovery. Thailand.2. 15. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. 24) (Figure 7. and islands of the South Pacific. While the chemical detection methods are most promising for the future. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). and has also been evaluated by a number of EU laboratories. as well as other parts.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. Most intoxication events have occurred in South East Asia. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. TTX most commonly occurs in many species of puffer fish. 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. 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.

hypotension. and even newts. 7. An oral dose of 1 . respiratory failure and coma. 27). 26). Severe poisoning causes paralysis and respiratory failure.2 Toxicology Like the PSP toxins. Typically this has been accomplished using HPLC followed by post 145 . All of the observed toxicity is secondary to the action potential blockade. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. raising speculation that TTX is produced by a bacterial source. Structure of tetrodotoxin 7. and in mild cases results in distal muscle weakness. the blue-ringed octopus. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. and once again the mouse bioassay has been used to detect the presence of the toxin. and more recently by an actinomycete Norcardiopsis dassonvillei (26). it is served in speciality restaurants with qualified chefs to properly prepare the fish.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions.FISH AND SHELLFISH TOXINS in the liver. horseshoe crabs. 7.2. and blocks diffusion of sodium through the sodium channel. nausea and vomiting. where puffer fish of the genus Fugu are a delicacy. Chemical methods for TTX detection have also been investigated and are mainly based on its smooth conversion by heating in an alkaline solution to a detectable fluorescent (C9) base. molluscs. Fig. The onset of symptoms usually occurs rapidly. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. gonads and roe (24). 25. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. A review of TTX detection methods has been published (12.2 mg of purified toxin can be lethal. 23.3.3. Since then. In Japan. preventing depolarisation and propagation of action potentials in nerve cells. and ingestion of larger amounts leads to cardiovascular effects including bradycardia.

Currently 12 isomers of DA have been reported (see Figure 7.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). 7.FISH AND SEAFOOD column oxidation to the fluorescent derivatives. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide.4. a polar glutamate agonist. but it is known that such congeners are less toxic than DA itself (30).3 for examples). Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. Structure of domoic acid and three naturally occurring photoisomers 7. and while promising. Fig. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28).4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). Immunoassay methods have also been investigated. However this method has been criticised for being complex. 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).3. but all of them are considerably less toxic than DA itself (29). but 146 . These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. 7. need further development (24).

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

dinophysistoxin-1 (DTX-1). DTX-2 was discovered in contaminated Irish shellfish (46). In the context of this chapter. 48. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. australis. 7.3 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.4). as can LC/MS. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). However. and these methods have been reviewed (40. although in the latter study the probes for Pseudonitzschia pungens. 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. Since then.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). DA can be detected by the mouse bioassay but the regulatory limit for shellfish is set below this level at 20 µg/g (20 mg/kg) shellfish tissue. and an immunosensor method has also been reported (4). DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. and consequently were often overlooked as the real source of the toxins. produce a variety of other 148 . An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. The benthic Prorocentrum spp. but just over 15 years ago a third toxin. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). 41). the heterotrophic dinoflagellates Dinophysis spp.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). though this method is most commonly used to analyse plankton and seawater (42). The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. A rapid immunodiagnostic kit is now available to detect DA in phytoplankton and is reported to work satisfactorily in this application in the field (43). To further complicate matters. and dinophysistoxin-2 (DTX-2) (Figure 7. and this is typically the method of choice..4. Alternatively. are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. 49). Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. in Monterey Bay. California.. Dinophysis spp.FISH AND SEAFOOD 7.

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

A regulatory level of 80 . In recent years.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. most progress in DSP toxin detection has been using new or improved LC/MS methods and a detection limit of 10 µg/kg has been reported. although a comparative study of in vitro phosphatase inhibition with mouse bioassay data has shown that OA is about twice as potent as DTX-2 (51). Due to the lipophilic nature of the toxins. DSP-Check is a commercially 150 . and a detection limit of 100 µg/kg has been reported (12). and DTX-2. Human symptoms include diarrhoea. various European countries.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. nausea. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). Since the late 1970s. 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. DSP episodes have been reported from Japan. 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. and abdominal pain. DSP episodes have been reported worldwide and are included in many monitoring programs (47). 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. even with the minimal of clean up steps (48). Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1.5. vomiting. 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). and while this provides critical seafood safety information. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. This regulatory level includes the possibility of LSPs in the DSP fraction being tested.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. 7. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. which can lead to an underestimation of the total DSP content in a sample.5.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . it does not provide information on the DSP toxin profile. 7. New Zealand and Chile.3 days.

5. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). Some representative examples of both groups of toxins are shown in Figures 7. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. 7. Indeed. 58). In yet another approach. If a European collaborative study of the fluorometric protein phosphatase inhibition method is successful it may eventually be approved for regulatory purposes in the EU.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity.6 Lipophilic Shellfish Toxins (LST) Lipophilic extraction of toxic shellfish for DSP toxins often results in the simultaneous extraction of other lipophilic polyethers such as the pectenotoxins (PTXs) and yessotoxins (YTXs). with a claimed detection limit of 20 µg/kg.6. 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 . for many years these compounds were included in the DSP toxin complex. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). 55). Within this group. with good agreement with the mouse bioassay and LC–based methods (54. and another rapid antibodybased test kit for DSP toxins has also been reported (53). 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. However.5 and 7. Fig. 7.

eastern Canada. 7.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). YTX and homo-YTX 7.6. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. 57. many parts of Europe and the Mediterranean. 58).FISH AND SEAFOOD Fig. the toxin content and profile may vary among different strains. 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.6. As is typical for many algal species. 7. Structure of the two parent yessotoxins. While 152 . The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum.1 Origins and distribution It is now well recognised that PTXs are co-produced with DSP toxins by various strains of the dinoflagellate Dinophysis and hence the distribution of both families of toxins in shellfish is similar (49).6. Japan. New Zealand and South America (49. PTX-1 and PTX-2 are frequently the most common derivatives to occur and PTX-2 is the most potent following intraperitoneal (IP) injection (230 µg/kg body weight). which may co-occur in blooms with other organisms.

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

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

Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. and an action level has been set at 800 µg/kg (or 80 µg PbTx-2 equivalents/100 g or 20 MU/100 g or 4 µg/mouse) of shellfish tissue to trigger regulatory action. The mean time to onset of symptoms is around 3 hours.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. The human health implications following exposure to NSPs have been reviewed (69). A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. In this application. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). the competitive ELISA method correlated most closely with the mouse bioassay results. 66. with hot spots in the coral reef areas of the Caribbean and French Polynesia. and in severe cases seizure. The mouse bioassay is used as the primary detection method.7. 7. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 .7. 7. and diarrhoea. which can result in a cascade of associated events in a cell (66). pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. and mammalian body fluids without pretreatment or dilution (72). Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. this family of toxins has proven to be a challenge both to identify and monitor. with a mean duration of symptoms of around 1 day. 69). While the mouse bioassay continues to be used. shellfish homogenates. malaise. vertigo.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). Ingestion of brevetoxins results in a variety of symptoms including parathesia. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). 7.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. nausea.

These include species belonging to the genera Ostreopsis and Coolia. a group of closely related polyethers that are considered the source of ciguatoxins. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin. green and brown seaweeds and also occur in sediments and coral debris. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7.8). lipid soluble polyethers. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red.8. CTXs are large. A detailed comprehensive review of ciguatoxins has recently been published (74). The diversity of benthic phytoplankton that exist on reefs further complicates the picture.8.1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins. but more are suspected to exist. complex. Fig.FISH AND SEAFOOD transferred through the food chain to the larger carnivores. Distribution through the food chain begins with 156 . although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. and only a few structures have been identified. 7.

3 Detection methods As a consequence of their potency. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. Like the brevetoxins. pruritus. These include paresthesias (numbness and tingling of the extremities). Local island populations attempt to avoid poisoning by catching only smaller fish. which in turn are consumed by carnivores such as snappers. and there are suggestions that such incidents are increasing and spreading worldwide (74).76). they bind to voltage-dependant sodium channels in the cell membrane causing them to open. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. only trace amounts of CTXs can cause illness following consumption of contaminated fish. barracudas. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. Even when suspect samples are available. and temperature sensation reversal. all of which can be linked to the effect of CTX on excitable membranes. The recommended method to extract fish tissue has been described in detail (74) and the initial warm acetone extract is partitioned against hexane and diethyl ether where the CTXs are concentrated. arthralgia.8. Another symptom that can develop over time is extreme fatigue. there are neither standards nor an official testing program. weeks or even months (74 . The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. where cold objects feel hot and vice versa. which can return following the consumption of alcohol and certain foods such as oily fish (74.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. and moray eels. Patients have reported symptoms lasting weeks and even months. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area.8. although not all CTXs are equipotent. 78). 75. and although the gastronomical problems may dissipate within a day or so. The onset of symptoms can occur rapidly within 24 hours. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . 7. myalgia.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. and the sensation of loose teeth (77). and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. the neurological symptoms can persist for days. 75). which can be confused with chronic fatigue syndrome. Other symptoms include a metallic taste. and it is estimated that ingestion of as little as 0. 7.1 µg of ciguatoxin can result in human illness (74. Globally. groupers. sharks. and in fact CTX poisoning is often only identified after consumption of contaminated fish.

high-throughput screen (74). specific for CTX activity. almost 20 additional derivatives have been reported (83 . Recent reviews of this new group of toxins have appeared (83. only a few laboratories are able to undertake such analyses. and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue.g. A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine.80%). followed by clean-up on silica and aminopropyl cartridges.9). and only a few structures (AZA 1 . and following synthesis of the molecule some time later. minor revisions of a few stereocenters were made. Since the original discovery and identification of the toxin. Acetone extracts of fish tissue are subjected to solvent partitioning. following consumption of contaminated mussels Mytelius edulis (82). and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. which features characteristic fragment ions corresponding to sequential losses of three molecules of water. In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). Chemical detection methods are severely compromised by the lack of standards.30%) and AZA3 (5 . another in vitro sodium channel-based assay. and another dealing with the chemistry and detection methods has been published (85). Usually the major AZA derivatives found in shellfish are AZA1 (50 . Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. The original structure proposed revealed an entirely new class of shellfish toxin. Immunoassays may be an alternative detection method (79). but this only has application in a clinical setting (81).25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. 84). The majority of these have been proposed based on LC/MS analytical data. and the corrected structure was published in 2004 (86) (Figure 7. although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). which are based on an LC-MS/MS approach. Consequently. Recently.85).9). has been developed. This is then subjected to LCMS/MS analysis. with AZA2 (10 . 7.FISH AND SEAFOOD Lethal Dose (MLD) 0.20%) present in lesser amounts (Figure 7. at C-3 and C-23).9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. 158 .

and to date this has been the only source from which these toxins have been found at toxic levels. However. Structure of the three principal azaspiracids found in shellfish 7. Portugal. Ostrea edulis). and some time after the original episode in Ireland. and to date no long-term effects have been recorded (83. 84. Mytilus galloprovincialis). cassipes produces AZAs in culture. 7. scallops (Pecten maximus). vomiting. which are ingested by P. In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. Donax spp. cockles (Cerastoderma edule).). which can persist for 2 . 90). 89). These symptoms are not unlike those experienced following the consumption of DSP toxins. and most recently in a crustacean (Cancer pagurus). AZA toxins do 159 . The bivalves affected include mussels (Mytilus edulis. oysters (Crassostrea gigas.9.9.FISH AND SHELLFISH TOXINS Fig.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish.3 days. Scandinavia. While this result is not necessarily in debate. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). and off the coast of Morocco (83.9. but current data indicates that unlike the DSP compounds. cassipes (83). Following the original episode. these molluscs were never considered to be the biogenetic source. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. AZA occurrence has been reported by several European countries including France. 7. 88. it has been impossible to establish that P. diarrhoea and stomach cramps. Ensis siliqua. clams (Tapes philipinarum.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. as in other cases of shellfish toxins. There has been considerable debate over the source of azaspiracids.

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

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

This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. and this is the detection method of choice.FISH AND SEAFOOD 7.900 µg/kg).100).11). Based on this diminished oral toxicity. abdominal breathing.1. 7. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. They were originally discovered in extracts of mussels (M. Since their original discovery which resulted in the characterisation of spirolides A-F (99. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode.to10-fold reduction of toxicity (LD50 600 . Oral administration of the toxin to mice by gavage resulted in about an 8. finishing with 100% methanol. A later study using purified toxin established an LD50 dose (Lethal Dose. paralysis of the hind quarters and eventually complete immobility. However. almost a dozen derivatives in total have been identified to date (94. 95) (Figure 7.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). and so spectroscopic detection by UV methods is not favoured.3 Detection methods The characteristic “fast-acting. The time lapse between ingestion and death is between 5 and 15 minutes. 7. Most cyclic imines lack an extended chromophore. together with a lack of evidence of human intoxication. Whether isolated from dinoflagellate cells directly or from shellfish tissue. 95). In particular. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine.10. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. There is a rapid onset of symptoms including respiratory distress.1. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. 50%) of 96 µg/kg. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94.10. Reduction of the imine group eradicated the toxicity.10. it has been stated that regulation against gymnodimine is not required (98).2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. 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 lack of responsiveness to stimuli.

7.10.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. 95). or by oral ingestion (381 . They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. All data reported here are for desmethyl spirolide C. Interestingly. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 .FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). While oral toxicity is considerably less than by IP injection.11.707 µg/kg. fasted mice).2. the western coast of Scotland. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). Fig. 7.8 µg/kg). southern Ireland.11. those spirolides possessing the vicinal dimethyl grouping (see Figure 7. Structure of four spirolide toxins A-D 7. in fjords in Denmark and Norway.2. fasted mice).166 µg/kg. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. by gavage (87 . their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102).2 Toxicology The spirolides are the most studied group of the cyclic imines. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . 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.10.

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. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. While the mechanism of action remains to be clearly defined. 95. 7. Three members of this group have been reported.10.B contain a spiro imine moiety similar to that found in the spirolides. which includes two diastereomers. The compounds are zwitterionic and therefore somewhat more polar than the spirolides.3 Pinnatoxins and pteriatoxins Pinnatoxins A . and in fact these compounds share a number of other structural features.3 Detection methods As discussed for gymnodimine.FISH AND SEAFOOD (100). The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7.10.12). the mouse bioassay can be used to detect these fast acting toxins. 7. consistent with modulation of acetyl cholinergic receptors (103). To date.2. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C. A number of methods following this procedure have been described (94. 7. Structure of the pinnatoxin group and the closely related pteriatoxins 164 .12. 102). However. four members of the group have been identified – two of them being the C-34 diastereomers. Fig.

and more advanced means of detection. With increasing knowledge of the chemistry of toxins. There is strong circumstantial evidence to suspect that these toxins.10.180 µg/kg). Only the acute toxicity in mice following IP injection has been reported.2 Toxicology There is little information describing the toxicology of these compounds (98).1 Origins and distribution Shellfish of the genus Pinna. which occupy tropical waters around Okinawa and the south east coast of China. However.3. 7. While this is recognised as a real possibility for a wide range of toxins. Coupled with this. This is generally a result of increased monitoring and continually improving detection technologies and methods. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. while pinnatoxin D was essentially inactive. 7.10. arise from an algal source . This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104).FISH AND SHELLFISH TOXINS 7. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. which in many cases are now found to be more widespread than previously believed. but this remains to be conclusively established (106). 99%) values for pinnatoxins A-C are in the range (22 . Despite this progress.3 Detection methods No formal detection methods for these compounds have been reported.most likely a dinoflagellate. have long been suspected of causing human illness after ingestion.3. new toxin groups such as the azaspiracids and cyclic imines have also been reported. 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. The LD99 dose for the pteriatoxins is reported to be in the range (8 . to date action or 165 .11 Conclusion There has been considerable progress in the detection of known marine toxins.100 µg/kg).3. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins.10. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). there is no compelling evidence to suggest that they have resulted in human toxicity (98). The LD99 (Lethal Dose. like the gymnodimines and spirolides. 7.

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

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

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

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

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

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

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4. procedure. 8. operation or stage in the food chain including raw materials.A point. Validation . procedures. from primary production to final consumption. The team should also include 177 .3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. food chemistry or engineering. in addition to monitoring to determine compliance with the HACCP plan. Establish a system to monitor control of the CCP. 5. quality assurance or control. Establish documentation concerning all procedures and records appropriate to these principles and their application.The application of methods.Obtaining evidence that the elements of the HACCP plan are effective. hygiene managers. 6. Figure 8. 3. Establish the critical limit(s). 2.1 Assemble HACCP team HACCP requires management commitment of resources to the process. 7.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . 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. Each stage is discussed below in detail. ingredient and packaging buyers. food microbiology. Determine the CCPs. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. 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.3. Establish procedures for verification to confirm that the HACCP system is working effectively. tests and other evaluations. Conduct a hazard analysis. Verification . 8.1 is a flow diagram illustrating this 12-stage logic sequence.

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

8.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e.3. In addition.2 Describe the product It is important to have a complete understanding of the product.3. Vulnerable groups of the population.g. etc. there may be species-specific hazards related to certain fish and shellfish. which should be described in detail.g. naturally present on fish from warm waters (> 15 °C).HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.). etc. storage and distribution conditions and instructions for use. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . aquaculture drug residues and potentially pathogenic Vibrio spp. composition. packaging. The description should include information such as the product name. including practices onboard fishing vessels.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. fermentation. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments.3. is a cooking process required?). pH. 8. physical and chemical structure (including water activity (aw). It is also important to identify the consumer target groups. shelf life. These factors are likely to have a significant effect on some potential hazards. such as enteric viruses in shellfish. or in aquaculture operations. 8. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. If the processing does not include a kill step. freezing.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. may need to be considered specifically.). marine toxins associated with particular fish and shellfish species. 8.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora. However. Expertise on such inherent hazards.1. contaminated fish and shellfish should not be used.3.3.3. processing conditions (e. such as children or the elderly. heat treatment.

The operation should be observed at all stages. ingredient specifications. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working.FISH AND SEAFOOD product. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. 8. 8.3. It may include details of the factory and equipment layout. 8.3.4.3. Ideally. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards.5 On-site confirmation of the flow diagram The HACCP team should confirm that the flow diagram matches the process that is actually being carried out. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. It is essential that the diagram is accurate.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. 180 ● ● .6 List all potential hazards associated with each step. and therefore a detailed description of the procedure applied must be available. However. 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. 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. features of equipment design. it should also include details of CCP steps. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. If HACCP studies are applied to proposed new process lines or products. survival or multiplication of pathogenic microorganisms. time/temperature data. cleaning and hygiene procedures and storage conditions. For example. conduct a hazard analysis. the qualitative and/or quantitative evaluation of the presence of hazards. it is essential that the details of each step are fully appreciated and recorded. or circumstances of any reclaim or rework activity. because the hazard analysis and decisions regarding CCPs are based on these data. once determined.

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

then the process must be modified to include a control measure. it can be said that effective control measures are likely to include the following: • • • • • 8.3.7. However. application of the decision tree should be flexible. Where possible. However.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8.3.2) to provide a logical. Some examples relevant to fish products are: • • • Time and temperature limits for pasteurised and cooked products Temperature limits for chilled fish Brine concentration in cured fish 182 . It is prudent to set stricter limits (often-called target or process limits/levels) to ensure that any trend towards a loss of control is noted before the critical limit is exceeded. If a significant hazard has been identified at a step where control is required for safety. critical limits should be specified and validated for each CCP. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. it is necessary to specify both time and temperature for a thermal process. 8.3. biological or sensory parameters. More than one critical limit may be defined for a single step.1 Fish and seafood Again.8. but for which no control exists at that step or any other. For example. structured approach to decision making. chemical. and the reader is referred to the appropriate product chapter.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. 8.3. and its use may not always be appropriate. it is not possible to generalise on likely CCPs. Criteria used to set critical limits must be measurable and may include physical.FISH AND SEAFOOD 8. given the enormous variety of fish products and processes in use.

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

and conditions of raw materials.9. transport and delivery vehicles requirements. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . to personnel requirements. the existence of microbiological standards alone cannot protect consumer health. with the intention of introducing consistency and clarity throughout the food production chain from primary production. from structural conditions in the factory or process facility. under which food is produced to optimise public health and to prevent. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials. 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. EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr. 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. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. The general food hygiene Directive 93/43/EEC and other Directives on the hygiene of foodstuffs and the health conditions for the production and placing on the market of certain products of animal origin intended for human consumption. and one that plays an important role in most countries’ food legislation. however. to sale or supply to the final consumer. final product specifications. The new legislation was designed to establish conditions. A number of standards are provided in food legislation. Hygiene is a general concept that covers a wide subject area. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. including microbiological criteria. eliminate or acceptably control pathogen contamination of food.

2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. Subsequently. and sets out what those enforcing the provisions have to do.2 Legislative Structure From 1 January 2006. European Communities L 268.1991. as last amended by Regulation (EC) No 806/2003. p. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. N. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. 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. 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. 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. processing. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No.7. 24. handling and offering for sale or supply to the consumer. J. storing. 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. 9. p.1991.15). 24. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. 41).B. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. 190 • • .1992.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. packaging. 7.9.9. as last amended by Regulation (EC) No 806/2003. A number of more detailed implementing and transitional measures have been adopted at EC level. manufacturing. p. transportation.1). distribution.

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

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

4. (iii) Additionally. the mark must be oval shaped and include the abbreviation CE. EC. Section I of Regulation (EC) 853/2004 are given as follows: 9.4. (iv) If the identification mark is applied in an establishment within the Community.2 Fishery product requirements 9. EF. when a product's packaging and/or wrapping is removed or is further processed in another establishment. a new mark must be applied to the product. EÜ.2.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. An identification mark can only be applied provided the relevant requirements as stated above in relation to Regulation (EC) 852/2004 have been met in particular for the registration and approval of establishments.1. (ii) However. for other codes. the new mark must show the approval number of the establishment where this occurred. Extracts of the requirements of Annex II. ES. EG. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. In this case. Section I of Regulation (EC) 853/2004. the food business operator may apply the same identification mark to both types of food. the mark must show the approval number of the establishment. and the characters easily decipherable and be clearly displayed. For the UK. EB or WE.1 Identification marking An identification mark needs to be applied following the requirements of Annex II.2 Form of the identification mark (i) The mark must be legible and indelible.FISH AND SEAFOOD 9. the code would be UK. (iii) Food business operators must have in place systems and procedures to identify food business operators from whom they have received and to whom they have delivered products of animal origin.4.4.2. EY. 198 . EK.1. EK. (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. please refer to Regulation (EC) 853/2004. EO.2. 9.

wrapping or packaging in another establishment. and fishery products carried in bulk.4. it applies to those animals when not placed on the market live. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. The requirements of this section accompany those in Regulation (EC) 852/2004. echinoderms. (v) When the mark is applied directly to products of animal origin.1. the approval number of the establishment and the abbreviation for the community where appropriate. including vessels. Therefore. and are intended for further handling. Section VIII of Regulation 853/2004.2.4.LEGISLATION 9. its wrapping or the packaging. processing. (ii) For products of animal origin that are placed in transport containers or large packages. Extracts of the requirements of Annex III. or be printed on a label affixed to the product. Part A and Chapter V apply to retail. granulate and powdered products of animal origin carried in bulk. it is sufficient to apply the mark to the exterior of that package only. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . 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 wrapping or the packaging.3 Method of marking (i) The mark is to be applied directly to the product. With the exception of Chapters I and II. (iii) In the case of liquid.2.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. Chapter IV. the mark may be applied to the outer surface of the container or packaging. This section does not apply to bivalve molluscs. tunicates and marine gastropods when placed on the market live.e. i. Chapter III. the colourings used must be EC authorised colours. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. C and D. reference should be made to the actual Regulation. 9. Parts A. as amended. for full requirements. a) requirements for establishments. specifically relating to fishery products are given below. they would need to be obtained following Section VII on live bivalve molluscs.

refrigeration and wrapping. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. this supplements the requirements of Annex II. bleeding. (a) primary production covers the farming.e.1 Chapter I .2.4. i. 2. if a vessel has water coming in that is used with fishery products. or to handle or process them after harvesting. it must be placed somewhere that will not contaminate the water supply.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.. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 . heading.FISH AND SEAFOOD c) for the water supply. There is a derogation from point a. The definition of fishery products covers primary production as well as its associated operations. Chapter VII of Regulation 852/2004. A. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. clean seawater may be used for the handling and washing of fishery products. removing fins. gutting. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking. fishing and collection of live fishery products with a view to their being placed on the market. from the place of production to the first establishment of destination.2. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter.) 9. 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. Structural and equipment requirements refer to the following points 1.

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

Requirements during and after landing 9. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. landed as soon as possible. c) Fishery products. (ii) In the case of food business operators responsible for auction and wholesale markets etc.4.2. provided no damage occurs to the flesh. if this is not possible. where fishery products are displayed for sale. 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. e) In cases where whole fish intended for canning is frozen in brine. 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 .2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable.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. 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. B. must be chilled or.2. chilled and wrapped. Handlers may use spiked instruments to move large fish or fish that might injure them. other than those kept alive. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. The brine must not be a source of contamination. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption. Chapter II . b) Factory vessels on board which crustaceans and molluscs are cooked. the product must be kept at no more than -9 °C. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. and products washed thoroughly.

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

(iii) sprat. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. (iii) A document from the manufacturer. D. stating the type of process they have undergone.2. Once produced.Requirements for certain processed fishery products A.4 Chapter IV . 9. Requirements concerning parasites (i) The fishery products below must be frozen at no more than .2. and whole fish must be gutted and washed beforehand. (iv) (wild) Atlantic and Pacific salmon. must accompany fishery products referred to above when placed on the market. cooling must continue until a temperature 204 . if they are to undergo a cold smoking process in which the internal temperature of the fishery product is not more than 60 °C: (i) herring. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. or a stabilising treatment. they must be cooled rapidly. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. if the processing is insufficient to destroy nematode larvae.20 °C in all parts of the product for at least 24 hours (this treatment must be applied to the raw product or the finished product): (a) fishery products to be consumed raw or almost raw.4. raw materials must have no guts. except when supplied to the final consumer. (ii) mackerel. and (c) marinated and/or salted fishery products. (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. (b) fishery products from the following species.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. If no other method of preservation is used.

B. 205 . 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. 2074/2005. the following conditions need to be met: A. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No.2. 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.2. (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. processing. 2073/2005.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. 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. C. 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. pressing. separation. and the raw material is processed within 36 hours after loading. Shelling/shucking is to be done hygienically.LEGISLATION approaching that of melting ice is reached. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII.5 Chapter V . 2073/2005). Organoleptic properties of fishery products An organoleptic examination of fishery products must be carried out to ensure that fishery products comply with any freshness criteria (such criteria have yet to be set). and after this. 2074/2005).4. centrifugation. 9. raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII.

prepared and processed fishery products belonging to the family Gempylidae. and when fishery products are wrapped on board fishing vessels. Diodontidae and Canthigasteridae.4.6 Chapter VI . in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. Fresh.2. The scientific name must accompany the common name on the label.2.4.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. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product.Storage of fishery products (i) Fresh fishery products. E. 9. 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. Fishery products contaminated with parasites may not be placed on the market for human consumption.FISH AND SEAFOOD D. it should be kept at no more than -9 °C. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption.2. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. thawed unprocessed fishery products. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. Molidae. 206 .2. food business operators must ensure that wrapping material is not contaminated and is stored in such a way that it is not exposed to contamination and is easy to clean where necessary. 9. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. In the case of whole frozen fish in brine intended for the manufacture of canned food.7 Chapter VII .

(b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar.4. provided that the journey is short and the competent authority allows this. Helicolenus dactylopterus. The temperature of frozen fishery products must not fluctuate above -15°C. 9. (ii) If fishery products are kept under ice.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. 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. 2074/2005. species belonging to the Merlucciidae family.Transport of fishery products The above points for Chapter VII apply in relation to transport. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. 9.2.).4. Sebastichthys capensis. species belonging to the Gadidae family.2.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. melt water must not remain in contact with the products. (d) 60 mg of nitrogen/100 g of whole fishery products used directly for the preparation of fish oil for human consumption as referred to in the second sub-paragraph of Part B(1) of Chapter IV of Section VIII of Annex III to 207 .2.8 Chapter VIII . The 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. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them.

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

Transport needs to allow sufficient drainage for best survival conditions. collection from: . 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. 9. B. 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.2 Chapter II . (ii) Provided the requirements of chapter V are met. C.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority. Requirements for harvesting and handling following harvesting This includes requirements that any further handling should not cause contamination or excess damage to shells/tissues of the live bivalve molluscs that might affect their suitability for purification.Class A means they can be placed directly on the market .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. poles or other fixed means are to identify the boundaries of the sites. processing or relaying.Hygiene requirements for the production and harvesting of live bivalve molluscs A.3.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market . 209 . Requirements for production areas (i) The competent authority classes (classed A.

Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements.4.3. requirements if there are many batches of live bivalve molluscs. Requirements for dispatch centres Handling should not result in contamination to any degree. 160 micrograms of okadaic acid equivalents per kilogram. and there are requirements for tanks and water storage containers. B. (iv) yessotoxins.4. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. 210 . 800 micrograms per kilogram. 9. for: (i) paralytic shellfish poison (PSP).3. (ii) amnesic shellfish poison (ASP).3.4 Chapter IV .3 Chapter III .FISH AND SEAFOOD 9. 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.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc.5 Chapter V .Hygiene requirements for purification and dispatch centres A. Before dispatch. the levels of marine biotoxins should not exceed the following limits. dinophysistoxins and pectenotoxins together.4. the purification system to allow them to resume /maintain filter-feeding activity. 1 milligram of yessotoxin equivalent per kilogram. Requirements for purification centres There are requirements for live bivalve molluscs to be washed. containers and a label to identify purification.

160 micrograms of azaspiracid equivalents per kilogram.6 Chapter VI .4.4. Requirements of Chapters III and IV are to be complied with. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’. 9.3.Other requirements (i) The temperature at which live bivalve molluscs are stored/ transported in should be at one that does not adversely affect food safety or their viability. Also.4.7 Chapter VII . (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9. 9.4. 211 . The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction. Registration document requirements also apply.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.3. the species of bivalve mollusc and date of packaging (at least the date and month) are required. dispatch centre or processing establishment. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre.9 Chapter IX .1). Finally.3. 9.4.8 Chapter VIII . the label.Identification marking and labelling In addition to general identification marking (contained in 9. (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.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.3. including the identification mark. they cannot be re-immersed in or sprayed with water. must be waterproof. by way of derogation from directive 2000/13/EC.LEGISLATION (v) azaspiracids.2.

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

Monitoring of classified relaying and production areas This is to ensure there is no malpractice. In the remaining 10 % of samples.5. that microbiological criteria and chemical contaminants amongst other requirements are met.5. Regulation (EC) 854/2004. A summary is given as follows: 9.000 E. Live bivalve molluscs from Class B areas must not exceed.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. chapter V is reinforced.600 E. three dilutions MPN test specified in ISO 16649-3. section VII. coli per 100 g of flesh and intravalvular liquid.2. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. - - - - B. live echinoderms. Annex II 9. 4. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated.1 Chapter II . three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. live bivalve molluscs must not exceed 46. Classification of production and relaying areas: This section reinforces that specified in Annex III. live tunicates and live marine gastropods.Official controls concerning live bivalve molluscs from classified production areas A. coli per 100 g of flesh and intravalvular liquid. The reference method for this analysis is the five-tube. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met.2 This states official controls for the production and placing on the market of live bivalve molluscs. coli per 100 g of flesh and intravalvular liquid. 213 . in 90% of the samples. Also. The reference method for this analysis is the five-tube. Live bivalve molluscs from Class C areas must not exceed 46. as previously stated. Sampling plans and frequency are required. 000 E.

E. F. the competent authority is to close the production area. Additional monitoring requirements This concerns the competent authority in relation to monitoring of classified production areas where bivalve mollusc harvesting is forbidden and of relaying and production zones to ensure there is no malpractice etc. establishes microbiological criteria for a range of foods.6 Regulation (EC) No. Decisions after monitoring There are specific requirements whereby if sampling results indicate that health standards for molluscs are exceeded or there is a risk to human health. boundaries or class of a production area or its closure. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. the Member States may permit the import of bivalve molluscs and fishery products from the countries listed respectively in Annex I and Annex II to regulation (EC) 2076/2005. 9. 214 . Procedures for re-opening a closed production area are stated D. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No.2. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. opening or closing a production area.2 Chapter III . 9. 9.FISH AND SEAFOOD C. 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.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. 2073/2005.5. which has applied since 1 January 2006.5.

I.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. a batch of foodstuffs or a process. based on the absence. area or batch.6. The criteria are outlined in Table 9. and the legislation applies to all businesses involved in food production and handling. presence or number of microorganisms.LEGISLATION The aim of this legislation is to complement food hygiene requirements. and/or on the quantity of their toxins or metabolites. Two kinds of criteria have been established: food safety criteria. ensuring that foods being placed on the market do not pose a risk to human health. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. volume.6. 9. per unit(s) of mass. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. 9.II. 215 . The relevant criteria are outlined in Table 9. with chapter 2.4 referring to fishery products.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria.

TABLE 9. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.3 Ready-to-eat foods unable to support the growth of L.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life .2 Ready-to-eat foods able to support the growth of L. monocytogenes. 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. 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. 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. monocytogenes 1.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L.

tunicates and gastropods 9 (18) 2 E.25 Live bivalve molluscs and live echinoderms.17 Live bivalve molluscs and live echinoderms. coli (15) 1.27 Fishery products which have undergone enzyme maturation treatment in brine. manufactured from fish species associated with a high amount of histidine (17) Histamine .Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1. tunicates and gastropods 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 1.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1.

The most recent edition of the standard shall be used. The operator may fix intermediate limits during the process that must be low enough to guarantee that the limit of 100 cfu/g is not exceeded at the end of shelf life. Dervin C. m = M.. 79. 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. excluding sprouted seeds. 1996. wine.92.1. Scombresosidae. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . Malle P. 5 This criterion shall apply if the manufacturer is able to demonstrate. — fresh. Single samples may be taken at retail level. subject to scientific justification. Bouquelet S. monocytogenes. live bivalve molluscs. References: 1. when he is not able to demonstrate. uncut and unprocessed vegetables and fruits. E. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. including cocoa and chocolate products. biscuits and similar products. according to which the whole batch is to be deemed unsafe.4 or aw ≤ 0. Bouquelet S. Coryfenidae. Valle M. A pooled sample comprising a minimum of 10 individual animals. AOAC Internat. 43-9. J.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. — — bottled or packed waters.. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. products with pH ≤ 5. Malle P. Particularly fish species of the families: Scombridae. products with a shelf-life of less than five days shall be automatically considered to belong to this category. c = number of sample units giving values between m and M For point 1. Assay of biogenic amines involved in fish decomposition. soft drinks.94. 2. Engraulidae.. cider. Duflos G. shall not apply.25.0 and aw ≤ 0.. coli is used here as an indicator of faecal contamination. This criterion shall apply to products before they have left the immediate control of the producing food business operator. that the product will not exceed the limit 100 cfu/g throughout the shelf life. — bread. to the satisfaction of the competent authority. Other categories of products can also belong to this category. products heat treated in their final package). Clupeidae. honey and confectionery. sugar. to the satisfaction of the competent authority. when recontamination is not possible after this treatment (for example. beer. Products with pH ≤ 4.. Pomatomidae. spirits and similar products.1 .

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

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

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

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

1). Regulation (EEC) 2136/89. Regulation (EC) No. 1662/2006.2006. 22) as amended by Regulation (EC) Nos.6. 223 . L338. Regulation (EC) No. L226. 1688/2005. 206). 30. 2074/2005. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. 9. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. Commission Regulation (EC) No. L283. as read with the corrigenda at OJ No. 2073/2005 on microbiological criteria for foodstuffs (OJ No. 83) as amended by Regulation Nos 882/2004. 1441/2007. Regulation (EC) No.2004. 14. 853/2004 is now set out in a Corrigendum (OJ No. 30.2004.10 Further Reading Regulation (EC) No.LEGISLATION 9.4. 1. L139. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 2076/2005.2004. 3) as amended by Regulation (EC) No. 62) as amended by Regulation (EC) No.4. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. Regulation (EC) No.10. 25. 30. 2074/2005.6. 854/2004 is now set out in a Corrigendum (OJ No. 2074/2005. The revised text of Regulation (EC) No.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. L226. namely.4. 55). 2076/2005. 852/2004 is now set out in a Corrigendum (OJ No. 2074/2005 and Regulation (EC) No. L139.12.2004. L139. 2075/2005 and Regulation (EC) No. 2076/2005.2004.2005.6. The revised text of Regulation (EC) No. 25. Regulation (EC) No. L226. Regulation(EC) No. these are not be discussed here. 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. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. as amended. 1663/2006. The revised text of Regulation (EC) No. 2076/2005. 22.2004. 1019/2008 and as read with Regulation 2073/2005. laying down common marketing standards for preserved sardines.

as amended by Commission Regulation (EC) No 1022/2008 OJ L 338. (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004 ( OJ L 338.2001. 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 278.2000. 21.1. 6) 224 .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. 23. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004.12.12.2005. 22.10. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.2005. derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004. 22.

under optimal incubation temperatures.5 µm 10.3. The optimum growth temperature is 28 °C (2).1.000-fold increase in number is observed after one week in food systems (1).5.1 Aeromonas spp. a 10 to 1. 225 .3 .1. 10.3 minutes. 0.1. Morphology Gram-negative rods. 10. respectively (1).10 (3).4 Heat resistance Aeromonads are sensitive to moderately high temperatures. and 2.1.2 Oxygen requirements Facultative anaerobe.1. PATHOGEN PROFILES 10.5 minutes. At 5 °C. 10.3 Temperature Aeromonas grows at temperatures of 2 .45 °C (1). and D-values at 45 and 51 °C have been reported as 29.1 10. At pH 4.0 x 1. Some death will occur at this pH if the temperature is 4 °C. no growth is observed (1). Aeromonas has the potential to grow over a pH range of ca 4 . The minimum pH value for growth is thought to be pH 5.10.1.5.5 pH Under otherwise optimal conditions.0 .

botulinum grow reasonably well in the temperature range of 20 45 ºC.4 Heat resistance The vegetative cells of C.2. at 85 ºC for 5 minutes. All type A strains are proteolytic and type E strains are usually non-proteolytic.7 .22. E and.even as low as 3 ºC (7). botulinum is an obligate anaerobe. or at 90 ºC for a few seconds. forming at least seven different toxins.FISH AND SEAFOOD 10.0 µm. to a lesser extent.3 Temperature All strains of C. types B and F can be either. 10. botulinum are known. but Group II strains are psychrotrophic.2.2% sodium chloride (NaCl) (1). 10.4 x 1.2. botulinum types produce heat labile toxins.2 Clostridium botulinum Seven different types of C. being capable of slow growth and toxin production at low temperatures . 10. 226 . and is sensitive to > 4. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . which may be inactivated by heating at 80 ºC for 20 . but the interior of the food may do so.30 minutes.2. spore-forming rods. 10. Group I will not grow at temperatures of 10 ºC or less.1 Morphology Gram-positive. and groups I and II are those responsible for the botulism cases.5 . A to G. 0. but the spores of this organism are more so. botulinum is prevented. F are the types that are responsible for most cases of human botulism (5. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. Thus an aerobically packed product may not support the growth of the organism on the surface.2.5% NaCl (4). 6). Types A.1. There are four main groupings of the organism.2 Oxygen requirements Although C. All C. 10. botulinum are not particularly heat resistant. B.

parcooked meals with pH value > 5.97) (10.2. botulinum depends on solute. grow on the surface of foods unless they are vacuum or gas-packed. the minimum aw for growth may be lower.3 . 9). These values have been established under carefully controlled laboratory conditions.7 Characteristics of C. botulinum spores The most heat resistant spores of Group I C.21 minutes) (9). 10.2 Oxygen requirements C.97 for Group II (5). The organism will grow well in the centre of meat or poultry dishes. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. 10. vacuum packed.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e.is an obligate anaerobe.like other clostridia .5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. particularly by cooking.1 Clostridium perfringens Morphology Gram-positive spore-forming rods. they may survive mild heat treatments (70 .3. Note also that if humectants other than salt are used.6 and pH 5. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. 227 .0 . 0.3 10. In commercial situations. therefore.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C.2.0. pH and temperature.0 µm.10.2. safety margins must be introduced to allow for process variability.1. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.g.9 x 2. 10. It will not.PATHOGEN PROFILES 10. 11)). However. perfringens . These concentrations correspond to limiting aw of 0. 10.3. where oxygen levels are reduced.0 and aw > 0. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains.94 for Group I and 0. respectively (8.

the enterotoxin is not heat resistant . (It is unusual for spores to be formed in foods after the growth of this organism (14)). although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance.3. will survive greater extremes of pH (and aw) (12. perfringens is not tolerant of low water activities.FISH AND SEAFOOD 10. perfringens has the potential ability to grow within the temperature range 15 . It grows best at pH values between 6 and 7 (the same pH as most meats).5 pH C. perfringens in roast beef at 60 ºC is 14.5 minutes (17). 10. perfringens is not a tolerant organism with respect to pH. perfringens in vacuum packed beef (18).3.3. perfringens appears to be 0. At cold temperatures.3.3 Temperature The most significant characteristic of C. Salt concentrations of 6 . vegetative cells die rapidly (12). perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. the limits for aw are affected by temperature. In addition. 18). perfringens spores The spores of C.6 Aw/Sodium chloride C. depending on strain and other conditions. 13).97 (glycerol and sucrose respectively) depending on the solute used to control aw (13.it is destroyed by heating at 60 ºC for 10 minutes (14. Some studies indicate that the presence of 3% NaCl delays growth of C. The D-value for C. Under otherwise ideal conditions. pH. The lowest aw recorded to support the growth of C. 10. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. 10. 13).45 ºC. however. lower concentrations may be effective in combination with other factors.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. although C. 16). perfringens.3. 15. and their heat resistance is also affected by the heating substrate.3. Its optimum temperature for growth is 43 . 10. As in the case of other factors limiting the growth/survival of this organism.8% inhibit growth of most C. perfringens can vary quite considerably in their heat resistance. etc.7 Characteristics of C. Spores. While some growth can occur at 50 ºC. type of solute.10 ºC. Recorded heat 228 . very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. 0 . perfringens strains.93 and 0.50 ºC.

Under ideal conditions. The upper temperature limit for the growth of L. and temperature.4.2.4 . 10.3 .19 minutes.3 is the lowest value where growth has been recorded. being potentially capable of growing .especially at refrigeration temperatures.6 . monocytogenes is not a particularly heat resistant organism.4.5 . or even below 0 ºC.1 Listeria spp. 1.0 minutes for heat resistant spores to 1. however. so can be destroyed by pasteurisation. and where acetic acid is used as an acidulant. pH < 5. 0. -0.0 µm. 60 and 65 ºC were reported as 10.98 and 0.5 pH The ability of Listeria to grow at different pH values (as with other bacteria) is markedly affected by the type of acid used. In foods. Its optimum growth temperature.4. 10.2 has been suggested as the lowest working limit (22). 229 . 10.at refrigeration temperatures down to.4 ºC is probably a more likely minimum for growth in foods (19). the lowest limit for growth is likely to be considerably higher . Morphology Gram-positive short rods. is between 30 and 37 ºC.albeit slowly . using hydrochloric acid as an acidulant).64.2 Oxygen requirements Aerobe or microaerophilic. D-values for L.8 minutes for heat sensitive spores (12).3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. however. growth at low temperatures can be very slow. respectively (21). the organism is able to grow at pH values well below pH 5 (pH 4.4 Heat resistance L.4. requiring days or weeks to reach maximum numbers.4.2.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. 10. monocytogenes is reported to be 45 ºC (20). it is not a sporeformer.23. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.0. monocytogenes in crawfish tail meat at 55.5 x 0.4 10. However. 10.

3 Temperature Plesiomonas can grow in the temperature range of 8 . It is likely to survive.5 pH Plesiomonas has a pH range for growth of pH 4.83 (20).FISH AND SEAFOOD 10. or even grow. 10.5.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).5 .12% NaCl or more).0. peritrichous flagella.8.5 (24. the optimum being 30 ºC (23). 10.5 .5.45 ºC. The bacterium may survive for long periods at aw values as low as 0. monocytogenes is quite tolerant of high NaCl/low water activities. at salt levels found in foods (10 . 0. It grows best at aw of ≥ 0. but has been shown to be able to grow at an aw level of 0.5.5 10.1. Morphology Gram-negative short rods.6 10. 10.1 Plesiomonas Morphology Gram-negative.0 . 25).3 µm.3.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24. 0.6 Aw/Sodium chloride L. 230 .0 x 1 .1 Salmonella spp.5. 10.6.7 x 1. 10.8 .90. 10. 10.5.0 µm.2 Oxygen requirements Facultative anaerobe.97.5.4. 25).

6.PATHOGEN PROFILES 10. depending on the type of acid and temperature.37 ºC.3 Temperature Salmonellae can grow in the temperature range of 7 . 10. under otherwise ideal conditions.6 Aw/Sodium chloride Where all other conditions are favourable.6. temperature and pH. the optimum being 35 .6. and with an appropriate acid. Salmonellae are quite resistant to drying.5 ºC. depending on serotype. Salmonella senftenberg 775W is about 10 . Some death will occur at pH values of less than about 4. Salmonella has the potential to grow at aw levels as low as 0. The growth of Salmonella is generally inhibited by the presence of 3 . Growth is slow at temperatures below about 10 ºC. 231 .9. pasteurisation and equivalent treatments will destroy the organism under normal circumstances.5.8 .20 times more heat resistant than the average strain of Salmonella at high aw (27). substrate. However.2 Oxygen requirements Facultative anaerobe.5.6.48 ºC. although salt tolerance increases with increasing temperature (28).0.4 Heat resistance Salmonella is not a spore-forming organism. a heat resistant organism.4% NaCl. 10. 10. Salmonellae are quite resistant to freezing.5 . high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. or possibly 0. 10.945. strains vary in their ability to withstand heating.6. with a z-value of 4 . It is not. D60 ºC values normally range from about 1 to 10 minutes. some strains are able to grow at temperatures as low as 4 °C (26).5 pH Salmonella has a pH range for growth of pH 3. However. Furthermore. The optimal pH for Salmonella growth is between 6.93 (as reported in dried meat and dehydrated soup). and appropriate heat treatments must be determined experimentally for low-aw foods.7. therefore.

FISH AND SEAFOOD 10.3 Temperature Under otherwise ideal conditions. fat content.7.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. aureus viability. with an optimum of 35 . Most strains of staphylococci can grow within the pH range 4.0. 31). In meat macerate.2 Oxygen requirements Facultative anaerobe.20 minutes.7 10. It can survive well at low temperatures.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.). The following relate to limits for growth only. NB: The growth of Staph.5 ºC. Under most circumstances. etc. The limits for toxin production are also narrower than for growth.5). but may cause some cell damage (30). Freezing and thawing have little effect on Staph.7.1 Staphylococcus aureus Morphology Gram-positive cocci.7. aw and temperature (sensitivity to acid increases with temperature).3 (optimum 7.7. depending on aw.9 µm diameter. stressed cells can also be less tolerant of heating.2 to 9. under otherwise ideal conditions (29.7. aureus is more limited under anaerobic than aerobic conditions. aureus can grow within the temperature range 7 – 48. aw. the D60 ºC value is 2 .7. the organism is heat sensitive and will be destroyed by pasteurisation. 232 . 10. Staph.37 ºC (29). 10. As is the case with other bacteria. 0. however. aureus than citric acid).0 . 10.7 . 10.

0 . 10.86 is the generally recognised minimum in foods (32).8 10. but certain strains can grow in 20% NaCl. strain. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.3 Temperature The optimum temperature for growth has been reported as 30 ºC. atmosphere.5) (29.9. In general.4 .87 and > 0.6 Aw/Sodium chloride Staph.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5.7.83 . heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. pH. 34).2 .7.5 .99 little or no toxin production in anaerobically packed foods.99 aerobically under otherwise optimal conditions.PATHOGEN PROFILES 10.10% NaCl.2.8 x 1. an aw of 0. 233 . 31) between 0. In general. and solute.>0. Staph.7.8.43 ºC (35).2 Oxygen requirements Facultative anaerobe.0. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.8. 10. 10.7 Limits permitting toxin production 10 . However. with a temperature range for growth of 10 . * dependent on temperature.8. Staphylococci are more resistant to salt present in foods than other organisms. An effect of increasing salt concentration is to raise the minimum pH of growth. It can grow over the aw range 0.0 (optimum 7. aureus can grow in 7 .6 µm 10. 0.1 Vibrio cholerae Morphology Gram-negative curved rods.

cholerae in crab meat homogenate at 60 ºC is 2. with an optimum of 37 ºC (39). there is evidence that V.65 minutes (35). 37. are 5 and 43 ºC.5 pH This organism is tolerant of high pH values.8.1 . and it is sensitive to refrigeration temperatures (0 .9.0% NaCl (35). but not acid conditions.998.9.8. 10. It can grow in the presence of 0. cholerae can grow over the aw range 0. 10. 234 . V. 38). but only moderately sensitive to freezing (40).0. it has a pH range for growth of pH 5 .1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. declining in number during storage.4.6 (35. The D-value for V. under otherwise favourable conditions. 10. parahaemolyticus is easily destroyed by drying. respectively.97 .3 Temperature The minimum and maximum temperatures reported for growth in laboratory media. 10. cholerae may persist on frozen fish for >180 days (36). cholerae is sensitive to heat at temperatures of >45 ºC. 10.4 Heat resistance V.6.9 10. with optimum at 7. with an optimum of 0. 10.9.8.FISH AND SEAFOOD Also.984 (35).5 ºC).2 Oxygen requirements Facultative anaerobe.6 Aw/Sodium chloride V.9.

5 minutes.4 Heat resistance V. 235 . respectively (39).29 minutes. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).4 Heat resistance V.3 Temperature The optimum temperature for growth has been reported at 37 ºC. vulnificus is heat sensitive. but the optimum concentration is 2 . 10. The organism can survive for long periods at freezer temperatures (42). respectively.9.10.11. with a temperature range for growth of 8 . with an optimum pH of 7.1 Morphology Gram-negative curved rods.43 ºC (42).10. depending on solute.8.10. and 2. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.922. 41).0. parahaemolyticus are not heat resistant.9.02 .8 . 10.PATHOGEN PROFILES 10. V. 10. 10. The D-values at 55 ºC for clam and crab homogenate are 0.6 . 10. parahaemolyticus is unable to grow in the absence of salt.6 (41).10. The optimum concentration of NaCl equates to aw of approximately 0.5% NaCl.2 Oxygen requirements Facultative anaerobe. and are readily destroyed by cooking.98 (35.6 Aw/Sodium chloride V. parahaemolyticus will grow at aw levels as low as 0.5 pH The pH range for growth is 4.10 Vibrio vulnificus 10. 10.4% (39. 41).9.

with an optimum of 2. 2003. Lund B.J.10. Marcel Dekker.E. Betts G. 279-303.. New York.H. in Food Microbiology: Fundamentals and Frontiers. Kim J. Diseases Caused by Bacteria.W. 5-19. in Foodborne Disease Handbook. Eds..P. Gorham J. 97-131.O.. 1989. Aeromonas hydrophila group.H.M. in Clostridium botulinum: Ecology and Control in Foods. in Foodborne Bacterial Pathogens. 4. Volume 1: Diseases Caused by Bacteria.L. Murrell K.. Bhunia A. Aeromonas species.H.R... Dodds K. Marcel Dekker. 2005. with an optimum pH of 7. New York.6 Aw/Sodium chloride V. Eds. 1-27. New York. Clostridium botulinum.5 .5 pH The pH range for growth is 5 . Marcel Dekker. Blackie.L. Gaze J.N. Marcel Dekker Inc..D. 2.P. Food Control. Faris A. Notermans S. 7. in Clostridium botulinum: Ecology and Control in Foods. 57-63.. Miliotis M. in Microorganisms in Foods. Kovacek K.0% NaCl. 6.L. Foegeding P. 1989. Fratamico P. 10. Abeyta C.W.M. Bier J. Great Britain.. 8. ASM Press. Hauschild A. Principles of control. Doyle M. Doyle M. Eds. 6 (1).. Austin J. in Foodborne Bacterial Pathogens. 1995.. Hui Y. Eds. Washington D. Caister Academic Press. 121-76.5% (42). Stelma G. Marcel Dekker. Cliver D.R. Hui Y. Doyle M. Aeromonas.W.. Volume 5: Microbiological Specifications of Food Pathogens. Palumbo S.. Stelma G.5. Clostridium botulinum.K. Beuchat L. Hauschild A. Hauschild A. New York. 9. in International Handbook of Foodborne Pathogens.. 1993.8 (42). 1993.D..11 1. Cliver D. Microbiology and Molecular Biology. 10.C.O. 383-408. in Foodborne Disease Handbook.N. Novak J. Eds.R.P. Ed. Clostridium botulinum and Clostridium perfringens. Ed. 236 . International Commission on Microbiological Specifications for Foods. New York.H.. vulnificus grows over the range 0. 357-67. 1996. 11. 1994. 10. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.D. New York.10.FISH AND SEAFOOD 10. Murrell K. Ed.W.. Montville T.A.. 3. Potential hazards associated with REPFEDS.. Smith J. 5. Eds.W. Dodds K. Dodds K. References International Commission on Microbiological Specifications for Foods. in Foodborne Pathogens. Eds. Peck M. Johnson E. Marcel Dekker.. 2001. 329-49.D. Volume 1..H.L. Aeromonas hydrophila. 1-19. Marcel Dekker.10. Clostridium botulinum..H. 111-89.M. Gorman J. 1994.. Juneja V. London. New York.

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

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

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

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

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

uk Institute of Food Research (IFR) Norwich Research Park Colney lane Norwich NR4 7UA United Kingdom Tel: + 44 (0) 160 3255000 Fax: + 44 (0) 160 3507723 Web site: www.gov.co.nz Web site: www.org Web site: www.seafood.ifr.fdf.org.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.ac.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.org.uk Web site: www.dh.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: generalenquiries@fdf.csiro.uk/en/index.gsi.hpa.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.uk Web site: www.com Web site: www.bsi-global.uk Institute of Food Science and Technology (IFST) 5 Cambridge Court 210 Shepherds Bush Road London W6 7NL United Kingdom TeI: + 44 (0) 207 6036316 Fax: + 44 (0) 207 6029936 Email: info@ifst.com/ 243 .ifst.cmar.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.co.gov.org.au Web site: www.CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: contact@nfi.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.

asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.int/ SIK .uk/index.int Web site: http://www.euro.sik.uk Web site: www.hps.who.euro.nhs.co.php Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: admin@sofht.de 244 .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.uk Web site: www.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/en/ WHO Regional Office for Europe Scherfigsvej 8 DK-2100 Copenhagen Ø Denmark Tel: + 45 39 171717 Fax: + 45 39 171818 Email: postmaster@euro. jsp Food Safety 10 .int/foodsafety World Health Organization (WHO) Headquarters CH .int Web site: www.se Web site: www.scot. Web site: www.The Swedish Institute for Food and Biotechnology Box 5401 SE-402 29 Göteborg Sweden Tel: + 46 31 3355600 Fax: + 46 31 833782 Email: info@sik.codexalimentarius.int.co.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.sfam.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: foodsafety@euro.se/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: fao-who-cc@bgvv.who.scot.who.sofht.who.who.uk/scieh.nhs.org Web site: www.net/web/indexen.org.

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

FISH AND SEAFOOD 246 .

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

indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang. 226 Breading process 36-8 Brevetoxins. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. of recipe fish meals 43 Cholera. 131 Carbon dioxide. 137. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni. of fish – effects on microflora 5-6 Carbohydrate.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. as cause of spoilage of MAP fish 15 Calciviruses. 68-9 Capture and handling. smoked and dried fish 113-6 hazard in mild-processed marinated products 101 hazard in sun-dried products 102 non-proteolytic – risk of growth in fish recipe meals 44 248 . 136. in crustacean shellfish 86-7 Campylobacter coli. caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. 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. 113. preserved by marinating 100 Chemical detection methods. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. for PSP toxins 144 Chemical indicators. importance in lactic acid fermentation in fish products 130. as autochthonous bacterial flora of fish 3 Clostridium botulinum. associated with consumption of fish/fish products 16. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. of fresh fish spoilage 11 Chemical pollutants. 98-9 Brochothrix thermosphacta. in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage. fish sauce 123 Basipetospora halophila.

of prepared fish products 44 Escherichia coli. 106 Contacts 241-5 Conway method. detection methods 148 implicated in amnesic shellfish poisoning 146 in crustacean shellfish 87 toxicology 147 Dried fish 93-121 Dry salting 97-8 Dry/wet salting 99 Drying. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). adverse condition on poorly preserved fish 110 EC grading scheme. smoked and dried fish 114 Enzymic spoilage. forms for purchase 80 Crustacean shellfish 79-92 Cured fish 93-121 Cyclic imines 160-1 detection methods 162 origins and distribution 161 toxicology 162 Degradation. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp.INDEX pathogen profile 226-7 Clostridium perfringens. to detect NSP toxins 155 ELISA kit (Biosense). as contaminants in scampi 36. to determine fresh fish spoilage 11 Eels. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. to determine fresh fish spoilage 11 Corynebacteria. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. as autochthonous bacterial flora of fish 3 Crabs. 152 Diphyllobothrium. growth and survival in crabmeat 84 growth and survival in cured. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . 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. processing 334 Cockles. for detection of DA 148 ELISA kit (DSP-Check). hot-smoking 107 ELISA assay. 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. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. of fish – different processes 94 Dun. 149-50. source of azaspiracids 159 source of DSP toxins 148-9.

processing 35 Fish component. of fish – effects on microflora 5-6 Hepatitis (non-A. concerning crustacean shellfish 87-8 European Grading Scheme. as source of ciguatoxins 156 Garlic. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. preserved by marinating 101 Gravlaks. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. 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. for evaluation of fish quality 9 Eurotium. non-B) viruses. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes.FISH AND SEAFOOD EU legislation. as source of inulin in lactic acid fermentation in fish products 134 Garos. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. member of cyclic imine group 161 Gymnodinium catenatum. source of PSP toxins 142 Gyoshoyu. 102 Gymnodimines. definition of terms 176-7 implementation and review of plan 184-6 in fish and seafood product manufacture 175-88 seven stages of system 17784 Halobacterium spp. mould causing spoilage of dried fish 110 Evaluation. specific EU hygiene rules 196-224 Fresh fish. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. of prepared fish products 44 Gambiertoxins. fermented fish product 126 Green cured split cod 97. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. 130-5 Fish-spoilage bacteria 12 Fishtester. causing pink condition on fish surface 110 in fish sauce 127 Handling. fish sauce 123 HACCP. 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. of fish at sea 5 Fatty fish. to measure fish skin resistance 10 Flavobacterium.

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

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

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

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

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

growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. processing for seafood recipe meal centres 39 Vibrio spp. 62 Yersinia spp. 152-3 256 . growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 67 pathogen profile 235-6 Vibrionaceae. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in fermented fish 136 growth and survival in molluscan shellfish 66-67 pathogen profile 234-5 Vibrio vulnificus. of shellstock 61. 68 Vibrio cholerae non-01. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses.

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