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


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

vi .

2 Initial Microflora 1.6 Pathogens: Growth and Survival 2.6 Published Microbiological Criteria 4.5 Factors Affecting Fresh Fish Spoilage 1. 4.5 Pathogens: Growth and Survival 3.7 Published Microbiological Criteria 2.6 Published Microbiological Criteria 3.8 References MOLLUSCAN SHELLFISH 3.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.2 Initial Microflora 4.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.5 Spoilage 2. CHILLED AND FROZEN RAW FISH 1.7 Published Microbiological Criteria 1.8 References 1.7 References 3. vii .3 Processing and its Effects on the Microflora 4.8 Further Reading CRUSTACEAN SHELLFISH 4.1 Definitions 1.6 Pathogens: Growth and Survival 1.1 Definitions 3. 3.5 Pathogens: Growth and Survival 4.4 Spoilage 3.1 Definitions 4.4 Spoilage 1.3 Processing and its Effects on the Microflora 1.4 Spoilage 4.4 Processing and its Effects on the Microflora 2.7 References 4.2 Initial Microflora 3.2 Definitions 2.3 Initial Microflora 2.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.1 Introduction 2.3 Processing and its Effects on the Microflora 3.

2 Legislative Structure 190 9.1 Definitions 6.5 Pathogens: Growth and Survival 5.6 Published Microbiological Criteria 5.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.5.8 Guidance 222 9.1 Introduction 7. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption 212 9. S.3 Processing and its Effects on the Microflora 5. 14 (Hygiene requirements specific to the UK) 221 9.9 Azaspiracids 7. EU FOOD HYGIENE LEGISLATION 189 9.4 Spoilage 5.10 Cyclic Imines 7.1 Introduction 8. 2006 No. 7.5 Pathogens: Growth and Survival 6.3 Regulation (EC) No.4 Implementation and Review of the HACCP Plan 8.11 Conclusion 7. CURED.7 References 5.2 Definitions 8. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.6 References 6.3 Stages of a HACCP Study 8.1 Definitions 5.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.5 Regulation (EC) No. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.2 Initial Microflora 5.6 Regulation (EC) No.3 Tetrodotoxin 7.2 Initial Microflora 6.6 Lipophilic Shellfish Toxins (LST) 7.9 Other Relevant Legislation 223 9.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.10 Further Reading 223 viii .3 Processing and its Effects on the Microflora 6.5 References 93 93 95 96 108 113 116 117 120 123 123 126 127 135 136 137 140 141 141 142 144 146 148 151 153 155 158 160 165 166 175 175 176 177 184 186 6.1 Introduction 189 9. SMOKED AND DRIED FISH 5.I.7 Further Reading FISH AND SHELLFISH TOXINS 7. 8.4 Spoilage 6.8 Further Reading FERMENTED FISH 6. 852/2004 on the General Hygiene of Foodstuffs 191 9. 9.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.4 Regulation (EC) No.8 Ciguatoxins 7.7 Food Hygiene (England) Regulations 2006.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.

4 Listeria spp. 10.10 Vibrio vulnificus 10.1 Aeromonas spp.9 Vibrio parahaemolyticus 10.10 PATHOGEN PROFILES 10.3 Clostridium perfringens 10.6 Salmonella spp.5 Plesiomonas 10. 10.2 Clostridium botulinum 10.8 Vibrio cholerae 10.7 Staphylococcus aureus 10. 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .


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

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

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

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

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

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

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

Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. 105 . 4 . Escherichia coli Yersinia enterocolitica 101 . Plesiomonas shigelloides 3 . In addition. estuarine environments.>106 Salmonella spp. birds.26 °C 10 . 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. and risk associated with the consumption of seafood is low (2.106 cells/g. sewage.102 101 .109 cells/g) * Some sources considered L. vulnificus) 5 .1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.14 .II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.37 °C Unknown Unknown TABLE 1.1 . However.103 High (107 .37 °C High for most species.19 µg/kg bodyweight from < 102 . and mud Ubiquitous. 4). stream water.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00.FISH AND SEAFOOD TABLE 1.0. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp.106 cells/g (exception: V. Toxin levels 0.35 °C 8 .

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

ferric ion (Fe3+)). and minimising handling will all reduce microbial cross-contamination. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. However.). but isolate the food in an environment different from the bulk atmosphere. Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. if there is some oxygen demand from the product. for example. nitrogen. 1. allowing more growth of spoilage organisms. however. These changes can be deleterious. but if applied properly should extend the life of the product. The atmospheric conditions surrounding a product may be passively or actively altered. fish have a higher post mortem pH.2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. in time. Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. they are usually process contaminants. partial fermentation of sugars occur leading to lower pH. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. 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 (6.3. often in the absence of adequate refrigeration. Packaging changes the intrinsic and extrinsic parameters affecting a product. These containers are also used for displaying the fish during auction at the quayside. use of effective detergents and disinfectants. What is more. Anaerobic atmospheres have less effect on fresh fish shelf life. Physical barriers not only protect from physical damage. not part of the normal flora of the meat animals (8). By vacuum packing foods. cleaning of the work environment. In MAP. and to limit the loss of quality (5).FISH AND SEAFOOD can be substantial. and of the quality of the water used for ice and to wash fish. oxygen etc. and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO). by actively altering the composition of the 6 . Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. from water activity (aw) through to physical damage. As with all foods. will result in fully anoxic conditions. 7). a reduction in the oxygen tension is achieved which. Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. In the absence of oxygen.

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

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

Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . 18) and are probably the most widely used. or for that matter evaluation of the taste of the fish. so lower carbon dioxide concentrations may have better preservative effects. A. 17. the gills from pink and shiny. and this pattern is the basis of freshness grading schemes. Over time. and metabolic activities of microorganisms present in the fish. Taste panel evaluation of cooked fish may be used to score or grade colour. Degradation of whole fresh fish stored in ice generally follows a set pattern. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. the skin turns from iridescent to dull and bleached with bacterial slime. these are summarised in Table 1.4 1. reactions caused by activities of the fish’s own enzymes.CHILLED AND FROZEN RAW FISH off-odours. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. with no smell. 1. 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.III. Loss of freshness and spoilage cannot be separated as processes. Various methods for whole fish freshness evaluation by a trained panel have been proposed. The eyes turn from convex and clear to concave and opaque. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids). B. When working with fillets. and the flesh turns from bright and elastic to dull and soft (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). TABLE 1. as traditional spoilage patterns of the external organs are typically very distinct. texture. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation.4. smell and 9 . but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). to brown and slimy with an intense off-odour.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16.

all marine fish use/have TMAO (21). the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. ADP. sea-weedy taste. texture becomes soft and watery or hard and dry. iv) Enhancement of the spoilage characteristic of phase iii. AMP. The process of ATP breakdown is used as an indicator of fish freshness (24). bacterial reduction of TMAO to TMA. During the initial storage. chemical. However. this will generally correlate with the length and severity of rigor. 19). To a varying extent. development of sickly sweet. As the fish degrades. rigor resolves. 23). a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. which increases from 0% towards 100% with time. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. These processes are strongly influenced by the physical conditions of storage.g. is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. the conductance generally increases. an intense odour compound. sulphurous and/or rancid smells. HxR and Hx) gives a quantity (K-value. ii) Neutral taste.8 to 6.5 (18. By measuring the concentrations of the six components. TMAO permits a high Eh to remain in the muscle tissue. ammoniacal. After death. 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. Because fish tend to have relatively little residual glycogen compared with mammals. the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). the muscles relax again and the fish returns to a flaccid state. post-rigor values are typically in the range of pH 5. spoiled and putrid. the Torrymeter. fruity and/or bitter off-flavours. RT Freshmeter or Fishtester). or simply a waste product present due to bioaccumulation (22). or its remaining shelf life (1). an anti-freeze. Generally. the pH drop of the muscle is correspondingly less. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical. as little endogenous reduction of TMAO occurs (18). the Eh of fish muscle remains relatively high (20). After death. little flavour. Changes in the resistance of the fish skin after death are used as the basis for tests that employ an electrode measurement of skin resistance (e. The pH of the muscle will drop after death depending upon the amount of residual glucose or glycogen that is reduced anaerobically to lactate with the coproduction of ATP. After some time. %). reduction in ATP and increase 10 . cabbage-like. rigor mortis is the first noticeable change in the fish. possibly slightly metallic. thus. IMP. an osmoregulator. iii) Traces of sour. biochemical and microbiological actions. From being flaccid.FISH AND SEAFOOD taste of cooked fish.

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

For example. 1.3 Recognised specific spoilage organisms (SSOs) The degree of spoilage leading to sensory rejection of fish is partly dependent on the perception of the consumer. careful temperature control during storage is not only important in terms of quality loss.5. 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.5 1. the results of altering one parameter may have far-reaching consequences on others. 1.1 Factors Affecting Fresh Fish Spoilage Temperature By far the most effective method of reducing the rate of whole fresh fish spoilage is temperature control (1. For fresh fish. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. a minority are often associated with the majority of the spoilage. The concept of specific spoilage organisms (SSOs) is not new.FISH AND SEAFOOD 1. biochemical and microbiological reactions taking place within and on the fish. 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 slower the spoilage processes proceed (within limits). Also.5. the lower the temperature of storage.and temperate-water fish spoilage rates may be many. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks. by altering the gas atmosphere (extrinsic).5. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. The factors are interactive and cannot be completely isolated. 34). This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. but also crucial for the assurance of consumer safety. For fresh fish iced immediately after capture and continually stored in ice. Fish spoil as a result of the chemical. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. The sum of these changes may result in a different spoilage profile. The reasons for the differences between tropical.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). the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH.

They are generally characterised as Gram-negative rods. catalase-positive. aldehydes. oxidase-positive. obligate respiratory bacteria. motile with polar flagella. processed (e. it is often isolated as about 1 . It is able to grow as fast as. therefore. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1.10% of the total flora of fresh fish from temperate marine waters. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. 37). putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s.5. which produced the characteristic spoilage compounds of the fish (30. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36.CHILLED AND FROZEN RAW FISH started in the 1940s. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. As previously noted. although only since the late 1960s has it been realised that specific metabolites of Sh. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms. and may play some role in the spoilage of freshwater fish. The importance of Sh. putrefaciens spoilage of fish is due to its biochemical action on muscle. Sh.g. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp. It is thus an important spoilage organism of gadoid fish such as cod. mediated spoilage is characterised by ‘fruity’. Pseudomonas spp. on fish are diverse and in many cases species-specific. 35). frozen) fish products. During the 1970s. 36). i.1 Pseudomonas spp.3.2 Shewanella putrefaciens Sh.5. the spoilage of a product may be strongly influenced by the conditions under which the product is held. produce hydrogen sulphide (H2S) from cysteine. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine. It is also present in fresh water. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. 1. These organisms are described as potential spoilers.e. or faster than the rest of the flora of ice-stored fresh marine fish (9).3. ‘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. 13 . plus other characteristic compounds of the species responsible for spoilage. more especially. its ability to reduce TMAO to TMA. Members of the genus are able to produce pigments. but only if or when they reach numbers capable of producing sufficient spoilage compounds to effect rejection do they become the SSOs of the product (9. putrefaciens growth may be used as indicators of spoilage (30). (9) and they are considered SSO of iced freshwater fish (35). and proteolytic and lipolytic enzymes that may affect the quality of fresh and. 35.

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

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

there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. is small.g. parahaemolyticus outbreaks. which can cause human gastroenteritis. and proper cooking (boiling for 1 minute. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control.2005). there is no specific guideline that describes a minimum level of V.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. Southeast Asia.3 °C) will eliminate all risk. Currently. 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. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. vulnificus is associated with warm marine and estuarine waters. 1978 . Other lightly or semi-preserved (e. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. the risk posed to the consumer of fresh fish. the majority are associated with products not cooked immediately before consumption. parahaemolyticus is a marine organism. 1.6. parahaemolyticus cases have increased across the world. parahaemolyticus O3:K6 is responsible for many of the recent V. V. Only about 1% of marine isolates produce a thermostable haemolysin. This strain has been responsible for 50 to 80% of all V. smoked. salted or pickled) fish have also been associated with botulism. sushi) (5). I-sushi. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . 11). including epidemics in India. accounted for the majority of cases (1.g. a Japanese boiled and semi-dried sardine dish. Despite the ubiquitous nature of type E C. due to the cultural preference for raw fish dishes. 5). botulinum in the marine environment (isolated from around 90% of marine and environmental samples from northern European waters and its consequent presence in seafood (a surveyed incidence of as high as 65%) (2. Russia. V. storage and preparation (11). which was probably contaminated from an uncooked food source or an excreting foodhandler. the majority of outbreaks are caused by consumption of raw fish products (e. again. USA. were associated with finfish between 1978 and 1998 (46). whether stored aerobically or in modified-atmospheres. and when in contact with contaminated water (no CDC reported cases from fish. and North America.2 Vibrio parahaemolyticus and other vibrios V. which is believed to be required for virulence. The bacterium causes primary septicaemia. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). A unique clone of V. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. Since 1996. Proper chilling. In Japan. Generally. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). Proper temperature control (<3. The first report of the organism being a foodborne agent was with shirasu. Japan. France.

although lack of data concerning infectious doses and the possibility that foodborne virulence is linked to the immune state of the host mean that full risk assessment is impossible. in assessing the risk of fishborne infection by P. 17).CHILLED AND FROZEN RAW FISH disorders).. Despite this. monocytogenes incidence has been between 0. there is no risk from the consumption of properly handled and cooked fresh fish. their potential to produce disease-causing exotoxin in food has not been fully examined (48). no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp.20 °C. in its habitat. proper chill storage will eliminate any risk associated with this organism. possibly due to under-reporting because of their likeness to E. 1. Similar problems as those encountered for Aeromonas spp.3 Aeromonas Aeromonas spp. monocytogenes has been well documented as a foodborne human pathogen. are present on fish and can grow to significant numbers during storage (1. P. L.000 17 . It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. but proper cooking should substantially reduce risk of infection. 5. V. Again.6. with minimum growth temperatures around 0 °C and optima of 15 . In the US. and especially estuarine environment. and proper cooking will substantially reduce risk. is historically associated with faecally contaminated water. There is no risk associated with the consumption of properly chilled and cooked fresh fish. listeriosis cases have been reportable to the CDC. Chill storage may not eliminate growth. The major virulence factor appears to be the production of toxins with enterotoxic.55 cases per 100. and especially Aeromonas hydrophila. others are psychrotrophic mesophiles (49). Although the psychrophiles are unable to grow at body temperature. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis.26 to 0. 1. sodium channel blocking and haemolytic activity (48). are associated with human diarrhoeal illnesses.4 Listeria monocytogenes L. shigelloides is similar to Aeromonas spp. Most cases have been sporadic rather than associated to large outbreaks. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. however. but the bacterium is known to survive and grow in the shallow marine. cytotoxic. They are aquatic organisms with an epidemiology that is yet to be fully understood.6. Some isolates are true psychrophiles. the causative agent of cholera. coli on isolation media (50). shigelloides are encountered. and fish and shellfish are probably the major reservoirs for the organism (50). which is often fatal (>50%). It is known that Aeromonas spp. infection. cholerae O1. Since 2000.

It is regularly isolated from seafoods (51)... AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. Clostridium spp. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. full risk analysis is not possible (52). D60 of 1. The most frequently reported parasites of human importance in fish are round worms of the genera Anisakis and Pseudoterranova and tapeworms of the genus Diphyllobothrium.6. Like other toxins. Being a psychrotroph. neonates. at risk groups . If fish are not properly refrigerated. but cooking will significantly reduce the risk of infection. 1. and Lactobacillus spp. Infections are mostly associated with the consumption of raw or 18 . it is not apparent to the consumer and it cannot be destroyed by cooking. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. Fortunately.6.48 minutes in salmon have been reported (53).FISH AND SEAFOOD persons. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 .. there is a possibility for growth of the organism on chill-stored fish.2002 (55)) and is also common worldwide. leads to the production of histamine. but proper cooking will reduce any risk. Klebsiella pneumoniae and Hafnia alvei. The organism poses a serious risk in chilled products not cooked before consumption. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). Photobacterium spp.pregnant women. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. 1. alcoholics. especially pickled fish. are exceedingly difficult to produce within the legal limits of histamine as the temperatures used during production can lead to rapid growth of histidine decarboxylating bacteria. 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. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). some Vibrio spp. Lightly preserved products. foetuses.98 minutes in cod and D60 of 4. but more especially by Morganella morganii. 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.

1997. parahaemolyticus Staph. long storage under refrigeration or one of the former prior to freezing. Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease. Faecal coliform counts may be used instead of E. 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. For fish from inshore or inland waters of doubtful microbiological quality. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. In summary. TABLE1. it may be desirable to test for Salmonella and V. parahaemolyticus (see Table 1. Rome.H. which in many cases will eliminate the risk. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF). risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low.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.7 Published Microbiological Criteria Microbiological criteria for food defines the acceptability of a product or a food lot based on the absence or presence or number of microorganisms. 1997) Huss H. EC. No.. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it. especially in warm-water areas and where fish are to be consumed raw.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi.IV) (57). coli Case 1 4 Plan 3 3 n 5 5 c 3 3 m 5 x 105 11 M 107 500 Additional tests to be carried out when appropriate Salmonella V. area or lot (CAC. 1. Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. FAO. aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . Ababouch L. coli counts where this method is preferred. 444.. including parasites and/or quantity of their toxins / metabolites per unit of mass. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). 2003. Gram L. Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers. volume. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration.

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

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

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

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

Villemure G. 1989.D. Technomic Publishers.. Handling and processing of scad. 1986. Fisheries and Food. Microbiological quality and shelf life prediction of chilled fish.1 Further Reading Processing Anon.FISH AND SEAFOOD 1. Houston N. International Journal of Food Microbiology. 1985.2 Spoilage Shewan J. Application of predictive microbiology to assure the quality and safety of fish and fish products. 317 (Suppl. Gram L. 271-85.9. 35.S. Olley J..G. Akiba M.. Torry Research Station of the Ministry of Agriculture. 127-34.M. International Journal of Food Microbiology.M. 13-32. Journal of Food Science. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. 1988.H. Jones Y. 1.H. Trolle G. Ed. Changes in microflora and chemical compounds in mackerel stored by partial freezing. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. Huss H. 1997.. Huss H. Torry Research Station of the Ministry of Agriculture... Ed. Simard R.. Aberdeen. in Fish as Food. Taoukis P.H. Gibson D. Huss H.. Ogden I. Florida. 4. 73-89. Labuza T.E. Torry Research Station. Kitts D. Food and Agriculture Organisation. 55. 1984.C.. 317-21. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. Rome..).. 15. Cann D. Sumner J. 24 . Bulletin of the Faculty of Fisheries (Hokkaido University).. in Torry Advisory Note No 93. 6. Borgstrom G. Eds. 1. in FAO Fisheries Report No. Lancaster. 62-70. Aberdeen. 1.M.1983. Estimation of the bacteriological quality of fish by automated conductance measurements....9. 1992. 1961.D. 1984.. Academic Press. 51.. 65-72. Fisheries and Food.A. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. Hobbs G. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. McMeekin T.9 1. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches. 1992. 741-50. The Microbiology of Seawater Fish. International Journal of Food Microbiology. Food and Agriculture Organisation of the United Nations. Magno-Orejana F. Dalgaard P. Picard G.. in Seafood Science and Technology. 1989. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Journal of Food Protection. Ravn Jørgensen B. Smith G. Shinano H. 295-307.. Cho Y.L.. Gibson D.P. Shahidi F. International Journal of Food Microbiology. Ross T.

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


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

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

and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. be flavoured with herb dips or peppercorns. Fish steaks can be cut from a natural fillet or loin (e. Again. 29 . A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes.5%.g. Fresh fillets are sold from wet fish counters.5% in the aqueous phase. the ingredients premixed. or pre-packed frozen with added glaze. Pâtés are prepared with raw fish. tuna steaks) or band-sawn from a frozen block (e. natural fillets of fish such as herring. Cold smoking is essentially a flavouring process. 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. mackerel. VP products) because of the high salt content acting as a barrier to spoilage.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze.2 days) longer than that of non-prepacked fish.g. haddock. mackerel. and the product then shaped and heat-processed to cook the product before being chilled. 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.2. or pre-packed in MAP or vacuum packaging (VP). salmon. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. which is approximately 30% (or 1 . Hot-smoked fish are prepared from brined. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product. cod).. 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. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. Added colours can be included with the brine solution to meet specific requirements for products e. in addition. and salmon. in VP or through mail order. The expected shelf life of these products is up to 8 days (or longer for some chilled. a protective glaze may be added to the frozen steak. but may be increased (up to 1 month) by packaging methods like MAP or VP. Some products may be salted for flavour only (e. sawdust with other woods adjusted for flavour profile and burn duration).5 . annatto with smoked haddock. This process in effect cooks the product and reduces or eliminates microbial contaminants. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. with a minimum residual salt content of 3. The shelf life for MAP fillets is typically up to 5 days. etc. They are sold from wet fish counters. which may.g. swordfish steaks). Cold-smoked natural fillets are prepared from a range of cold water fish including herring. The result is fish with a typical residual salt content of 1.g.

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

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

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

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

On arrival at the processing plant.3.1. after landing. frozen in a nitrogen blast freezer and weighed into sacks for later processing.Fish Cakes). quickfrozen.1A . Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product.4.1B . Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process.1. by mincing and mixing with a binding agent (as described in section 2. Frozen cores are usually batter-enrobed.2. 2. 2. It is then size-graded.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). Raw material preparation stages for chilled or frozen coated fish products 34 . with the coating heat-fixed. 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.1 (2. drained and then refrozen for storage until required for further processing. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.7 days. 2.4.1C . The “blown” meat is graded. The processing of three examples types are described in Figure 2.3. are placed into plastic boxes and covered in ice to preserve their condition. may not rise above 0 ºC during processing. chilled or sold chilled on defrost with an expected shelf life of 6 .2). Fig.Scampi.1.Shaped Fillets. the scampi is washed in potable water to remove foreign bodies.FISH AND SEAFOOD processed.

quickfrozen. or those where the trimmings. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. The remaining dried ingredients are mixed with the fish. once the dry components have been rehydrated.. with an expected shelf life of 6 . are handled and stored as any other food product.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. spoil more rapidly than the dry ingredients. The mix is shaped and formed ready for packing or for enrobing.g. 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. as the physical structure of the material has been destroyed. from potato flake.3. ii) Fish-filleting waste . Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e.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. iii) Fresh/fillet/minced fish block. and water is added to rehydrate the components. such as aerobic spore formers and Staphylococcus spp. Waste material produced from this type of processing is often reused in the third category (fish cakes). pathogens such as Salmonella and Listeria. that are to be reused for other foods. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish.7 days.4. or promptly processed to avoid further quality loss. This is especially the case with block-sawn dust. Fish cake cores may be batter-enrobed. those that use frozen blocks of whole fillets. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . The fish component of the cake will. 35 .2 Shaped products This type of product falls into two categories. chilled. 2. fish fingers). or sold as a chilled product after thawing. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or.block-sawn dust.3.4.1. unless it is poorly handled. with the coating heat-fixed. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss.off-cuts or belly flaps. because of its nature. material will need to be transferred swiftly to a chiller.1. additional spoilage organisms may be introduced. It is for this reason that all by-products from primary processing. but. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. Therefore. more seriously.

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

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

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

2. Raw material is typically fish fillet. For premium products. and reduce spoilage organisms and potential pathogens.3 log reduction in bacterial numbers on the surface of vegetables. i.lactobacilli.4.5. Although the processes are claimed to be superior to using unmodified hypochlorite for washing. Washing with a biocide will at best achieve a 2 . In an attempt to improve washing and reduce chlorine taint. However. or breadcrumbs.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. any processing stage. control is more onerous. coryneforms. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. frozen block or canned fish. and E. the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. coli O157. The majority are products made from various types of whitefish (cod. (7. spore-formers. For example. Any assumptions regarding the application of good agricultural practice to produce should be made with caution. haddock. not just as accompaniments. will disperse the saprophytic flora of vegetables . with the accompanying release of additional nutrients by processing. Products must be washed thoroughly to remove pests and soil. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. However. for example. they should be stored under temperature-controlled conditions. encourage more rapid decay and loss of structural integrity. but also as components in recipe meals. if the process is not appropriately controlled. loss of quality. 8) will. 39 . treated as a highrisk ingredient. all-year-round fresh vegetables are preferred. Salmonella typhi. rice or pasta. The opportunity for growth of L. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components.2 Seafood ready meals (frozen or chilled) These differ from seafood recipe meal centres in that they contain a source of carbohydrate such as a potato topping. A number of examples of direct faecal contamination of vegetables exist in the literature (8). and may include molluscs or prawns that may or may not have undergone a minimal heat process. For example. Once vegetables have been blanched. pastry. and protected from any post-processing contamination. Blanching of vegetables can be an effective means of reducing or removing surface contamination. chopping and mixing of material. coliforms and pseudomonads throughout the mix. even those with pH-modified dressings. with cases of foodborne disease traced to Listeria monocytogenes. The presentation of vegetables has a significant effect on the product quality. colour and water will occur.e. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. and may include sauce accompaniments. and mixed whitefish). Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp.

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

suspended in an acidified sauce with an oil component.2. that of a dairy-based sauce. Figures 2. the sauce component will spoil rapidly and the possibility of vegetative pathogens from post-process contamination or raw components growing to significant numbers cannot be ruled out. Marinade-type products are becoming more popular. Materials prepared with regard to good hygienic practice need only be assembled in a tray or pouch before chilling or freezing in readiness for distribution. flour products. respectively.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig.herbs. and handling of processed sauce can be less rigorous than. etc. It is expected that this product format would have a restricted shelf life to limit potential for growth of psychrotrophic bacilli or any survival of enteric pathogens (12. 13). Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. The format uses flavour compounds . spices. Without barriers. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. for instance.4.4 and 2. 41 .5 illustrate the recipe dish ingredients preparation and assembly and packaging.

FISH AND SEAFOOD Fig. The oxygen-free conditions will however allow other anaerobic bacteria such as C. Flow diagram outline assembly and packaging of recipe dish 2. botulinum to grow.7.7. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. 2.4. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues.2 Modified-atmosphere packaging Originally developed for the red meat industry. These include: 2. which may restrict either microbial growth or enzyme activity. unless the temperature is strictly controlled to below 5 °C throughout the supply chain. as it dissolves in the surface moisture to form a weak acid.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth.4.4. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow.5. 42 . and tuna loins are packed by placing the product in a plastic pouch and removing all the air.2. the method replaces air in the sealed packaging with a defined mixture of gases. generally carbon dioxide and nitrogen.

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

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

histamine intoxication. and many raw or processed foods (26). perfringens contaminating seafood will depend on the source of raw material.6. are dealt with elsewhere in this book.3 Clostridium perfringens A study examining C. agricultural run-off or sewage can contaminate in-shore species.104/ g should be treated with some concern (26). Inappropriate holding times and temperatures will. Cases quoting contamination of seafood with organisms such as S. 2. etc. These are C. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. with pathogenic microorganisms or viruses. permit rapid growth of B. botulinum when held in favourable conditions for growth (28) i. and Campylobacter are recorded in filter feeding molluscs such as mussels. or a heat shock followed by holding at a temperature suitable for growth.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. 27). water. air. 2. Counts of between 103 . 45 . cereus. spices and herbs and vegetable shoots have been reported (12.6. Germination of spores requires favourable conditions. soil. Therefore. Additional hazards from marine bio-toxins. Hazards attributable to the non-seafood components are listed below and this information. botulinum type E and Vibrio parahaemolyticus (25). In addition. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). potato.. in some cases. may be of benefit within any risk assessment for chilled and frozen prepared fish products. A number of other Bacillus spp.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. 2. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. farmed or free-living crustacea (30). typhi.6. have been linked with food poisoning and should also be considered.1 Bacillus cereus This is a common contaminant of dust. Incidents in such diverse materials as cream. Vibrio spp. and some effects related to product type and time to toxin production have been noted. 13. low oxygen or anaerobic conditions. along with previous discussion. and this issue may be of greater consequence for warmwater. any risk of C. especially molluscan filter feeders.e.

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

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

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

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

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

A. 60 (4).A. Shapton N. 788-801. Microbiological Criteria.A. McCarthy S. 41. 36. 38. U.L. Cambridge.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. Journal of Food Protection. Chilled Food Association. CRC Press. Woodhead Publishing Ltd. Psychrotrophic Bacteria in Foods: Disease and Spoilage. 40. Journal of Food Protection.H.uk/cday/posters2008/npd6. 2006.2. Chilled Food Association. Wolfe Publishing.. London. 34. U.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology. Development and Use of Microbiological Criteria for Foods. Foodborne Pathogens an Illustrated Text. Smith J. 1999. 39. British Retail Consortium. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs.pdf Shapton D. http://www.europa. 1991. http://www. Kraft A. Varnam A.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33.campden.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. Phil Voysey.F. 372-6.K. Institute of Food Science and Technology. London. http://eur-lex. Principles and Practices for the Safe Processing of Foods. 1998. 1997.K. 1992.eu/LexUriServ/ LexUriServ. 35. Shigella as a foodborne pathogen. Evans M. 51 . 50 (9).G..chilledfood. Gloucestershire. Campden & Chorleywood Food Research Association.co. 1987. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. 37.


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

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

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

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

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

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

2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding. and the environment. Live: in shell. 3. Raw and partially cooked shellfish. For example sessile and benthic shellfish dwelling on the bottom or burrowed in marine sediment are especially vulnerable to pollutants (sewage and/or wastewater) introduced into the shellfish water beds. Canned: in own juice. in sauce. Vinegar cured: bottled in malt vinegar after brining. 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. 3. Whatever contaminants are in the water will eventually get into the shellfish (7).2. pH. Because molluscan shellfish are filter feeders. Fresh: meats removed from the shell by boiling. they accumulate pathogenic microorganisms from polluted waters. and butter. which affects the temperature.1. are vectors of disease transmission (8). The following pathogenic microorganisms causing illness in humans are characteristically associated with molluscan shellfish: Vibrio parahaemolyticus Vibrio cholerae O1 Vibrio fluvialis Vibrio hollisae Vibrio vulnificus Vibrio cholerae non-O1 Vibrio mimicus Vibrio furnissii 59 . Salted: meats either lightly or heavily dry salted. also bottled in brine. Canned: in brine. depending on length of journey. mostly in oysters. nutrient concentration and pollutants of the water column. clams and mussels.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves.MOLLUSCAN SHELLFISH Frozen: shelled meats. salinity. The microflora are dependent upon a number of factors such as season.

have been observed microscopically in the digestive tract of Eastern and Pacific oysters. Campylobacter coli Bacillus cereus Hepatitis non-A. the initial flora composition and concentration of bivalves are highly variable. They are termed viable non-culturable microorganisms. that have never been cultured on laboratory media. 11..1 Shellstock 3.. or by 60 .FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp.1 and 3.1 Processing and its Effects on the Microflora Oysters. 12.1.. (9.3..1. but have never been cultured (14. shellstock (live) which is shellfish in the shell..3 3. 13). Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. Water temperatures affect the concentrations of microflora in the water. from which one or both shells have been removed. mechanical harvesters. 3.. 3. There are microorganisms. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. Enterococci spp. Temperature. Vibrios. Flavobacterium spp. and shucked shellfish that is shellfish. Other microorganisms found in lesser numbers are the Achromobacter spp. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora.. spirochetes of the genus Cristispira and Saprospira. nutrients.1. Moraxella spp. and cockles The processing of bivalves takes two forms. Micrococcus spp.3. The largest groups found are the Pseudomonads. 10. salinity. and Aeromonads. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. In summary. At harvest. mussels.. clams.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. Alcaligenes spp.3.. dredges. Acinetobacter spp. Pseudomonas spp.000 /g. rakes. 15). observed microscopically. whole or in part. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. Bacillus spp.2. responding to the environment in which they are harvested. and yeast cells of Rhodotorula rubra and Trichosporon spp. For example. Flow diagrams of the major processing steps of molluscs are presented in Figures 3.

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

including V.2 ºC (45 ºF) or below (19). Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. without adequate refrigeration. 26). Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. coli. 24.FISH AND SEAFOOD 3. shellstock are not refrigerated on the harvest boat. Shellfish may contain higher levels of pathogens than are found in the water in which they grow.1. Furthermore. synthetic seawater for the purpose of de-sanding and salt uptake. 17). Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. involving 691 cases was attributed to oysters contaminated in a wet storage area (27).1. are prevented. 23.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. however. handling. with prolonged storage the microbiological quality of the bivalve decreases. bivalves tightly close their shells thus trapping water and microflora. Generally. hydrophila (18). Over a period. 22). Klebsiella. The most important factor in controlling bacterial growth in shellstock is temperature. vulnificus. and Citrobacter species) and Vibrios. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. 21. 16. If long delays occur before reaching the processing facility and/or refrigeration. Water used for shellstock washing should be of good sanitary quality. At this temperature the multiplication of faecal coliforms (E. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). with the exception of A. there is evidence that in the absence of adequate refrigeration. Generally. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. some pathogenic organisms will survive and multiply in shellfish (20. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8.3. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . Harvest conditions are another factor that influences the growth of bacteria in shellstock. the bacterial counts in the meats will increase. and temperature storage of shellstock to point of sale. for a considerable length of time. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. Enterobacter. and thus contaminating the shucked meat at the point of sale. The washing of sediment and detritus soon after harvesting will prevent further contamination. At harvest. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. An outbreak of infectious hepatitis in Sweden in 1956. to avoid possible contamination of the shellstock. and the sanitary practices applied during harvesting and shipping. when it reaches the processing facility. shellstock are placed in refrigerated trucks for transport or placed in wet storage. 25).

3. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. Shuckers routinely use either cotton or rubber gloves during the shucking operation. washing. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. and packing shellfish into containers (Figure 3. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station.2 Shucked shellfish 3.3. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. through which the washed meats are discharged for rinsing and then packed in containers.2 Effects of microflora during processing The same considerations as previously discussed. To remove the shellfish attached to the remaining valve. In addition. Under these conditions if the shuckers enter the packing room. 3. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). the tank is opened through a drain gate and chute.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. If shellfish are not reasonably clean at the time of processing. Studies have shown the use of rubber gloves reduces the contamination level.1. thus contributing to high bacterial counts in the final product (28).1. 3. Thus. After washing of meats. washing and culling shellfish prior to storage is essential. 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). and fish.1.2. are applied to the shucking operation of shellfish. Containers are placed in packing boxes for shipping at refrigerated temperatures. shellstock is subjected to contamination when stored on the floor in standing water. removal of shellfish (shucking). Due to the nature of the shucking operation. the finished product may become contaminated.2). clothing becomes very soiled.2. In the blower-washer. whereas cotton become wet and 63 . Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported.1 Processing description The processing of shucked shellfish is limited to washing of shellstock. lobsters. or subjected to splash from foot traffic. for the effects of microflora during the processing of shellstock.3. The process of shucking involves forcing a knife between the valves and cutting the adductor muscle of one valve freeing the shellfish from one valve. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank.

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

Temperature abuse will result in high bacterial counts. All equipment coming in contact with the shellfish such as colanders. at the turn of the century. thus producing acid and gas (38). Achromobacter and Flavobacterium levels decrease during storage. where there is a breakdown of temperature control or prolonged storage of shellfish. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. clean gloves. Controls that should be applied include using clean packing equipment.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. 33. in oysters.2 ºC or less. and yeasts. The environment has an effect on the bacterial population found in shellfish. abundant gas production. As described previously. 31. The initial flora found in shellfish is the major factor of spoilage in bivalves. 36). 35. For example. Alcaligenes. blowers and other utensils that have cracked. Proteus.47 to 6. a carbohydrate. rough. Earlier studies. Shucked shellfish are an excellent medium for the growth of bacteria. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. Micrococcus. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. Much of the work has been done with oysters and clams. 32). 34. glycogen levels range from 0. particularly oysters stored at 7 ºC. Lactobacilli seem to be the predominant microflora in shellfish. are an excellent medium for growth of bacteria. This process changes the bacterial profile to microorganisms favouring low pH. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. Glycogen. is stored in the bivalve as a source of energy.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. Glycogen is hydrolysed by the initial flora found in shellfish. particularly shucked meats. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. streptococci. 65 . and packing the shellfish in a timely manner. Flavobacterium. anywhere during the process of shellfish from harvesting to the point of sale. The spoilage process in shellfish may be divided into three stages: increase of acidity. Shucked shellfish are held and transported at temperatures of 7.8% (37). Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. and proteolysis (10). These organisms may play a significant role in the spoilage processes. 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. shucking pails.2 ºC. whereas bacterium such as Pseudomonas. spoilage will occur. Pseudomonas. showed that organisms such as Achromobacter. Shellfish. 3. In summary. these include lactobacilli.

5 cases/100.1 Vibrio parahaemolyticus V. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). during processing. Sporadic cases are frequent along the coast of the United States. primarily in brackish or saltwater. Between May and September 1997. and very common in Japan.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish.48 hours with abdominal pain. Major outbreaks are associated with the warmer months. One of 250 cases reported involved shucked products with low levels. V. and estimates based from a hospital study show less than 0. It is a common marine bacterium. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. 66 . although non-hemolytic strains have been associated with illness (43. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. Diseases caused by Vibrios are enteric in nature. parahaemolyticus possibly producing illnesses (40). These organisms can be present in the shellfish from the point of harvesting. widespread in both polluted and unpolluted waters. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. parahaemolyticus causes gastroenteritis lasting 24 . headache and fever. Products involved were from the same processor. parahaemolyticus.000 population/year (46). and to the point of sale. 3.e. ranging from epidemic cholera to sporadic cases of diarrhoea. 41). there is a recent report of low levels of V. only a small percentage of strains in the marine environment are potentially pathogenic (45). however.1. 3. nausea.FISH AND SEAFOOD 3. 44). prevention of cross-contamination. of V. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. V. where outbreaks occur regularly. lot and retail outlet. and harvesting of shellfish in the cooler months. approximately 23 cfu/g. shellstock and packing of shellfish in jars) (29).5.5. The incidence of V. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. from harvesting to finished product (i. Fortunately. parahaemolyticus from shellfish is low. coastal areas).1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. They can be introduced to the shellfish from the environment or added during processing. This bacterium is present in all the major processing steps of a shellfish plant. diarrhoea.

parahaemolyticus growth in shellfish meats is temperature dependent.5. In one study. They are sensitive to cold. Hazards from this bacterium can be controlled by thorough cooking of shellfish. V. these individuals are advised not to eat raw shellfish. those who have liver disease. 46. Again. which have been implicated in shellfish-related disease outbreaks (31). 56). 49).II (8.MOLLUSCAN SHELLFISH Control of V. The bacterium is an estuarine species that causes wound infections. cholerae. cholerae non-01 has been associated with septicaemia. 58). however. abdominal cramps. 48).48 hours of onset. the organism has been found in the Gulf of Mexico. 3. 3. or primary septicaemia.4 . 57. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8.3 Vibrio cholerae non-01 Illness caused by V. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. cirrhosis. vulnificus are fever.1.0. and rapidly become inactive at low storage temperatures.7 days. It is one of the most severe foodborne infectious diseases. All isolates are considered pathogenic. The occurrence of V. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). vulnificus. Control of V. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52.2 Vibrio vulnificus The symptoms of V. like V. Primarily. fever. gastroenteritis. and preventing cross contamination once the product is cooked. and nausea within 24 . are particularly susceptible to primary septicaemia. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. It causes diarrhoea. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. cholerae non-01 is ubiquitous in the estuarine environment. vulnificus is by rapid refrigeration during warm weather months. or immunosuppression. 46. 54. nausea. Oysters are the most hazardous food associated with V. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). The organism attaches to the intestine and release of a toxin is suspected. Healthy individuals are susceptible to gastroenteritis. lasting 6 . Onset of symptoms occurs in 48 hours and can be severe. chills. cholerae non-01 (51). diabetes.000 population/year (8.1. Inadequate refrigeration will accelerate the growth of V. and vomiting. leukaemia. parahaemolyticus the estimated annual incidence rate is very low. V. High-risk individuals. 23 of 24 major USA-West Coast estuaries were positive for V. 53. Control of oyster-associated 67 . parahaemolyticus. in the range of 0.5.8 case/100. 55. with a fatality rate of 50% for individuals with septicaemia. Death has been reported to occur within 36 hours from onset. V.

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

70. infection occurred among participants of a firemen's banquet. although their role as a pathogen is highly controversial and circumstantial. 3.3 Other organisms of concern 3. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. To reduce the risk of shellfish-borne campylobacteriosis. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. Recently. V.3. 76. vulnificus. have been isolated from shellfish beds throughout the world (67.500 cells/gram of Campylobacter may cause illness (64. 69.5. ranging from 0. cholerae 01 and non-01. They have been implicated in limited shellfishassociated gastroenteritis cases (8.5.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. In one case. jejuni were recovered. jejuni does not multiply in shellstock at refrigerated temperatures. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. Campylobacter spp. 68. 77. Human feeding studies suggest that about 400 . jejuni begins in the harvest of shellstock from approved waters. Campylobacter appear to survive well in oysters during storage.MOLLUSCAN SHELLFISH TABLE 3. 83. 85). V. and their immediate proper refrigeration. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. of Incidents 4 279 0 1 0 No. 66). Although.II Molluscan shellfish associated outbreaks caused by Vibrio spp. The seasonality of 69 . hydrophila group (75. V. and V. shigelloides (79. however. control of C. C. of Cases 4 362 0 1 0 a Includes V. 80) are ubiquitous in the marine environment. 71). C. hollisae Infective dose varies with host susceptibility and virulence of the strain. parahaemolyticus. from Pacific shellstock oysters purchased from a retail grocery store (74). 72. 81. they survive well at refrigerated temperatures. whereas at 10 ºC Campylobacter survive only for 5 days (71). fluviallis. V. mimicus. 84.4 to 114 cells. This finding is significant in that C. 65. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). 78) and P. 82. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. 73).a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. however.

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

K. USA.E. Wiley-Blackwell. American Journal of Public Health. 1936.M. Liston J. Applied and Environmental Microbiology. 1970. Lovelace T. 1991.D. New York. Hackney C. 105-8. Ray S. Effects of harvesting waters and storage conditions on yeast population in shellfish. 26. 42. Ward D.S. 13. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay.F. Microbiology of bivalve molluscan shellfish. 447-52. Thompson C. Lee J. Schmidt C.R. 419-25. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). Jr. U.S. Rippey S. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. Bernard F. Bacterial flora of the market oyster..C. 633-40. Villee C.K.. 13. 14. Veliger. Vanderzant C. 64. Applied Microbiology. 7. 8. 46.. Government Printing Office. 71-6. 11. 245-7. Biology of the oyster in relation to sanitation.. in Multilingual Dictionary of Fish and Fish Products. Smith F. 1995. 1981. 10. 2. 33643. Eds. 71 . 1960.7 1..R. 8.. 1964. 58. Proceedings of the National Shellfisheries Association. Journal of Food Protection.R. 1926. Tall B. References The Phylum Mollusca. 15. 104-9. water and sediment from Galveston Bay. Saunders Company. 30. Hood M. Walker W. Eds. A typhoid fever epidemic caused by carrier bootlegging oysters.C. Organisation for Economic Co-operation and Development. Clinical Microbiology Review. 19-34.R. Eliot C.MOLLUSCAN SHELLFISH 3. 72-8.A. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. 33-6. 9. Fleet G. Van Nostrand Reinhold.. in Microbiology of Marine Food Products.B. Pfeifer D. The American Oyster.N. 1940. Washington D. 231-51. 1963.. Fish and Wildlife Service.R.W. W. Ed. 7.A. Tubiash H.. 6. 1975.S. 44. 29. Viability and release Salmonella charity and Escherichia coli from oyster faeces. 6. Journal of the Mississippi Academy of Sciences. 755-76. Applied and Environmental Microbiology. Gill S. 3. 1973. Cook D. Old H.L. Fate of enteric bacteria in estuarine sediments and oyster faeces.W. American Journal of Public Health. 4. 1984.H. 17. Colwell R. Philadelphia. Cook D. Galtsoff P.J. 1983. American Journal of Hygiene. Microbiological characteristics of Dungeness crab (Cancer magister).S.A. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). Journal of Milk and Food Technology. Nauman R. 12. Infectious diseases associated with molluscan shellfish consumption. in Fishery Bulletin 1964 No. 82-7. Colwell R... Galtsoff P. 16...E. 36. 30. 1994. 1982. Rowse A. 5. in General Zoology. Jr. 544-8.R. Applied Microbiology. 1968.

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

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

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

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

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

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


CRUSTACEAN SHELLFISH Linda Nicolaides. lobsters.5 kilo pack (boxes or bags). jointed limbs and a chitinous exoskeleton. M.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. Such products are usually only partially cooked before freezing. the front pair ends in claws. 79 .g.. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption.5 and 1. Parapeneopsis spp. Peeled and cooked shrimp are ready-to-eat after controlled thawing.5 and 1.0 kg polythene bags. and are usually aquatic.0 kg polythene bags. Shrimps. Peeled ready-to-cook shrimp are raw. with shells and heads on or off are usually packed. crayfish and shrimp (1). lobsters and crabs have ten legs..4. and frozen in 1 and 2. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. They possess a segmented body. and available in 0.. Pandalus borealis (deep-water shrimp). e. Species eaten from tropical waters include Penaeus spp. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi).Ph. Pandalus montagui and Crangon crangon (inshore shrimp). packed and frozen in 0. crabs. with additional water to form a glaze. (deep-water shrimp). peeled product. and Trachypenaeus spp. Crayfish.

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

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

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

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

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

the pathogen can grow at temperatures as low as 3 °C. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. as part of the natural bacterial flora. aureus on their hands. 4. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. 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. especially in products that have been vacuum packaged. cholerae is divided into two groups.4 Clostridium botulinum C. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru.5.70% of healthy individuals carry toxin-producing strains of Staph. combined with the degree of handling of these types of product. which can become colonised by V. particularly crabmeat (18. was associated with seafood consumption. although contamination by types A and B may occur from the environment. botulinum type E is commonly associated with fish and fishery products. The main source of contamination is the hands of processing personnel. about 50 . pregnant women and the immuno-compromised. botulinum type E at 10 °C but not at 4 °C (20). e. V. the causative agent of cholera.5. 22) as well as from undercooked crustacea when harvested from contaminated waters (23. cholerae non-O1. therefore. Once the competing microflora has been removed by a heat treatment. 25). Researchers have demonstrated that inoculated shrimp supported toxin production by C. The pathogen has resistant spores.g. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. cholerae. To date.5.CRUSTACEAN SHELLFISH old. so is able to survive heat treatments that eliminate vegetative cells. aureus is not a member of the intrinsic flora associated with crustacean shellfish. Crabs are covered in a chitin exoskeleton. V. whilst boils and cuts are also common sources of the pathogen. The former. pasteurisation and cooking. no incidence of foodborne illness caused by L. is widely distributed in aquatic environments.5 Vibrio spp. An epidemic that began in South America in 1991. aureus because of the ubiquity of this organism in the processing environment. 19). 85 . monocytogenes has been attributed to crustaceans. Control should be achieved by promoting GHP and staff training programmes. Vibrio cholerae. cholerae O1 and V. Crustaceans have been linked with food intoxications attributed to Staph. 4. 4.3 Staphylococcus aureus Staph. However. 24.

to date. V.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. Japan. Astroviruses and non-A. occurs in crustaceans from warm environments.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.g. South America. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates.5. coli.30 MPN/g) (27).5.6 Campylobacter jejuni and Campylobacter coli C. particularly trematodes (6). non-B Hepatitis viruses (31). It is more commonly implicated in countries where crustacean shellfish are eaten raw.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. however. 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. a marine halophile. the potential hazard exists as C. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). However. e. 4. 4. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. hydrophila and A. Quantitative levels were below limits of detection in all cases (<0. parahaemolyticus.5. V. Caliciviruses.8% of samples contained C. been implicated in a case of foodborne infection associated with crustaceans. hydrophila has been isolated from prawns (29). Of these. A. However. 31. 32). sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture.5. whilst 5.7 Aeromonas hydrophila and Aeromonas sobria A. in a survey carried out in the United States between 1977 and 86 . but has the potential to cause fatal foodborne illness (26). 4. hydrophila is able to grow well at refrigeration temperatures (0 . or freezing at a temperature of -23 °C for 7 days (30. although A. Parasitic infections are controlled by either cooking infected material prior to consumption. Hepatitis A virus presents the most serious hazard. vulnificus is an invasive and lethal pathogen associated with wound infections. jejuni has not.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. 4. Norovirus.

produced or imported into the European Union (EU). 2073/2005. 4. which results in a persistent and apparently permanent loss of short-term memory (36).CRUSTACEAN SHELLFISH 1981. The toxin causes amnesic shellfish poisoning (ASP) when consumed. A summary of microbiological criteria for crustacean shellfish is listed below. 39). although it is one of the European Union (EU) legislated allergens.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. 87 . 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. or chlorine concentrations similar to those found in drinking water. Norwalk viruses can also withstand heating to 60 °C. They can survive low pH levels. Selected parts from this regulation is presented below. only 0. 4.11 Chemical pollutants and biotoxins Researchers have demonstrated that crabs feeding in waters contaminated with algae do have potentially high levels of algal toxin domoic acid (DA) in their viscera.5. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34).10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. risk-based system based upon the seven principles of Hazard Analysis and Critical Control Point (HACCP) supported by pre-requisite or good practices programmes at all stages of the food chain.6. No foodborne incidents have been attributed to this source to date. 4.5.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33).6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. 38. The risk of such hazards is far higher with fin fish and molluscan shellfish. consistent with that in the stomach. from farm to fork (37. 4. are included in the Commission Regulation (EC) No. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control.

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.


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

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

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

6 g of salt or more per litre of brine) may cause salt burn in this way. sometimes the salt crystallises onto the surface of the fish. only the most common processes can be described here. TABLE 5. When working with salting. processes and producers’ private recipes.2 105.I). Different brine strengths are required according to the curing processes and/or the type of product. it is important to state the temperature. Salt in the water phase links the two conditions of moisture and saltiness.FISH AND SEAFOOD 5. Salt concentration means the percentage of salt in the water contained in the fish. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat.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. the fish is usually salted with a 70 .4 52. a 90º or above brine (237. This should be distinguished from the salt content.2 237. it causes swelling because the fish absorbs water from the salt solution (1). At 18 °C a 100º brine is produced when the solution contains 26.0 .80º brine (184.6 264.0 158. etc. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. When a milder brine. The strength of the solution is described as degrees of saturation (brineometer degrees). about 50º (132. When a 100º brine is used. % 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.8 79. Therefore.8 211. Thus.6 132. it is impossible to establish standardised processing stages in the curing of fish.3 Processing and its Effects on the Microflora Owing to the wide variety of products.8 211.0 g of salt per litre of brine).4 184. or salt in the water phase. a 0º brine saturation means pure water. smoking or marinating processes. is employed.2 g of salt per litre of brine). For smoking salmon. all using salting steps. it is important to understand the concept of salt concentration.4% salt (Table 5. and salt content. The dissolution of salt in water is dependent on the temperature.

mainly in tropical conditions. See Figure 5.1.15% calcium and magnesium.CURED. outgrowth of halophilic bacteria may occur.15 . but the concentration of copper and iron must be kept under 0. ‘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. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. are likely to cause hydroscopicity of the product and. and a ‘putty’ fish product is also obtained (1). ‘Green cured’ split cod with about 60% of the backbone removed. caught in North Atlantic waters. it tends to lead to a slight yellowing and softening of the product. low in calcium and magnesium.1 Dry salting / Kench curing Usually. For a light salting. respectively. the fish is restacked with fresh salt after about a week. and such delays. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. Usually it is followed by other preservation techniques such as drying and/or smoking. The calcium and magnesium impurities. SMOKED AND DRIED FISH 5. The most common fish used in the northern hemisphere is cod (Gadus morhua). This condition is known as ‘putty fish’ (4). which include calcium and magnesium. are recommended to help make the product whiter. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. This is because fatty fish turn rancid quickly. when the air humidity is high. Also. producing characteristic pink patches. and salt can speed this oxidation process. the fish is mixed with one part of salt per eight parts fish.1. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). addition of levels of 0. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. Salting can be done in a number of different ways. However. if very pure sodium chloride is used for curing purposes.35% and 0.1 Salting Salting of fish for preservation purposes has been used since pre-history.3. having a short section at the end of the tail. for which there is a high demand in Mediterranean countries. and essentially free from iron and copper (3). When pure salt is used. 5. can be dried further to produce ‘klipfish’ or ‘bacalão’. however. It takes about 3 weeks until proper salt penetration and moisture loss are achieved.3. The salt used should be of food-grade quality. and the salt has penetrated throughout the fish (3). respectively (1). On the other hand. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. 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. 97 . lean white fish is used for this purpose.0.1 ppm and 10 ppm. and for a heavy salting a ratio of 1:3 salt:fish is used (1).

eviscerated and/or split). when a light brine is used.2 Wet salting or brining As referred to above. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole.1.90 are rarely achieved.3. such as Staphylococcus aureus could survive and grow at such water activities in the product. Fig. such as 98 . Where fish are salted in their whole form. so that the risk of spoilage is high.1. Only small fish can be salted whole. although toxin production would be unlikely (4). Food-poisoning organisms. the process is mainly for flavouring purposes prior to the use of other preservation processes.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. it appears that water activities much less than 0. Processing of dried salted cod 5. For large fish it is advisable to split it before salting. The greatest barriers to salt penetration are the skin and scales.5.

0 4. For preservation purposes.25 5.49 4. The fish are kept immersed in their own brine near saturation point for several days.90 (1). It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.47 6.II.70 3. the liquid that comes out of the fish forms a brine. with pressure being applied to the top of the container to help the brine to drain away.51 3.76 4. limiting the amount of rancidity that can develop.46 4.85 3.76 4.80 5. For example. with a shelf life of about 4 weeks (water activity (aw) about 0.66 5.17 5.22 6. and a red burgundy colour will develop in the product.0 3. Anchovies are the main fish used for these purposes. can be cured in a saturated brine solution for 6 days and.3.70 6.3 days.66 5.26 2.86 2.II Salt in the water phase (after Ian Doré (5)) Moisture (%) 40 45 50 55 60 65 70 75 80 85 90 Percentage salt in the flesh (by weight) 2.5 10. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.84 1.11 4.00 3.61 3.78 2. See Table 5.26 3.76 4.85 3.5 3.23 3.16 7.60 2.09 8. the fish is dry-salted.17 4.5 3. This is why it is the preferred method for fatty fish.0 2.1.44 2.45 3.96 1. herrings and other fatty fish can be used. after being drained and pressed at 465 kg/m2 for 15 hours.04 5.03 2. Here. the brine is then replaced over the fish. There are several variations of this process. SMOKED AND DRIED FISH smoking.78) (1). a saturated brine is used. they can be stored at 28 °C. the product obtained is called gaspé-cured (5). Fresh sardines.61 3.98 6.41 5.51 5.11 9.73 1. the fish enzymes play an important role by maturing or ripening the product.26 4.5 4.98 6.06 4.30 2.99 2.41 6.65 2.26 7. and can be collected and replaced again over the fish. but mackerel.23 8.33 5. TABLE 5.26 1. if split fish is dry-salted. 5.76 99 .23 2.76 4.41 4.74 9.19 3.25 5.56 6.11 3.91 2.09 8. The main objective of the brining method of salting fish is to limit air penetration during processing.44 2.78 6. sardines. Histamine formation is of importance when using scombroid and some clupeidean species.05 7.CURED. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .95 3.98 5. headed and gutted. thus.41 3.35 4.03 4. but as it is placed in containers.3 Dry / wet salting In this process.64 4.54 5.76 4. Histamineforming bacteria do not generally grow at water activities below 0.23 2.10 1.88 5.

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

marinated products will keep for several months at a temperature of 4 °C.CURED. split and dried gadoid species) are traditionally made in cold climates. gravad fish. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. under a light pressure. Stockfish is extensively used in Portugal. it will last for many years (5). the fish is dried after salting or brining. In Scandinavian countries. Usually. mixed with salt. It takes about 2 days to marinate completely. and a fine product is obtained. South America and the Caribbean. some fish species such as moray eel and varieties of dog fish. such as squid. bacterial toxins and parasites (7). Foodborne pathogens. used as an appetiser.) where no acid is added to the process. They are an important dried product. which are placed in a mixture of salt. These can be considered as ‘lightly preserved’ products with a salt content <6% sodium chloride (w/w) in the water phase and low acid content (pH >5. In the first case. after frying. The most common method used for fish products is the air-drying process. a very fine delicacy for Oriental food lovers. and the flesh will break down. jellyfish. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). Stockfish products have a golden brown colour and are very hard. the fish is dried naturally in a relatively cold environment. 101 . Dried shrimps are widely used in Asia.3. Stockfish (dressed. such as Listeria monocytogenes and Clostridium botulinum.There are several products named marinades (matjes. It is important to assure the quality of the raw material. air flows naturally. Shark pectoral fins. At pH levels below 4. etc. Fresh dill is placed between the layers of fish.0) (7). and the stacks are placed in the refrigerator. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. sugar and spices. In some coastal areas of Portugal. the temperature and the amount of pressure applied (5). Provided the product is kept in dry and fairly cool conditions. Matjes are made of fat young herring. are sundried and. Dried octopus is also a very common dish in the interior of the country. sea cucumbers and seaweeds. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins.4 Air drying This is probably the oldest method of fish preservation. are generally air-dried at sea. Other products. are commonly dried in Asian countries. Some bacteria and enzymes are still active in the presence of acid and salt. but sometimes it is dried without prior salting. 5. by using either solar heaters or mechanical dryers. gutted. Gravad fish is a Scandinavian product made of salmon fillets. depending on the thickness. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). are likely to survive in these mild-processed products. Italy and Spain (5).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). still made without prior salting on a large scale.

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

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

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

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

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

and substances unconnected with smoke but with a smoky flavour or smell. including Eurotium repens and Eurotium rubrum. 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). Vacuum packaging retards the onset of rancidity when fatty fish is used. but mechanical drying is used instead of sun drying.3. Natural smoke extracts. (Penicillum cyclopium and Penicillium puterellii) (19). Aspergillus oryzae. but seems. such as yeast derivatives. Hot-smoked products made from white fish will usually keep better than those from fatty fish. They are generally not cooked prior to consumption. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. Eels. 5. 18). Aspergillus ostianus. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. apart from the improved appearance. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. 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). Katsuobushi. 17. Growth of these moulds on the tuna results in the extraction of moisture. a very expensive and fashionable smoked product in the Netherlands. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. are usually hot-smoked after being immersed in 80º brine or dry-salted (6).3½ hours using the typical temperature set for hot-smoking referred to above (12). to have no further advantage over other packing methods for other fish (12). resulting in a characteristic flavour (20. rewetted and remoulded to give the desired shape and consistency. The production method is similar to masmeen. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). When refrigerated at 3 °C. are usually 80º brined for 5 . break down of fats and the moderation of smoke flavour. similar to masmeen. 21. 22.5. are used. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. cooking in brine. Mackerel. including ‘umami’ (16.CURED. white fish about 8 days. Aspergillus tamarii) and several Penicillium spp. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). The product is removed from the drying cycle. then hot smoking and finally sun drying it. In terms of safety. Boiled or steamed clams. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. synthetic smoke flavours. chilled or freeze-thawed. moulding it into long thick sections. The classical liquid smoke is 107 . they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12).3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. 23). They will keep well frozen. and many closely related Aspergilli (Aspergillus ochraceus.6 hours and smoked for 3 . fatty fish will remain in good condition for 6 days.

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.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. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. The synergistic effect of liquid smoke with other preservatives could reduce the amount of salt in the product when light diets are necessary whilst still meeting the necessary safety criteria (26). It has been shown that liquid smoke can reduce the formation of PAHs (3). both of which are considered potential carcinogens.8% and still provide protection for 7 days (25).6 to 2.4 Spoilage Brining. 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. or applying it as an atomised spray within a modern automatic smoke kiln. In experiments comparing traditionally smoked fish with liquid smoke. nutritional neutrality and antioxidant activity (27).3. Liquid smoke was considered to provide advantages of safety. Fresh fish spoilage may be measured by 108 .FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). drying. According to some recent research.03 μg/kg of benzopyrene. When liquid smoke was used. the salt level could effectively be reduced from 4. 5. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. so as to impart an unnatural red colouration to the flesh. 5. and less than 0. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25).06 μg/kg of benzoanthracene in the final foodstuff (28). or smoke extracts in organic solvents. applying it as a dip after brining. smoking. 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. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine.5. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. injecting it into the fish flesh. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). consistency of flavour and colour. The European Scientific Commitee for Food.

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

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

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

IV Spoilage of salted. halophila Halophilic spoilage organisms Halophilic spoilage organisms. boiled.variable.not used/unchanged. 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. LAB.FISH AND SEAFOOD TABLE 5. . sebi and B.W. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. + to +++ strength of preservative effect 4 Post-process contaminants 112 . smoked and dried Citrus marinade Dried Brined or salted then liquid or cold smoked Brined or salted then liquid or hot smoked 1 2 +++ Jellied eels + Ikan kaju Ceviche Stockfish Coldsmoked fish Hotsmoked fish +++ +++ ++ +/++ + + ++ ++ - ++ + + + ++ ++ ++ ++ + ++ + ++ Antimicrobial compounds added Refrigeration applied 3 v. Dun. and Halobacterium spp. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted.

under optimal conditions. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C.75 0.8 1. aureus Yeasts Moulds Lactic acid bacteria pH 5.5 0-11 3.86 0.8 0. The concentration required to prevent growth at room temperature can vary from as low as 3. botulinum type E. smoked and dried. the importance of cold-chain integrity must be emphasised.5 5. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). 9 and 8 of the outbreaks. the products were heavily salted.5-8. with this change. Before. In the United States. could not grow and produce toxin.5% to 5%.9 0. and as botulism is a serious hazard in foods. the fermented products being the most dangerous group (7). SMOKED AND DRIED FISH TABLE 5. salted and pickled products accounted for 10.5 4. botulinum Staph. However.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. types A and B are not affected by 3 .0-9. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. but if the fish products are stored at temperatures <10 °C. the non-proteolytic type E strains can grow in up to 5% sodium chloride.97 0. The presence of salt in cured products has a great effect on the growth of the bacteria. respectively. when present in the raw material. although the combination of the two factors. pressure on the traditional methods of curing. and C. It has been shown that. popular demand for healthy foods has led to a change in the tastes preferred by the public and.5. sodium 113 .5-10 4. botulinum type E Staph.97 0.5 5.CURED. botulinum. Since neither the smoking nor the drying parts of this curing process are particularly severe. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E.5-8. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). Examination of 165 outbreaks of botulism caused by fish products showed that smoked.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13).5-10.61 Organisms Most Pseudomonadaceae C. botulinum type E and production of toxin.

5. if storage temperatures are over 10 °C. they do not grow well in the raw material. if it contains sodium nitrite ≥100 ppm.0 . more importantly. and the minimum pH for growth is 4. but. However. when the bacterial load of the product is lower. In foodstuffs.5. The organism is mesophilic with a minimum growth temperature of 7 °C. the organism may contaminate the product. the organism can survive most of the light curing processes. has been reported (42). It is a halotolerant bacterium able to grow at water activities as low as 0. the risk of infection can be eliminated by proper cooking. 5.4 Listeria monocytogenes The psychrotrophic L. after processing.5% salt in the water phase. coli all occur on fish products as a result of contamination from animal/human reservoirs. throat and skin. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. growth and toxin formation may occur.94 (7). as they require more handling. gills and guts as well as in several lightly preserved fish 114 . 5.or cold-smoked fish should have at least 3. but. As with other organisms. if the person handling the product is a carrier of Staph.86 and 10% sodium chloride.5. but requires higher temperatures for toxin production (>10 °C).5. aureus growth. if recontamination occurs and storage conditions are favourable. aureus. Sliced smoked fish products. 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.3 Enterobacteriaceae The pathogens Salmonella. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). it should contain not less than 3. Infection by Shigella sonnei has increased in the UK. As they are poor competitors. The FDA established that hot. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. Usually. Shigella and E.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. Once the product is contaminated. related to the consumption of a raw smoked fish meal.30% of the population positive for enterotoxin-producing strains (41). Thus.4.5.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. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . the human nose. An outbreak of Shigella in an Italian cruise ship.0% salt in the water phase (30). are particularly susceptible to contamination and. they will grow rapidly (7).

a short period of freezing either of the raw material or of 115 . when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). monocytogenes in levels at or above 102 per gram (46).8 . 5. Some LAB. SMOKED AND DRIED FISH products such as cold-smoked fish. Its limiting conditions for growth are 0 . The use of LAB in fish preservation is a topic that has been studied in projects (UP. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. and in CT95-1207FAIR). a number of fish products. Unlike some European countries.92 . have been shown to have an anti-listerial activity in many food products. and other local traditional products (7). cold-smoked fish. The Health Protection Agency (HPA) in the UK considered that it was unacceptable for readyto-eat foods (including all the products that are consumed without prior cooking) to contain any serogroup of L. The round worms. which. in combination with curing agents and refrigerated storage. marinades (ceviche) (43) and gravad fish (44). They have complicated life cycles.5 Parasites Despite the presence of parasites in fish being very common. Of microbiological concern in the northern hemisphere are two parasites . from freshwater fish.45 °C. which include intermediate hosts.5. do not survive 1 min at 60 °C.9. lightly salted caviar. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. the USA established ‘zero tolerance’ for the organism. or nematodes. and the tape worm Diphyllobothrium spp.12%. monocytogenes’ in 25 g of the finished product. gravad fish. the CT 3162-FAIR program.CURED. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. such as ‘lightly preserved’ fish products. the only concern to public health is when eating raw or uncooked fish products. Until 1997. which means that the process must conform to a level of ‘no detectable L. This includes matjes herring. both being detected in cold-smoked salmon. favours growth of psychrotrophic LAB. Pseudoterranova dicipiens. Thus. Two outbreaks in the United States have been reported to be linked to salmon (10).2. a round worm. As they are heat-sensitive. are considered unsafe. in the EU Fisheries R&D-FLAIR program. there were no reported outbreaks related to cured fish. salt levels of 8 . widely used as starter cultures in food processes.0.95 (45). which considered L. has also been detected in cod (7). most of them are of little concern to public health. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. monocytogenes pathogenic for only specific segments at risk.the round worm Anisakis spp. from seawater fish.514. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48).6 and water activities below 0. pH 4.

These foods fall into category 5.<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 >100 N/A Pathogens Salmonella spp. coli O157 & other VTEC Vibrio. Acceptability is based on appearance.<100 10 .FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). † Guidelines for aerobic colony counts may not apply to certain fermented foods.VI Guidelines for the microbiological quality of ready-to-eat smoked fish.<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . E. (not L. perfringens B. monocytogenes Staph. the organism should be identified # N/A denotes not applicable 116 .<100 20 .<104 100 . smell.<104 104 .6 Published Microbiological Criteria TABLE 5.<104 20 . salami.<100 >104 >100 N/A N/A <20 20 . ‡ On occasions some strains may be pathogenic. soft cheese. for example. texture. cereus and Bacillus subtilis# <20 <20** <20 <10 20 . cholerae Vibrio parahaemolyticus L. If the Bacillus counts exceed 104/g. taramasalata and cooked shellfish sampled at the point of sale (adapted from (11)) Criterion Satisfactory Borderline limit of acceptability (cfu/g unless stated) Unsatisfactory Unacceptable/ potential hazard* (cfu/g unless stated) (cfu/g unless stated) Aerobic plate count† (30 °C. coli (total) Listeria spp. and the levels or absence of indicator organisms or pathogens. aureus C.<107 ≥107 N/A <100 <20 100 .<100 200 . 5.<100 20 . and unpasteurised yoghurt. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E.<103 N/A 100 . monocytogenes) (cfu/g unless stated) <106 106 . not detected in 25 g Campylobacter spp.

Torry Advisory Note No. Fernandes R. 1984. in Micro-Facts: The Working Companion for Food Microbiologists. FAO Fisheries Technical Paper. 3273. Huss H.. 117 . Torry Advisory Note No. Leatherhead Publishing. Martin R. Marinades. LT or ST positive Presence of the organism Presence of the organism Positive for enterotoxin or ≥104 (Most Probable Number) Presence of toxigenic 01 or non-01 ≥ 1 x104 (Kanagawa positive or negative) Presence of the organism Presence of viable spores or vegetative cells in products that will support their growth. coli (ETEC) L. in Fish Processing Technology. 7. Flick G. Eds. 4. or presence of toxin 0.. Eds. 1997. Leatherhead. 1991. parahaemolyticus Vibrio vulnificus C. Doré I. 1993. Moody M. New York. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. Wareing P. salting and smoking).M. USA. aureus * Vibrio cholerae V. Regenstein C. New York. 1990. 1994. 8. 37(revised). 381-406. Bannerman A.. botulinum * Limit of tolerance 1x 103 ETEC/g. Horne J. Wareing P. 2. except in the viscera of dungeness crab.M. Hall G. 6. Ed. Preservation of fish by curing (drying.A. McLay R.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. Smoked. Chapman & Hall. Fernandes R. cured and dried fish. References Regenstein J. 9. 120-38. Introduction to Fish Technology. Flick G.CURED.F. Assurance of Seafood Quality. Van Nostrand Reinhold. monocytogenes Salmonella spp. 1982. 334. London.W.H. The Smoked and Cured Seafood Guide.. 3.E. Food spoilage fungi. Torry Advisory Note No..F. Horner W. SMOKED AND DRIED FISH TABLE 5. 56.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. Catching. Urner Barry Publications Inc. 1979. 82 (revised). in The Seafood Industry.. 254-82 5. Handling and Processing Eels. * Staph. 2007. Van Nostrand Reinhold.7 1. Birnie K. Hot Smoking of Fish.

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

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

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

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


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

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. where the bacterial production of lactic acid is a major feature.II. This is well exemplified by some of the fish/salt products of Thailand described in Table 6. In Europe. made from fish livers in Greece. a variety of products can be produced within a single country. made from whole fish in France. 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. Fermented fish products are of very minor importance outside Asia. 124 . who had adopted them from the Greeks. salty taste.1. may be direct descendants.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. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. A selection of Asian products is presented in Table 6. and pissala. to products analogous to other lactic fermented foods such as salami.FISH AND SEAFOOD TABLE 6. although garos. 6.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. These have long since fallen from use.III.

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

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

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

1. 6. Belacan (Condiment) Fig. Fig. 6.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 .2. Patis (Condiment) 128 .12).

Ika-Shiokara (side dish) Fig.4. thought to be largely the result of autolytic breakdown of the fish tissues. the 129 . In fish sauce production. eviscerated fish are sometimes used to slow the autolytic process. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are.3. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. however.FERMENTED FISH Fig. 14). 6.6. 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.

6.5. Some examples of products and their formulation are presented in Table 6.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.V and Figures 6. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur.8. 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. volatile fatty acids and nucleotides. particularly glutamate. snack) 130 . for example. filtered and bottled.3.6 – 6.5). In contrast to free-swimming fish.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. which is eventually separated from the residual fish tissues. Fig. Hoi-Malangpu-Dong (side dish. Figure 6. 6. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens.

NE. C NE. central. C. C NE. south of Thailand. 3:1:0. N.FERMENTED FISH TABLE 6. In these cases.2-4 Fish (pieces): salt:khaomark (fermented rice). north. N.12 days 3 . the prefix 'pla' is replaced by 'kung'.12 months Main dish 2 Pla-jao C.6.12 months Shelf life Use 1 .4 are also produced using shrimps. N S 5 . Kung-Chom (staple food. NE Formulation Fish (whole or pieces): salt: roasted rice. N.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 . side dish) 131 .V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . Fig. NE 10 .25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. S.7 days 5 .5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0.03 Preparation time 6 . Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus.6. north-east. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0.5-1.

If acidity is a desired sensory property of the product.salt and lactic acid.10%. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. This is readily explained by the susceptibility of LAB to elevated salt concentrations. these products are salted at lower levels than the fish sauces and pastes. giving a reduced rate of acid generation.VI).3.7. the salt is of far greater importance in determining the product’s long-term stability. but there is considerable variation in this. of the two factors contributing to shelf life .FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. typically in the range 2 . then this will take longer to develop than if lower salt levels are used. It also indicates that. Generally.6.2. Red burong isda 132 . (from Arroyo et al. 2.1 The use of higher salt levels results in a longer production phase but a better keeping quality product. 106) Fig. 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.. The LAB isolated from these products are those with more marked salt tolerance but. Phillipines Journal of Food Science and Technology. their growth and metabolism are nonetheless slowed. while they are able to grow at high salt levels.

2 7.2 4.8 2.4.9 4.6.0 Shelf life 3 weeks 6 .5 .5.4.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.8 .9.FERMENTED FISH (from Arroyo et al.1 4. 106) Fig.0 .5 3.5. White balao-balao TABLE 6.5 .9.9 .9 4.2 4.3 5. 2.8 .8 pH 4.3 . 133 . Philippines Journal of Food Science and Technology.5.4 .6 3.5.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).6.6.12 months 1 .8.1 .7 .0 .17.0 .5.3 years 1 ..

a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. of course. 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 . Garlic.3. For example.3. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. 21). This results in a more active lactic fermentation and a lower pH.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi.5) despite other factors. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. Rice is the most common carbohydrate source used. Addition of carbohydrate will. inulin.FISH AND SEAFOOD 6.0 . efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. When added as boiled or roasted rice alone.4. In several products.VII).9) compared with the white product (pH 4.1 .2 Inclusion of more carbohydrate gives a faster fermentation and a stronger acid taste Increasing the proportion of carbohydrate in the formulation decreases the product’s buffering capacity and provides the LAB with more substrate on which to act. such as salt content. 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). Where they are used. TABLE 6. also lower the unit cost of the product and this is probably a significant factor in many cases. being similar (18. Although the ability to ferment starch is not widespread in the LAB. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. although the final population of LAB may be similar. a common ingredient in these products. also serves as a valuable source of the more readily fermented carbohydrate. alternatively. the rice may be added in a partially saccharified form such as kao-mark (Thailand). 19).2. 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.

Traditional packing techniques used with lactic fermented fish products. markedly extending the shelf life (25). The mixture is allowed to ferment for 2 .5. Clearly.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). presumably correlating with the salt content. but they are likely to behave similarly to lactic fermented products produced from other commodities. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24).5:9:3. such as wrapping in banana leaves.9%. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others.FERMENTED FISH Sikhae.2 at the end of fermentation.7 to below 4. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB.8 .3 days. reporting the product’s pH to range from 4. in turn. Some authors have claimed that the function of the roasted rice added during pla-ra production is to improve texture and slow the fermentation rather than to serve as a substrate for lactic fermentation (22). The growth of yeasts and moulds could. Results of a product survey were consistent with this. 6.2. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage.3 weeks at around 20 °C. the level of salt employed is on the high side for fish/salt/carbohydrate products. 135 . The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). Generally.5 in the first 2 . is something of an exception since millet is used along with garlic as the carbohydrate source.7 up to 6. and there is a marked regional variation in the preferred type. around 12%. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. Excessive growth and acid production by the LAB could render products too sour for acceptability. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. the Korean fermented fish product. There is little published information on the factors that limit the shelf life of the low-salt products. In a successful fermentation. will not reliably exclude air. allowing other spoilage organisms to grow. thereafter declining to about 4.17. lead to an increase in pH. the pH drops from 6. which ranged from 7. 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.

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

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

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

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

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

the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. 5). It remains to be seen if this approach gains momentum in the future. physiology and detection of marine and freshwater toxins has also appeared (8). FISH AND SHELLFISH TOXINS Jeffrey L. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. Nevertheless. Although most of these antibody-based test kits have yet to receive international regulatory approval. a number of new toxins and toxin families have been identified. Another book dealing with the clinical effects of a variety of poisons and toxins. Scientific and Cultural Organisation) has been updated (7). An emerging technology is the use of immunosensors (1 . Ph. Increasingly sophisticated instrumentation (e. several scientific reports have described their efficacy and potential. Since the original chapter in this series was published (6). and a second updated edition dealing with the pharmacology. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. The manual on harmful marine algae and their products. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins.M. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue.C.D. While some of these toxins only occur in seafood from a restricted geographical region.. most notably the azaspiracids and the cyclic imines as well as some additional polyethers.. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7. C. Wright.g. including the well known marine toxins 141 .1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins. known and unknown.3) and some practical applications for detecting shellfish toxins have been reported (4.7. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational.

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. 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. All PSPs degrade rapidly in alkaline conditions.g. Many of these species are extremely common and are found throughout the world in a variety of cold. The Ncarbamoyl sulfates are readily hydrolysed to STX. can become contaminated with these toxins. The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13).1). Around twenty PSP toxins are known. Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. some are the result of biotransformation processes by the filter-feeding host. Gymnodinium catenatum. and a tropical species Pyrodinium bahamense (16). Consequently. In addition. even crustaceans such as lobsters. all of which contain a tetrahydropurine skeleton (Figure 7. 7. a great variety of shellfish from all latitudes. dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). 7. temperate. Saxitoxin Group instead of the Paralytic Shellfish Poisons). 142 . A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). with intoxications occurring in many countries even today (15). PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. and a review on the risk assessment of marine toxins has also been produced (14). and sub-tropical waters and habitats (15). and another. Not all the toxins found in shellfish extracts are microalgal products.FISH AND SEAFOOD has been produced (9). neoSTX and Gonyautoxins (GTX) 1-4.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium.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. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10).

Human symptoms begin with tingling and numbness of the lips. and fingers within 5 . 7.2. tongue. However.1. PSPs bind to site 1 of sodium channels reversibly blocking sodium conductance in nerve and muscle membranes by binding to a specific receptor site located on the outside surface of the membrane close to the ion channel opening. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis.30 minutes of consumption. and humans seem to be particularly susceptible. Neurophysiological studies have shown that PSP toxins all possess the same mode of action though some derivatives such as STX and neoSTX are considerably more potent than the C-toxins (15). 143 .FISH AND SHELLFISH TOXINS STX GTX2 GTX3 GTX5 (B1) C1 C2 NEO GTX1 GTX4 GTX6 (B2) C3 C4 R1 H H H H H H OH OH OH OH OH OH R2 H H OSO3 H H OSO3 H H OSO3 H H OSO3 - R3 H OSO3 H H OSO3 H H OSO3 H H OSO3 H R4 H H H SO3 SO3 SO3 H H H SO3 SO3 SO3 - Fig.2 Toxicology PSP toxins affect the central nervous system of mammals. this is never sufficient to render the seafood safe to consume. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7.

and has also been evaluated by a number of EU laboratories. Since its discovery. 17). and while simple and relatively inexpensive to operate. After some development and various trials. Two approaches have been taken here. China. Taiwan. Most intoxication events have occurred in South East Asia. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. 23. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19).3. and islands of the South Pacific.2). 24) (Figure 7.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. 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. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0.2. However it remains as the monitoring method of choice in many countries. other derivatives have been reported. including Japan. Alternative HPLC methods have subsequently reduced this to a single HPLC run. 15. specifically 144 . While the chemical detection methods are most promising for the future.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised.FISH AND SEAFOOD 7. TTX most commonly occurs in many species of puffer fish. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). a criticism has been that such methods are time consuming. and over ten analogs are known (22. 7. The mouse bioassay has been in place for decades and the method has been standardised. the detection limit (4 µg/g) of the method is close to the safe level and does not provide any information on the profile of PSP toxins present in the extract.8 µg/g) of shellfish or seafood tissue. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). as well as other parts. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. 7. a pre-column method has received Association of Analytical Communities (AOAC) approval (18). Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. Thailand. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives.

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

FISH AND SEAFOOD column oxidation to the fluorescent derivatives. 7.3 for examples). need further development (24). However this method has been criticised for being complex. 7. and while promising. a polar glutamate agonist.3. 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). Immunoassay methods have also been investigated. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule.4. but it is known that such congeners are less toxic than DA itself (30). but all of them are considerably less toxic than DA itself (29).4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). Fig. 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). Currently 12 isomers of DA have been reported (see Figure 7. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. but 146 . Structure of domoic acid and three naturally occurring photoisomers 7. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28).

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

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

are also recognised as a threat to human health. which include the pectenotoxins and the yessotoxins. Dinophysis caudata. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. In this case Dinophysis was suspected as the culprit organism. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). Fig. Structure of okadaic acid and the two related DSP toxins 7.5. 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. 7. 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. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. Today over ten Dinophysis spp. and include Dinophysis acuminata.4. 149 .FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. and Dinophysis norvegica among others (47. In comparison. such as Prorocentrum lima..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. Dinophysis acuta. Dinophysis fortii. DSP toxins produced by benthic Prorocentrum spp. This was probably not the first DSP episode. Lately these compounds. and Prorocentrum belizeanum. are recognised as DSP toxin producers. Prorocentrum maculosum. 49). The parent acids are the only DSP congeners to be found in shellfish tissue. DTX-5. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. While these esters are not toxic in vitro.

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

However. Fig.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . Indeed.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). 7.5. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. for many years these compounds were included in the DSP toxin complex. Some representative examples of both groups of toxins are shown in Figures 7.5 and 7. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). Some doubt remains as to the significance of these compounds as marine or shellfish toxins with respect to human health since the PTXs for example are co-extracted with the DSP toxins. and another rapid antibodybased test kit for DSP toxins has also been reported (53). but with the increased availability of pure toxins permitting detailed toxicology studies it is now recognised that both these groups of compounds possess a different toxic profile from the DSP toxins. 7. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. with good agreement with the mouse bioassay and LC–based methods (54. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. 58). with a claimed detection limit of 20 µg/kg.6. If a European collaborative study of the fluorometric protein phosphatase inhibition method is successful it may eventually be approved for regulatory purposes in the EU. In yet another approach. 55). and the latter group are usually found to be responsible for any reported shellfish toxicity (59). Within this group.

While 152 . 7. which may co-occur in blooms with other organisms.6. YTX and homo-YTX 7. eastern Canada. Japan. PTX-1 and PTX-2 are frequently the most common derivatives to occur and PTX-2 is the most potent following intraperitoneal (IP) injection (230 µg/kg body weight). many parts of Europe and the Mediterranean. 58).2 Toxicology PTXs show varied toxicity in the mouse bioassay (59).6. As is typical for many algal species. 7. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. the toxin content and profile may vary among different strains.FISH AND SEAFOOD Fig. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum.1 Origins and distribution It is now well recognised that PTXs are co-produced with DSP toxins by various strains of the dinoflagellate Dinophysis and hence the distribution of both families of toxins in shellfish is similar (49). 57. Structure of the two parent yessotoxins. New Zealand and South America (49.6. 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.

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

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

this family of toxins has proven to be a challenge both to identify and monitor. and mammalian body fluids without pretreatment or dilution (72). While the mouse bioassay continues to be used. The mean time to onset of symptoms is around 3 hours. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73).7. Ingestion of brevetoxins results in a variety of symptoms including parathesia. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. vertigo. nausea. 66. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. the competitive ELISA method correlated most closely with the mouse bioassay results.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. which can result in a cascade of associated events in a cell (66). In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. malaise. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). 69). A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater.7. and diarrhoea.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. with hot spots in the coral reef areas of the Caribbean and French Polynesia. 7.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). In this application. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . shellfish homogenates. The human health implications following exposure to NSPs have been reviewed (69). 7. with a mean duration of symptoms of around 1 day. and in severe cases seizure. and an action level has been set at 800 µg/kg (or 80 µg PbTx-2 equivalents/100 g or 20 MU/100 g or 4 µg/mouse) of shellfish tissue to trigger regulatory action. The mouse bioassay is used as the primary detection method. 7.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68).

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

there are neither standards nor an official testing program. Patients have reported symptoms lasting weeks and even months. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . the neurological symptoms can persist for days. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. barracudas. 75). Local island populations attempt to avoid poisoning by catching only smaller fish. all of which can be linked to the effect of CTX on excitable membranes. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. and there are suggestions that such incidents are increasing and spreading worldwide (74).1 µg of ciguatoxin can result in human illness (74. arthralgia. myalgia. 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. where cold objects feel hot and vice versa. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. Other symptoms include a metallic taste. which in turn are consumed by carnivores such as snappers.8. only trace amounts of CTXs can cause illness following consumption of contaminated fish. although not all CTXs are equipotent. which can return following the consumption of alcohol and certain foods such as oily fish (74. and although the gastronomical problems may dissipate within a day or so. The onset of symptoms can occur rapidly within 24 hours. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue.8. and it is estimated that ingestion of as little as 0. 7. and the sensation of loose teeth (77). groupers. Even when suspect samples are available. which can be confused with chronic fatigue syndrome. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. and temperature sensation reversal. 75.76). Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. These include paresthesias (numbness and tingling of the extremities).3 Detection methods As a consequence of their potency.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. Like the brevetoxins. 78). and in fact CTX poisoning is often only identified after consumption of contaminated fish. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. Another symptom that can develop over time is extreme fatigue. pruritus. and moray eels. sharks.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. weeks or even months (74 . Globally. 7. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective.

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

oysters (Crassostrea gigas. cockles (Cerastoderma edule). Ensis siliqua. and to date this has been the only source from which these toxins have been found at toxic levels. AZA occurrence has been reported by several European countries including France. Portugal. There has been considerable debate over the source of azaspiracids. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). diarrhoea and stomach cramps. Structure of the three principal azaspiracids found in shellfish 7. as in other cases of shellfish toxins. cassipes (83).9.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. cassipes produces AZAs in culture. Following the original episode. scallops (Pecten maximus).9. clams (Tapes philipinarum. However. and to date no long-term effects have been recorded (83. 84. 7. which can persist for 2 . Scandinavia. These symptoms are not unlike those experienced following the consumption of DSP toxins. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp.9. and some time after the original episode in Ireland. 90). In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. Donax spp. 7. and off the coast of Morocco (83. but current data indicates that unlike the DSP compounds. Mytilus galloprovincialis). it has been impossible to establish that P. The bivalves affected include mussels (Mytilus edulis.FISH AND SHELLFISH TOXINS Fig. and most recently in a crustacean (Cancer pagurus). which are ingested by P.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea.). Ostrea edulis). While this result is not necessarily in debate. AZA toxins do 159 .3 days. these molluscs were never considered to be the biogenetic source. 88. 89). vomiting.

Importantly these pathological outcomes were reported as being different from those caused by DSP. AZAs also display cytotoxicity towards various mammalian cell lines (83. 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. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . Once again. Based on IP minimum lethal dose data. and necrosis of epithelial cells on the microvilli (91). 91. though the possibility exists that they may inhibit other phosphatases.10 – 7.10 Cyclic imines These toxins are represented by several distinct structural types. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). abnormalities in the thymus and spleen. the pinnatoxins. 85). Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. 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. 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. The mouse bioassay has been used to monitor shellfish extracts. 7. but all contain a spirocyclic or cyclic imine function within their structure. which are particularly amenable to detection by positive ion electrospray mass spectrometry. 7. 92). and closely related ptierotoxins (Figures 7. PSP. and erosion and bleeding in the stomach. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. However. parenchymal cell pyknosis in the pancreas.12). it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. and ASP. the spirolides. 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. 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).9. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. Using the same approach.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue.

Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94. Subsequent taxonomic studies revised this again to a new dinoflagellate K. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis).520 Da (94.10.1. At the time of the initial observation of gymnodimine.10). but they are included here for completeness. they are sometimes referred to as emerging toxins of uncertain human health concern. cockles.10. 95). 161 . Fig. 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. selliformis (97).1 Origins and distribution The original observation of toxicity in oysters led to a wider survey of other shellfish species in New Zealand when it was discovered that the toxin was present in a variety of bivalve molluscs including mussels.10. 7.FISH AND SHELLFISH TOXINS health. In only two cases (gymnodimines and spirolides) has a planktonic source been identified. Some time later. Structure of the cyclic imine gymnodimine 7. Cyclic imines are often referred to as fast acting toxins due to their “all or nothing” effect in the mouse bioassay over a very narrow dose range. 95). a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin. it was proposed that a dinoflagellate identified as Gymnodinium spp. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7.. scallops. two additional congeners. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . surf clam and New Zealand abalone. Pacific oyster.

and there was a further ten-fold reduction in toxicity when the toxin was supplied with food.900 µg/kg).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. 50%) of 96 µg/kg. However. Oral administration of the toxin to mice by gavage resulted in about an 8. Based on this diminished oral toxicity.FISH AND SEAFOOD 7. finishing with 100% methanol.11). The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. paralysis of the hind quarters and eventually complete immobility. Whether isolated from dinoflagellate cells directly or from shellfish tissue.10. and this is the detection method of choice. it has been stated that regulation against gymnodimine is not required (98).100). 7. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. together with a lack of evidence of human intoxication.3 Detection methods The characteristic “fast-acting. 95) (Figure 7.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. The time lapse between ingestion and death is between 5 and 15 minutes.1. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. Since their original discovery which resulted in the characterisation of spirolides A-F (99.1. There is a rapid onset of symptoms including respiratory distress. Most cyclic imines lack an extended chromophore. and so spectroscopic detection by UV methods is not favoured. 95). 7. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. They were originally discovered in extracts of mussels (M. Reduction of the imine group eradicated the toxicity. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction.10. and lack of responsiveness to stimuli. A later study using purified toxin established an LD50 dose (Lethal Dose. abdominal breathing. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. almost a dozen derivatives in total have been identified to date (94.to10-fold reduction of toxicity (LD50 600 .2 Toxicology A review of the toxicology of all cyclic imines has been published (98). The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . In particular.

the gulf of Maine. fasted mice).707 µg/kg.10. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101).10.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. 7. 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. 95). southern Ireland. Structure of four spirolide toxins A-D 7. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. or by oral ingestion (381 . these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 .8 µg/kg). Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. fasted mice). 7. Fig. the western coast of Scotland.166 µg/kg.2.11. Interestingly. While oral toxicity is considerably less than by IP injection. in fjords in Denmark and Norway.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). All data reported here are for desmethyl spirolide C. by gavage (87 .2. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98).11.2 Toxicology The spirolides are the most studied group of the cyclic imines. Structural differences among the various spirolides result from variations in the number of methyl groups and the sequence of spiroketal rings 6-5-5 in one group and 6-6-5 in the other.

The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7.3 Detection methods As discussed for gymnodimine. 7. consistent with modulation of acetyl cholinergic receptors (103). 102). which includes two diastereomers.B contain a spiro imine moiety similar to that found in the spirolides.10. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry.12).10.FISH AND SEAFOOD (100).2. A number of methods following this procedure have been described (94. Three members of this group have been reported. four members of the group have been identified – two of them being the C-34 diastereomers. 95.3 Pinnatoxins and pteriatoxins Pinnatoxins A . However. While the mechanism of action remains to be clearly defined. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . 7. To date. 7. Fig. and in fact these compounds share a number of other structural features. the mouse bioassay can be used to detect these fast acting toxins.12. 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. The compounds are zwitterionic and therefore somewhat more polar than the spirolides.

new toxin groups such as the azaspiracids and cyclic imines have also been reported. which occupy tropical waters around Okinawa and the south east coast of China.10.10.3. However.most likely a dinoflagellate. and more advanced means of detection.10. With increasing knowledge of the chemistry of toxins. but this remains to be conclusively established (106). 7. Coupled with this.FISH AND SHELLFISH TOXINS 7. which in many cases are now found to be more widespread than previously believed.3.2 Toxicology There is little information describing the toxicology of these compounds (98).11 Conclusion There has been considerable progress in the detection of known marine toxins.1 Origins and distribution Shellfish of the genus Pinna. like the gymnodimines and spirolides. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). Only the acute toxicity in mice following IP injection has been reported. there is no compelling evidence to suggest that they have resulted in human toxicity (98). The LD99 (Lethal Dose. This is generally a result of increased monitoring and continually improving detection technologies and methods.3. 7. While this is recognised as a real possibility for a wide range of toxins. arise from an algal source . The LD99 dose for the pteriatoxins is reported to be in the range (8 . while pinnatoxin D was essentially inactive.180 µg/kg). but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. research and certified standards of the key toxins are often unavailable or in short supply and in the future considerable logistical effort will be required to address this issue. 7. There is strong circumstantial evidence to suspect that these toxins.3 Detection methods No formal detection methods for these compounds have been reported. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). have long been suspected of causing human illness after ingestion. Despite this progress.100 µg/kg). to date action or 165 . and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. 99%) values for pinnatoxins A-C are in the range (22 .

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

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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.


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

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

processing conditions (e. naturally present on fish from warm waters (> 15 °C). aquaculture drug residues and potentially pathogenic Vibrio spp. If the processing does not include a kill step. etc. storage and distribution conditions and instructions for use. 8.3. 8.3. may need to be considered specifically. physical and chemical structure (including water activity (aw). 8.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. In addition. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . Vulnerable groups of the population. However. pH. or in aquaculture operations. such as enteric viruses in shellfish. packaging. The description should include information such as the product name.).g. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation.). heat treatment. These factors are likely to have a significant effect on some potential hazards.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.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. 8. Expertise on such inherent hazards. It is also important to identify the consumer target groups. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. contaminated fish and shellfish should not be used. shelf life. marine toxins associated with particular fish and shellfish species.1.3.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.3.g. etc. fermentation.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments. there may be species-specific hazards related to certain fish and shellfish. composition. freezing. is a cooking process required?). for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. such as children or the elderly.3. including practices onboard fishing vessels. which should be described in detail.3.2 Describe the product It is important to have a complete understanding of the product.

If HACCP studies are applied to proposed new process lines or products. 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. 180 ● ● . then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. it is essential that the details of each step are fully appreciated and recorded. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects.3. It is essential that the diagram is accurate. It may include details of the factory and equipment layout. For example.3.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. 8. ingredient specifications.6 List all potential hazards associated with each step.3. or circumstances of any reclaim or rework activity.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. and therefore a detailed description of the procedure applied must be available. 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. The operation should be observed at all stages. conduct a hazard analysis.FISH AND SEAFOOD product. cleaning and hygiene procedures and storage conditions. 8. because the hazard analysis and decisions regarding CCPs are based on these data. However. features of equipment design. it should also include details of CCP steps. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. Ideally. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. 8. time/temperature data. once determined. Many fish-processing operations have relatively few steps and the flow diagrams appear simple.4. the qualitative and/or quantitative evaluation of the presence of hazards. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. survival or multiplication of pathogenic microorganisms.

Salmonella spp. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. Vibrio spp. Hazards introduced during processing of fish products depend very much on the characteristics of the process.3.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products. 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. For example.g. any food safety objectives or manufacturer’s food safety requirements. 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. Note: it is important at this stage that no attempt is made to identify CCPs. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin.). Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. or desirable.6. the number of consumers potentially exposed and their vulnerability. and there are an equally varied range of potential hazards associated with them.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. or may originate from polluted water or from post-capture contamination (e. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. botulinum. 181 . The incidence of psychrotrophic. and viruses). and the potential growth of Listeria monocytogenes on smoked fish. since this may interfere with the analysis. One control measure may be relevant to several process steps. For example. 8. Therefore it is not possible. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage. The HACCP team should then determine what control measures exist that can be applied for each hazard. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards. where a hazard is repeated. Pathogens may be part of the resident microflora of the living animal (e. the hurdle effect.g. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual.

More than one critical limit may be defined for a single step.1 Fish and seafood Again. it is necessary to specify both time and temperature for a thermal process.8.7. application of the decision tree should be flexible. 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 . Where possible.3.8 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.2) to provide a logical. and the reader is referred to the appropriate product chapter. and its use may not always be appropriate. then the process must be modified to include a control measure.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8.FISH AND SEAFOOD 8.3. Criteria used to set critical limits must be measurable and may include physical. If a significant hazard has been identified at a step where control is required for safety. given the enormous variety of fish products and processes in use. biological or sensory parameters. chemical. It is prudent to set stricter limits (often-called target or process limits/levels) to ensure that any trend towards a loss of control is noted before the critical limit is exceeded. structured approach to decision making. critical limits should be specified and validated for each CCP. 8. but for which no control exists at that step or any other. For example.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits.3. it can be said that effective control measures are likely to include the following: • • • • • 8. However. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. it is not possible to generalise on likely CCPs. 8. However.3.


• • • 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.


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

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

1991. Subsequently. 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. 9. p.2 Legislative Structure From 1 January 2006. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No.B. transportation. 24. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. 7. A number of more detailed implementing and transitional measures have been adopted at EC level. storing. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No.9.7. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. European Communities L 268.9. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004.1991. handling and offering for sale or supply to the consumer. as last amended by Regulation (EC) No 806/2003. 41). N. Council Directive 91/493/EEC of 22 July 1991 laying down the health conditions for the production and the placing on the market of fishery products (OJ L 268. as last amended by Regulation (EC) No 806/2003. J. p. existing hygiene Directives including those below were repealed: • Council Directive 91/492/EEC of 15 July 1991 laying down the health conditions for the production and the placing on the market of live bivalve molluscs (Off. p.1992. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. processing.1). packaging. manufacturing. 24. distribution. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No.15). 190 • • . 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.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. and sets out what those enforcing the provisions have to do.

19. 191 . the legislation is directly applicable law. e. Wales and Northern Ireland). and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer.g. penalties and powers of entry. 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. 2006 No. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No.11) The EU hygiene regulations apply to all stages of food production including primary production.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. etc. of stock farming.) in England. 1 . p. The Food Hygiene (England) Regulations 2006 (S. and equivalent legislation in Scotland.I. for example. 2073/2005. Additionally the requirements of EC Regulation 853/2004 must be complied with. domestic preparation or handling. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. which will be covered later in this chapter. Wales and Northern Ireland. There are some exemptions. national legislation in the form of a Statutory Instrument (S. and of hunting and fishing. with primary production. revocation of existing implementing legislation. As regulations. food storage that is for private or domestic consumption. and binding in its entirety on all member states from the date of entry into force. is required to give effect to the EU regulations.1993. 9. 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. 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. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. Primary products are defined as products of primary production including products of the soil. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland.I. right through to the implementation of procedures based on HACCP principles. setting offences. 852/2004. Although the regulations have the force of law.14.7.3 Regulation (EC) No. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production.



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:


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

EO. For the UK.2. Extracts of the requirements of Annex II. (iv) If the identification mark is applied in an establishment within the Community. EÜ. Section I of Regulation (EC) 853/2004.2.2. the food business operator may apply the same identification mark to both types of food. 198 . the mark must be oval shaped and include the abbreviation CE. Section I of Regulation (EC) 853/2004 are given as follows: 9. please refer to Regulation (EC) 853/2004.4.FISH AND SEAFOOD 9. 9.4. the new mark must show the approval number of the establishment where this occurred. for other codes.4. EK.1. An identification mark can only be applied provided the relevant requirements as stated above in relation to Regulation (EC) 852/2004 have been met in particular for the registration and approval of establishments.1.2 Form of the identification mark (i) The mark must be legible and indelible. a new mark must be applied to the product. and the characters easily decipherable and be clearly displayed. EK. EF.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. the code would be UK. (iii) Additionally. EY. (ii) However.2 Fishery product requirements 9. EC. In this case.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. ES. EB or WE. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. (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. when a product's packaging and/or wrapping is removed or is further processed in another establishment.4. (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 show the approval number of the establishment. EG.

4. the wrapping or the packaging. With the exception of Chapters I and II.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. tunicates and marine gastropods when placed on the market live. 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.1.2. as amended. they would need to be obtained following Section VII on live bivalve molluscs. the approval number of the establishment and the abbreviation for the community where appropriate. it is sufficient to apply the mark to the exterior of that package only.LEGISLATION 9. i. (ii) For products of animal origin that are placed in transport containers or large packages. its wrapping or the packaging. specifically relating to fishery products are given below. or be printed on a label affixed to the product. including vessels. The requirements of this section accompany those in Regulation (EC) 852/2004. wrapping or packaging in another establishment. This section does not apply to bivalve molluscs. echinoderms. C and D.2. Part A and Chapter V apply to retail. (iii) In the case of liquid. Parts A. it applies to those animals when not placed on the market live. Chapter IV.e. Chapter III. granulate and powdered products of animal origin carried in bulk. and fishery products carried in bulk. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . Therefore. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. the colourings used must be EC authorised colours. for full requirements. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. processing. Extracts of the requirements of Annex III. the mark may be applied to the outer surface of the container or packaging. a) requirements for establishments.3 Method of marking (i) The mark is to be applied directly to the product. and are intended for further handling.4. Section VIII of Regulation 853/2004. 9. (v) When the mark is applied directly to products of animal origin.

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

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

c) Fishery products.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. chilled and wrapped. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption. Handlers may use spiked instruments to move large fish or fish that might injure them. other than those kept alive. and products washed thoroughly. the product must be kept at no more than -9 °C. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. 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. provided no damage occurs to the flesh. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. b) Factory vessels on board which crustaceans and molluscs are cooked. 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 . landed as soon as possible. e) In cases where whole fish intended for canning is frozen in brine. where fishery products are displayed for sale. The unloading and landing process is to be done quickly to avoid contamination (but at the same time not causing bruising to the fish) and to ensure fishery products are placed in a protected environment at the temperatures stated in Chapter VII.2.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. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. The brine must not be a source of contamination. if this is not possible.2.4. B.Requirements during and after landing 9. must be chilled or. Chapter II . (ii) In the case of food business operators responsible for auction and wholesale markets etc.

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

if they are to undergo a cold smoking process in which the internal temperature of the fishery product is not more than 60 °C: (i) herring. (b) fishery products from the following species. D. if the processing is insufficient to destroy nematode larvae. cooling must continue until a temperature 204 . they must be cooled rapidly. (iii) sprat. (iv) (wild) Atlantic and Pacific salmon. and whole fish must be gutted and washed beforehand.2.2. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. Once produced. (ii) mackerel. except when supplied to the final consumer. 9.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . (iii) A document from the manufacturer. stating the type of process they have undergone. raw materials must have no guts. (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.20 °C in all parts of the product for at least 24 hours (this treatment must be applied to the raw product or the finished product): (a) fishery products to be consumed raw or almost raw. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked.4. If no other method of preservation is used.Requirements for certain processed fishery products A. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. must accompany fishery products referred to above when placed on the market.4 Chapter IV . or a stabilising treatment. and (c) marinated and/or salted fishery products.

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

Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. thawed unprocessed fishery products.2. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. Molidae. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption.7 Chapter VII . Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market.2. 206 . and when fishery products are wrapped on board fishing vessels. The scientific name must accompany the common name on the label.FISH AND SEAFOOD D.2. 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. Fresh.2.4. it should be kept at no more than -9 °C. Diodontidae and Canthigasteridae. prepared and processed fishery products belonging to the family Gempylidae. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. E. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. 9.4.6 Chapter VI .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. In the case of whole frozen fish in brine intended for the manufacture of canned food. Fishery products contaminated with parasites may not be placed on the market for human consumption. food business operators must ensure that wrapping material is not contaminated and is stored in such a way that it is not exposed to contamination and is easy to clean where necessary. 9.Storage of fishery products (i) Fresh fishery products.

(d) 60 mg of nitrogen/100 g of whole fishery products used directly for the preparation of fish oil for human consumption as referred to in the second sub-paragraph of Part B(1) of Chapter IV of Section VIII of Annex III to 207 .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.). Helicolenus dactylopterus. 2074/2005. (ii) If fishery products are kept under ice. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. The following in relation to TVB-N limits is extracted from the regulation: Unprocessed fishery products shall be regarded as unfit for human consumption where organoleptic assessment has raised doubts as to their freshness and chemical checks reveal that the following TVB-N limits are exceeded: (a) 25 mg of nitrogen/100 g of flesh for the species: Sebastes spp. species belonging to the Merlucciidae family. melt water must not remain in contact with the products.2. species belonging to the Gadidae family. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. 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. Sebastichthys capensis.Transport of fishery products The above points for Chapter VII apply in relation to transport. The temperature of frozen fishery products must not fluctuate above -15°C.8 Chapter VIII . 9.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. provided that the journey is short and the competent authority allows this. 9. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated.

(b) and (c) of Part B(1) of that Chapter. The sample must consist of about 100 g of flesh.General requirements for the placing on the market of live bivalve molluscs Live bivalve molluscs can only be put on the market for retail sale if done through a dispatch centre.4. (b) In the case of other operations. VI.1 Chapter I . for full requirements. Chapters V. please refer directly to the regulation. 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. as amended. The information required in the registration document is given in point 4 and includes the health status of the production area. Extracts of the requirements of Annex III. they supplement the requirements of Annex II to that Regulation. however. specifically relating to bivalve molluscs are given below. 9. this Section also applies to live echinoderms.3 The regulation details specific hygiene requirements for bivalve molluscs. Live bivalve mollusc requirements - 9. taken from at least three different points and mixed together by grinding. VIII and IX. and point 3 of Chapter VII applies to retail. shellfish species etc.4. Chapter IX applies to pectinidae harvested outside those areas. they supplement the requirements of Annex I to that Regulation. The requirements of this Section accompany those laid down in Regulation (EC) No 852/2004: (a) In the case of operations that take place before live bivalve molluscs arrive at a dispatch or purification centre. reference should be made to the actual regulation. tunicates and marine gastropods. Section VII of Regulation 853/2004.FISH AND SEAFOOD Regulation (EC) No 853/2004. Apart from the provisions on purification. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. The copy of 208 . 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.

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

Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. 9. Before dispatch. 160 micrograms of okadaic acid equivalents per kilogram. the purification system to allow them to resume /maintain filter-feeding activity.5 Chapter V .Hygiene requirements for purification and dispatch centres A. (iv) yessotoxins. for: (i) paralytic shellfish poison (PSP). (ii) amnesic shellfish poison (ASP).4. Requirements for dispatch centres Handling should not result in contamination to any degree. Requirements for purification centres There are requirements for live bivalve molluscs to be washed. 210 . 800 micrograms per kilogram.4.4.3. and there are requirements for tanks and water storage containers. dinophysistoxins and pectenotoxins together.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. requirements if there are many batches of live bivalve molluscs. 1 milligram of yessotoxin equivalent per kilogram. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. the levels of marine biotoxins should not exceed the following limits.3 Chapter III . B. shells of live bivalve molluscs are to be washed throughout with clean water and the molluscs are to come from one of the following: (i) Class A production area (ii) Relaying area (iii) Purification centre (iv) Another dispatch centre 9.FISH AND SEAFOOD 9.4 Chapter IV .3.3. containers and a label to identify purification.

9.4.3. 211 . must be waterproof. Finally.8 Chapter VIII . including the identification mark. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. dispatch centre or processing establishment. 9.4. Registration document requirements also apply. 9.9 Chapter IX . (v) azaspiracids.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.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.Identification marking and labelling In addition to general identification marking (contained in 9.6 Chapter VI .3.4.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. the species of bivalve mollusc and date of packaging (at least the date and month) are required. 160 micrograms of azaspiracid equivalents per kilogram.7 Chapter VII . (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 requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’. by way of derogation from directive 2000/13/EC. Also.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.3. they cannot be re-immersed in or sprayed with water.

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

600 E. Live bivalve molluscs from Class C areas must not exceed 46. coli per 100 g of flesh and intravalvular liquid. three dilutions MPN test specified in ISO 16649-3.2.000 E.5.1 Chapter II . Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. coli per 100 g of flesh and intravalvular liquid. Regulation (EC) 854/2004. that microbiological criteria and chemical contaminants amongst other requirements are met.5. In the remaining 10 % of samples.2 This states official controls for the production and placing on the market of live bivalve molluscs. The reference method for this analysis is the five-tube. Live bivalve molluscs from Class B areas must not exceed. The reference method for this analysis is the five-tube. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. as previously stated. live tunicates and live marine gastropods. 4. live bivalve molluscs must not exceed 46. in 90% of the samples. - - - - B. live echinoderms. 000 E. 213 . section VII. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004.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. A summary is given as follows: 9. Annex II 9. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. Sampling plans and frequency are required. coli per 100 g of flesh and intravalvular liquid. Also. chapter V is reinforced.Official controls concerning live bivalve molluscs from classified production areas A. Classification of production and relaying areas: This section reinforces that specified in Annex III.

214 . 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. the competent authority is to close the production area. 2073/2005. E. 9.5. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004.2 Chapter III . dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. boundaries or class of a production area or its closure. 9. 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) 2076/2005.6 Regulation (EC) No.2. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. Procedures for re-opening a closed production area are stated D. opening or closing a production area.5. Additional monitoring requirements This concerns the competent authority in relation to monitoring of classified production areas where bivalve mollusc harvesting is forbidden and of relaying and production zones to ensure there is no malpractice etc. F. establishes microbiological criteria for a range of foods. 9. 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.FISH AND SEAFOOD C. which has applied since 1 January 2006.

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

monocytogenes. other than those intended for infants and for special medical purposes EN/ISO 11290-1 Products placed on the market during their shelf life Before the food has left the immediate control of the food business operator.I Food safety criteria Sampling plan(1) n 10 5 Absence in 25 g EN/ISO 11290-1 c m M Products placed on the market during their shelf life Limit(2) Analytical reference method(3) Stage where the criterion applies Food category Microorganisms 1. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1.2 Ready-to-eat foods able to support the growth of L. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1. other than those intended for infants and for special medical purposes(4) (8) L.TABLE 9. monocytogenes. monocytogenes 1.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L. monocytogenes L.3 Ready-to-eat foods unable to support the growth of L.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life .

17 Live bivalve molluscs and live echinoderms. coli (15) 1. manufactured from fish species associated with a high amount of histidine (17) Histamine . 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 1.27 Fishery products which have undergone enzyme maturation treatment in brine.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1.Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.25 Live bivalve molluscs and live echinoderms. tunicates and gastropods 9 (18) 2 E.

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

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

coli 5 shucked products of cooked crustaceans and molluscan shellfish 2 100 cfu/g 1000 cfu/g EN/ISO 6888-1 or 2 FISH AND SEAFOOD Coagulasepositive staphylococci 5 Improvements in production hygiene 220 .II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2.TABLE 9.4.1 Shelled and E.

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

Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions. 2006 No. 14) do not apply to any food covered by EU Regulation 853/2004 on hygiene of products of animal origin or any food business operation carried out on a ship or aircraft. 2006 No. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. such as that below.I. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. the Food Standards Agency has published guidance notes on the requirements of the EU hygiene and microbiological criteria regulations which can be accessed at the following link: http://www.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . was kept at above 8 °C or the recommended temperature for an unavoidable reason. For other food.I. 3).K. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing.8 Guidance In the U. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment. 9. for a limited period consistent with food safety. and was kept at above 8 °C or the recommended temperature. or.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. 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. S.food. 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. it would need to be proved that the food was being transferred to or from a vehicle used for the activities of a food business. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. Alternatively. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S.S.gov. 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.

1688/2005. as read with the corrigenda at OJ No. as amended. L226.4. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 1663/2006. 9. 2074/2005.12. 2076/2005. L139.6. 22. L226. Regulation (EEC) 2136/89. 1662/2006. 2076/2005. 2075/2005 and Regulation (EC) No. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption (OJ No. Regulation (EC) No.2004. these are not be discussed here. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. 853/2004 is now set out in a Corrigendum (OJ No.2004. L338. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No.2004. Regulation (EC) No. Regulation (EC) No.4. 2073/2005 on microbiological criteria for foodstuffs (OJ No. 3) as amended by Regulation (EC) No.9 Other Relevant Legislation The following regulations about fish designations need to be considered: • Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products Regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products • Note: Requirements for preserved fish are laid down elsewhere.6. laying down common marketing standards for preserved sardines. Commission Regulation (EC) No. 223 . L283. The revised text of Regulation (EC) No. L226. 83) as amended by Regulation Nos 882/2004.2004. 2076/2005. namely. 2076/2005. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41.2006.6.10 Further Reading Regulation (EC) No.2004. The revised text of Regulation (EC) No. 22) as amended by Regulation (EC) Nos. 30. 1). 25.2004. 206). 25. 2074/2005. 2074/2005. 14.4. 854/2004 is now set out in a Corrigendum (OJ No. L139. 62) as amended by Regulation (EC) No.LEGISLATION 9. Regulation (EC) No. 1441/2007. 2074/2005 and Regulation (EC) No. 1. L139. 55). 30. 25.2005. 1019/2008 and as read with Regulation 2073/2005. 852/2004 is now set out in a Corrigendum (OJ No. The revised text of Regulation (EC) No. Regulation (EC) No. 30.10. Regulation(EC) No.

12.2000.2001. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17. 21.1.2005. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.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. derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004. 22) Commission regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products as amended by Commission Regulation (EC) No 1792/2006 of 23 October 2006 (OJ L 278. 6) 224 . (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. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. 22. 22.

and 2.3 Temperature Aeromonas grows at temperatures of 2 .1 10.5 pH Under otherwise optimal conditions.3 . no growth is observed (1). 0. Some death will occur at this pH if the temperature is 4 °C. 10. 10. The minimum pH value for growth is thought to be pH 5.1.0 . At 5 °C.5 minutes. a 10 to 1.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.10 (3).1.000-fold increase in number is observed after one week in food systems (1).5.1. under optimal incubation temperatures.3. and D-values at 45 and 51 °C have been reported as 29. 225 .45 °C (1).5. 10.1.0 x 1.3 minutes.1 Aeromonas spp. Morphology Gram-negative rods.10. At pH 4.5 µm 10.2 Oxygen requirements Facultative anaerobe. PATHOGEN PROFILES 10.1. respectively (1).1. The optimum growth temperature is 28 °C (2). Aeromonas has the potential to grow over a pH range of ca 4 .

2. F are the types that are responsible for most cases of human botulism (5. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. botulinum are not particularly heat resistant. being capable of slow growth and toxin production at low temperatures .5 .22.3 Temperature All strains of C. E and. and is sensitive to > 4.2.30 minutes.0 µm. 226 .1.2. A to G. forming at least seven different toxins. 10.5% NaCl (4). and groups I and II are those responsible for the botulism cases. to a lesser extent. B. botulinum grow reasonably well in the temperature range of 20 45 ºC. but the interior of the food may do so. botulinum is an obligate anaerobe. 10. or at 90 ºC for a few seconds. Group I will not grow at temperatures of 10 ºC or less. types B and F can be either. 6).6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . 10. botulinum types produce heat labile toxins. botulinum is prevented.even as low as 3 ºC (7). many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C.4 Heat resistance The vegetative cells of C. but the spores of this organism are more so.2.4 x 1.FISH AND SEAFOOD 10. Types A. botulinum are known.7 . at 85 ºC for 5 minutes. but Group II strains are psychrotrophic.2. There are four main groupings of the organism.2 Oxygen requirements Although C. spore-forming rods. 0. 10. All C.2 Clostridium botulinum Seven different types of C. 10. All type A strains are proteolytic and type E strains are usually non-proteolytic.1 Morphology Gram-positive. Thus an aerobically packed product may not support the growth of the organism on the surface.2% sodium chloride (NaCl) (1). which may be inactivated by heating at 80 ºC for 20 .

2.1 Clostridium perfringens Morphology Gram-positive spore-forming rods.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. 10. The organism will grow well in the centre of meat or poultry dishes.6 and pH 5. 11)). grow on the surface of foods unless they are vacuum or gas-packed.PATHOGEN PROFILES 10.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. 9). 10.is an obligate anaerobe.2. particularly by cooking. parcooked meals with pH value > 5. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. botulinum spores The most heat resistant spores of Group I C. where oxygen levels are reduced. the minimum aw for growth may be lower. 0.0 and aw > 0. However. perfringens .1.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. 227 .21 minutes) (9).2. These concentrations correspond to limiting aw of 0. These values have been established under carefully controlled laboratory conditions. vacuum packed.94 for Group I and 0. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I.2 Oxygen requirements C.7 Characteristics of C. Note also that if humectants other than salt are used. pH and temperature. therefore. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains.10.3 . they may survive mild heat treatments (70 .0 µm.97 for Group II (5).3 10.0 .3. It will not.g. 10.97) (10. respectively (8.9 x 2.3. In commercial situations. 10.like other clostridia . botulinum depends on solute. safety margins must be introduced to allow for process variability.0.

10. Under otherwise ideal conditions. Recorded heat 228 . Its optimum temperature for growth is 43 . perfringens in vacuum packed beef (18). (It is unusual for spores to be formed in foods after the growth of this organism (14)).4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. 15. 16). type of solute.8% inhibit growth of most C. 13). the limits for aw are affected by temperature.3. perfringens is not tolerant of low water activities.FISH AND SEAFOOD 10.3 Temperature The most significant characteristic of C. As in the case of other factors limiting the growth/survival of this organism. lower concentrations may be effective in combination with other factors. will survive greater extremes of pH (and aw) (12. the enterotoxin is not heat resistant . Spores. perfringens can vary quite considerably in their heat resistance. perfringens spores The spores of C. In addition. pH. however. and their heat resistance is also affected by the heating substrate. depending on strain and other conditions.6 Aw/Sodium chloride C. although C. 13).45 ºC.5 minutes (17).3. etc.3. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures.3. It grows best at pH values between 6 and 7 (the same pH as most meats). At cold temperatures. Salt concentrations of 6 .7 Characteristics of C. perfringens strains. The lowest aw recorded to support the growth of C. perfringens is not a tolerant organism with respect to pH. 18). perfringens has the potential ability to grow within the temperature range 15 . While some growth can occur at 50 ºC.3. perfringens appears to be 0. 10. perfringens.93 and 0. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance.10 ºC. perfringens in roast beef at 60 ºC is 14. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12.it is destroyed by heating at 60 ºC for 10 minutes (14.50 ºC. vegetative cells die rapidly (12). 0 . The D-value for C. 10.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. Some studies indicate that the presence of 3% NaCl delays growth of C.5 pH C. 10.3.

and temperature. Morphology Gram-positive short rods.8 minutes for heat sensitive spores (12). is between 30 and 37 ºC. requiring days or weeks to reach maximum numbers.4.4 .4. -0. 229 .at refrigeration temperatures down to.3 . It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.4. 10.23.2 has been suggested as the lowest working limit (22). the lowest limit for growth is likely to be considerably higher .albeit slowly .98 and 0. 1. being potentially capable of growing . monocytogenes in crawfish tail meat at 55. Its optimum growth temperature. growth at low temperatures can be very slow.1 Listeria spp.6 . or even below 0 ºC. 10. 10. D-values for L. The upper temperature limit for the growth of L.4 Heat resistance L. monocytogenes is reported to be 45 ºC (20).19 minutes. However.4 ºC is probably a more likely minimum for growth in foods (19). In foods. 0.2. 10.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. Under ideal conditions. it is not a sporeformer.especially at refrigeration temperatures.0 minutes for heat resistant spores to 1.5 . and where acetic acid is used as an acidulant.0 µm. the organism is able to grow at pH values well below pH 5 (pH 4.4.3 is the lowest value where growth has been recorded. however.64. however.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic.4 10.2 Oxygen requirements Aerobe or microaerophilic.5 x 0. so can be destroyed by pasteurisation. pH < 5. respectively (21). monocytogenes is not a particularly heat resistant organism. 60 and 65 ºC were reported as 10.5 pH The ability of Listeria to grow at different pH values (as with other bacteria) is markedly affected by the type of acid used.0.4. using hydrochloric acid as an acidulant). 10.2.

97. peritrichous flagella. 10. 10.7 x 1. 10. The bacterium may survive for long periods at aw values as low as 0.2 Oxygen requirements Facultative anaerobe.5 10.1 Salmonella spp.1.3 Temperature Plesiomonas can grow in the temperature range of 8 .5 . 25). 230 .5 pH Plesiomonas has a pH range for growth of pH 4.6.5 (24.0 µm.8 . 10.6 monocytogenes is quite tolerant of high NaCl/low water activities.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).12% NaCl or more).3 µm. 10.8. at salt levels found in foods (10 . 0. 10. Morphology Gram-negative short rods.5 .83 (20). or even grow. It grows best at aw of ≥ 0.90.45 ºC.5.4. but has been shown to be able to grow at an aw level of 0.6 Aw/Sodium chloride L. 25).1 Plesiomonas Morphology Gram-negative. 0.0 x 1 .5.FISH AND SEAFOOD 10.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24. 10. It is likely to survive.5. the optimum being 30 ºC (23).5.3.0 .

93 (as reported in dried meat and dehydrated soup). D60 ºC values normally range from about 1 to 10 minutes. although salt tolerance increases with increasing temperature (28). 10. 10.6.7. However. Furthermore.3 Temperature Salmonellae can grow in the temperature range of 7 . the optimum being 35 . some strains are able to grow at temperatures as low as 4 °C (26).5. and appropriate heat treatments must be determined experimentally for low-aw foods.20 times more heat resistant than the average strain of Salmonella at high aw (27). The optimal pH for Salmonella growth is between 6. Salmonella senftenberg 775W is about 10 .PATHOGEN PROFILES 10.5 ºC. strains vary in their ability to withstand heating.945. substrate.8 . high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. under otherwise ideal conditions. a heat resistant organism. Salmonella has the potential to grow at aw levels as low as 0.6 Aw/Sodium chloride Where all other conditions are favourable.5 pH Salmonella has a pH range for growth of pH 3. or possibly 0.6. Salmonellae are quite resistant to drying.37 ºC. Growth is slow at temperatures below about 10 ºC. temperature and pH.0. However. with a z-value of 4 .2 Oxygen requirements Facultative anaerobe.5 . It is not. depending on the type of acid and temperature. The growth of Salmonella is generally inhibited by the presence of 3 .6. depending on serotype.4% NaCl.6. 231 . and with an appropriate acid. 10. therefore.9. 10. Some death will occur at pH values of less than about 4. pasteurisation and equivalent treatments will destroy the organism under normal circumstances.6.4 Heat resistance Salmonella is not a spore-forming organism.5.48 ºC. Salmonellae are quite resistant to freezing.

7. however. the organism is heat sensitive and will be destroyed by pasteurisation. the D60 ºC value is 2 . fat content.7 10. 0. The limits for toxin production are also narrower than for growth. with an optimum of 35 . etc. 10.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. NB: The growth of Staph.9 µm diameter. Staph.20 minutes.0 .3 (optimum 7.1 Staphylococcus aureus Morphology Gram-positive cocci. aureus than citric acid).7.7. Most strains of staphylococci can grow within the pH range 4. aw. depending on aw. 232 .2 Oxygen requirements Facultative anaerobe. aureus viability.7. 31). 10. stressed cells can also be less tolerant of heating. 10.37 ºC (29).7 . aw and temperature (sensitivity to acid increases with temperature). aureus can grow within the temperature range 7 – 48. As is the case with other bacteria.).5 ºC.2 to 9. under otherwise ideal conditions (29.7.3 Temperature Under otherwise ideal conditions. In meat macerate. It can survive well at low temperatures. Freezing and thawing have little effect on Staph.5). Under most circumstances.7.5 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.FISH AND SEAFOOD 10. 10. but may cause some cell damage (30). aureus is more limited under anaerobic than aerobic conditions.0. The following relate to limits for growth only.

an aw of 0.4 . * dependent on temperature.8. It can grow over the aw range 0. pH. atmosphere.99 little or no toxin production in anaerobically packed foods. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.7.>0. with a temperature range for growth of 10 .83 . and solute.8. 0.99 aerobically under otherwise optimal conditions.5 . but certain strains can grow in 20% NaCl.6 µm 10.86 is the generally recognised minimum in foods (32). especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.7 Limits permitting toxin production 10 .2 .87 and > 0. In general.7.0 (optimum 7.2. 10.PATHOGEN PROFILES 10. An effect of increasing salt concentration is to raise the minimum pH of growth.2 Oxygen requirements Facultative anaerobe.7. 10.9. Staphylococci are more resistant to salt present in foods than other organisms. However. Staph.5) (29. 34).3 Temperature The optimum temperature for growth has been reported as 30 ºC.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5.8 10.0.0 . heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. 233 . strain. 10. 31) between 0.8 x 1. In general.10% NaCl.6 Aw/Sodium chloride Staph.43 ºC (35).1 Vibrio cholerae Morphology Gram-negative curved rods.8. aureus can grow in 7 .

2 Oxygen requirements Facultative anaerobe.998. respectively.6 (35. 234 .65 minutes (35).9 10. there is evidence that V.FISH AND SEAFOOD Also. It can grow in the presence of 0. cholerae may persist on frozen fish for >180 days (36). 38).9. cholerae can grow over the aw range 0. 10. with an optimum of 37 ºC (39).8. 10. declining in number during storage. 10.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media. under otherwise favourable conditions. 10.9. V.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. it has a pH range for growth of pH 5 .1 . and it is sensitive to refrigeration temperatures (0 . are 5 and 43 ºC. but only moderately sensitive to freezing (40).6 Aw/Sodium chloride V.9.4 Heat resistance V. with an optimum of 0.0% NaCl (35).8.984 (35).5 ºC). with optimum at 7.97 .9.0. 10.5 pH This organism is tolerant of high pH values. 37. 10. The D-value for V.6. parahaemolyticus is easily destroyed by drying.8. cholerae is sensitive to heat at temperatures of >45 ºC. but not acid conditions. cholerae in crab meat homogenate at 60 ºC is 2.4.

0. but the optimum concentration is 2 . 10. respectively.PATHOGEN PROFILES 10.9.4% (39. with an optimum pH of 7. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0. and 2.922.5% NaCl. 41). respectively (39).4 Heat resistance V. parahaemolyticus will grow at aw levels as low as 0.2 Oxygen requirements Facultative anaerobe. 10.5 pH The pH range for growth is 4. V.02 .6 (41).5 minutes. The organism can survive for long periods at freezer temperatures (42). it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).10. The D-values at 55 ºC for clam and crab homogenate are 0.9.11. 10. The optimum concentration of NaCl equates to aw of approximately 0.8 . depending on solute. 10. (35. and are readily destroyed by cooking.10 Vibrio vulnificus 10. 235 . parahaemolyticus are not heat resistant.1 Morphology Gram-negative curved rods. 41). with a temperature range for growth of 8 .4 Heat resistance V. 10.6 Aw/Sodium chloride V.6 .3 Temperature The optimum temperature for growth has been reported at 37 ºC.29 minutes.43 ºC (42).10.10. parahaemolyticus is unable to grow in the absence of salt.8. vulnificus is heat sensitive.

Ed.W. Marcel Dekker..M. 1-27.0% NaCl. Ed. in Foodborne Bacterial Pathogens. Eds. New York. Miliotis M. 1994. Beuchat L. Aeromonas species. Principles of control.. Abeyta C.11 1. 2005. 1989..W. Smith J. Ed. New York. Eds.W. Faris A. Gorman J. Aeromonas. Cliver D. Kim J.. 279-303. 9. 2001. Montville T.5 pH The pH range for growth is 5 . 383-408. 121-76. Dodds K. 3. 10.W. 57-63. Eds. 111-89. 6 (1). Bhunia A... Gaze J. Peck M.D. 2. 329-49. Blackie.R.8 (42).H.. ASM Press. 10.D... Diseases Caused by Bacteria. 1994. Caister Academic Press. 7. Lund B. 11.. 8. Marcel Dekker. Clostridium botulinum. New York. Hauschild A. Dodds K.. Doyle M.. Eds. 1-19. Bier J. International Commission on Microbiological Specifications for Foods.K.H. Volume 5: Microbiological Specifications of Food Pathogens. in Foodborne Bacterial Pathogens. New York. 10. Clostridium botulinum. References International Commission on Microbiological Specifications for Foods. Juneja V. 1989. Eds. Hauschild A. Hui Y. Potential hazards associated with REPFEDS. Murrell K. 6. London. New York. Food Control.M. Doyle M.L. Kovacek K.6 Aw/Sodium chloride V. Doyle M.5% (42). Hauschild A.P. 5-19. Aeromonas hydrophila group. in Foodborne Disease Handbook. Marcel Dekker. 5.E. in Food Microbiology: Fundamentals and Frontiers. in Clostridium botulinum: Ecology and Control in Foods.H. 1993.W.. in Microorganisms in Foods. Stelma G. 97-131. 1996. 1993.C. 4. 2003. Aeromonas hydrophila. Marcel Dekker.J..L. Murrell K.P. Stelma G.L. Eds.10.R. Palumbo S. Cliver D. Gorham J. vulnificus grows over the range 0.H.10. Marcel Dekker.10..P.D.. in International Handbook of Foodborne Pathogens. Great Britain. Eds.. New York.H. in Foodborne Disease Handbook. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.N. Foegeding P. Washington D. Novak J.A. Microbiology and Molecular Biology. 357-67. Volume 1..M.D. Clostridium botulinum and Clostridium perfringens. Notermans S.O. with an optimum pH of 7.O. Betts G.. Dodds K. with an optimum of 2. Clostridium botulinum.. New York. Marcel Dekker. 1995. Johnson E. Fratamico P. Hui Y. Marcel Dekker Inc. in Clostridium botulinum: Ecology and Control in Foods. 236 .L. in Foodborne Pathogens..R.FISH AND SEAFOOD 10.N...H. Austin J. Volume 1: Diseases Caused by Bacteria.5 .5.

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

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

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


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

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

nz/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.gov.co.seafood.org.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.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.bsi-global.hpa.cmar.uk Web site: www.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: generalenquiries@fdf.csiro.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.fdf.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.gsi.gov.ac.ifst.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: reception-cmar-hobart@csiro.org Web site: www.com Web site: www.au Web site: www.uk/en/index.ifr.CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: contact@nfi.org.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.co.uk Web site: www.nz Web site: www.com/ 243 .dh.org.

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

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


as autochthonous bacterial flora of fish 3 Aeromonas spp. source of PSP toxins 142 Alexandrium ostenfeldii. 69-70 Aeromonas hydrophila. to test microbiological criteria 19 Aeromonadaceae. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. 102 Bacillus spp. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. in crustacean shellfish 87 Alteromonas spp. preserved by dry/wet salting 99 Anisakis spp. in crustacean shellfish 86-7 ATP. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. 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 . causing spoilage of dried fish 110. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. growth and survival in crustacean shellfish 86 Aeromonas sobria. 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.INDEX Achromobacter spp. as autochthonous bacterial flora of fish 3 as spoilage organisms in oysters 65 as spoilage organisms in temperate crustacean shellfish 80 as spoilage organisms in tropical crustacean shellfish 81 in molluscan shellfish 60 Aerobic plate counts. 111 Astroviruses. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies.

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

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

causing pink condition on fish surface 110 in fish sauce 127 Handling. 102 Gymnodimines. as source of inulin in lactic acid fermentation in fish products 134 Garos. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. of prepared fish products 44 Gambiertoxins. 130-5 Fish-spoilage bacteria 12 Fishtester. 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. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. mould causing spoilage of dried fish 110 Evaluation. as source of ciguatoxins 156 Garlic. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning.FISH AND SEAFOOD EU legislation. for evaluation of fish quality 9 Eurotium. of fresh fish spoilage 911 Evisceration. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes. to measure fish skin resistance 10 Flavobacterium. concerning crustacean shellfish 87-8 European Grading Scheme. member of cyclic imine group 161 Gymnodinium catenatum. as autochthonous bacterial flora of fish 3 in molluscan shellfish 60 spoilage organism in oysters 65 Food hygiene legislation 189-224 Food of animal origin. specific EU hygiene rules 196-224 Fresh fish. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. processing 35 Fish component. non-B) viruses. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . preserved by marinating 101 Gravlaks. of fish at sea 5 Fatty fish. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. fermented fish product 126 Green cured split cod 97. source of PSP toxins 142 Gyoshoyu. of fish – effects on microflora 5-6 Hepatitis (non-A. fish sauce 123 HACCP.

cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. as source of yessotoxins 152 Lipid content. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured. in fish fillet 1 Lipid oxidation. to reduce rate of fish spoilage 12 Ikan kaju. outbreak caused by gravid 115 Lobster. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. preserved by salt-boiling 100 Karenia brevis. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. as autochthonous bacterial flora of fish 3 as spoilage organisms in oysters 65 as spoilage organisms in tropical crustacean shellfish 81 in fish sauce 127 Microflora. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. definition 1 Legislation. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149. enabled by carbohydrate source in fish products 130. hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. as autochthonous bacterial flora of fish 3 Lean fish. concerning crustacean shellfis 88 Jellied eels. 135 I-zushi. food hygiene 189-224 Lingulodinium polyedrum. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 . smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. 100-1 Masmeen 107 Matjes. as source of botulism 136 International legislation. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. preserved by marinating 101 Microbiological criteria. preserved by salt-boiling 100 Ionising radiation. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. for microbial inactivation in fish products 127. for fresh and frozen fish (Table) 20 Micrococcus spp.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage.

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

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

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

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

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

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