Leatherhead Food International Leatherhead Food International

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

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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

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


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

vi .

CHILLED AND FROZEN RAW FISH 1.5 Pathogens: Growth and Survival 3.2 Initial Microflora 1.2 Definitions 2. vii .7 Published Microbiological Criteria 2.5 Pathogens: Growth and Survival 4.3 Initial Microflora 2.8 Further Reading CRUSTACEAN SHELLFISH 4.2 Initial Microflora 3.8 References MOLLUSCAN SHELLFISH 3.6 Published Microbiological Criteria 3.5 Factors Affecting Fresh Fish Spoilage 1.2 Initial Microflora 4. 4.3 Processing and its Effects on the Microflora 1.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.4 Spoilage 3.4 Spoilage 1.8 References 1.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.4 Spoilage 4.7 References 4.6 Pathogens: Growth and Survival 2.1 Definitions 4.7 Published Microbiological Criteria 1.1 Definitions 1.7 References 3.3 Processing and its Effects on the Microflora 4.5 Spoilage 2.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.3 Processing and its Effects on the Microflora 3.6 Pathogens: Growth and Survival 1.1 Introduction 2.4 Processing and its Effects on the Microflora 2.1 Definitions 3. 3.6 Published Microbiological Criteria 4.

3 Regulation (EC) No.5 Pathogens: Growth and Survival 6.3 Stages of a HACCP Study 8.2 Paralytic Shellfish Poisoning (PSP) Toxins 7. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.3 Processing and its Effects on the Microflora 5. CURED.1 Definitions 5.1 Definitions 6.2 Definitions 8. 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 9.2 Legislative Structure 190 9.I.7 Further Reading FISH AND SHELLFISH TOXINS 7.5 Regulation (EC) No.6 Regulation (EC) No.4 Spoilage 5.10 Cyclic Imines 7.6 References 6.9 Azaspiracids 7. 852/2004 on the General Hygiene of Foodstuffs 191 9.3 Tetrodotoxin 7.8 Further Reading FERMENTED FISH 6.7 Food Hygiene (England) Regulations 2006.10 Further Reading 223 viii .5 Pathogens: Growth and Survival 5. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.2 Initial Microflora 5. 7.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.8 Guidance 222 9.3 Processing and its Effects on the Microflora 6. EU FOOD HYGIENE LEGISLATION 189 9. 8.4 Spoilage 6.1 Introduction 7.4 Implementation and Review of the HACCP Plan 8.6 Published Microbiological Criteria 5. S. 14 (Hygiene requirements specific to the UK) 221 9.1 Introduction 8.7 References 5.4 Regulation (EC) No.9 Other Relevant Legislation 223 9.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.5 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. SMOKED AND DRIED FISH 5.11 Conclusion 7.1 Introduction 189 9.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. 2006 No.5.6 Lipophilic Shellfish Toxins (LST) 7.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7. 9.8 Ciguatoxins 7.2 Initial Microflora 6.

10.8 Vibrio cholerae 10.6 Salmonella spp.2 Clostridium botulinum 10.10 PATHOGEN PROFILES 10.3 Clostridium perfringens 10.9 Vibrio parahaemolyticus 10.5 Plesiomonas 10.1 Aeromonas spp. 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix . 10.4 Listeria spp.7 Staphylococcus aureus 10.10 Vibrio vulnificus 10.


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

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

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

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

The lipid fraction is the component showing the greatest variation. Fish flesh is dominated by the abundance of white muscle in relatively short segments. typically fluctuates between 0.2 . typically 3 and 15% of total weight. and spawning season. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. offering less than 0. In this chapter. “fish” will be used for fresh and seawater finfish.5% (1). but typically present a low fat percent when compared to other meats. and other forms of aquatic animal as part of their ‘fish’ definition.21%. giving it its characteristically flaky structure. In addition. and is mostly interspersed between the muscle fibres. 1 . Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. which can be up to 67%. most fish contain omega 3-fatty acids and other essential nutrients. However.20%. crustaceans. Fish are an important part of a healthy diet since they contain high quality protein. The connective tissue content of fish is also lower than that found in meat.1.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. CHILLED AND FROZEN RAW FISH Associate Prof. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. Muscle composition and structure of fish also differ from those found in other meat. migratory patterns. fins and a streamline body. The lipid content. lean fish are those fish that store most of their fat in the liver. Fish fillet composition can vary significantly within the same species due to feed intake. it shows a typical season pattern especially in migratory species such as herring or mackerel. and excludes molluscs and Crustacea. Fish can be divided into fatty and lean fish. In addition. while fatty fish have fat cells distributed along their bodies. respectively (1). ‘fish’ also refers to the flesh of such animals used as food. This super class of vertebrates includes all the bony and cartilaginous finfish. Protein content in fish fillet varies typically from 16 . Fish fillets are a poor source of carbohydrates.

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

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

However. and mud Ubiquitous.37 °C Unknown Unknown TABLE 1. numbers of pathogenic bacteria in raw fish tend to be low. 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.35 °C 8 .102 101 . Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. and risk associated with the consumption of seafood is low (2.FISH AND SEAFOOD TABLE 1.106 cells/g.109 cells/g) * Some sources considered L.14 . Plesiomonas shigelloides 3 .I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00.1 . 105 . Escherichia coli Yersinia enterocolitica 101 . 4). Toxin levels 0. estuarine environments.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.37 °C High for most species. 4 .103 High (107 . vulnificus) 5 .II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.>106 Salmonella spp. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 .26 °C 10 .0. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp.106 cells/g (exception: V. birds. sewage. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.19 µg/kg bodyweight from < 102 . In addition. stream water.

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

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

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

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

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

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

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

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

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

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

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

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

1. It is known that Aeromonas spp. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. and especially Aeromonas hydrophila. coli on isolation media (50). 1. Although the psychrophiles are unable to grow at body temperature. Chill storage may not eliminate growth. Most cases have been sporadic rather than associated to large outbreaks. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. proper chill storage will eliminate any risk associated with this organism. The major virulence factor appears to be the production of toxins with enterotoxic. in assessing the risk of fishborne infection by P. are present on fish and can grow to significant numbers during storage (1. with minimum growth temperatures around 0 °C and optima of 15 .6. in its habitat. Despite this. but the bacterium is known to survive and grow in the shallow marine. cytotoxic. 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. Some isolates are true psychrophiles.26 to 0. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. There is no risk associated with the consumption of properly chilled and cooked fresh fish. and proper cooking will substantially reduce risk. shigelloides are encountered. and especially estuarine environment. possibly due to under-reporting because of their likeness to E. Similar problems as those encountered for Aeromonas spp. are associated with human diarrhoeal illnesses. V. listeriosis cases have been reportable to the CDC. but proper cooking should substantially reduce risk of infection.CHILLED AND FROZEN RAW FISH disorders). sodium channel blocking and haemolytic activity (48). cholerae O1. monocytogenes incidence has been between 0. others are psychrotrophic mesophiles (49). They are aquatic organisms with an epidemiology that is yet to be fully understood. P. there is no risk from the consumption of properly handled and cooked fresh fish. Again.6.000 17 .4 Listeria monocytogenes L. Since 2000..3 Aeromonas Aeromonas spp. infection. 5. shigelloides is similar to Aeromonas spp. 17). which is often fatal (>50%). L. In the US. and fish and shellfish are probably the major reservoirs for the organism (50). however. the causative agent of cholera. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. their potential to produce disease-causing exotoxin in food has not been fully examined (48). is historically associated with faecally contaminated water.55 cases per 100. monocytogenes has been well documented as a foodborne human pathogen.20 °C.

especially pickled fish. leads to the production of histamine.pregnant women. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54).6. Lightly preserved products.. some Vibrio spp. Klebsiella pneumoniae and Hafnia alvei. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6).. It is regularly isolated from seafoods (51). but cooking will significantly reduce the risk of infection. at risk groups . full risk analysis is not possible (52). Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. 1.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. Infections are mostly associated with the consumption of raw or 18 . D60 of 1. it is not apparent to the consumer and it cannot be destroyed by cooking. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. If fish are not properly refrigerated. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. Like other toxins. Being a psychrotroph. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. 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. but more especially by Morganella morganii. and Lactobacillus spp. alcoholics. there is a possibility for growth of the organism on chill-stored fish. foetuses.FISH AND SEAFOOD persons. The organism poses a serious risk in chilled products not cooked before consumption. 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. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . 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..98 minutes in cod and D60 of 4. Clostridium spp. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. neonates. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. Photobacterium spp.48 minutes in salmon have been reported (53). 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. Fortunately.6.2002 (55)) and is also common worldwide. but proper cooking will reduce any risk. 1.

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

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

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

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

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

A. in Torry Advisory Note No 93.2 Spoilage Shewan J. Huss H. Technomic Publishers. Torry Research Station of the Ministry of Agriculture. McMeekin T. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. Olley J. Lancaster.G.. Borgstrom G. 271-85. Cho Y. Food and Agriculture Organisation of the United Nations. in Seafood Science and Technology. in Fish as Food. Sumner J.E. Gibson D. 1.. 317 (Suppl. Torry Research Station. Estimation of the bacteriological quality of fish by automated conductance measurements. International Journal of Food Microbiology...C.S. 1992. 62-70. Ed. 1986. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Trolle G. Villemure G. Smith G. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging.. Ravn Jørgensen B. The Microbiology of Seawater Fish. 1984. 24 . Gibson D. Handling and processing of scad. 1997.. Aberdeen.FISH AND SEAFOOD 1. Ogden I. Magno-Orejana F. Taoukis P. 1985. 1989. Changes in microflora and chemical compounds in mackerel stored by partial freezing. Kitts D. 127-34.. 6.). 1992. Journal of Food Science. in FAO Fisheries Report No. International Journal of Food Microbiology. 1. Labuza T. Torry Research Station of the Ministry of Agriculture. 51. Houston N.. Journal of Food Protection.. Huss H.1 Further Reading Processing Anon.P.1983.. Shahidi F. International Journal of Food Microbiology.. Cann D. Akiba M. 1989. Microbiological quality and shelf life prediction of chilled fish.L.. 1988. Shinano H. Eds.. Food and Agriculture Organisation. 4. Ross T.9..H. 55.D.H. Jones Y. 317-21..H.. Application of predictive microbiology to assure the quality and safety of fish and fish products. 73-89. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. Huss H. 1961. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. International Journal of Food Microbiology.. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. Fisheries and Food.D. 741-50.M.. Academic Press. 295-307. Dalgaard P.9 1. 13-32. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches. Simard R.9. 35. Rome. Bulletin of the Faculty of Fisheries (Hokkaido University). Gram L. Florida.M. 15. Hobbs G. Ed.M. 1. 1984.. Fisheries and Food. Picard G. Aberdeen.. 65-72.

International Journal of Food Microbiology..4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. 322-6. Klausen N. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. in Quality Assurance in the Fish Industry.H. in Fish processing technology. Effect of culture age. 59.H. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. 1992. Huss H. 25 . London.9. 1997. Otero A. Huss H.K.eu/LexUriServ/LexUriServ. Applied and Environmental Microbiology.europa. 1987. 1993. 4.CHILLED AND FROZEN RAW FISH Einarsson H.M. Elsevier Science. Modified atmosphere packaging of fish and fish products.L. Blackie Academic & Professional. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Liston J. A rapid method for detection of histamine producing bacteria. International Journal of Food Microbiology... Hall G. 137-46.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. 1992.R. Journal of Applied Bacteriology. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. Trolle G. http://eur-lex. 2197-203. Eds. 1. 65-72. Huss H. 1987. Amsterdam. Gram L. 72. Garcia M. 20023.. Gram L.H. 479-88.. 1.3 Pathogens Condon S..J.. Jacobsen M. Sala F. 5.9. Ed..


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

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

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

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

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

Moulds Enterobacteriaceae Salmonella. product assembly. histamine Cryptosporidium. coryneforms. shelf life determination. enteric viruses 2. yeasts Lactobacilli. Bacillus subtilis B. cereus B. cereus Bacillus licheniformis. Aeromonas. coliforms. protozoa and viruses B. 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. and so are the potential microbial contaminants associated with them. for chilled products. Clostridium perfringens. spore-formers. moulds Bacillus Bacillus. Table 2. pseudomonads.I provides an overview of common raw materials. B. Clostridium botulinum. 32 . intermediate processing. and. Shigella.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. Fusarium.3.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. moulds. mushrooms. be considered at an early stage during new product development. Clostridium Lactobacilli. In addition. streptococci.4. Aspergillus. Listeria. moulds Bacillus. by the HACCP team or by a hazard and risk assessment programme. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter. anaerobic spore-formers. Staph. Enterobacteriaceae. nuts and fruit Associated microflora Lactobacilli.FISH AND SEAFOOD 2. Staphylococcus. cream. Bacillus cereus. moulds Bacillus. and associated spoilage and pathogenic microorganisms. TABLE 2. toxin. 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.4 2.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. Escherichia coli. yeasts Pathogenic microflora/toxins Salmonella. Vibrio cholerae. staphylococcal toxin. cereus Staphylococcus aureus. raw materials. however. and milk Cheese Water/Ice Bacillus. Pseudomonas. Penicillium.

but will also need to ensure that the optimum shelf life can be safely obtained. and potential opportunities for growth of foodpoisoning bacteria. the core temperature. Band-sawn materials or fish dice are normally promptly further processed. line extensions and process reviews. to evenly raise the temperature of the entire frozen product from -20 °C or lower.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. It is recommended that the principles of HACCP are applied to novel processes. as the full complexity of the processes used for specific products cannot be covered in a general text.4. but it is assumed that any band sawing or dice process will introduce heat from blade friction. or the use of microwave technology. 2. this may be an incidental effect.2. defined holding time at a chill temperature (0 – 3 °C). 2. up to -5 °C so as to improve the cutting and handling properties during processing.4. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes.4.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. All have the same intention. and contamination from equipment to clean fish muscle. Figures 2.1 . there are a number of distinct methods used. Examples of tempering techniques include the use of heated air. 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. as a heat process is often required for functional or organoleptic reasons. Where the raw materials are frozen. 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.3. Generic examples have been used to illustrate only the major points. depending on the size of the fillet 33 . Rapid controlled cooling of heat-processed components will help to minimise spoilage. but it also poses a risk of cross-contaminating other products undergoing tempering. Effects on the microflora are minimal during tempering (7).3 Preparation of fish component of chilled and frozen recipe dishes 2. For some components.

7 days. the scampi is washed in potable water to remove foreign bodies.1C . drained and then refrozen for storage until required for further processing. chilled or sold chilled on defrost with an expected shelf life of 6 .1A . 2.FISH AND SEAFOOD processed.Scampi. by mincing and mixing with a binding agent (as described in section 2. frozen in a nitrogen blast freezer and weighed into sacks for later processing.1B .1 Whole-fillet / scampi Scampi (Nephrops norvegicus). 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. It is then size-graded.1 (2.Shaped Fillets.4. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product.1.2). 2. 2.1. The “blown” meat is graded.2. may not rise above 0 ºC during processing.4. Fig. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.3. The processing of three examples types are described in Figure 2. are placed into plastic boxes and covered in ice to preserve their condition.1. after landing.3.Fish Cakes). with the coating heat-fixed. Raw material preparation stages for chilled or frozen coated fish products 34 . quickfrozen. Frozen cores are usually batter-enrobed. On arrival at the processing plant.

off-cuts or belly flaps. Therefore. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. but. or those where the trimmings.7 days. The fish component of the cake will. because of its nature. The mix is shaped and formed ready for packing or for enrobing.. once the dry components have been rehydrated.block-sawn dust. ii) Fish-filleting waste . Herbs or seasonings used in the recipe may contribute additional spoilage organisms or.3. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. with an expected shelf life of 6 . such as aerobic spore formers and Staphylococcus spp.g. Fish cake cores may be batter-enrobed. additional spoilage organisms may be introduced. material will need to be transferred swiftly to a chiller. from potato flake.3.2 Shaped products This type of product falls into two categories. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. those that use frozen blocks of whole fillets. chilled. more seriously. Waste material produced from this type of processing is often reused in the third category (fish cakes). off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. iii) Fresh/fillet/minced fish block.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. or promptly processed to avoid further quality loss. are handled and stored as any other food product.4. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . pathogens such as Salmonella and Listeria. that are to be reused for other foods. unless it is poorly handled. 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.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.1. It is for this reason that all by-products from primary processing. 2. 35 . quickfrozen. and water is added to rehydrate the components.1.4. as the physical structure of the material has been destroyed. The remaining dried ingredients are mixed with the fish. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. This is especially the case with block-sawn dust. spoil more rapidly than the dry ingredients. fish fingers). or sold as a chilled product after thawing. with the coating heat-fixed.

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

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

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

encourage more rapid decay and loss of structural integrity. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. treated as a highrisk ingredient. or breadcrumbs. Raw material is typically fish fillet. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. and reduce spoilage organisms and potential pathogens. any processing stage. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components. i. will disperse the saprophytic flora of vegetables . but also as components in recipe meals. Once vegetables have been blanched. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. they should be stored under temperature-controlled conditions. and may include sauce accompaniments.3 log reduction in bacterial numbers on the surface of vegetables. all-year-round fresh vegetables are preferred.e. 39 . with cases of foodborne disease traced to Listeria monocytogenes. Although the processes are claimed to be superior to using unmodified hypochlorite for washing. if the process is not appropriately controlled. 2. loss of quality.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. However. coli O157. The presentation of vegetables has a significant effect on the product quality. not just as accompaniments.5.4. Products must be washed thoroughly to remove pests and soil. A number of examples of direct faecal contamination of vegetables exist in the literature (8). 8) will. However. the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. In an attempt to improve washing and reduce chlorine taint. Salmonella typhi. The majority are products made from various types of whitefish (cod. coliforms and pseudomonads throughout the mix. rice or pasta. frozen block or canned fish. control is more onerous. For example. For example. and E.lactobacilli. and mixed whitefish). (7. for example.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. spore-formers. even those with pH-modified dressings. Washing with a biocide will at best achieve a 2 . and may include molluscs or prawns that may or may not have undergone a minimal heat process. For premium products. coryneforms. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. haddock. Any assumptions regarding the application of good agricultural practice to produce should be made with caution. and protected from any post-processing contamination. The opportunity for growth of L. Blanching of vegetables can be an effective means of reducing or removing surface contamination. pastry. with the accompanying release of additional nutrients by processing. chopping and mixing of material. colour and water will occur.

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

spices. respectively. suspended in an acidified sauce with an oil component. 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 (12.5 illustrate the recipe dish ingredients preparation and assembly and packaging. that of a dairy-based sauce. 13).herbs. and handling of processed sauce can be less rigorous than. 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.4 and 2.2. The format uses flavour compounds . Without barriers. 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. for instance.4.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. 41 . Marinade-type products are becoming more popular. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. Figures 2. flour products.

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

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

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

and Campylobacter are recorded in filter feeding molluscs such as mussels.6. Vibrio spp.6. In addition. with pathogenic microorganisms or viruses. 2. may be of benefit within any risk assessment for chilled and frozen prepared fish products.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. have been linked with food poisoning and should also be considered. and this issue may be of greater consequence for warmwater. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). Cases quoting contamination of seafood with organisms such as S. permit rapid growth of B. along with previous discussion. Inappropriate holding times and temperatures will. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. 13.104/ g should be treated with some concern (26).6.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. botulinum type E and Vibrio parahaemolyticus (25). especially molluscan filter feeders. 45 . botulinum when held in favourable conditions for growth (28) i. water. air. etc. Incidents in such diverse materials as cream. perfringens contaminating seafood will depend on the source of raw material. low oxygen or anaerobic conditions. spices and herbs and vegetable shoots have been reported (12. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. any risk of C. farmed or free-living crustacea (30). in some cases. histamine intoxication. Additional hazards from marine bio-toxins. and some effects related to product type and time to toxin production have been noted.3 Clostridium perfringens A study examining C. These are C. soil. 2. typhi.. Counts of between 103 . are dealt with elsewhere in this book.1 Bacillus cereus This is a common contaminant of dust. cereus. A number of other Bacillus spp. 27). 2.e. potato. and many raw or processed foods (26). Therefore. Hazards attributable to the non-seafood components are listed below and this information. agricultural run-off or sewage can contaminate in-shore species. Germination of spores requires favourable conditions.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. or a heat shock followed by holding at a temperature suitable for growth.

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

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

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. L. coli GMP* <102 Maximum 104 TABLE 2. cereus Staph. 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. coated fish. aureus V. TABLE 2.II IFST Guidelines for part-cooked foods – e. monocytogenes B.g. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp.III IFST Guidelines for processed foods . parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E.FISH AND SEAFOOD the microbial criteria for other products (rice. fish fingers. pasta.fish products. 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.

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

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

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


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

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

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.

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

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

Fresh: shelled meats (often only the adductor muscle and the roe are eaten.2. In some species.) Dried: peeled.FISH AND SEAFOOD 3. scallops possess a single large adductor muscle. 3. Fresh: cooked meats. gutted.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. Salted: cooked meats bottled in brine. or packed with jelly. brine. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. Live: whole with shells. by interlocking tufts of hair-like cilia. or filaments. scallops can swim or leap by a series of rapid closings and openings of the valves. in which the individual branches. in mussel liquor. mussel paste. cooked meats packed in dry salt for transport. Canned: cooked meats. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves. 58 . The familiar marine mussel is asymmetrical. vinegar solution. Smoked: meats may be canned in edible oil. are united. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). The adductor muscle is the only edible part of the animal. Bait: used live extensively for baiting fish hooks for line fishing. and held apart.1.3 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. Unlike other sessile bivalve molluscs. The shells are fluted and round with a broad.1. mussel in butter sauce. and the anterior end smaller and pointed. Internally. also with other ingredients. boiled. Semi-preserves: cooked meats with vinegar-acidified brine and spices. with the posterior ends of the shells (valves) broad and rounded. sauces. The common scallop is about 5 cm long. They have filibranch gills. smouldered and afterwards dried (Japan). flattened ear or wing at the hinge area.

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

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

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

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

perforated tray in which shucked shellfish are spray washed and/or drained) or in a blower-washer (a tank-like device for immersion washing of shucked shellfish). Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab.2. Due to the nature of the shucking operation.1.1. Thus. Under these conditions if the shuckers enter the packing room.2. and fish. Shuckers routinely use either cotton or rubber gloves during the shucking operation. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. or subjected to splash from foot traffic. After washing of meats.2 Shucked shellfish 3. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). clothing becomes very soiled. 3. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. 3.3. In addition. washing and culling shellfish prior to storage is essential. In the blower-washer. Studies have shown the use of rubber gloves reduces the contamination level. thus contributing to high bacterial counts in the final product (28). removal of shellfish (shucking). whereas cotton become wet and 63 . To remove the shellfish attached to the remaining valve. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported. Containers are placed in packing boxes for shipping at refrigerated temperatures. If shellfish are not reasonably clean at the time of processing.1 Processing description The processing of shucked shellfish is limited to washing of shellstock. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation.3.2). Meats are collected in either perforated or non-perforated buckets for delivery to the washing station.2 Effects of microflora during processing The same considerations as previously discussed. are applied to the shucking operation of shellfish. 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.1. the finished product may become contaminated. through which the washed meats are discharged for rinsing and then packed in containers. for the effects of microflora during the processing of shellstock. and packing shellfish into containers (Figure 3. shellstock is subjected to contamination when stored on the floor in standing water. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. washing. the tank is opened through a drain gate and chute. lobsters.3.

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

clean gloves. glycogen levels range from 0. The high incidence of proteolytic bacteria and other types capable of fermenting glucose in the natural flora may be responsible in postmortem spoilage of shellfish. skimmers. The spoilage process in shellfish may be divided into three stages: increase of acidity. is stored in the bivalve as a source of energy. Shucked shellfish are an excellent medium for the growth of bacteria. whereas bacterium such as Pseudomonas. rough. Achromobacter and Flavobacterium levels decrease during storage. Alcaligenes.47 to 6.2 ºC. in oysters. streptococci. Lactobacilli seem to be the predominant microflora in shellfish. This process changes the bacterial profile to microorganisms favouring low pH. spoilage will occur. and proteolysis (10). Shucked shellfish are held and transported at temperatures of 7. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10.2 ºC or less. where there is a breakdown of temperature control or prolonged storage of shellfish. blowers and other utensils that have cracked. 32). abundant gas production. showed that organisms such as Achromobacter. and yeasts. 33. shucking pails. Temperature abuse will result in high bacterial counts. In summary. The environment has an effect on the bacterial population found in shellfish. The initial flora found in shellfish is the major factor of spoilage in bivalves. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. Earlier studies. Glycogen is hydrolysed by the initial flora found in shellfish. thus producing acid and gas (38). Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. are an excellent medium for growth of bacteria. As described previously. particularly shucked meats. 65 . or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. All equipment coming in contact with the shellfish such as colanders. 31. 35. and packing the shellfish in a timely manner. Much of the work has been done with oysters and clams. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. particularly oysters stored at 7 ºC.8% (37). These organisms may play a significant role in the spoilage processes. Proteus. Micrococcus. Shellfish. For example. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. anywhere during the process of shellfish from harvesting to the point of sale. 34. at the turn of the century. a carbohydrate. Flavobacterium. Controls that should be applied include using clean packing equipment. 36). these include lactobacilli. Pseudomonas. Glycogen. 3.

48 hours with abdominal pain. nausea.1 Vibrio parahaemolyticus V. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. however. during processing. and harvesting of shellfish in the cooler months. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. One of 250 cases reported involved shucked products with low levels.FISH AND SEAFOOD 3.000 population/year (46). V. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). 66 . approximately 23 cfu/g. They can be introduced to the shellfish from the environment or added during processing.5. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). 44). V.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. Diseases caused by Vibrios are enteric in nature. and to the point of sale.5 cases/100. 3. These organisms can be present in the shellfish from the point of harvesting. Major outbreaks are associated with the warmer months. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. from harvesting to finished product (i.5. headache and fever. primarily in brackish or saltwater. 3. This bacterium is present in all the major processing steps of a shellfish plant. lot and retail outlet. and estimates based from a hospital study show less than 0. parahaemolyticus from shellfish is low. parahaemolyticus possibly producing illnesses (40). Between May and September 1997. of V. Sporadic cases are frequent along the coast of the United States. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. coastal areas). 41). Fortunately. parahaemolyticus. ranging from epidemic cholera to sporadic cases of diarrhoea.1. only a small percentage of strains in the marine environment are potentially pathogenic (45). prevention of cross-contamination. shellstock and packing of shellfish in jars) (29). where outbreaks occur regularly. The incidence of V. Products involved were from the same processor. there is a recent report of low levels of V. although non-hemolytic strains have been associated with illness (43.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. diarrhoea. It is a common marine bacterium. and very common in Japan. parahaemolyticus causes gastroenteritis lasting 24 . widespread in both polluted and unpolluted waters.e.

and preventing cross contamination once the product is cooked.2 Vibrio vulnificus The symptoms of V. parahaemolyticus growth in shellfish meats is temperature dependent. Control of V. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50).8 case/100. All isolates are considered pathogenic. chills. Healthy individuals are susceptible to gastroenteritis. V.II (8. 54. the organism has been found in the Gulf of Mexico. or immunosuppression. are particularly susceptible to primary septicaemia.48 hours of onset. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. and rapidly become inactive at low storage temperatures. They are sensitive to cold. Again.7 days. cholerae non-01 is ubiquitous in the estuarine environment. cholerae non-01 has been associated with septicaemia. Hazards from this bacterium can be controlled by thorough cooking of shellfish. 57. which have been implicated in shellfish-related disease outbreaks (31).5. leukaemia. 48). The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. Onset of symptoms occurs in 48 hours and can be severe. parahaemolyticus. It is one of the most severe foodborne infectious diseases. The organism attaches to the intestine and release of a toxin is suspected. 3. 55.1. Illnesses are usually associated with the consumption of raw oysters as described in Table 3.5. however. lasting 6 . with a fatality rate of 50% for individuals with septicaemia. vulnificus are fever. The occurrence of V. 49). vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. those who have liver disease. or primary septicaemia. and vomiting.MOLLUSCAN SHELLFISH Control of V. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. and nausea within 24 . diabetes.000 population/year (8. Oysters are the most hazardous food associated with V. these individuals are advised not to eat raw shellfish. cholerae non-01 (51). 23 of 24 major USA-West Coast estuaries were positive for V. like V. nausea.0. It causes diarrhoea. parahaemolyticus the estimated annual incidence rate is very low. 58). cholerae. Death has been reported to occur within 36 hours from onset. High-risk individuals. 53. 46. Control of oyster-associated 67 .1. 46. gastroenteritis.4 . V. V. vulnificus. 3. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. The bacterium is an estuarine species that causes wound infections. 56). the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). abdominal cramps.3 Vibrio cholerae non-01 Illness caused by V. vulnificus is by rapid refrigeration during warm weather months. In one study. cirrhosis. in the range of 0. Inadequate refrigeration will accelerate the growth of V. fever. Primarily.

processors should know the product source. V. Poor sanitation and contaminated water supplies will spread the disease. cholerae 01.5%) and 2 . widespread in both polluted and unpolluted waters.10% carbon dioxide for optimal growth conditions. Illness can include abdominal cramps. fluvialis. including V. and illness lasts 7 .4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools.S. They are common marine bacteria. particularly in oysters. nausea. (8. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). cholerae non-01 illnesses is by cooking seafood thoroughly. 3. however. There are several other Vibrio spp. There is evidence of strains of V. cholerae 01 is transmitted by faecal contamination of food or water. respectively. vulnificus. and nausea.g. Illnesses are associated with the warmer months. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. acidity and heat. V. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. vomiting. nausea. low pH. parahaemolyticus and V. 61. cholerae 01 establishing in the U.5. The control of V. In developing countries. 3. undercooked. and the transmission of the disease by consumption of raw. preventing crosscontamination of cooked seafood.5. Onset of disease is 6 hours to 5 days. dehydration. Gulf coast environment. V. jejuni requires reduced levels of oxygen (3 . e. The presence of these strains appears to be in low numbers and non-virulent (59).5.FISH AND SEAFOOD V.1.10 days. 62. imported product from a country experiencing an epidemic. and southern Asia including India and Bangladesh.1. It is considered a fragile and sensitive organism to environmental stresses such as oxygen.5 Other Vibrios spp. In general. or cross-contaminated shellfish (57. salinity. It causes diarrhoea. and V. In addition.2 Campylobacter jejuni C. diarrhoea. 60). drying. hollisae. abdominal pain. mimicus. 63). and shock.48 hours with abdominal pain. Death may occur after severe fluid and electrolyte loss. Control of infection from these organisms is no different from the other Vibrios. the pathogenic severity is lower than V. is the same as previously discussed for the non-01 strains. cholerae 01 in molluscan shellfish. 68 . Onset of illness is from 3 to 5 days after ingestion of contaminated food. The frequency of infection of these organisms is no different from other Vibrios. headache and fever. these organisms cause gastroenteritis lasting 24 . fever. associated with shellfish-borne illness outbreaks. 3.

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

FDA/ Administrative Guidelines.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. coli Salmonella V. 2 Source (90). They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. Control of these organisms in shellfish products is limited. They are easily destroyed with mild cooking temperatures.000/gram (29). 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. Similar results were seen by other investigators (86. 70 . non-dairy frozen desserts. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. Caution should be exercised by individuals with haematological malignancies (88). coli Enterotoxigenic E. aureus C. 3.0001 negative for the presence3 Under review4 Source (89). FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. Pathogen monitoring of Selected High Risk Foods Domestic including: milk.000 cfu/g1 <230/100 g2 <1000 per gram. Note: Although molluscan shellfish are not included in the selected high risk food.600. FDA/Administrative Guidelines 3 Source (91). jejuni V. cholerae V. hydrophila group stored at refrigeration temperatures. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. In one study. Test Standard Plate Counts E.000 to 3. 87) of the outgrowth of A. parahaemolyticus Staph. dried milk/whey drink powders. vulnificus in molluscan shellfish. prepared sandwiches. being isolated more often during the warmer month. and where indicated are intended to be used for official control purposes. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. soy products.

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

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

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

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

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

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

Ed. Slavin J. Food and Drug Administration. Food and Drug Administration.8 Further Reading Ahmed F. 91. Ed.Pathogen monitoring of selected high risk foods (FY 88/89).H.S. Food and Drug Administration. 1997. 3.S.E. Baird-Parker T. 90. 2000. 77 .R. Food and Drug Administration. 7. 1995..MOLLUSCAN SHELLFISH 88. Inc.S. Gaithersburg..C. and HACCP: A Global Focus. Food and Drug Administration Compliance Policy Guides.S.12). Fish Inspection.A. Burdge D. Lund B. The Microbiological Safety and Quality of Food.S. Technomic Publishing Co..L. U. 1995. U. 560. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. Food and Drug Administration. Collette R. Aspen Publishers Inc.E.600 (CPG 7119.030. U. in U. Part I Sanitation of Shellfish Growing Areas.C. 63-8. fresh or frozenadulteration by bacteriological contamination Sec.C. in U.. Food and Drug Administration Compliance Program Guidance Manual. U.W. U. 1991. Quality Control. Program 7303. U. Seafood Safety. Martin R.C. Washington D.. Food and Drug Administration. oysters.W. Gould G. Smith J... Food and Drug Administration. Chapter 5: Foods. 89. Food and Drug Administration.S. Government Printing Office . 1987.1987. Sherlock C.S. Lancaster. mussels. National Academy Press. Ed. Clams.. in National Shellfish Sanitation Program Manual of Operations. Pittsurgh.. Washington D.S. Washington D.M.


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

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

These animals can survive out of water if the surrounding environment is kept cool and damp. whether GHP are in place. chilled 81 . They are then taken to market alive in either holding tanks. gills and thorax.for example. especially if they are stored in ice or iced water.1). from the time of capture and processing. or cooked. corynebacteria. or are found in brackish water around the shores.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. 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. or with the shell and central vein removed. Shrimp will then be glazed and frozen in a raw state.3.1. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected. graded according to size. After capture and transport to the processing plant. lobsters and crayfish are harvested from cages or pots containing bait. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. glazed and frozen (see Figure 4. Hence. The claws are usually bound together to prevent the animals fighting and causing damage to each other. Micrococcus spp. Vibrio spp.1 Prawns and shrimps An example of the process used for preparing frozen. In contrast. 4.10 days for temperate water species (3). The rate of spoilage will be a function of time and temperature. 4. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. chilled sea water. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots. uncooked prawns and shrimp is presented in Figure 4. thus. When the shrimp arrive at the processing plant they are washed and then sorted into size. they are washed... A large proportion of shrimps are produced by aquaculture. or transported directly to processing plants.. the rate of deterioration of the crustacean shellfish by these organisms is reduced. Pseudomonas spp. so they are either sold on the local market or are processed in local processing plants. As with finfish. the genera associated with tropical species include Enterobacteriaceae. bacterial contamination and spoilage are inevitable. Shrimp might be further processed with the shell on. however. with “head on” or “head off” whereby the edible tail is removed from the head. and Achromobacter spp. peeled and cooked. Alcaligenes. In comparison shrimps die quickly after capture.3 Processing and its Effects on the Microflora Crabs. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . up to 16 days for tropical species compared with 8 . Shrimps can be processed whole.

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

indicative of protein degradation and the growth of the principal spoilage bacterium Sh. If the tail drops. In comparison. or packaged and processed. there is no indication of life. wet and ‘soapy’ or ‘jammy’ to the touch. whilst boiled crustaceans have a pleasant slightly sweet smell. with the growing demand for ready-to-eat products from crab and lobster meat. 4.4.4 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. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. or in hermatically sealed cans that have undergone an appropriate heat treatment.4. the meat is either repacked and sold in the shells. putrefaciens and Pseudomonas spp. Heat is produced if the shrimps decompose in a confined space. With spoiled crabs and lobsters. viruses or parasites. bacteria of public health significance. the joint between the tail and carapace is open and the limbs hang loosely. However. there is no associated spoilage problem. An offensive odour is also produced. slightly iodine smell. and a strong ammoniacal odour is produced.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. immediately prior to boiling. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. they include Sh. contaminating microflora may include enteric bacteria. Once removed and cleaned. for crabs.2 Cooked crabs and lobsters As indicated earlier. 4. dull and sticky. 83 . If hygienic procedures are not followed.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. 4.CRUSTACEAN SHELLFISH 4. The shells have a crisp. spoiling prawns and shrimps have shells that are soft.3. and have a sweetish. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. The shell will be soft. bright and dry appearance. putrefaciens. since crabs and lobsters are traditionally consumed immediately after cooking. If boiled.

Hence. as well as for Salmonella. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . nearly 30 years elapsed before the potential hazard of L.cooked prior to consumption.5 4. cooked and smoked fish. shrimp and shellfish (11). Salmonella cannot be completely removed from raw shrimp during processing. 6. 4. The presence of L. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. Although foodborne illness had not been associated with the product.5. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . An import alert was put in place in June 1987. in domestic/imported shrimps (15). Aquatic birds and the hands of workers have also been identified as sources of the pathogen. 5.1 Pathogens: Growth and Survival Salmonella spp. shrimps and cockles sold (17). monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). monocytogenes was absent from 40 samples of cooked prawns. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. Further surveys identified Listeria spp. The above results prompted the FDA to develop a compliance programme for the pathogen. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). However. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11).5% of frozen raw shrimps from different countries (13). even if GMP is followed. Moreover. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). 7). monocytogenes in cooked ready-to-eat foods. a common pond fertiliser used in aquaculture. Following this. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals.5. and usually for both.2 Listeria spp.FISH AND SEAFOOD 4. 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. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. animal manure. fish fingers. 16). later surveys in 1990 recovered the pathogen from retail raw. although efficient cooking will quickly eliminate it (6). in 24.

The main source of contamination is the hands of processing personnel. Once the competing microflora has been removed by a heat treatment. Crustaceans have been linked with food intoxications attributed to Staph. the pathogen can grow at temperatures as low as 3 °C. Crabs are covered in a chitin exoskeleton. whilst boils and cuts are also common sources of the pathogen. 4. botulinum type E at 10 °C but not at 4 °C (20). GMP should be followed in the processing plant to ensure that the final product is held at 3 °C.5. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21.70% of healthy individuals carry toxin-producing strains of Staph. 85 . cholerae O1 and V. pasteurisation and cooking. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. Vibrio cholerae. so is able to survive heat treatments that eliminate vegetative cells. 22) as well as from undercooked crustacea when harvested from contaminated waters (23.4 Clostridium botulinum C. was associated with seafood consumption.CRUSTACEAN SHELLFISH old. the causative agent of cholera. cholerae non-O1. combined with the degree of handling of these types of product. V. cholerae is divided into two groups. therefore. An epidemic that began in South America in 1991. V.5 Vibrio spp. aureus on their hands. no incidence of foodborne illness caused by L. 4. aureus is not a member of the intrinsic flora associated with crustacean shellfish. However. is widely distributed in aquatic environments. monocytogenes has been attributed to crustaceans. 24. 19). especially in products that have been vacuum packaged. botulinum type E is commonly associated with fish and fishery products. 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. 4.5.g.5. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. Control should be achieved by promoting GHP and staff training programmes. The former. cholerae. pregnant women and the immuno-compromised.3 Staphylococcus aureus Staph. aureus because of the ubiquity of this organism in the processing environment. which can become colonised by V. 25). Researchers have demonstrated that inoculated shrimp supported toxin production by C. particularly crabmeat (18. To date. about 50 . as part of the natural bacterial flora. The pathogen has resistant spores. e. although contamination by types A and B may occur from the environment.

V. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. Astroviruses and non-A.30 MPN/g) (27). parahaemolyticus. 4. a marine halophile. but has the potential to cause fatal foodborne illness (26). vulnificus is an invasive and lethal pathogen associated with wound infections. 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. Quantitative levels were below limits of detection in all cases (<0. or freezing at a temperature of -23 °C for 7 days (30. however. in a survey carried out in the United States between 1977 and 86 . However. non-B Hepatitis viruses (31).FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States.7 Aeromonas hydrophila and Aeromonas sobria A. occurs in crustaceans from warm environments.8% of samples contained C. hydrophila and A. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. A.5.g. jejuni has not. 4. Norovirus. although A. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture.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. e. been implicated in a case of foodborne infection associated with crustaceans. V. 4. Japan. coli. 31.5.5. particularly trematodes (6). Caliciviruses. 32).9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. It is more commonly implicated in countries where crustacean shellfish are eaten raw.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. Of these. the potential hazard exists as C. whilst 5. 4.5. hydrophila has been isolated from prawns (29). they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). hydrophila is able to grow well at refrigeration temperatures (0 . Hepatitis A virus presents the most serious hazard. South America.6 Campylobacter jejuni and Campylobacter coli C. However. Parasitic infections are controlled by either cooking infected material prior to consumption. to date.

The toxin causes amnesic shellfish poisoning (ASP) when consumed.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive.5. The risk of such hazards is far higher with fin fish and molluscan shellfish. from farm to fork (37. 4. produced or imported into the European Union (EU). although it is one of the European Union (EU) legislated allergens. They can survive low pH levels. 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.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33).6.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish.11 Chemical pollutants and biotoxins Researchers have demonstrated that crabs feeding in waters contaminated with algae do have potentially high levels of algal toxin domoic acid (DA) in their viscera. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). No foodborne incidents have been attributed to this source to date. only 0. 2073/2005. A summary of microbiological criteria for crustacean shellfish is listed below. 39). Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. are included in the Commission Regulation (EC) No.CRUSTACEAN SHELLFISH 1981. 4. which could be transferred to the meat during unskilled removal from the shell (35). Provisions laid down concerning the use of additives would also apply to these types of products.5. which results in a persistent and apparently permanent loss of short-term memory (36). Selected parts from this regulation is presented below. 4. Norwalk viruses can also withstand heating to 60 °C. 87 . or chlorine concentrations similar to those found in drinking water. consistent with that in the stomach.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. 4. 38.

FISH AND SEAFOOD Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 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. 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



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. 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.

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.




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.





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.


28. 29.


31. 32.


34. 35. 36.


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.



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.


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

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

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

it is important to understand the concept of salt concentration.6 132. etc.6 264. all using salting steps.0 . it is important to state the temperature. processes and producers’ private recipes. it causes swelling because the fish absorbs water from the salt solution (1). The strength of the solution is described as degrees of saturation (brineometer degrees). the fish is usually salted with a 70 . and salt content.4 184.4% salt (Table 5.80º brine (184.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. Salt in the water phase links the two conditions of moisture and saltiness. or salt in the water phase. TABLE 5.8 79. This should be distinguished from the salt content. A 100º brine solution means that the solution is saturated and no more salt can be dissolved.3 Processing and its Effects on the Microflora Owing to the wide variety of products. sometimes the salt crystallises onto the surface of the fish.FISH AND SEAFOOD 5. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.2 g of salt per litre of brine).2 105. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. When working with salting. about 50º (132.8 211.4 52.I). a 90º or above brine (237. When a 100º brine is used. smoking or marinating processes. When a milder brine. Therefore. For smoking salmon.8 211. The dissolution of salt in water is dependent on the temperature. At 18 °C a 100º brine is produced when the solution contains 26.0 158.0 g of salt per litre of brine).2 237. Different brine strengths are required according to the curing processes and/or the type of product. it is impossible to establish standardised processing stages in the curing of fish.6 g of salt or more per litre of brine) may cause salt burn in this way. only the most common processes can be described here. Thus. Salt concentration means the percentage of salt in the water contained in the fish. is employed. % Salt in the water phase = % salt content % salt content + % moisture x 100 Another important point to remember when wet salting is used is the concentration or strength of a brine. a 0º brine saturation means pure water.

can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. the fish is restacked with fresh salt after about a week.3. However. can be dried further to produce ‘klipfish’ or ‘bacalão’. When pure salt is used. 97 . respectively (1). Usually it is followed by other preservation techniques such as drying and/or smoking. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. and such delays. the fish is mixed with one part of salt per eight parts fish. outgrowth of halophilic bacteria may occur.1. SMOKED AND DRIED FISH 5.0. which include calcium and magnesium. and for a heavy salting a ratio of 1:3 salt:fish is used (1). ‘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.1 Salting Salting of fish for preservation purposes has been used since pre-history. having a short section at the end of the tail. ‘Green cured’ split cod with about 60% of the backbone removed. low in calcium and magnesium. and a ‘putty’ fish product is also obtained (1).1 Dry salting / Kench curing Usually. are likely to cause hydroscopicity of the product and. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). when the air humidity is high. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. producing characteristic pink patches. 5. it tends to lead to a slight yellowing and softening of the product. if very pure sodium chloride is used for curing purposes. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. 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. are recommended to help make the product whiter. For a light salting. but the concentration of copper and iron must be kept under 0.15% calcium and magnesium. Also.CURED. and the salt has penetrated throughout the fish (3). and essentially free from iron and copper (3). which consequently inhibits the diffusion of the water from the bulk of the product to the surface.15 . The salt used should be of food-grade quality. The calcium and magnesium impurities. lean white fish is used for this purpose.35% and 0. for which there is a high demand in Mediterranean countries.3. however. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. addition of levels of 0. On the other hand. respectively.1 ppm and 10 ppm. Salting can be done in a number of different ways. mainly in tropical conditions. See Figure 5. This is because fatty fish turn rancid quickly. The most common fish used in the northern hemisphere is cod (Gadus morhua).1. and salt can speed this oxidation process. caught in North Atlantic waters. This condition is known as ‘putty fish’ (4).

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

78 6.CURED.16 7. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 . they can be stored at 28 °C.23 2. sardines.80 5.76 4.5 3.45 3.78 2.41 5.56 6. herrings and other fatty fish can be used. but mackerel.90 (1). headed and gutted.10 1.00 3.1. limiting the amount of rancidity that can develop. Histamine formation is of importance when using scombroid and some clupeidean species. the brine is then replaced over the fish.25 5.23 2.11 4. Histamineforming bacteria do not generally grow at water activities below 0.78) (1).64 4.23 3.30 2.76 99 .17 4.03 4. TABLE 5.26 1. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance. after being drained and pressed at 465 kg/m2 for 15 hours. 5.51 3. There are several variations of this process.70 3. the fish enzymes play an important role by maturing or ripening the product.41 3.3 Dry / wet salting In this process.22 6.44 2.74 9.66 5.09 8.70 6.17 5.5 3. Anchovies are the main fish used for these purposes.91 2.47 6.3.86 2.85 3.0 3. can be cured in a saturated brine solution for 6 days and. Here.88 5.33 5.25 5.76 4.03 2.23 8. and a red burgundy colour will develop in the product. For preservation purposes.04 5. if split fish is dry-salted.76 4. with pressure being applied to the top of the container to help the brine to drain away.98 5. the fish is dry-salted. a saturated brine is used.0 2.95 3.99 2. See Table 5.06 4. the liquid that comes out of the fish forms a brine.5 10.3 days.09 8.98 6.11 3.60 2. This is why it is the preferred method for fatty fish.26 7.19 3.65 2.44 2.51 5. and can be collected and replaced again over the fish.46 4.26 3.61 3. with a shelf life of about 4 weeks (water activity (aw) about 0.26 4.84 1.73 1.41 4.85 3. the product obtained is called gaspé-cured (5). but as it is placed in containers.35 4.54 5. thus. For example. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.11 9. SMOKED AND DRIED FISH smoking.II.66 5.49 4.0 4. The main objective of the brining method of salting fish is to limit air penetration during processing.61 3. The fish are kept immersed in their own brine near saturation point for several days.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) 2.76 4.5 4.05 7.76 4.96 1.26 2.41 6. Fresh sardines.98 6.

flavouring (sugar. it can be repacked in milder brine (1 . and packed (3). In a second step.3. only the nonspore-forming organisms are eliminated in the process and. then packed in barrels with a 70º brine. whereas the latter method preserves the fish for only a few days.4% of salt) for distribution and sale (1). is made of different varieties of fish. underlines the need for careful control (4).30 minutes. which is dipped into a boiling brine for 10 . After this. salt.2% acetic acid and 2 .2 months. and 6% salt. Another salt-boiled Indonesian product. containing 1 kg of salt for every 25 kg of fish. Other preservation techniques can be used for extending the shelf life of these products. even if the product is hermetically sealed.6 days without refrigeration (1). or can be boiled by being placed in a basket. because of the salt content of the product. they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. The first process can give a longer shelf life. Traditional jellied eels are a product where pieces of fish are cooked in boiling water. However. ‘ikan kaju’. As referred to above. until the flesh is soft. outgrowth of bacterial spores after processing may occur (4).5% 120grain distilled vinegar. Afterwards. Another variation uses sour cream instead of the second brine step.7%) for as long as 3 weeks. and spices) is added to the brine. They will have a salt content >6% (w/w) in the water phase. they are packed for consumption within 2 weeks at chill temperature (6). drying and salt-boiling steps until a hard translucent block is obtained (4). The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. The fish can be soaked in a high concentration of vinegar (about 4 . placed in a marinade of lime juice. usually requiring refrigeration below 10 °C for longer-term storage. Usually. a Peruvian product. 5. the occasional implication of pindang in cases of foodborne illness. and lasts for 3 . has a shelf life of several years.FISH AND SEAFOOD 5. Ceviche.3. herring or other fatty species are used. or a pH <5 and they are stabilised for a limited period. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. Once cooled.3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. up to 1 . giving a cooked appearance to the product. usually flavoured with onions and hot chillies. Herring can also be cured for several days in an 80 to 90º brine with 2.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. and even death. A much firmer product is obtained when the fish is salted prior to acidification. There is epidemiological evidence that these products have been the cause of some 100 . The lime juice plays the role of the vinegar. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. but it has further smoking. All these products are ‘semi-preserves’.

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

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

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

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

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

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

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

Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine.8% and still provide protection for 7 days (25). drying. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25).FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. consistency of flavour and colour. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. 5.5. When liquid smoke was used.4 Spoilage Brining. The synergistic effect of liquid smoke with other preservatives could reduce the amount of salt in the product when light diets are necessary whilst still meeting the necessary safety criteria (26). 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. so as to impart an unnatural red colouration to the flesh. smoking. Liquid smoke was considered to provide advantages of safety. nutritional neutrality and antioxidant activity (27). In experiments comparing traditionally smoked fish with liquid smoke. 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. injecting it into the fish flesh.03 μg/kg of benzopyrene. or applying it as an atomised spray within a modern automatic smoke kiln. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. or smoke extracts in organic solvents. 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. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). The European Scientific Commitee for Food. 5. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0.06 μg/kg of benzoanthracene in the final foodstuff (28). both of which are considered potential carcinogens. According to some recent research. applying it as a dip after brining. the salt level could effectively be reduced from 4. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish.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.6 to 2. Fresh fish spoilage may be measured by 108 . and less than 0.3. It has been shown that liquid smoke can reduce the formation of PAHs (3).

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

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

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

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

5-10 4. The concentration required to prevent growth at room temperature can vary from as low as 3. respectively.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades.5% to 5%. the non-proteolytic type E strains can grow in up to 5% sodium chloride. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). under optimal conditions. However.CURED.97 0. smoked and dried. botulinum type E. and as botulism is a serious hazard in foods. but if the fish products are stored at temperatures <10 °C. SMOKED AND DRIED FISH TABLE 5.86 0. and C. the fermented products being the most dangerous group (7).5-8. popular demand for healthy foods has led to a change in the tastes preferred by the public and. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. It has been shown that. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. Since neither the smoking nor the drying parts of this curing process are particularly severe. aureus Yeasts Moulds Lactic acid bacteria pH 5.5-10. botulinum type E Staph.5 5. botulinum type E and production of toxin.5 4. In the United States. botulinum.5.8 1.75 0.61 Organisms Most Pseudomonadaceae C. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. with this change. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. although the combination of the two factors. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. botulinum Staph. The presence of salt in cured products has a great effect on the growth of the bacteria.97 0. sodium 113 . types A and B are not affected by 3 .5-8.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13).0-9. when present in the raw material. Before. the products were heavily salted. Examination of 165 outbreaks of botulism caused by fish products showed that smoked.5 5. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). pressure on the traditional methods of curing.8 0.5 0-11 3.9 0. could not grow and produce toxin.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. the importance of cold-chain integrity must be emphasised. salted and pickled products accounted for 10. 9 and 8 of the outbreaks.

Shigella and E. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0.or cold-smoked fish should have at least 3.94 (7). after processing. if it contains sodium nitrite ≥100 ppm. As they are poor competitors. the risk of infection can be eliminated by proper cooking.86 and 10% sodium chloride. are particularly susceptible to contamination and. An outbreak of Shigella in an Italian cruise ship. they do not grow well in the raw material. as they require more handling. has been reported (42). the organism may contaminate the product. the human nose.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.5. the organism can survive most of the light curing processes. aureus. if recontamination occurs and storage conditions are favourable. Thus.0 . It is a halotolerant bacterium able to grow at water activities as low as 0. when the bacterial load of the product is lower.4. Once the product is contaminated. 5. but.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and.5% salt in the water phase.3 Enterobacteriaceae The pathogens Salmonella. throat and skin. The organism is mesophilic with a minimum growth temperature of 7 °C. it should contain not less than 3. 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. if storage temperatures are over 10 °C.5. aureus growth. 5.5. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). Sliced smoked fish products. In foodstuffs. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. more importantly. Infection by Shigella sonnei has increased in the UK. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene.5. As with other organisms. The FDA established that hot. gills and guts as well as in several lightly preserved fish 114 . related to the consumption of a raw smoked fish meal. if the person handling the product is a carrier of Staph. growth and toxin formation may occur. but requires higher temperatures for toxin production (>10 °C). and the minimum pH for growth is 4. Usually. but. they will grow rapidly (7).4 Listeria monocytogenes The psychrotrophic L. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 .30% of the population positive for enterotoxin-producing strains (41). However.0% salt in the water phase (30). 5. coli all occur on fish products as a result of contamination from animal/human reservoirs.

CURED. marinades (ceviche) (43) and gravad fish (44). which. monocytogenes in levels at or above 102 per gram (46). 5. both being detected in cold-smoked salmon. This includes matjes herring. Unlike some European countries. and other local traditional products (7).5. and in CT95-1207FAIR). As they are heat-sensitive. Its limiting conditions for growth are 0 . Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. Some LAB. Until 1997.the round worm Anisakis spp. lightly salted caviar. has also been detected in cod (7). researchers are looking for other ways of preventing the pathogen’s occurrence or growth. in the EU Fisheries R&D-FLAIR program. which means that the process must conform to a level of ‘no detectable L.12%.9. Two outbreaks in the United States have been reported to be linked to salmon (10). or nematodes. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. do not survive 1 min at 60 °C. 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. the only concern to public health is when eating raw or uncooked fish products. most of them are of little concern to public health. a number of fish products. Pseudoterranova dicipiens. from freshwater fish. gravad fish.0. They have complicated life cycles. the USA established ‘zero tolerance’ for the organism. The round worms. monocytogenes’ in 25 g of the finished product.5 Parasites Despite the presence of parasites in fish being very common. a round worm. SMOKED AND DRIED FISH products such as cold-smoked fish. there were no reported outbreaks related to cured fish. the CT 3162-FAIR program. The use of LAB in fish preservation is a topic that has been studied in projects (UP. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48).92 . in combination with curing agents and refrigerated storage.45 °C. widely used as starter cultures in food processes. which include intermediate hosts. pH 4.8 . a short period of freezing either of the raw material or of 115 . As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47).2. and the tape worm Diphyllobothrium spp. are considered unsafe. which considered L. favours growth of psychrotrophic LAB. salt levels of 8 .6 and water activities below 0.514. monocytogenes pathogenic for only specific segments at risk.95 (45). cold-smoked fish. have been shown to have an anti-listerial activity in many food products. Thus. such as ‘lightly preserved’ fish products. Of microbiological concern in the northern hemisphere are two parasites . from seawater fish.

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

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

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

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

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

. Applied Microbiology. 394-6. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. Keusch A. in Seafood Quality Determination. Incidence of Listeria in hot and cold-smoked fish. 1992.A.C. Letters in Applied Microbiology. Slade A. solute.. and salt concentration on growth and survival of Vibrio parahaemolyticus. Journal of Food Protection. Food Microbiology. 1994.. Kramer D. Bayliss A. Applied Microbiology.. Histamine production by psychrophilic flora. Marine toxins of microbial origin. 11. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. Ratnam S. 1995.W..R. 309-16.. 293301. International Journal of Food Microbiology. Liston J.J. 1974. Jemmi T. Eklund M.N. Dallas H. Fuchs R.T. 1973. in a salmon slaughterhouse and smoked salmon processing plant. International Journal of Food Microbiology. 121 . 1992. temperature.K. Roberts T. International Journal of Food Microbiology. 1987. Food Technology. 1075-80. Beuchat L.. Journal of Food Safety. Prevalence of Listeria in smoked fish..S. Interacting effects of pH. pH and sodium chloride.P.A. 25 (5). SMOKED AND DRIED FISH Sutherland J. Contamination pattern of Listeria monocytogenes and other Listeria spp.. 55 (11).R. 1991. 25. 21 (3).. Eds. Pelroy G. Taylor S. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. Rørvik L. 14. 309-18. 23 (1). 58 (5). Presence. van Spreckens K. 1994. Beuchat L. Poindexter C.A. Davies A. New York. Caugant D.E.. Poysky F. Ben Embarek P. 1988. Tran T. 27 (6). Hudson J. 42. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. Elsevier. 19. Journal of Food Protection. 844-6.. Dillon R.. 1734.. Combined effects of water activity. Patel T. Competition of food bacteria with Listeria monocytogenes during enrichment culture. 4389-95. 1994.178-80. detection and growth of Listeria monocytogenes in seafoods: a review. Nicolaides L. Lashbrook L. 217-36. 1994...CURED. Hitchins A. Letters in Applied Microbiology. Paranjpye R.R. Yndestad M.. Letters in Applied Microbiology. 1995. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. 11. 94-8.. and temperature on the growth of Vibrio parahaemolyticus. 19-27. Peterson M. 1995. 61 (12). 866-70. 502-8.


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

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

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

126 .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. which has been increasing in popularity over the last 50 years. the most popular varieties of which are rakørret. paddy ponds beside rice fields used for collecting fish when the field has dried up. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. 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. however. in Norway and surstrømming. and a large local freshwater reservoir (2). made from herring. for example. 6. Table 6. The diversity of fish used in fermented products (see. freshwater fish are the usual raw material. and the initial microflora is likely to be considerably larger in these cases. There are also more heavily fermented products.IV Fish species used in lactic fermented fish products of the Philippines Product names Burong ayungin Burong bangus Burong dala Burong gurami Burong hito Burong kanduli Burong tilapya Balao balao Burong hipon Local Ayungin Bangus Dalag Gurami Hito Kanduli Tilapya Tagunton Suwahe English Silver perch Milkfish Mudfish Goramy Catfish Sea catfish Tilapia Shrimp Shrimp Scientific name Therapon plumbeus Chanos chanos Ophicephalus striatus Osphronemus goramy Clarias batrachus Arias manillensis Tilapia nilotica Macrobrachium spp. some fermented fish products remain popular. A modern account of their production. These are often described as fish/salt products. For example.FISH AND SEAFOOD In Northern Europe.the flooded rice fields. in a survey of fermented fish products in north-east Thailand. Penaeus indicua Many of the fish used in fermentation are uneviscerated. malt or flour (1). fermented trout.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. relatively mild-tasting product. which employ lower levels of salt than Asian products. and old descriptions of their production often refer to the addition of whey. especially in fish/salt products. rakefisk or surfisk. TABLE 6. states that sugar is often included with the salt. in Sweden. and their microflora tends to reflect their local environment more than that of marine species. or buried salmon. Here. and counts ranging from below 104 /g to in excess of 107 have been reported. is a traditional. it was found that several different sources were used for the fish . Norwegian gravlaks.

In another microbiological study of four fish sauces. When this is a simple carbohydrate source such as sugar. Hansenula. More recently. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7).FERMENTED FISH Where other ingredients are used.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. Endomycopsis and Saccharomyces . and studies 127 .3.4.3 6. mean that microorganisms have been generally held to play little or no role in the process that follows.1 – 6. and the anaerobic conditions that prevail in the bulk of the material. were found to be the predominant isolates. however. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. (3. or boiled or roasted rice. Similar results were seen with patis. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). however. 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.74 or below. angkak (Philippines and Thailand) and look-pang (Thailand) are used. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. after one month. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. 6. traditional starter culture preparations such as koji (Japan). either directly or as the partially saccharified rice they produce. principally species of the genera Rhizopus. they can contribute to the initial microflora. Bacillus spp. Generally. These preparations contain a mixed microflora of moulds and yeasts. The amount of salt used is often only vaguely defined but is generally sufficient to reduce the aw below levels at which most bacteria associated with the raw material will grow and.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. In some cases. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). 4). suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. which included nam-pla and patis. This. sufficient to saturate the water phase present and produce an aw of 0. Other halophilic species have been identified from fish/salt products. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. in many cases. the Philippine fish sauce. It also demonstrated their significant proteolytic activity during the first month of fermentation. Mucor.

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

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

6 – 6.V and Figures 6.5. for example. 6. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. 6.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. In contrast to free-swimming fish. Fig. filtered and bottled. which is eventually separated from the residual fish tissues. Some examples of products and their formulation are presented in Table 6. snack) 130 . particularly glutamate. volatile fatty acids and nucleotides.3. Hoi-Malangpu-Dong (side dish.5).8. Figure 6. shellfish such as mussels often have appreciable levels of carbohydrate in their tissues and in these cases addition of carbohydrate is not necessary for a lactic fermentation to occur (see.

In these cases.FERMENTED FISH TABLE 6. N.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. C NE. north. Kung-Chom (staple food. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. 3:1:0. NE. NE 10 . N S 5 .6.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C. north-east. N. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus.12 months Shelf life Use 1 . south of Thailand. the prefix 'pla' is replaced by 'kung'. NE Formulation Fish (whole or pieces): salt: roasted rice.12 months Main dish 2 Pla-jao C.4 are also produced using shrimps. central.7 days 5 . C NE.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .12 days 3 . side dish) 131 .3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 .6.5-1.03 Preparation time 6 . Fig.2-4 Fish (pieces): salt:khaomark (fermented rice). N.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. S. C.

This is readily explained by the susceptibility of LAB to elevated salt concentrations. It also indicates that. 106) Fig. but there is considerable variation in this.2..salt and lactic acid. Phillipines Journal of Food Science and Technology. (from Arroyo et al. The LAB isolated from these products are those with more marked salt tolerance but. Generally. Red burong isda 132 . typically in the range 2 .7.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6.3. while they are able to grow at high salt levels. the salt is of far greater importance in determining the product’s long-term stability. then this will take longer to develop than if lower salt levels are used. If acidity is a desired sensory property of the product.VI). 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. giving a reduced rate of acid generation.10%. these products are salted at lower levels than the fish sauces and pastes.6. 2. of the two factors contributing to shelf life . their growth and metabolism are nonetheless slowed.1 The use of higher salt levels results in a longer production phase but a better keeping quality product.

Philippines Journal of Food Science and Technology.9 .1 .6 3.2 4.8 .5 . 133 .6.4. 106) Fig. 2.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).8 2.17.3 .5.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.4 .9 months 1 .5.5 .4.9 FISH (from Arroyo et al.5 3.1 4.8 .8 pH 4.0 .6.0 .3 5.0 Shelf life 3 weeks 6 .7 .2 7..6. White balao-balao TABLE 6.9.0 .3 years 1 .2 4.

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

3 days. The mixture is allowed to ferment for 2 . A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. Excessive growth and acid production by the LAB could render products too sour for acceptability. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). presumably correlating with the salt content. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. reporting the product’s pH to range from 4. such as wrapping in banana leaves. 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. and there is a marked regional variation in the preferred type.5 in the first 2 .8 . There is little published information on the factors that limit the shelf life of the low-salt products. allowing other spoilage organisms to grow. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products. 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).17.4 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).3 weeks at around 20 °C. which ranged from 7. the Korean fermented fish product. In a successful fermentation. but they are likely to behave similarly to lactic fermented products produced from other commodities. Results of a product survey were consistent with this. around 12%. 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). Generally. will not reliably exclude air. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases.5:9:3.2. is something of an exception since millet is used along with garlic as the carbohydrate source.2 at the end of fermentation. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. the pH drops from 6. the level of salt employed is on the high side for fish/salt/carbohydrate products.9%.FERMENTED FISH Sikhae. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. 135 . but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. lead to an increase in pH. markedly extending the shelf life (25).7 up to 6.7 to below 4. Traditional packing techniques used with lactic fermented fish products. Clearly. 6. in turn.5. thereafter declining to about 4. The growth of yeasts and moulds could.

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

31. Mahidol University.. References Riddervold A. whale meat. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. Raw materials subject to this process include seal flipper. Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. 2. Dollar A. Kasemsarn B. and salmon heads and eggs. they could act as an important vehicle of transmission. Journal of Food Science. though 27% survival was still recorded (33). Lutefisk. salt content was found to have the greatest impact on the viability of larvae. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. One study with the trematode Haplorchis taichui. With this particular organism. Institute of Nutrition. and warm weather can result in rapid growth of C.. Many of these products are described as fermented. Oslo. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. botulinum. botulinum and the production of toxin. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada. Dhanamitta S. Salting levels are often insufficiently low to prevent growth of C. while increasing the acetic acid content of the formulation had little or no effect (32). Fermented Fish Products Study (Thailand) Phase 1. 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). beaver tail and paw. Microbiology and chemistry of fermented fish. 105-10.O. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28).FERMENTED FISH botulism have become less frequent in recent years. Rakefisk and Herring in the Norwegian Tradition. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. 1966. Where these products are consumed without final cooking.. giving an alkaline fermentation. Liston J.. although the absence of any source of fermentable carbohydrate means that the process is putrefactive.. Novus Press. which is generally consumed raw (29). 1990. Greenland and Alaska. affecting an estimated 40 million people. walrus nose. which will persist into the product. Baseline Study (1987-1988). 1989.6 1. Foodborne trematode infections are emerging as a major public health problem. 3. 6.M. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31). Thailand. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. 137 . and these are often used in the production of fermented fish products. Puwastien P. Saisithi P. particularly in south-east Asia and the western Pacific regions (30). Yhoung-Aree J.

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

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

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

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

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

Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7.2. and fingers within 5 . 143 .2 Toxicology PSP toxins affect the central nervous system of mammals. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. However. tongue.FISH AND SHELLFISH TOXINS STX GTX2 GTX3 GTX5 (B1) C1 C2 NEO GTX1 GTX4 GTX6 (B2) C3 C4 R1 H H H H H H OH OH OH OH OH OH R2 H H OSO3 H H OSO3 H H OSO3 H H OSO3 - R3 H OSO3 H H OSO3 H H OSO3 H H OSO3 H R4 H H H SO3 SO3 SO3 H H H SO3 SO3 SO3 - Fig. Human symptoms begin with tingling and numbness of the lips.30 minutes of consumption.1. this is never sufficient to render the seafood safe to consume. and humans seem to be particularly susceptible. 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. 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). Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis.

other derivatives have been reported. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. 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. Since its discovery.2. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). 7. TTX most commonly occurs in many species of puffer fish. and islands of the South Pacific.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. 15.3. 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. Thailand. and over ten analogs are known (22. China. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). Most intoxication events have occurred in South East Asia. a criticism has been that such methods are time consuming. However it remains as the monitoring method of choice in many countries. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. 24) (Figure 7. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. 7. Two approaches have been taken here. including Japan. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. Alternative HPLC methods have subsequently reduced this to a single HPLC run. 17).8 µg/g) of shellfish or seafood tissue. as well as other parts. After some development and various trials. While the chemical detection methods are most promising for the future. and while simple and relatively inexpensive to operate. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. a pre-column method has received Association of Analytical Communities (AOAC) approval (18).2). The mouse bioassay has been in place for decades and the method has been standardised.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes.FISH AND SEAFOOD 7. 23. Taiwan. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation).3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. specifically 144 . and has also been evaluated by a number of EU laboratories.

23. and once again the mouse bioassay has been used to detect the presence of the toxin. it is served in speciality restaurants with qualified chefs to properly prepare the fish. horseshoe crabs. respiratory failure and coma. where puffer fish of the genus Fugu are a delicacy. and even newts. Severe poisoning causes paralysis and respiratory failure. raising speculation that TTX is produced by a bacterial source. 7. gonads and roe (24). 27). An oral dose of 1 .2 mg of purified toxin can be lethal. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. 26). This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. nausea and vomiting. and blocks diffusion of sodium through the sodium channel. 7. A review of TTX detection methods has been published (12. molluscs. Structure of tetrodotoxin 7.3. Since then. and more recently by an actinomycete Norcardiopsis dassonvillei (26). and in mild cases results in distal muscle weakness. 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.FISH AND SHELLFISH TOXINS in the liver. preventing depolarisation and propagation of action potentials in nerve cells.2.3. In Japan. the blue-ringed octopus. All of the observed toxicity is secondary to the action potential blockade. Typically this has been accomplished using HPLC followed by post 145 . The onset of symptoms usually occurs rapidly.2 Toxicology Like the PSP toxins. Fig. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. 25. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. hypotension.

but 146 . both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. 7. Structure of domoic acid and three naturally occurring photoisomers 7. and while promising.3 for examples). Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31).4.3. a polar glutamate agonist. need further development (24). 7. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. Fig. Immunoassay methods have also been investigated. Currently 12 isomers of DA have been reported (see Figure 7. However this method has been criticised for being complex. but all of them are considerably less toxic than DA itself (29).4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28).FISH AND SEAFOOD column oxidation to the fluorescent derivatives. but it is known that such congeners are less toxic than DA itself (30). and more rapid LC-MS methods [(M+H)+ m/z 320] have been reported for the detection and identification of TTX and its various derivatives (27).

has been reported as a medium-level producer of DA (33).10% of the administered dose. disorientation and coma.. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. When DA was identified as a new marine toxin it was quickly determined to be a glutamate agonist that binds to a sub-class of neuronal receptors called kainite receptors. and this has led to ASP toxicity in birds. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample.FISH AND SHELLFISH TOXINS only three strains (P. To a great extent. 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. Victims display a variety of symptoms including headache. as does impaired renal function which increases serum concentration and residence time of the toxin. Pseudonitzschia australis. and in locations around New Zealand (32. 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 . but any impairment of the blood brain barrier results in additional risk. southern and western portions of Ireland. 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. 33). The oral absorption of DA is around 5 . multiseries. Indeed. Okinawa and more northerly parts of Japan. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). another diatom species Nitzschia navisvaringica. and clams. scallops. found in the Pacific around Vietnam.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. the Mediterranean. and sea animals such as sea lions and even whales (35). For example. Levels of DA in the other strains are generally <1 pg/cell. the wide distribution of toxin-producing strains of the diatom have resulted in the frequent occurrence of DA in many filter feeding organisms such as mussels.39).4. Penetration of the blood brain barrier is poor in healthy subjects. the Philippines. and review of the toxicologic pathology of DA has been published (36). which has been well documented in studies along the west coast of the US. seizures. Another factor is the movement of the toxin through the food chain. and is mainly distributed in the blood. 147 . nausea. Recently. 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. cormorants and pelicans (34). 7.

and these methods have been reviewed (40. 41). Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them.4.. although in the latter study the probes for Pseudonitzschia pungens. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 .. The benthic Prorocentrum spp. The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. as can LC/MS. However. australis. are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. DTX-2 was discovered in contaminated Irish shellfish (46). though this method is most commonly used to analyse plankton and seawater (42). Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). 49).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. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. Alternatively. the heterotrophic dinoflagellates Dinophysis spp. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. In the context of this chapter. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). To further complicate matters. 7.4). produce a variety of other 148 . and dinophysistoxin-2 (DTX-2) (Figure 7.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). shellfish tissue can be efficiently extracted with aqueous methanol (1:1). Since then. 48. Fortunately DA is conveniently detected by a variety of LC-linked analytical methods and this is the recommended approach for DA monitoring in EU countries. but just over 15 years ago a third toxin. Dinophysis spp. and consequently were often overlooked as the real source of the toxins. California. dinophysistoxin-1 (DTX-1). 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. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. in Monterey Bay.FISH AND SEAFOOD 7. and this is typically the method of choice. A rapid immunodiagnostic kit is now available to detect DA in phytoplankton and is reported to work satisfactorily in this application in the field (43). and an immunosensor method has also been reported (4).

The parent acids are the only DSP congeners to be found in shellfish tissue. DSP toxins produced by benthic Prorocentrum spp. Structure of okadaic acid and the two related DSP toxins 7. Today over ten Dinophysis spp. Dinophysis caudata. 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. which include the pectenotoxins and the yessotoxins. are also recognised as a threat to human health. 149 .4. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. and Dinophysis norvegica among others (47.1 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. 7. In comparison. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. Dinophysis fortii. This was probably not the first DSP episode. Fig. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). Prorocentrum maculosum. In this case Dinophysis was suspected as the culprit organism. 49). DTX-5. and include Dinophysis acuminata. such as Prorocentrum lima. and Prorocentrum belizeanum.5.. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. 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. While these esters are not toxic in vitro. are recognised as DSP toxin producers. Dinophysis acuta. Lately these compounds.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms.

All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1.5. and DTX-2. The 7-O acyl derivatives (DTX-3) of OA that can form in shellfish also show poor cross reactivity and this further exacerbates the problem.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. DSP episodes have been reported from Japan. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. DSP-Check is a commercially 150 . vomiting.3 days. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. which can lead to an underestimation of the total DSP content in a sample. and abdominal pain. Due to the lipophilic nature of the toxins. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. 7.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. New Zealand and Chile. 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. it does not provide information on the DSP toxin profile. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). In recent years. Human symptoms include diarrhoea.5. and a detection limit of 100 µg/kg has been reported (12).FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. and while this provides critical seafood safety information. even with the minimal of clean up steps (48). The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 .2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. A regulatory level of 80 . 7. nausea. DSP episodes have been reported worldwide and are included in many monitoring programs (47). 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). At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. 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. various European countries. 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. Since the late 1970s. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. 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).

for many years these compounds were included in the DSP toxin complex. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. Some representative examples of both groups of toxins are shown in Figures 7. with a claimed detection limit of 20 µg/kg. 55). and the latter group are usually found to be responsible for any reported shellfish toxicity (59).6. with good agreement with the mouse bioassay and LC–based methods (54.6 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). In yet another approach.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. Fig. 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. 7. 7. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. and another rapid antibodybased test kit for DSP toxins has also been reported (53). fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. 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.5 and 7. Indeed. Within this group. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). 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. 58). However.5.

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). 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. While 152 . but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. the toxin content and profile may vary among different strains.6. New Zealand and South America (49.6. 7. Japan. many parts of Europe and the Mediterranean.6. 58). eastern Canada.FISH AND SEAFOOD Fig. 57. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. 7. Structure of the two parent yessotoxins. YTX and homo-YTX 7. As is typical for many algal species.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). which may co-occur in blooms with other organisms.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59).

7. There are no clear cases of human illness resulting from ingestion of PTXs (59). While LC/UV detection is possible for PTXs. Because of the number and complexity of the YTX group. a regulatory level of 1 mg/kg shellfish tissue has been set. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7. 58). Some of the PTXs are potent cytotoxins in vitro (60). Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. some of which are found as glycosides and sulfates. 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. Their toxicology and mechanism of action remain uncertain. the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. which aids detection by LC/UV.6.3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue. it may be compromised by other biological materials in the matrix. and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. The YTX group has not been reported to cause human illness. For many years it was believed that 153 . but a fluorescent derivative has been prepared for others. though its cell effects may be linked to modulation of calcium channels (62). 64). 7. The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. The structures and chemical data including the molecular weight of each member have been collated (56). However. which is further compounded by differences in the experimental design. The crude extract may be used directly in the mouse bioassay. Many YTX toxins contain a conjugated diene system.7). the PTXs show considerably less toxicity when administered orally. but less so following oral dosing (61). chemical analysis is best carried out by LC/MS and several approaches have been described (57. Estimates of acute toxicity of the parent compound following IP injection vary considerably. but various congeners are acutely toxic in the mouse bioassay following IP injection. Although there is no evidence that yessotoxins are responsible for human illness.FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds.

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

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

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

Patients have reported symptoms lasting weeks and even months. 75. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 .76). and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. which in turn are consumed by carnivores such as snappers. weeks or even months (74 . 7. and moray eels. Globally. arthralgia. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. and the sensation of loose teeth (77). Even when suspect samples are available.3 Detection methods As a consequence of their potency. all of which can be linked to the effect of CTX on excitable membranes. and although the gastronomical problems may dissipate within a day or so. sharks. Local island populations attempt to avoid poisoning by catching only smaller fish. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. where cold objects feel hot and vice versa. and it is estimated that ingestion of as little as 0.8. 75). there are neither standards nor an official testing program. These include paresthesias (numbness and tingling of the extremities).2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. 7. although not all CTXs are equipotent. pruritus. which can be confused with chronic fatigue syndrome. only trace amounts of CTXs can cause illness following consumption of contaminated fish. The onset of symptoms can occur rapidly within 24 hours. which can return following the consumption of alcohol and certain foods such as oily fish (74.1 µg of ciguatoxin can result in human illness (74. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. Like the brevetoxins. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. groupers. 78). The recommended method to extract fish tissue has been described in detail (74) and the initial warm acetone extract is partitioned against hexane and diethyl ether where the CTXs are concentrated. and temperature sensation reversal. and there are suggestions that such incidents are increasing and spreading worldwide (74). myalgia. Other symptoms include a metallic taste. barracudas. and in fact CTX poisoning is often only identified after consumption of contaminated fish. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs.8. the neurological symptoms can persist for days. Another symptom that can develop over time is extreme fatigue.

and following synthesis of the molecule some time later.30%) and AZA3 (5 . and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge.20%) present in lesser amounts (Figure 7.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. 158 . 84). following consumption of contaminated mussels Mytelius edulis (82). minor revisions of a few stereocenters were made. Consequently. Recently.g. Since the original discovery and identification of the toxin. high-throughput screen (74). Usually the major AZA derivatives found in shellfish are AZA1 (50 . which are based on an LC-MS/MS approach. Recent reviews of this new group of toxins have appeared (83. with AZA2 (10 .9). Chemical detection methods are severely compromised by the lack of standards.85). The majority of these have been proposed based on LC/MS analytical data. In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). only a few laboratories are able to undertake such analyses. which features characteristic fragment ions corresponding to sequential losses of three molecules of water. 7. has been developed. Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. Acetone extracts of fish tissue are subjected to solvent partitioning. A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine.FISH AND SEAFOOD Lethal Dose (MLD) 0. almost 20 additional derivatives have been reported (83 .25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. specific for CTX activity. and only a few structures (AZA 1 .9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. followed by clean-up on silica and aminopropyl cartridges. Immunoassays may be an alternative detection method (79). but this only has application in a clinical setting (81). The original structure proposed revealed an entirely new class of shellfish toxin. although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). and the corrected structure was published in 2004 (86) (Figure 7. another in vitro sodium channel-based assay. at C-3 and C-23).80%). and another dealing with the chemistry and detection methods has been published (85). This is then subjected to LCMS/MS analysis.9).

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

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

selliformis (97). Some time later. 95). 7. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). two additional congeners. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. 7.1 Origins and distribution The original observation of toxicity in oysters led to a wider survey of other shellfish species in New Zealand when it was discovered that the toxin was present in a variety of bivalve molluscs including mussels. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin.10. Structure of the cyclic imine gymnodimine 7.1 Gymnodimines The observation of neurotoxic symptoms in the mouse bioassay of lipophilic extracts of dredge oysters (Tiostrea chilensis) collected from around the South Island of New Zealand. At the time of the initial observation of gymnodimine. 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.10.FISH AND SHELLFISH TOXINS health. Subsequent taxonomic studies revised this again to a new dinoflagellate K. scallops. 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. 95). Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94. but they are included here for completeness. they are sometimes referred to as emerging toxins of uncertain human health concern.520 Da (94.1. it was proposed that a dinoflagellate identified as Gymnodinium spp. surf clam and New Zealand abalone. cockles. 161 .10).10. Fig. Pacific oyster.

extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. 95) (Figure 7. Since their original discovery which resulted in the characterisation of spirolides A-F (99. abdominal breathing. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. and this is the detection method of choice. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. 50%) of 96 µg/kg. However. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. A later study using purified toxin established an LD50 dose (Lethal Dose.10. 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.900 µg/kg).10. and so spectroscopic detection by UV methods is not favoured. 95).2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. and lack of responsiveness to stimuli. it has been stated that regulation against gymnodimine is not required (98).1. They were originally discovered in extracts of mussels (M.10. Reduction of the imine group eradicated the toxicity. finishing with 100% methanol. In particular. Based on this diminished oral toxicity. together with a lack of evidence of human intoxication. Most cyclic imines lack an extended chromophore.to10-fold reduction of toxicity (LD50 600 . paralysis of the hind quarters and eventually complete immobility. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. Oral administration of the toxin to mice by gavage resulted in about an 8.1.100).2 Toxicology A review of the toxicology of all cyclic imines has been published (98).FISH AND SEAFOOD 7. Whether isolated from dinoflagellate cells directly or from shellfish tissue. 7.3 Detection methods The characteristic “fast-acting. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . The time lapse between ingestion and death is between 5 and 15 minutes. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. There is a rapid onset of symptoms including respiratory distress. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay.11). 7. almost a dozen derivatives in total have been identified to date (94.

7.707 µg/kg. fasted mice). The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . those spirolides possessing the vicinal dimethyl grouping (see Figure 7.10. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). Structural differences among the various spirolides result from variations in the number of methyl groups and the sequence of spiroketal rings 6-5-5 in one group and 6-6-5 in the other. by gavage (87 . Interestingly. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). the gulf of Maine.11.2.8 µg/kg). fasted mice). in fjords in Denmark and Norway.2. While oral toxicity is considerably less than by IP injection. 7. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. All data reported here are for desmethyl spirolide C. southern Ireland. or by oral ingestion (381 .2 Toxicology The spirolides are the most studied group of the cyclic imines. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101).10.166 µg/kg. 95). Fig.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). Structure of four spirolide toxins A-D 7.11. the western coast of Scotland.

four members of the group have been identified – two of them being the C-34 diastereomers. 7. To date. 102). 95.12).10. Fig. 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.3 Pinnatoxins and pteriatoxins Pinnatoxins A .3 Detection methods As discussed for gymnodimine.12. Three members of this group have been reported.FISH AND SEAFOOD (100). 7. the mouse bioassay can be used to detect these fast acting toxins. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. 7. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7.2. While the mechanism of action remains to be clearly defined. A number of methods following this procedure have been described (94. and in fact these compounds share a number of other structural features. which includes two diastereomers. consistent with modulation of acetyl cholinergic receptors (103). However. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C.B contain a spiro imine moiety similar to that found in the spirolides.10.

most likely a dinoflagellate. new toxin groups such as the azaspiracids and cyclic imines have also been reported. However. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins.11 Conclusion There has been considerable progress in the detection of known marine toxins.1 Origins and distribution Shellfish of the genus Pinna. there is no compelling evidence to suggest that they have resulted in human toxicity (98). 99%) values for pinnatoxins A-C are in the range (22 .3.2 Toxicology There is little information describing the toxicology of these compounds (98). There is strong circumstantial evidence to suspect that these toxins. to date action or 165 . which in many cases are now found to be more widespread than previously believed. Despite this progress. Coupled with this. The LD99 dose for the pteriatoxins is reported to be in the range (8 . The LD99 (Lethal Dose. This is generally a result of increased monitoring and continually improving detection technologies and methods. While this is recognised as a real possibility for a wide range of toxins. 7. With increasing knowledge of the chemistry of toxins. Only the acute toxicity in mice following IP injection has been reported.10. 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.180 µg/kg). arise from an algal source . but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. which occupy tropical waters around Okinawa and the south east coast of China. but this remains to be conclusively established (106). while pinnatoxin D was essentially inactive. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105).3. 7.3 Detection methods No formal detection methods for these compounds have been reported. and more advanced means of detection. like the gymnodimines and spirolides. have long been suspected of causing human illness after ingestion.10. 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. 7.10.100 µg/kg). once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures.3. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other.

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

Lacaze J-P. Smith E. AOAC International.. Petrie J. Naoki H. Rome. 14. 40. Eds.. 2002. Shoji Y.. 2008. 2001.F. 1995. 391 (1).A. Murata M. CRC Press. 22. Arakawa O. 15. 5-52. Fukuyo Y. 2008.06. 2008. 2005. Chemical analysis... Botana L. 349-71... Wang D-Z. 2006. Stobo L. 177-96. Yasumoto T. Interspecies distribution and biogenetic origin of tetrodotoxin and its derivatives. Yotsu-Yamashita M. Food and Agriculture Organisation.H. Gaithersburg. Mackintosh F. 3-20. Ed.C. Scott A.M.M. Eds. CRC Press..V. 23.L. Rapid detection methods for marine toxins: needs and limits. in parallel with the regulatory AOAC mousse bioassay. Paris. 230-42.A. Barug D. Analytical and Bioanalytical Chemistry. Paralytic Shellfish Poisoning. Ben-Gigirey B. Ed. Boca Raton. 24.. a new rapid test.. Aune T. 283-317. Noguchi T..J. 61. Detection of paralytic shellfish poisoning (PSP) toxins in shellfish tissue using MIST Alert. in FAO Food and Nutrition Paper 80. Toxicon. 18. prechromatographic oxidation and liquid chromatography with fluorescence detection.A. Quilliam M. 1407-25. Taylor F..L.A. Villar-Gonzalez A. Barrett R. 51. Yasumoto T. Hallegraeff G. and liquid chromatography/tandem mass spectrometry.. Roberts R. 21. 505-8. Marine Biotoxins. Journal of Shellfish Research. 2008. 10-7.. 167 .. Boca Raton. Christian B.E. Wageningen Academic Publishers.M. Food and Agriculture Organisation of the United Nations. Surveillance of algal toxins in shellfish from Scottish waters. 21. Taxonomy of harmful dinoflagellates. Electrospray ionisation mass spectrometry of tetrodotoxin and its analogs: Liquid chromatography/mass spectrometry. Marine Drugs. Endo A. 455-60.M. Association of Official Analytical Chemists. 2002. Paralytic shellfish poisoning toxins in shellfish. 12. Laycock M... Toxicon. 635-48. 20. Pure and Applied Chemistry.. Evaluation of MIST Alert rapid test kits for the detection of paralytic shellfish poisoning toxins in shellfish. in Rapid Methods for Biological and Chemical Contaminants in Food and Feed..M. Turrell E. 19. Neurotoxins from marine dinoflagellates: A brief review. 1989. Miyazawa T.FISH AND SHELLFISH TOXINS 11. 290. 219-27. Cembella C.. Anderson D. Determination of marine biotoxins for regulations: from the mouse bioassay to coupled LC-MS methods. Risk assessment of marine toxins. Larsen J. 16.. 2008. in Manual on Harmful Marine Microalgae. UNESCO. tandem mass spectrometry.. Luckas B. 117-34. in Methods of Analysis of the Association of Official Analytical Chemists. in Seafood and Freshwater Toxins. Jellet J.C. Krys S.. 6. 2008. 6. Marine Drugs. Analytical Biochemistry. Food and Agriculture Organisation of the United Nations. Ed.. Yotsu M. Van Amerongen A. 2004. in Seafood and Freshwater Toxins. Lauwaars A. Wageningen.. Botana L. 17.R. 13. method 2005. Tetrodotoxin – distribution and accumulation in aquatic organisms and cases of human intoxication.

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

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

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

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

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

80. 81.








89. 90.



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


94. 95.





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

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


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


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.


3.Obtaining evidence that the elements of the HACCP plan are effective. from primary production to final consumption. food microbiology. 2. The team should also include 177 . It is recommended by the Codex Alimentarius that the practical application of the HACCP principles be approached by breaking the seven principles down into a 12-stage logic sequence. Verification . Determine the CCPs. 6. Validation . in addition to monitoring to determine compliance with the HACCP plan. Each stage is discussed below in detail. Establish the critical limit(s). food chemistry or engineering. Establish a system to monitor control of the CCP. procedures.1 is a flow diagram illustrating this 12-stage logic sequence. 4. Establish the corrective action to be taken when monitoring shows that a CCP is not under control.1 Assemble HACCP team HACCP requires management commitment of resources to the process. ingredient and packaging buyers.3. quality assurance or control.The application of methods. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. operation or stage in the food chain including raw materials. Establish documentation concerning all procedures and records appropriate to these principles and their application.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . tests and other evaluations. 7.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. 8. Figure 8.A point. procedure. Establish procedures for verification to confirm that the HACCP system is working effectively. 5. Conduct a hazard analysis. 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. hygiene managers. 8.

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

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

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

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

1 Fish and seafood Again. 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. More than one critical limit may be defined for a single step. and its use may not always be appropriate. it is necessary to specify both time and temperature for a thermal process. 8.3.FISH AND SEAFOOD 8.3.2) to provide a logical. biological or sensory parameters. For example.8. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. 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 not possible to generalise on likely CCPs. 8. Where possible. then the process must be modified to include a control measure. Criteria used to set critical limits must be measurable and may include physical.3.3. it can be said that effective control measures are likely to include the following: • • • • • 8.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. critical limits should be specified and validated for each CCP. given the enormous variety of fish products and processes in use. chemical. and the reader is referred to the appropriate product chapter. structured approach to decision making. application of the decision tree should be flexible. However. but for which no control exists at that step or any other.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8.8 Careful selection of sources for raw materials Adequate temperature control Effective sanitation Food handler hygiene Prevention of cross-contamination Establish critical limits for each CCP Critical limits separate acceptable from unacceptable products. However. If a significant hazard has been identified at a step where control is required for safety.7.


• • • 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


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.



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



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.


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.


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

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

N. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. 41). p. 24. 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. 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. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No.1991. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004.B. transportation. European Communities L 268.9. p.1992. 7.1). p. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. 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 187.7. J. as last amended by Regulation (EC) No 806/2003. handling and offering for sale or supply to the consumer. and sets out what those enforcing the provisions have to do. 24. processing. Regulation 854/2004 relates to the organisation of official controls on products of animal origin.2 Legislative Structure From 1 January 2006. Subsequently. 9. distribution. 190 • • . A number of more detailed implementing and transitional measures have been adopted at EC level.1991.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No.15). as last amended by Regulation (EC) No 806/2003. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. packaging. storing.9. manufacturing.

I. 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. p. 1 .I. 9.7. of stock farming. the legislation is directly applicable law.1993. and of hunting and fishing.3 Regulation (EC) No. which will be covered later in this chapter. and equivalent legislation in Scotland. Additionally the requirements of EC Regulation 853/2004 must be complied with. 2006 No. and binding in its entirety on all member states from the date of entry into force. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. 852/2004. revocation of existing implementing legislation. 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.) in England. Wales and Northern Ireland). 19. right through to the implementation of procedures based on HACCP principles.g. is required to give effect to the EU regulations. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. etc.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. As regulations. for example. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. e. Primary products are defined as products of primary production including products of the soil. There are some exemptions. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. 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. food storage that is for private or domestic consumption. Wales and Northern Ireland. national legislation in the form of a Statutory Instrument (S. 191 .11) The EU hygiene regulations apply to all stages of food production including primary production. setting offences. Although the regulations have the force of law.14. domestic preparation or handling. penalties and powers of entry. The Food Hygiene (England) Regulations 2006 (S. with primary production.



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: 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.




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


- 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.

- 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.



FISH AND SEAFOOD 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.

- 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.

- 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.



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.


LEGISLATION 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. 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.

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





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:


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

In this case. (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. EO. EB or WE. the mark must show the approval number of the establishment.2. (ii) However.2. please refer to Regulation (EC) 853/2004.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. Section I of Regulation (EC) 853/2004. EK. An identification mark can only be applied provided the relevant requirements as stated above in relation to Regulation (EC) 852/2004 have been met in particular for the registration and approval of establishments. For the UK. EY. EF. 9. EG. EÜ.1. for other codes. Section I of Regulation (EC) 853/2004 are given as follows: 9. the food business operator may apply the same identification mark to both types of food.FISH AND SEAFOOD 9.4. EK. (iv) If the identification mark is applied in an establishment within the Community.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production.4.4. when a product's packaging and/or wrapping is removed or is further processed in another establishment. the new mark must show the approval number of the establishment where this occurred. EC.4.2 Form of the identification mark (i) The mark must be legible and indelible. 198 .2 Fishery product requirements 9. (iii) Additionally. ES. Extracts of the requirements of Annex II. a new mark must be applied to the product.2. the code would be UK.1. (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. the mark must be oval shaped and include the abbreviation CE. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. and the characters easily decipherable and be clearly displayed.

Part A and Chapter V apply to retail. granulate and powdered products of animal origin carried in bulk. processing.4. Chapter III. Chapter IV. the colourings used must be EC authorised colours. Therefore. Extracts of the requirements of Annex III. 9. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. This section does not apply to bivalve molluscs. and fishery products carried in bulk. Parts A. it applies to those animals when not placed on the market live. With the exception of Chapters I and II. as amended. (ii) For products of animal origin that are placed in transport containers or large packages. (v) When the mark is applied directly to products of animal origin.2. for full requirements. Section VIII of Regulation 853/2004. C and D. specifically relating to fishery products are given below. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. (iii) In the case of liquid. the approval number of the establishment and the abbreviation for the community where appropriate. echinoderms. the mark may be applied to the outer surface of the container or packaging.e. it is sufficient to apply the mark to the exterior of that package only.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products.LEGISLATION 9. the wrapping or the packaging. a) requirements for establishments.4.2. they would need to be obtained following Section VII on live bivalve molluscs. reference should be made to the actual Regulation. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. or be printed on a label affixed to the product. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . i. and are intended for further handling.1. The requirements of this section accompany those in Regulation (EC) 852/2004. its wrapping or the packaging. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. tunicates and marine gastropods when placed on the market live. wrapping or packaging in another establishment. including vessels.3 Method of marking (i) The mark is to be applied directly to the product.

) 9. removing fins.FISH AND SEAFOOD c) for the water supply. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 . this supplements the requirements of Annex II. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter. Chapter VII of Regulation 852/2004.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. 2.1 Chapter I . they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products. A. heading.2. refrigeration and wrapping.. fishing and collection of live fishery products with a view to their being placed on the market. The definition of fishery products covers primary production as well as its associated operations. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material.4. or to handle or process them after harvesting. Structural and equipment requirements refer to the following points 1. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. There is a derogation from point a. i. from the place of production to the first establishment of destination.e. bleeding. clean seawater may be used for the handling and washing of fishery products. (a) primary production covers the farming. it must be placed somewhere that will not contaminate the water supply.2. gutting. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. 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. if a vessel has water coming in that is used with fishery products.

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

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

Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed.LEGISLATION consumption. Food business operators need to co-operate with the relevant competent authorities so as to allow them to carry out official controls as laid down in Regulation (EC) No. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. C. (ii) Heading and gutting must be carried out hygienically. They should be wrapped. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . B. handling fishery products A. 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. 9. Fillets and slices must then be removed from work tables as soon as possible after preparation. C. and where necessary. (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. 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.2. Additionally. dispatched. points 1 and 2. prepared or processed immediately after arrival at an establishment on land. packaged and chilled as quickly as possible after preparation.4. 854/2004. part I. they must be stored under ice in appropriate facilities.Requirements for establishments. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. Chapter I.3 Chapter III .2. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. including vessels. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises.

(b) fishery products from the following species. (iv) (wild) Atlantic and Pacific salmon. (iii) A document from the manufacturer.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. (iii) sprat. or a stabilising treatment. D.4 Chapter IV .2. (ii) mackerel. they must be cooled rapidly.Requirements for certain processed fishery products A. and (c) marinated and/or salted fishery products. 9.2. 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. and whole fish must be gutted and washed beforehand. raw materials must have no guts. stating the type of process they have undergone. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. Requirements concerning parasites (i) The fishery products below must be frozen at no more than .20 °C in all parts of the product for at least 24 hours (this treatment must be applied to the raw product or the finished product): (a) fishery products to be consumed raw or almost raw. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. (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. (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. If no other method of preservation is used. except when supplied to the final consumer. Once produced. cooling must continue until a temperature 204 .4. must accompany fishery products referred to above when placed on the market.

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

FISH AND SEAFOOD D.4. it should be kept at no more than -9 °C. In the case of whole frozen fish in brine intended for the manufacture of canned food. 9.4.Storage of fishery products (i) Fresh fishery products. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. 206 .2. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. 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. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. thawed unprocessed fishery products. prepared and processed fishery products belonging to the family Gempylidae.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.6 Chapter VI .2. 9. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. and when fishery products are wrapped on board fishing vessels. Fresh. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing.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. Diodontidae and Canthigasteridae.7 Chapter VII . The scientific name must accompany the common name on the label. E.2. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption. Molidae. Fishery products contaminated with parasites may not be placed on the market for human consumption.

2.4.2. (ii) If fishery products are kept under ice. 9. Sebastichthys capensis.2. species belonging to the Merlucciidae family. 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.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. Helicolenus dactylopterus.). species belonging to the Gadidae family.3 Other information: Rules relating to visual inspections for the detection of parasites in fishery products are stated in Annex II (obligations on the competent authorities) of Regulation (EC) No. 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.4. The temperature of frozen fishery products must not fluctuate above -15°C. provided that the journey is short and the competent authority allows this. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. (d) 60 mg of nitrogen/100 g of whole fishery products used directly for the preparation of fish oil for human consumption as referred to in the second sub-paragraph of Part B(1) of Chapter IV of Section VIII of Annex III to 207 . (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp.8 Chapter VIII . 9. 2074/2005.Transport of fishery products The above points for Chapter VII apply in relation to transport. melt water must not remain in contact with the products.

they supplement the requirements of Annex II to that Regulation.FISH AND SEAFOOD Regulation (EC) No 853/2004. they supplement the requirements of Annex I to that Regulation. as amended. (b) In the case of other operations. reference should be made to the actual regulation. this Section also applies to live echinoderms. where the raw material complies with points (a). 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. Chapter IX applies to pectinidae harvested outside those areas. Apart from the provisions on purification. VI. shellfish species etc. taken from at least three different points and mixed together by grinding. 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. tunicates and marine gastropods. The sample must consist of about 100 g of flesh. 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.3. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. The copy of 208 . please refer directly to the regulation. however.4.4. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. Section VII of Regulation 853/2004.1 Chapter I . 9. The information required in the registration document is given in point 4 and includes the health status of the production area. and point 3 of Chapter VII applies to retail. VIII and IX. Extracts of the requirements of Annex III. for full requirements. Live bivalve mollusc requirements - 9.General requirements for the placing on the market of live bivalve molluscs Live bivalve molluscs can only be put on the market for retail sale if done through a dispatch centre. (b) and (c) of Part B(1) of that Chapter. specifically relating to bivalve molluscs are given below. Chapters V.

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

210 . containers and a label to identify purification.5 Chapter V . and there are requirements for tanks and water storage containers. (iv) yessotoxins.4. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. 1 milligram of yessotoxin equivalent per kilogram.3. Requirements for dispatch centres Handling should not result in contamination to any degree.4.3. Requirements for purification centres There are requirements for live bivalve molluscs to be washed. Before dispatch. for: (i) paralytic shellfish poison (PSP).3.4. shells of live bivalve molluscs are to be washed throughout with clean water and the molluscs are to come from one of the following: (i) Class A production area (ii) Relaying area (iii) Purification centre (iv) Another dispatch centre 9. requirements if there are many batches of live bivalve molluscs.Hygiene requirements for purification and dispatch centres A. 9.4 Chapter IV . B.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. the levels of marine biotoxins should not exceed the following limits. 800 micrograms per kilogram.3 Chapter III . the purification system to allow them to resume /maintain filter-feeding activity. dinophysistoxins and pectenotoxins together.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. (ii) amnesic shellfish poison (ASP).FISH AND SEAFOOD 9. 160 micrograms of okadaic acid equivalents per kilogram.

Identification marking and labelling In addition to general identification marking (contained in 9.2. 9.4. Also. by way of derogation from directive 2000/13/EC.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.3.3. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. 9. including the identification mark. 160 micrograms of azaspiracid equivalents per kilogram. 9. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.4. Requirements of Chapters III and IV are to be complied with. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9. the label.Specific requirements for pectinidae harvested outside classified production areas Requirements for harvesting and handling following harvesting and health standards for live bivalve molluscs must be met. the species of bivalve mollusc and date of packaging (at least the date and month) are required.7 Chapter VII . must be waterproof.1). (ii) Individual consumer-size packages of live bivalve molluscs must be closed and remain closed after leaving the dispatch centre and until presented for sale to the final consumer. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.4. Finally.LEGISLATION (v) azaspiracids. Registration document requirements also apply. 211 .4. dispatch centre or processing establishment.3.4.9 Chapter IX .6 Chapter VI .3.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.8 Chapter VIII . they cannot be re-immersed in or sprayed with water.

general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given. 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 Regulation (EC) 854/2004 gives requirements for official controls on products of animal origin and states requirements for those enforcing the provisions. Therefore the health standards for fishery products as previously stated in Annex III. Chapter V of Regulation 853/2004 apply. Section VIII. This refers to official controls or production and placing on the market of fishery products. echinoderms.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. they contain in their edible parts contaminants or residues in excess of the limits laid down in Community legislation or at levels where the calculated dietary intake would exceed the acceptable daily or weekly intake for humans. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. they derive from: (i) poisonous fish. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. chemical. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. or - - 212 .FISH AND SEAFOOD 9.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. Fishery products in particular must comply with the requirements of Annex III of the regulation.5. (ii) fishery products not complying with the requirement concerning biotoxins. In this regulation. 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. or (iii) bivalve molluscs.

Official controls concerning live bivalve molluscs from classified production areas A. live bivalve molluscs must not exceed 46. coli per 100 g of flesh and intravalvular liquid. Also. coli per 100 g of flesh and intravalvular liquid. 213 . section VII. in 90% of the samples.2.000 E. The reference method for this analysis is the five-tube. 4.600 E. live tunicates and live marine gastropods. The reference method for this analysis is the five-tube. chapter V is reinforced.1 Chapter II . three dilutions MPN test specified in ISO 16649-3. A summary is given as follows: 9. Live bivalve molluscs from Class B areas must not exceed. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. as previously stated. - - - - B. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. In the remaining 10 % of samples.LEGISLATION - the competent authority considers that they may constitute a risk to public or animal health or are for any other reason not suitable for human consumption. coli per 100 g of flesh and intravalvular liquid. that microbiological criteria and chemical contaminants amongst other requirements are met. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. live echinoderms. Classification of production and relaying areas: This section reinforces that specified in Annex III. Monitoring of classified relaying and production areas This is to ensure there is no malpractice.5.2 This states official controls for the production and placing on the market of live bivalve molluscs. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. Sampling plans and frequency are required.5. Live bivalve molluscs from Class C areas must not exceed 46. Annex II 9. 000 E. Regulation (EC) 854/2004.

2. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No.6 Regulation (EC) No. 9.2 Chapter III .5.5. E. boundaries or class of a production area or its closure. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. opening or closing a production area. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. Procedures for re-opening a closed production area are stated D.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. which has applied since 1 January 2006. 9. F. the competent authority is to close the production area. Decisions after monitoring There are specific requirements whereby if sampling results indicate that health standards for molluscs are exceeded or there is a risk to human health. establishes microbiological criteria for a range of foods.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. 2073/2005. the Member States may permit the import of bivalve molluscs and fishery products from the countries listed respectively in Annex I and Annex II to regulation (EC) 2076/2005. 9. 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. 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. 214 .FISH AND SEAFOOD C.

II.I. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. ensuring that foods being placed on the market do not pose a risk to human health. per unit(s) of mass. presence or number of microorganisms. 215 .4 referring to fishery products. and the legislation applies to all businesses involved in food production and handling. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. Two kinds of criteria have been established: food safety criteria. based on the absence. volume.LEGISLATION The aim of this legislation is to complement food hygiene requirements.1 Food safety criteria Chapter 1 of the regulation focuses on food safety criteria that cover foods such as ready to eat foods and fishery products and live bivalve molluscs. The relevant criteria are outlined in Table 9.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. with chapter 2.6. a batch of foodstuffs or a process. and/or on the quantity of their toxins or metabolites. 9. 9. The criteria are outlined in Table 9. area or batch.6.

monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 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. monocytogenes 1. monocytogenes.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life . who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1.3 Ready-to-eat foods unable to support the growth of L. monocytogenes L.2 Ready-to-eat foods able to support the growth of L. other than those intended for infants and for special medical purposes(4) (8) L.TABLE 9. monocytogenes.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.

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 1.25 Live bivalve molluscs and live echinoderms. 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 1.17 Live bivalve molluscs and live echinoderms. coli (15) 1.27 Fishery products which have undergone enzyme maturation treatment in brine. tunicates and gastropods 9 (18) 2 E.

Single samples may be taken at retail level. This criterion shall apply to products before they have left the immediate control of the producing food business operator. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. subject to scientific justification..0 and aw ≤ 0. Duflos G.. including cocoa and chocolate products.25. — fresh. shall not apply. Malle P. cider. The most recent edition of the standard shall be used. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. when he is not able to demonstrate. Bouquelet S.4 or aw ≤ 0. Other categories of products can also belong to this category. J. monocytogenes. Malle P. products heat treated in their final package). Scombresosidae. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . — bread. Bouquelet S.. Dervin C. 2. 43-9. biscuits and similar products. honey and confectionery.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. sugar. uncut and unprocessed vegetables and fruits. wine. 1996. soft drinks. 79. coli is used here as an indicator of faecal contamination. 5 This criterion shall apply if the manufacturer is able to demonstrate. 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. to the satisfaction of the competent authority. A pooled sample comprising a minimum of 10 individual animals. m = M.. Coryfenidae. AOAC Internat. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. c = number of sample units giving values between m and M For point 1. Products with pH ≤ 4. Pomatomidae.. products with pH ≤ 5. products with a shelf-life of less than five days shall be automatically considered to belong to this category. spirits and similar products. References: 1. beer. — — bottled or packed waters. 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.94.1. Valle M. Engraulidae. live bivalve molluscs. Clupeidae. E. Assay of biogenic amines involved in fish decomposition. Particularly fish species of the families: Scombridae. to the satisfaction of the competent authority. that the product will not exceed the limit 100 cfu/g throughout the shelf life. according to which the whole batch is to be deemed unsafe. excluding sprouted seeds. when recontamination is not possible after this treatment (for example.1 .92.

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

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 .1 Shelled and E.TABLE 9.4.

E. 2006 No. the defendant will need to prove that the food was on service or display. The regulations prescribe a chilled food holding temperature of 8 °C or less. 9.1 In the U. if a maximum of c/n values are between m and M.K. S. had not been 221 . if all the values observed are ≤ m. Schedule 4 of the Food Hygiene (England) Regulations 2006. with chill holding tolerance periods.7 Food Hygiene (England) Regulations 2006. and the rest of the values observed are ≤ m. Hot-held foods (food having been cooked or reheated and is for service or on display for sale) must not be kept below 63 °C. — unsatisfactory. and the rest of the values observed are ≤ m. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. — acceptable. 2006 No. 14 (Hygiene requirements specific to the U. tolerance periods for chill-holding of foods and hot-holding variations. c = number of sample units giving values between m and M The most recent edition of the standard shall be used. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. — unsatisfactory.I.II The limits given refer to each sample unit tested.7. if a maximum of c/n values are between m and M. 14 details temperature control requirements for foods in general. The test results demonstrate the microbiological quality of the process tested. which overrides the 8 °C requirement. if one or more of the values observed are > M or more than c/n values are between m and M.LEGISLATION 1 n = number of units comprising the sample. The regulations provide for defences in relation to upward variations of the 8 °C temperature.I. 2 Interpretation of the test results relating to Table 9. — acceptable.K) Temperature control requirements 9. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health. if all the values observed are ≤ m. For example. The defendant may be required to produce well-founded scientific proof to support his claims. S. if one or more of the values observed are > M or more than c/n values are between m and M.

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

30. 30. Regulation (EC) No.2004. 1688/2005. L226. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 25. Regulation (EC) No. 2076/2005.6.2004. The revised text of Regulation (EC) No.10. 83) as amended by Regulation Nos 882/2004. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption (OJ No. namely. Regulation(EC) No. Regulation (EC) No. 853/2004 is now set out in a Corrigendum (OJ No. 62) as amended by Regulation (EC) No. 22) as amended by Regulation (EC) Nos. 9. 1663/2006. 25.2004. 30. L139.2006. laying down common marketing standards for preserved sardines. 223 . 206). The revised text of Regulation (EC) No. 14. L226. 2074/2005. 1).4.6. 2075/2005 and Regulation (EC) No. 2074/2005. 2074/2005. 3) as amended by Regulation (EC) No. 25. L283. these are not be discussed here.12. Regulation (EC) No. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41.2004. L139. 2076/2005.10 Further Reading Regulation (EC) No.4. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 22.6. 2076/2005. Regulation (EC) No. 1662/2006. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. Commission Regulation (EC) No. Regulation (EEC) 2136/89. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito.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. as amended. L338.2004. L226. 2076/2005.LEGISLATION 9. 2073/2005 on microbiological criteria for foodstuffs (OJ No.4. L139. 1.2005. as read with the corrigenda at OJ No. 1441/2007. The revised text of Regulation (EC) No. 852/2004 is now set out in a Corrigendum (OJ No. 1019/2008 and as read with Regulation 2073/2005. 854/2004 is now set out in a Corrigendum (OJ No. 55).2004. 2074/2005 and Regulation (EC) No.

6) 224 .1.12. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.2001. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.12. 22) Commission regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products as amended by Commission Regulation (EC) No 1792/2006 of 23 October 2006 (OJ L 278.2005.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. 21.2005. 23. (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. 22. 22. 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 854/2004.2000.10.

respectively (1).5 minutes.10.1. The minimum pH value for growth is thought to be pH 5. Aeromonas has the potential to grow over a pH range of ca 4 .1.1.5 µm 10. a 10 to 1.1. 10.3 Temperature Aeromonas grows at temperatures of 2 . 225 . The optimum growth temperature is 28 °C (2).5 pH Under otherwise optimal conditions.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.5.1 Aeromonas spp. under optimal incubation temperatures.3. no growth is observed (1). PATHOGEN PROFILES 10. At pH 4.5. and D-values at 45 and 51 °C have been reported as 29. 10.3 minutes. 0.10 (3).1. Morphology Gram-negative rods. 10.0 .1 10. and 2.1.3 .0 x 1. Some death will occur at this pH if the temperature is 4 °C.2 Oxygen requirements Facultative anaerobe. At 5 °C.45 °C (1).000-fold increase in number is observed after one week in food systems (1).

2. and groups I and II are those responsible for the botulism cases.4 Heat resistance The vegetative cells of C.4 x 1.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 .7 .FISH AND SEAFOOD 10. or at 90 ºC for a few seconds.30 minutes. Types A.even as low as 3 ºC (7).2.5 . Group I will not grow at temperatures of 10 ºC or less.5% NaCl (4). All C. to a lesser extent. Thus an aerobically packed product may not support the growth of the organism on the surface. botulinum is an obligate anaerobe. B.2.22. botulinum types produce heat labile toxins. 10. being capable of slow growth and toxin production at low temperatures .2% sodium chloride (NaCl) (1). botulinum grow reasonably well in the temperature range of 20 45 ºC. 10.2 Oxygen requirements Although C. 10. botulinum is prevented. botulinum are not particularly heat resistant. 6). 0.1 Morphology Gram-positive. 10. 10.2. There are four main groupings of the organism. A to G. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different.0 µm. at 85 ºC for 5 minutes. botulinum are known.3 Temperature All strains of C. spore-forming rods. but the interior of the food may do so. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C. but Group II strains are psychrotrophic. 226 .2 Clostridium botulinum Seven different types of C.2.1. F are the types that are responsible for most cases of human botulism (5. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. E and. forming at least seven different toxins. types B and F can be either. All type A strains are proteolytic and type E strains are usually non-proteolytic. and is sensitive to > 4. but the spores of this organism are more so. which may be inactivated by heating at 80 ºC for 20 .

It will not.g.2. 10.3 10. The organism will grow well in the centre of meat or poultry dishes. botulinum spores The most heat resistant spores of Group I C. where oxygen levels are reduced.0 µm.3. grow on the surface of foods unless they are vacuum or gas-packed.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. parcooked meals with pH value > 5.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains.94 for Group I and 0. perfringens .9 x 2. 227 . 9). vacuum packed.97 for Group II (5). These values have been established under carefully controlled laboratory conditions. the minimum aw for growth may be lower. 10. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I.21 minutes) (9).0 and aw > 0. In commercial situations.3.2. 11)).like other clostridia .10. 10. therefore.6 and pH 5.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4.3 .0.2 Oxygen requirements C.is an obligate anaerobe.1.2. safety margins must be introduced to allow for process variability. 10. respectively (8. particularly by cooking.7 Characteristics of C. However. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. These concentrations correspond to limiting aw of 0. 0. pH and temperature.0 . botulinum depends on solute.97) (10. they may survive mild heat treatments (70 . Note also that if humectants other than salt are used.1 Clostridium perfringens Morphology Gram-positive spore-forming rods.PATHOGEN PROFILES 10.

3. 10. The lowest aw recorded to support the growth of C.3. While some growth can occur at 50 ºC. type of solute. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. perfringens in vacuum packed beef (18).3.3 Temperature The most significant characteristic of C. Spores. 0 .5 minutes (17). perfringens strains. the enterotoxin is not heat resistant . perfringens is not a tolerant organism with respect to pH. and their heat resistance is also affected by the heating substrate. 18). Under otherwise ideal conditions. etc. 10. perfringens spores The spores of C. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. will survive greater extremes of pH (and aw) (12. The D-value for C. 16). At cold temperatures. however. perfringens has the potential ability to grow within the temperature range 15 .3.3. As in the case of other factors limiting the growth/survival of this organism. 13).45 ºC. perfringens can vary quite considerably in their heat resistance. (It is unusual for spores to be formed in foods after the growth of this organism (14)). depending on strain and other conditions. In addition.5 pH C. perfringens.7 Characteristics of C.6 Aw/Sodium chloride C. Salt concentrations of 6 .8% inhibit growth of most C. perfringens is not tolerant of low water activities.10 ºC.FISH AND SEAFOOD 10.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. Some studies indicate that the presence of 3% NaCl delays growth of C. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8.3. the limits for aw are affected by temperature.50 ºC. Its optimum temperature for growth is 43 . although C. perfringens in roast beef at 60 ºC is 14.it is destroyed by heating at 60 ºC for 10 minutes (14.93 and 0. Recorded heat 228 . 13). perfringens appears to be 0.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. pH. It grows best at pH values between 6 and 7 (the same pH as most meats). 10. 15. 10. lower concentrations may be effective in combination with other factors. vegetative cells die rapidly (12).

10.2. using hydrochloric acid as an acidulant).6 . the lowest limit for growth is likely to be considerably higher . Its optimum growth temperature.2. so can be destroyed by pasteurisation. 1.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic.98 and 0. The upper temperature limit for the growth of L.5 . monocytogenes is not a particularly heat resistant organism. growth at low temperatures can be very slow. Morphology Gram-positive short rods.4 Heat resistance L. is between 30 and 37 ºC. monocytogenes is reported to be 45 ºC (20). respectively (21).4 10.4. the organism is able to grow at pH values well below pH 5 (pH 4. In foods.2 Oxygen requirements Aerobe or microaerophilic. However.4. it is not a sporeformer.4. D-values for L.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. 10. 10.4 . however.23. 10. 229 . however.0. and temperature. being potentially capable of growing .albeit slowly .4.19 minutes.at refrigeration temperatures down to. 10. -0.8 minutes for heat sensitive spores (12).4 ºC is probably a more likely minimum for growth in foods (19). or even below 0 ºC. monocytogenes in crawfish tail meat at 55.5 x 0.4.2 has been suggested as the lowest working limit (22).especially at refrigeration temperatures. 0. pH < 5.64. Under ideal conditions.0 µm.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. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens. requiring days or weeks to reach maximum numbers.0 minutes for heat resistant spores to 1.1 Listeria spp.3 . and where acetic acid is used as an acidulant. 60 and 65 ºC were reported as 10.3 is the lowest value where growth has been recorded.

1.3 µm. The bacterium may survive for long periods at aw values as low as 0. but has been shown to be able to grow at an aw level of 0.8 . 0.0. It is likely to survive.1 Plesiomonas Morphology Gram-negative.0 x 1 .2 Oxygen requirements Facultative anaerobe.5.0 .4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.12% NaCl or more).5 . or even grow. µm.45 ºC. It grows best at aw of ≥ 0.5.6 Aw/Sodium chloride L.5. 10.5. Morphology Gram-negative short rods.8. 25).5 (24.6. monocytogenes is quite tolerant of high NaCl/low water activities.FISH AND SEAFOOD 10.3 Temperature Plesiomonas can grow in the temperature range of 8 . 25).5 10.6 10.5 .97. 10. 10. 10. 0. 230 .5.1 Salmonella spp.4. at salt levels found in foods (10 .83 (20). 10.7 x 1. the optimum being 30 ºC (23).5 pH Plesiomonas has a pH range for growth of pH 4. peritrichous flagella. 10. 10.

6. 10. and with an appropriate acid. Growth is slow at temperatures below about 10 ºC.48 ºC. with a z-value of 4 .6. pasteurisation and equivalent treatments will destroy the organism under normal circumstances. strains vary in their ability to withstand heating.93 (as reported in dried meat and dehydrated soup). or possibly 0. a heat resistant organism. some strains are able to grow at temperatures as low as 4 °C (26).6 Aw/Sodium chloride Where all other conditions are favourable. substrate. Some death will occur at pH values of less than about 4. and appropriate heat treatments must be determined experimentally for low-aw foods.945.37 ºC. Furthermore.6. the optimum being 35 . 10. Salmonellae are quite resistant to freezing.PATHOGEN PROFILES 10.5. The growth of Salmonella is generally inhibited by the presence of 3 .9.8 . temperature and pH. 10. D60 ºC values normally range from about 1 to 10 minutes. Salmonella has the potential to grow at aw levels as low as 0. It is not.5. depending on serotype. The optimal pH for Salmonella growth is between 6.0. under otherwise ideal conditions.7.4% NaCl. although salt tolerance increases with increasing temperature (28). 10. Salmonella senftenberg 775W is about 10 . 231 .6.4 Heat resistance Salmonella is not a spore-forming organism.5 ºC.20 times more heat resistant than the average strain of Salmonella at high aw (27).5 . Salmonellae are quite resistant to drying. However.5 pH Salmonella has a pH range for growth of pH 3.6.2 Oxygen requirements Facultative anaerobe. therefore.3 Temperature Salmonellae can grow in the temperature range of 7 . However. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. depending on the type of acid and temperature.

). aureus is more limited under anaerobic than aerobic conditions.7.7 . but may cause some cell damage (30). the D60 ºC value is 2 . Freezing and thawing have little effect on Staph.9 µm diameter. under otherwise ideal conditions (29.7.0. 10. 10. 31). aureus viability. 10. aw and temperature (sensitivity to acid increases with temperature). In meat macerate.7. fat content. however.FISH AND SEAFOOD 10.5 pH The pH at which a staphylococcal strain will grow is dependent upon the type of acid (acetic acid is more effective at destroying Staph. It can survive well at low temperatures. depending on aw. with an optimum of 35 .4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. etc. stressed cells can also be less tolerant of heating. aureus can grow within the temperature range 7 – 48. The following relate to limits for growth only. The limits for toxin production are also narrower than for growth.5). Under most circumstances. aureus than citric acid). aw.0 .3 Temperature Under otherwise ideal conditions. NB: The growth of Staph. the organism is heat sensitive and will be destroyed by pasteurisation.2 to 9.1 Staphylococcus aureus Morphology Gram-positive cocci. Staph.3 (optimum 7. 232 .7 10.7. 0.2 Oxygen requirements Facultative anaerobe. 10. As is the case with other bacteria.37 ºC (29).7.20 minutes.5 ºC. Most strains of staphylococci can grow within the pH range 4.7.

with a temperature range for growth of 10 .46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5.86 is the generally recognised minimum in foods (32). 233 .8.4 .0.2 . * dependent on temperature.6 µm 10. 34).7.PATHOGEN PROFILES 10. Staph.1 Vibrio cholerae Morphology Gram-negative curved rods. an aw of 0. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant. An effect of increasing salt concentration is to raise the minimum pH of growth.10% NaCl.8 10. In general. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities. 31) between 0.99 aerobically under otherwise optimal conditions. 10.0 (optimum 7.8.87 and > 0. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. Staphylococci are more resistant to salt present in foods than other organisms. but certain strains can grow in 20% NaCl.0 .8. In general.43 ºC (35).>0. pH.7. and solute. It can grow over the aw range 0.99 little or no toxin production in anaerobically packed foods.5 . aureus can grow in 7 .5) (29.7 Limits permitting toxin production 10 .2 Oxygen requirements Facultative anaerobe.8 x 1.9. 10.2. However. strain.83 .6 Aw/Sodium chloride Staph.7. atmosphere. 0.3 Temperature The optimum temperature for growth has been reported as 30 ºC. 10.

0.9.5 ºC). The D-value for V. 10.97 . 38). but not acid conditions.9. with an optimum of 37 ºC (39). cholerae can grow over the aw range 0. V.8.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods.5 pH This organism is tolerant of high pH values.FISH AND SEAFOOD Also. and it is sensitive to refrigeration temperatures (0 .8. under otherwise favourable conditions.984 (35). cholerae may persist on frozen fish for >180 days (36).3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.9. there is evidence that V. 10. It can grow in the presence of 0.8. 10. but only moderately sensitive to freezing (40).9 10. it has a pH range for growth of pH 5 .6 (35.6 Aw/Sodium chloride V.2 Oxygen requirements Facultative anaerobe.65 minutes (35). are 5 and 43 ºC. 10. with an optimum of 0. 10. respectively. declining in number during storage.4 Heat resistance V.0% NaCl (35).998. with optimum at 7. cholerae in crab meat homogenate at 60 ºC is 2.1 . 37. parahaemolyticus is easily destroyed by drying. cholerae is sensitive to heat at temperatures of >45 ºC. 234 .

The D-values at 55 ºC for clam and crab homogenate are 0. 10.29 minutes.5 pH The pH range for growth is 4.5% NaCl.10 Vibrio vulnificus 10.2 Oxygen requirements Facultative anaerobe. V.6 (41).4% (39. and are readily destroyed by cooking. with an optimum pH of 7. 10. and 2.02 . respectively (39).10. 10. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).43 ºC (42).922. 41).98 (35. parahaemolyticus are not heat resistant. minutes.6 Aw/Sodium chloride V. with a temperature range for growth of 8 . 10.1 Morphology Gram-negative curved rods.9.10. 41). depending on solute.6 . 10.4 Heat resistance V.3 Temperature The optimum temperature for growth has been reported at 37 ºC. 235 . respectively. but the optimum concentration is 2 . parahaemolyticus is unable to grow in the absence of salt.4 Heat resistance V.8 . parahaemolyticus will grow at aw levels as low as 0.10. The organism can survive for long periods at freezer temperatures (42).PATHOGEN PROFILES 10. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.9. The optimum concentration of NaCl equates to aw of approximately vulnificus is heat sensitive.

Miliotis M.8 (42). in Clostridium botulinum: Ecology and Control in Foods. in Foodborne Disease Handbook. 10. Aeromonas hydrophila group. Hui Y. in Foodborne Pathogens. Palumbo S. in Food Microbiology: Fundamentals and Frontiers. ASM Press. Dodds K. London.5. Great Britain. Marcel Dekker. New York.6 Aw/Sodium chloride V. Clostridium botulinum. 2005.L. Aeromonas species. New York.. 1993. 10. 6.. Abeyta C.. Fratamico P. 1994. Cliver D. Volume 1: Diseases Caused by Bacteria.5 . Gorman J. Microbiology and Molecular Biology. Doyle M.W.L. Bier J.C. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’. Doyle M. Aeromonas hydrophila.W. Doyle M. in Foodborne Disease Handbook.L. Novak J. in Foodborne Bacterial Pathogens.D. Hauschild A. 10. Hauschild A. Eds.W. Stelma G.D. 9. Montville T.... Johnson E. 11. Cliver D. Marcel Dekker. 3.H.. Marcel Dekker Inc. Principles of control.M. New York. Potential hazards associated with REPFEDS. Ed. Notermans S. Eds. in Clostridium botulinum: Ecology and Control in Foods. New York. Volume 5: Microbiological Specifications of Food Pathogens. Kim J. Dodds K. References International Commission on Microbiological Specifications for Foods. Beuchat L.FISH AND SEAFOOD 10.. Clostridium botulinum. Foegeding P. in Microorganisms in Foods. in International Handbook of Foodborne Pathogens. 5-19.10. Marcel Dekker.11 1.. Marcel Dekker. Bhunia A. with an optimum of 2..H. 8.H. International Commission on Microbiological Specifications for Foods. Kovacek K.A. Ed. Hauschild A. Marcel Dekker.. 4. 6 (1). Volume 1. Food Control. 2003. Eds.N. 5. Gaze J.10. Smith J. Ed. Lund B. Betts G. 2001.. 1989. Murrell K.R. Austin J.0% NaCl. Dodds K.. Faris A... Stelma G.. 236 . 1989..O..H. Clostridium botulinum.W. Clostridium botulinum and Clostridium perfringens. Washington D.D. Marcel Dekker. Juneja V. 279-303.. 121-76. Eds.M. Peck M. in Foodborne Bacterial Pathogens.K..L. Blackie. 1994. 1-19. vulnificus grows over the range 0.P.P.R.D. Murrell K. with an optimum pH of 7.. Caister Academic Press. Hui Y. 329-49.J.N. 97-131. New York.H. Diseases Caused by Bacteria. 383-408. Eds. 2. 1995.P. 111-89. New York. 7. Gorham J. Eds. 1993.R.5% (42). 357-67. Eds.E.5 pH The pH range for growth is 5 . Aeromonas. 1996. 57-63. New York..M.O.H. 1-27.10..W.

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

. Smith J. Volume 5: Microbiological Specifications of Food Pathogens. Warner R. 523. in Microorganisms in Foods. Twedt R. Academic Press. R15764. Academic Press.C. in Foodborne Pathogens: Microbiology and Molecular Biology. International Commission on Microbiological Specifications for Foods.M. 32. Food and Environmental Sanitation. in Foodborne Pathogens. 2005.FISH AND SEAFOOD 27. International Commission on Microbiological Specifications for Foods. International Commission on Microbiological Specifications for Foods.J..A.O. Fratamico P. 273-84. Stewart G. Lee-Wong A. 238 . International Commission on Microbiological Specifications for Foods. Eds.M. London. Staphylococcal gastroenteritis. 642. 2005. Cliver D. Eds. Staphylococcal Food Poisoning. Marcel Dekker. Microbiological Specifications of Food Pathogens. in Foodborne Bacterial Pathogens. Salmonella.P. 327-445. Woodhead Publishing Ltd. Staphylococcal intoxications. 1996. Riemann H. Blackie.L.C. New York. Ed. Vibrio cholerae. 30. Bergdoll M. Crowcroft N. New York. Cleland D. Staphylococcus aureus. Loessner M. Loessner M. Gustafson J. 543-68. 1930. International Commission on Microbiological Specifications for Foods. 426-35. 1996. Bergdoll M. Volume 5. 1989.62. Vibrio parahaemolyticus. Wilkinson B. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures.O.. 545-66. Ed. 1990. 1993. International Commission on Microbiological Specifications for Foods. 37... London.S. Blackie. 40. 1994. in Foodborne Disease. Eds. Ed. 29.H. 35. Ed. 331-57. Japanese Journal of Experimental Medicine..M. Dairy. Golden D.. 31. Springer Science.M. D’Aoust J-Y.R. Cliver D. London. Ed. Reed G. 36. CDR Review. in Foodborne Infections and Intoxications. Marcel Dekker.. in Microorganisms in Foods.S.P. Oliver J. Blackie. in Microorganisms in Foods. in Understanding Pathogen Behaviour Virulence.J.. Vibrio parahaemolyticus. 4 (13). 34. 1996. 45 (3). 1983. 28. 1989. Salmonellae. London. Doyle M. Tohyama Y. 327-52. London. 13 (11). Applied and Environmental Microbiology.S. Doyle M. Jay J. 8. 33. Volume 5. New York. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment.A. Caister Academic Press. Griffiths M. 985-98.D. Stress Response and Resistance. 2005. Cambridge. Wymondham. 217-64..A. Golden D. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. 85-106. Bhunia A. 414-25. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). Ed. Cholera: current epidemiology. 39. 2005. 38. Microbiological Specifications of Food Pathogens.P. in Modern Food Microbiology. Jay J. Ed. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems.K.

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


org.gov.uk Web site: www.uk/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: helpline@defra.org.seafish.uk Web site: www.gov.uk Web site: www.scotland.uk Prepared Fish Products Association (PFPA) Federation House 6 Catherine Street London WC2B 5JJ United Kingdom Tel: + 44 (0) 208 78362460 The Scottish Government Fisheries Group Environment and Rural Affairs Department Pentland House 47 Robb's Loan Edinburgh EH14 1TY United Kingdom Tel: + 44 (0) 8457 741741 or + 44 (0)131 5568400 Fax: + 44 (0)1397 795001 Email: ceu@scotland.defra.ifm.org/ .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.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.co.gsi.uk or v.gov.org.holt@ifm.gsi.uk/Topics/Fisheries Seafish Industry Authority 18 Logie Mill Logie Green Road Edinburgh EH7 4HS United Kingdom Tel: + 44 (0)131 5583331 Fax: + 44 (0)131 5581442 Web site: www.

fiskeriforskning.uk/index/fisheriesfarming-and-food.matis.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.suite. Andersens Boulevard 37.uk Web site: www. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo.dardni. 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. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.eapo.fiskehandlerne.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.bim.fao. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.is/ 242 .gov.dk Web site: www.dk Danmarks Fiskehandlere H.com Web site: www.dk Web site: www.org Web site: www.dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .FISH AND SEAFOOD Department of Agriculture and Rural Development.C.ifpea@eircom.org/fishery/ European Association of Fish Producers’ Organisations H.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.ie/templates/homepage.is Web site: www.no Web site: www.difta.gov.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: post@fiskeriforskning.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.

uk Web site: www.dh.gov.ifr.org Other Sources of Information British Standards Institution 389 Chiswick High Road London W4 4AL United Kingdom Tel: + 44 (0) 208 9969001 Fax: + 44 (0) 208 9967001 Email: cservices@bsigroup.org.gov.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: generalenquiries@fdf.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.org.com/ 243 .ac.bsi-global.com Web site: www.seafood.hpa.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: reception-cmar-hobart@csiro.uk Web site: www.nz Web site: www.csiro.nz/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.uk/en/index.org Web site: www.au Web site: www.gsi.co.CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: contact@nfi.co.ifst.uk Institute of Food Research (IFR) Norwich Research Park Colney lane Norwich NR4 7UA United Kingdom Tel: + 44 (0) 160 3255000 Fax: + 44 (0) 160 3507723 Web site: www.fdf.au New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: info@seafood.org.cmar.

euro. Web site: www.int Web site: http://www.scot.uk Web site: www.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: foodsafety@euro.org.int/ SIK .uk/scieh.scot.int Web site: www.who.co.nhs.se Web site: www.uk Web site: www.net/web/indexen.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.sfam.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.org Web site: www.who.sik.who.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.php Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: admin@sofht.who.nhs.hps.int/foodsafety World Health Organization (WHO) Headquarters CH .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.co. jsp Food Safety 10 .uk/index.who.sofht.codexalimentarius.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.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.euro.

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


69-70 Aeromonas hydrophila. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. planktonic source of spirolides in shellfish 163 Allergens. to test microbiological criteria 19 Aeromonadaceae. preserved by dry/wet salting 99 Anisakis spp. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. causing spoilage of dried fish 110. as autochthonous bacterial flora of fish 3 Aeromonas spp. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. 111 Astroviruses. 102 Bacillus spp.INDEX Achromobacter spp. growth and survival in crustacean shellfish 86 Aeromonas sobria. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. 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. 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. in crustacean shellfish 87 Alteromonas spp. in crustacean shellfish 86-7 ATP.

indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang. fish sauce 123 Basipetospora halophila. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. as autochthonous bacterial flora of fish 3 Clostridium botulinum. 226 Breading process 36-8 Brevetoxins. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. as cause of spoilage of MAP fish 15 Calciviruses. of recipe fish meals 43 Cholera. 68-9 Capture and handling. preserved by marinating 100 Chemical detection methods. of fresh fish spoilage 11 Chemical pollutants. smoked and dried fish 113-6 hazard in mild-processed marinated products 101 hazard in sun-dried products 102 non-proteolytic – risk of growth in fish recipe meals 44 248 . of fish – effects on microflora 5-6 Carbohydrate. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. 136. 137. associated with consumption of fish/fish products 16. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. concern with respect to use of MAP 7 growth and survival in chilled and frozen prepared fish products 45 growth and survival in chilled and frozen raw fish 15-6 growth and survival in crustacean shellfish 84 growth and survival in cured. importance in lactic acid fermentation in fish products 130. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. 113. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. 131 Carbon dioxide. 98-9 Brochothrix thermosphacta. in crustacean shellfish 86-7 Campylobacter coli. for PSP toxins 144 Chemical indicators.

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

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

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

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

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

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

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

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

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