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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© 2009 Leatherhead Food International Ltd

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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


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

vi .

4 Spoilage 4.6 Pathogens: Growth and Survival 2. 4.7 Published Microbiological Criteria 2.3 Processing and its Effects on the Microflora 3.4 Spoilage 1. 3.8 References 1.2 Initial Microflora 4.6 Pathogens: Growth and Survival 1.7 References 3.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.4 Spoilage 3.1 Definitions 3.8 Further Reading CRUSTACEAN SHELLFISH 4.4 Processing and its Effects on the Microflora 2.2 Initial Microflora 3.5 Spoilage 2.3 Processing and its Effects on the Microflora 4.7 References 4.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.2 Definitions 2.5 Factors Affecting Fresh Fish Spoilage 1.3 Processing and its Effects on the Microflora 1.1 Definitions 1.2 Initial Microflora 1.1 Definitions 4.3 Initial Microflora 2.7 Published Microbiological Criteria 1. vii . CHILLED AND FROZEN RAW FISH 1.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.5 Pathogens: Growth and Survival 3.8 References MOLLUSCAN SHELLFISH 3.6 Published Microbiological Criteria 3.5 Pathogens: Growth and Survival 4.1 Introduction 2.6 Published Microbiological Criteria 4.

8 Further Reading FERMENTED FISH 6.5. 9.3 Regulation (EC) No.I.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.2 Initial Microflora 5.6 Lipophilic Shellfish Toxins (LST) 7.1 Introduction 7.4 Implementation and Review of the HACCP Plan 8. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption 212 9.2 Legislative Structure 190 9. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.1 Introduction 189 9.4 Regulation (EC) No.10 Further Reading 223 viii .7 Food Hygiene (England) Regulations 2006.5 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 6.8 Ciguatoxins 7.3 Processing and its Effects on the Microflora 5. 852/2004 on the General Hygiene of Foodstuffs 191 9.10 Cyclic Imines 7.1 Definitions 5.2 Definitions 8. 2006 No.2 Initial Microflora 6.5 Regulation (EC) No.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.7 References 5. 14 (Hygiene requirements specific to the UK) 221 9. CURED.1 Definitions 6.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.3 Processing and its Effects on the Microflora 6.4 Spoilage 6.8 Guidance 222 9.6 Regulation (EC) No.6 References 6.6 Published Microbiological Criteria 5.11 Conclusion 7.7 Further Reading FISH AND SHELLFISH TOXINS 7.9 Azaspiracids 7. S.5 Pathogens: Growth and Survival 6. EU FOOD HYGIENE LEGISLATION 189 9.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.5 Pathogens: Growth and Survival 5.3 Stages of a HACCP Study 8. 7.4 Spoilage 5. SMOKED AND DRIED FISH 5. 8.1 Introduction 8. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.9 Other Relevant Legislation 223 9.3 Tetrodotoxin 7.

6 Salmonella spp.10 PATHOGEN PROFILES 10.2 Clostridium botulinum 10.10 Vibrio vulnificus 10.3 Clostridium perfringens 10. 10.4 Listeria spp.8 Vibrio cholerae 10.9 Vibrio parahaemolyticus 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix . 10.1 Aeromonas spp. 10.7 Staphylococcus aureus 10.5 Plesiomonas 10.


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

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

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

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

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

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

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

35 °C 8 . In addition. and mud Ubiquitous. However. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . Plesiomonas shigelloides 3 .102 101 .19 µg/kg bodyweight from < 102 .14 . 105 . Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. and risk associated with the consumption of seafood is low (2.109 cells/g) * Some sources considered L. Toxin levels 0. Escherichia coli Yersinia enterocolitica 101 . sewage. stream water.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp.37 °C High for most species. numbers of pathogenic bacteria in raw fish tend to be low.0.1 .26 °C 10 .37 °C Unknown Unknown TABLE 1.106 cells/g (exception: V. estuarine environments. 4). vulnificus) 5 .>106 Salmonella spp. 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. 4 .1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.FISH AND SEAFOOD TABLE 1.106 cells/g.103 High (107 . birds.

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

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

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

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

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

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

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

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

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

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

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

Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. Generally. Since 1996. Japan. and North America.g. accounted for the majority of cases (1. 5). Proper temperature control (<3. V. sushi) (5). the risk posed to the consumer of fresh fish.6. salted or pickled) fish have also been associated with botulism. due to the cultural preference for raw fish dishes.2005). use of post harvest treatments and/or cooking of fresh seafood will reduce risks. The first report of the organism being a foodborne agent was with shirasu. Other lightly or semi-preserved (e. the majority of outbreaks are caused by consumption of raw fish products (e.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked.3 °C) will eliminate all risk. is small. parahaemolyticus cases have increased across the world. Only about 1% of marine isolates produce a thermostable haemolysin. which is believed to be required for virulence. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. a Japanese boiled and semi-dried sardine dish. USA. Russia. storage and preparation (11). In Japan. and when in contact with contaminated water (no CDC reported cases from fish. again. the majority are associated with products not cooked immediately before consumption. A unique clone of V.2 Vibrio parahaemolyticus and other vibrios V. France.g. This strain has been responsible for 50 to 80% of all V. Despite the ubiquitous nature of type E C. which was probably contaminated from an uncooked food source or an excreting foodhandler. Southeast Asia. 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. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. I-sushi. Proper chilling. Currently. 1. parahaemolyticus is a marine organism. V. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). It is generally undetectable in marine water below 19 °C but may grow in culture at temperatures as low as 5 °C and on food at 10 °C. The bacterium causes primary septicaemia. 11). vulnificus is associated with warm marine and estuarine waters. whether stored aerobically or in modified-atmospheres. were associated with finfish between 1978 and 1998 (46). but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . 1978 . and proper cooking (boiling for 1 minute. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). which can cause human gastroenteritis. parahaemolyticus outbreaks. parahaemolyticus O3:K6 is responsible for many of the recent V. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. smoked. there is no specific guideline that describes a minimum level of V. including epidemics in India.

26 to 0. the causative agent of cholera.CHILLED AND FROZEN RAW FISH disorders). The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. L. listeriosis cases have been reportable to the CDC. cytotoxic. There is no risk associated with the consumption of properly chilled and cooked fresh fish. are present on fish and can grow to significant numbers during storage (1.20 °C. in its habitat.000 17 . are associated with human diarrhoeal illnesses. but the bacterium is known to survive and grow in the shallow marine. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. They are aquatic organisms with an epidemiology that is yet to be fully understood. Some isolates are true psychrophiles. monocytogenes incidence has been between 0. proper chill storage will eliminate any risk associated with this organism. Although the psychrophiles are unable to grow at body temperature. shigelloides is similar to Aeromonas spp. their potential to produce disease-causing exotoxin in food has not been fully examined (48). but proper cooking should substantially reduce risk of infection. Similar problems as those encountered for Aeromonas spp. with minimum growth temperatures around 0 °C and optima of 15 . coli on isolation media (50). 1. in assessing the risk of fishborne infection by P. 17). 1. P. others are psychrotrophic mesophiles (49). Most cases have been sporadic rather than associated to large outbreaks. 5. is historically associated with faecally contaminated water. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. monocytogenes has been well documented as a foodborne human pathogen. which is often fatal (>50%). The major virulence factor appears to be the production of toxins with enterotoxic. V. Chill storage may not eliminate growth.3 Aeromonas Aeromonas spp. cholerae O1. and proper cooking will substantially reduce risk. and especially estuarine environment. Again.4 Listeria monocytogenes L.. 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. It is known that Aeromonas spp. Since 2000. infection. Despite this. In the US. sodium channel blocking and haemolytic activity (48). and especially Aeromonas hydrophila. shigelloides are encountered. and fish and shellfish are probably the major reservoirs for the organism (50). possibly due to under-reporting because of their likeness to E. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters.6. there is no risk from the consumption of properly handled and cooked fresh fish. however.55 cases per 100.6.

Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54).6. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection.48 minutes in salmon have been reported (53). it is not apparent to the consumer and it cannot be destroyed by cooking. Photobacterium spp. there is a possibility for growth of the organism on chill-stored fish. full risk analysis is not possible (52).pregnant women. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . Clostridium spp. Fortunately. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. Klebsiella pneumoniae and Hafnia alvei. 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. at risk groups . 1.98 minutes in cod and D60 of 4. foetuses.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. but proper cooking will reduce any risk.2002 (55)) and is also common worldwide.6.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity... The most frequently reported parasites of human importance in fish are round worms of the genera Anisakis and Pseudoterranova and tapeworms of the genus Diphyllobothrium. some Vibrio spp. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. especially pickled fish. and Lactobacillus spp.. Like other toxins. 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. Being a psychrotroph. alcoholics. leads to the production of histamine. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). If fish are not properly refrigerated.FISH AND SEAFOOD persons. Lightly preserved products. D60 of 1. but cooking will significantly reduce the risk of infection. Infections are mostly associated with the consumption of raw or 18 . It is regularly isolated from seafoods (51). 1. but more especially by Morganella morganii. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. The organism poses a serious risk in chilled products not cooked before consumption. neonates. 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.

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

11.. Kautter D..R.T. Gram L. 1992.S.. Feldhusen F. 1992. Critical Reviews in Food Science and Nutrition. 8. 5-7. 91-8. 2. Codex Alimentarius Commission. Rosnes J. 243-60.. 9. Ahmed F... 1993.8 1.. 12. 348. 19.A. 10. 2003. 37. 2001. 1997.J.. Food Control. Capell C. Microbes and Infection. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres.FISH AND SEAFOOD 1. Davey K. Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review. 12.H. References Huss H. 2. Food Control. Fletcher R. parasites. 1651-60. 12.A. Gibbs P. 5 (1). Davies A. International Journal of Food Microbiology.N. in FAO Fisheries Technical Paper No. Quality and quality changes in fresh fish. Gram L. Huss H. 27. 8 (2). 3. Davies A. 24. 1996. Sivertsvik M. Jeksrud W. 291-304. 1996. 1994. Microbiological spoilage of fish and fish products. 87121. 13. 7. 5.H. International Journal of Food Microbiology.H. Food and Agriculture Organisation. activities and safety. Huss H. Simpson B. Codex Alimentarius Commission. Food Control.J. Food and Agriculture Organisation of the United Nations. Ed. Italy.. 1989. and natural toxins.P. 36.H. 20 .. Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms. Journal of Food Safety. Huss H.R. Kirby R. 33. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). M. Rome. 2002. Incidence and growth of psychrotrophic Clostridium botulinum in foods.. 1994. Food and Agriculture Organisation. Incidence of foodborne pathogens in European fish. in FAO Fisheries Technical Paper No.H.. International Journal of Food Science and Technology.K. The role of seafood in bacterial foodborne diseases. Spoilage and shelf life extension of fresh fish and shell-fish.J. Assurance of seafood quality. 4.. 1995.E. 334. Control of indigenous pathogenic bacterial in seafood.A. Jehanno D. 283-94.K.. Dalgaard P. Nychas G. Ashie I. 14. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth. Reddy N. Rhodehamel E. International Journal of Food Microbiology. 107-27. 87-118. Rome. 67-71. Smith J. 121-37. Armstrong D. Ed.. 2000. Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef. Huss H. International Journal of Food Sciences and Techonology. 6..R.R. Food and Agriculture Organisation of the United Nations.E.

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

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

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

6. 1992.M. Cho Y.H. Houston N. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures.L. 317-21. International Journal of Food Microbiology. Akiba M.. 295-307. Huss H. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches. Ravn Jørgensen B. 317 (Suppl. 271-85. McMeekin T. in Fish as Food.2 Spoilage Shewan J. Gibson D. 55. Eds. International Journal of Food Microbiology. Food and Agriculture Organisation. 1984. Florida. International Journal of Food Microbiology.. International Journal of Food Microbiology.C.E.G. Rome.. 127-34..9. 1989. 1..M. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. 62-70. Torry Research Station of the Ministry of Agriculture... Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. 1989. Journal of Food Science. 1985. Bulletin of the Faculty of Fisheries (Hokkaido University). Kitts D..D.P. Application of predictive microbiology to assure the quality and safety of fish and fish products. Handling and processing of scad.FISH AND SEAFOOD 1. Ed.. Fisheries and Food. Technomic Publishers. Picard G. 24 .M. in Seafood Science and Technology. Trolle G. 1961. Huss H. Olley J. in Torry Advisory Note No 93. 1988. Changes in microflora and chemical compounds in mackerel stored by partial freezing. Lancaster. Jones Y. 15. Huss H.. 65-72.1 Further Reading Processing Anon.1983. Estimation of the bacteriological quality of fish by automated conductance measurements. Taoukis P..9 1. Food and Agriculture Organisation of the United Nations. 73-89. 1992. Ross T. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. in FAO Fisheries Report No. Gram L. 1984. 35. Sumner J. Microbiological quality and shelf life prediction of chilled fish. 741-50.S. Torry Research Station.. Torry Research Station of the Ministry of Agriculture. Shahidi F.). Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere.. 1986. 4. Cann D.. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Dalgaard P. Fisheries and Food. Journal of Food Protection.A. 1. Aberdeen.D.H. Borgstrom G. Ed. 1997. Shinano H.. Magno-Orejana F.. Simard R.. Academic Press. The Microbiology of Seawater Fish. Ogden I. 1.. Villemure G. Gibson D. Smith G. Hobbs G. Labuza T. 51.9..H. Aberdeen.. 13-32.

Huss H. Garcia M. 2197-203. 479-88. 72.9. 1993.M. 20023. 1997. London. Eds.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A.J. Huss H. in Quality Assurance in the Fish Industry. 1. Huss H.H. 137-46. 25 ...L. Trolle G. A rapid method for detection of histamine producing bacteria. 322-6.K. Liston J. 1987. 1987.H.. 1. Elsevier Science. Gram L.. Otero A. Modified atmosphere packaging of fish and fish products.9.H. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. 59. Sala F. Klausen N. in Fish processing technology. Effect of culture age.R. 1992. Blackie Academic & Professional.europa. 1992. 65-72. Jacobsen M. 5. International Journal of Food Microbiology.. Applied and Environmental Microbiology. 4. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures...CHILLED AND FROZEN RAW FISH Einarsson H. Ed. http://eur-lex.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. Amsterdam. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. Journal of Applied Bacteriology..3 Pathogens Condon S. International Journal of Food Microbiology. Hall G.eu/LexUriServ/LexUriServ. Gram L. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C.


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

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

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

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

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

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

It is recommended that the principles of HACCP are applied to novel processes. Examples of tempering techniques include the use of heated air. For some components. to evenly raise the temperature of the entire frozen product from -20 °C or lower. and contamination from equipment to clean fish muscle. there are a number of distinct methods used. or the use of microwave technology. Effects on the microflora are minimal during tempering (7). Generic examples have been used to illustrate only the major points. but will also need to ensure that the optimum shelf life can be safely obtained. but it also poses a risk of cross-contaminating other products undergoing tempering.1 . up to -5 °C so as to improve the cutting and handling properties during processing. defined holding time at a chill temperature (0 – 3 °C).4.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. the core temperature.3 Preparation of fish component of chilled and frozen recipe dishes 2.4. Band-sawn materials or fish dice are normally promptly further processed. 2.1 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. Figures 2.4. and potential opportunities for growth of foodpoisoning bacteria.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 .2. Where the raw materials are frozen.3. All have the same intention. line extensions and process reviews. Block-sawn dust that is required for use in fish cakes is swiftly transferred to a chiller to avoid water loss and spoilage from process contaminants. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. It is critical that the time and product temperature during tempering be controlled to avoid elevated temperatures (>5 °C) on the product surface allowing microbial growth or the onset of spoilage in raw material. as a heat process is often required for functional or organoleptic reasons. but it is assumed that any band sawing or dice process will introduce heat from blade friction.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. Rapid controlled cooling of heat-processed components will help to minimise spoilage. 2. as the full complexity of the processes used for specific products cannot be covered in a general text. this may be an incidental effect.

2. 2.3. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product.2).1. Frozen cores are usually batter-enrobed.Shaped Fillets. This material is transferred to a processing facility with the ice replaced as necessary to ensure that the product is maintained at the temperature of melting ice.7 days. On arrival at the processing plant. may not rise above 0 ºC during processing. Raw material preparation stages for chilled or frozen coated fish products 34 .1. after landing.1.4. chilled or sold chilled on defrost with an expected shelf life of 6 . quickfrozen.3. 2.4.1B .1 (2. The processing of three examples types are described in Figure 2.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen. by mincing and mixing with a binding agent (as described in section 2. with the coating heat-fixed. the scampi is washed in potable water to remove foreign bodies. frozen in a nitrogen blast freezer and weighed into sacks for later processing. drained and then refrozen for storage until required for further processing. are placed into plastic boxes and covered in ice to preserve their condition.Scampi.1A .Fish Cakes). Fig.1C . The “blown” meat is graded.FISH AND SEAFOOD processed. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. It is then size-graded.2.

such as aerobic spore formers and Staphylococcus spp. are handled and stored as any other food product. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. chilled. unless it is poorly handled.2 Shaped products This type of product falls into two categories.1.. from potato flake. 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. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp.off-cuts or belly flaps.4. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. material will need to be transferred swiftly to a chiller. This is especially the case with block-sawn dust. spoil more rapidly than the dry ingredients.1.3 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. and water is added to rehydrate the components. The mix is shaped and formed ready for packing or for enrobing. pathogens such as Salmonella and Listeria. more seriously. 2. fish fingers). The fish component of the cake will.7 days. because of its nature. The remaining dried ingredients are mixed with the fish.block-sawn dust. quickfrozen. additional spoilage organisms may be introduced. Waste material produced from this type of processing is often reused in the third category (fish cakes). Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. or sold as a chilled product after thawing.4. 35 . off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. or those where the trimmings. but. with the coating heat-fixed. ii) Fish-filleting waste .g.3.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. that are to be reused for other foods. iii) Fresh/fillet/minced fish block. Fish cake cores may be batter-enrobed. once the dry components have been rehydrated. with an expected shelf life of 6 . as the physical structure of the material has been destroyed.3. It is for this reason that all by-products from primary processing. Therefore. or promptly processed to avoid further quality loss. those that use frozen blocks of whole fillets.

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

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

as this could introduce off-flavours and reduce shelf life. The vegetables must withstand processing.5. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. to produce a specific product. and must not show evidence of disease.3. Fig. rice. but they are supplied without a source of carbohydrate e. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). There are two main categories. canned fish may be used in their preparation owing to it having wider availability and low wastage. A temperature no greater than 10 ºC for 12 hours cumulatively or 21.1 ºC for no more than 3 hours is recommended (1). 38 .2.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe.g. For example.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products.4. pasta or potato. Flow diagram outline for recipe dish ingredients preparation 2. Figure 2. 2. The protein source can be fish fillet or block.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal.4.

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

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

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

the method replaces air in the sealed packaging with a defined mixture of gases. Flow diagram outline assembly and packaging of recipe dish 2. 2.7. The oxygen-free conditions will however allow other anaerobic bacteria such as C. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow.7. These include: 2.4. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. which may restrict either microbial growth or enzyme activity.2 Modified-atmosphere packaging Originally developed for the red meat industry. and tuna loins are packed by placing the product in a plastic pouch and removing all the air. unless the temperature is strictly controlled to below 5 °C throughout the supply chain.4.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. botulinum to grow. generally carbon dioxide and nitrogen.4.5. 42 .1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions). as it dissolves in the surface moisture to form a weak acid.2.FISH AND SEAFOOD Fig.

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

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

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

growth may occur in foods (26). Rates of contamination with smoked seafood are much higher than those with unsmoked products. but under the appropriate conditions. L.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. at the present time.6. 2. infected food handlers. and will therefore cause “food poisoning” even in a product that is cooked before consumption.6. 2.5 Shigella spp. the restrictions on growth rate seen with seafood will not apply.6. are considered primarily to be waterborne organisms but epidemiology indicates that contamination of vegetables and possibly in-shore caught seafood could act as vehicles for infection by Shigella (34). 46 . 2. will colonise factory environments in a similar fashion except for possibly Vibrio spp. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). It is unclear. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. indigenous or other. However.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). but. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). and this is believed to be due to a tolerance to brine used in the smoking process (32). when introduced or inoculated into sterile components of chilled prepared recipe dishes. which is not affected by any subsequent cooking process. Sources of contamination are many because of the widespread distribution of this organism in the environment. if any other potentially pathogenic organisms. aureus (35). and inadequate cooking or holding within the temperature growth range (27). Shigella spp. These include inadequate refrigeration and preparing foods far in advance of planned service. An acceptance has now developed in the UK of the presence at low levels of L. These conditions will allow the production of the toxin. Chilling may help Shigella to survive. aureus enterotoxins in foods. monocytogenes as a contaminant in foods (31). L. monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32.

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

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

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

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

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


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

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

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

mostly marine bivalve molluscs. hard-shell or soft-shell clam. laterally compressed. geoduck. or siphons.2.1 Clam Clam is the common name of a large group of often edible. extended above the surface to maintain water currents needed for respiration and feeding. littleneck. Buried clams leave tubes. and other names. but 56 . cherrystone. 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 3.1. A mantle tissue encloses the body of the clam. The muscular foot is used to burrow in mud or sand. Different species are variously called quahog. Most clams are only a few centimetres in their maximum dimensions.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint.

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

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

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

Water temperatures affect the concentrations of microflora in the water.3 3.3. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. Vibrios. Alcaligenes spp. Enterococci spp. responding to the environment in which they are harvested. Acinetobacter spp. the initial flora composition and concentration of bivalves are highly variable. At harvest. spirochetes of the genus Cristispira and Saprospira.. In summary. There are microorganisms. and yeast cells of Rhodotorula rubra and Trichosporon spp.1 and 3. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp.3. Moraxella spp. 11.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. mussels. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. and cockles The processing of bivalves takes two forms. Flavobacterium spp. 10. that have never been cultured on laboratory media. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. shellstock (live) which is shellfish in the shell.. Temperature.1 Shellstock 3.1. clams. whole or in part. mechanical harvesters.3.2. 12.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. 13).. 15).1. observed microscopically. Other microorganisms found in lesser numbers are the Achromobacter spp. and Aeromonads.. rakes.1 Processing and its Effects on the Microflora Oysters.. The largest groups found are the Pseudomonads.. 3. Bacillus spp.000 /g. nutrients. salinity. Campylobacter coli Bacillus cereus Hepatitis non-A. For example. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. Pseudomonas spp. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. 3. (9. dredges. or by 60 . from which one or both shells have been removed. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves.1.. but have never been cultured (14. Micrococcus spp.. They are termed viable non-culturable microorganisms.. and shucked shellfish that is shellfish.

Shellstock are placed in drycooler storage containers in the facility. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock. If by boat. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. 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. or in bay-approved shellfish-growing waters prior to sale or processing. for transportation to the point of sale. There are three primary reasons for wet storage: first. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale.1.1). after harvesting. or immediately placed in refrigerated trucks. Fig. Water used for shellstock washing is from approved growing areas.3. The shellstock is transported dry. Shellstock may be wet-stored at the processing facility in tanks of clean seawater. in containers from which they can be retrieved easily. or from other sources approved by the State Shellfish Control Agency (SSCA). Secondly. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. Thirdly. usually in open-mesh containers and/or burlap sacks. Generic flow diagram of live shellfish from harvester to point of sale 61 .MOLLUSCAN SHELLFISH hand (Figure 3. by truck or boat.

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

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

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

8% (37). Pseudomonas. and packing the shellfish in a timely manner.2 ºC. shucking pails. 36). or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. Proteus. abundant gas production. spoilage will occur.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. blowers and other utensils that have cracked. these include lactobacilli. Shucked shellfish are an excellent medium for the growth of bacteria. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. 33. streptococci. and proteolysis (10). Glycogen is hydrolysed by the initial flora found in shellfish. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30.2 ºC or less. Achromobacter and Flavobacterium levels decrease during storage. For example.47 to 6. Glycogen. clean gloves. 65 . Temperature abuse will result in high bacterial counts. and yeasts. This process changes the bacterial profile to microorganisms favouring low pH. Controls that should be applied include using clean packing equipment. The initial flora found in shellfish is the major factor of spoilage in bivalves. glycogen levels range from 0. particularly shucked meats. All equipment coming in contact with the shellfish such as colanders. anywhere during the process of shellfish from harvesting to the point of sale. Shucked shellfish are held and transported at temperatures of 7. As described previously. These organisms may play a significant role in the spoilage processes. Much of the work has been done with oysters and clams. Flavobacterium. particularly oysters stored at 7 ºC. Shellfish. are an excellent medium for growth of bacteria. is stored in the bivalve as a source of energy. 31. whereas bacterium such as Pseudomonas. a carbohydrate. The spoilage process in shellfish may be divided into three stages: increase of acidity. in oysters. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. Alcaligenes.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. thus producing acid and gas (38). where there is a breakdown of temperature control or prolonged storage of shellfish. 3. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. Lactobacilli seem to be the predominant microflora in shellfish. In summary. 34. rough. 35. skimmers. Micrococcus. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. at the turn of the century. showed that organisms such as Achromobacter. The environment has an effect on the bacterial population found in shellfish. 32). 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. Earlier studies.

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

1. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. those who have liver disease.5. or primary septicaemia. and preventing cross contamination once the product is cooked. these individuals are advised not to eat raw shellfish.0. 57. High-risk individuals.4 . cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. Control of oyster-associated 67 . and vomiting. 46. with a fatality rate of 50% for individuals with septicaemia. The bacterium is an estuarine species that causes wound infections. gastroenteritis. leukaemia.MOLLUSCAN SHELLFISH Control of V. Control of V. vulnificus. 53. vulnificus are fever. chills. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). cholerae non-01 (51). 55. like V. cholerae non-01 has been associated with septicaemia. Inadequate refrigeration will accelerate the growth of V. It causes diarrhoea. parahaemolyticus the estimated annual incidence rate is very low. parahaemolyticus growth in shellfish meats is temperature dependent. 23 of 24 major USA-West Coast estuaries were positive for V. 54.3 Vibrio cholerae non-01 Illness caused by V. and rapidly become inactive at low storage temperatures. V.48 hours of onset. are particularly susceptible to primary septicaemia. 58). in the range of 0. Hazards from this bacterium can be controlled by thorough cooking of shellfish. The occurrence of V. The organism attaches to the intestine and release of a toxin is suspected. 48). diabetes. 3. lasting 6 . 49). Oysters are the most hazardous food associated with V. fever. vulnificus is by rapid refrigeration during warm weather months. 3. however.7 days. cholerae non-01 is ubiquitous in the estuarine environment. cirrhosis. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. All isolates are considered pathogenic.5. the organism has been found in the Gulf of Mexico. They are sensitive to cold.2 Vibrio vulnificus The symptoms of V. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52.000 population/year (8. Primarily. nausea. and nausea within 24 . The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. 46. V. cholerae. Again.II (8. In one study.8 case/100. which have been implicated in shellfish-related disease outbreaks (31). parahaemolyticus. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). 56). or immunosuppression. It is one of the most severe foodborne infectious diseases. Death has been reported to occur within 36 hours from onset. V. Onset of symptoms occurs in 48 hours and can be severe. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8.1. abdominal cramps. Healthy individuals are susceptible to gastroenteritis.

5. 3. 62.1. salinity. 60). low pH. 63). 3. vomiting. There is evidence of strains of V. mimicus. There are several other Vibrio spp. vulnificus.S. In addition. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. fever. fluvialis. cholerae 01 establishing in the U. or cross-contaminated shellfish (57. parahaemolyticus and V. these organisms cause gastroenteritis lasting 24 .5 Other Vibrios spp. undercooked. cholerae 01 is transmitted by faecal contamination of food or water. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). nausea. and southern Asia including India and Bangladesh. and shock. respectively. and illness lasts 7 . Illnesses are associated with the warmer months.10% carbon dioxide for optimal growth conditions. however. widespread in both polluted and unpolluted waters.5. They are common marine bacteria. Death may occur after severe fluid and electrolyte loss.FISH AND SEAFOOD V. Onset of disease is 6 hours to 5 days. cholerae 01. particularly in oysters. is the same as previously discussed for the non-01 strains. dehydration. preventing crosscontamination of cooked seafood. Onset of illness is from 3 to 5 days after ingestion of contaminated food. 3. and nausea. diarrhoea. e. The frequency of infection of these organisms is no different from other Vibrios. V. processors should know the product source. jejuni requires reduced levels of oxygen (3 . In general. Control of infection from these organisms is no different from the other Vibrios. nausea. In developing countries. V.5%) and 2 .4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. It causes diarrhoea.2 Campylobacter jejuni C. The presence of these strains appears to be in low numbers and non-virulent (59). Gulf coast environment. Illness can include abdominal cramps. 68 . and V. (8. the pathogenic severity is lower than V.g. headache and fever. including V.10 days. and the transmission of the disease by consumption of raw. drying. Poor sanitation and contaminated water supplies will spread the disease.48 hours with abdominal pain. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. V. cholerae 01 in molluscan shellfish. The control of V. associated with shellfish-borne illness outbreaks. 61. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. abdominal pain.5. hollisae.1. acidity and heat. cholerae non-01 illnesses is by cooking seafood thoroughly. imported product from a country experiencing an epidemic.

They have been implicated in limited shellfishassociated gastroenteritis cases (8. of Incidents 4 279 0 1 0 No. control of C. 69. mimicus. 3. V. 77. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. 83.3. cholerae 01 and non-01. To reduce the risk of shellfish-borne campylobacteriosis. from Pacific shellstock oysters purchased from a retail grocery store (74). of Cases 4 362 0 1 0 a Includes V. 78) and P. The seasonality of 69 . however. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. Campylobacter spp.MOLLUSCAN SHELLFISH TABLE 3. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. 70. shigelloides (79. In one case. C.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. parahaemolyticus. Recently. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). 71). the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. V. jejuni does not multiply in shellstock at refrigerated temperatures. they survive well at refrigerated temperatures. although their role as a pathogen is highly controversial and circumstantial. have been isolated from shellfish beds throughout the world (67. 66). Campylobacter appear to survive well in oysters during storage. hydrophila group (75. whereas at 10 ºC Campylobacter survive only for 5 days (71). This finding is significant in that C. V. vulnificus. hollisae Infective dose varies with host susceptibility and virulence of the strain. V. 85). fluviallis. and V.4 to 114 cells. 72.II Molluscan shellfish associated outbreaks caused by Vibrio spp. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. 73).1 Aeromonas hydrophila group and Plesiomonas shigelloides A. 65. infection occurred among participants of a firemen's banquet. 76. Although. 68. 81. 80) are ubiquitous in the marine environment. C.5. 84. and their immediate proper refrigeration.3 Other organisms of concern 3.5. jejuni were recovered. 82. Human feeding studies suggest that about 400 . ranging from 0.500 cells/gram of Campylobacter may cause illness (64. however. jejuni begins in the harvest of shellstock from approved waters.

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

1981. 755-76.S. A typhoid fever epidemic caused by carrier bootlegging oysters. 1960.D.. 42. U. Fish and Wildlife Service. 2. Walker W. 1983. American Journal of Public Health. Applied and Environmental Microbiology. 1982.N. Bernard F.H. Cook D.S. 1970.W. 1994..A. 6. water and sediment from Galveston Bay.R. Galtsoff P.. Vanderzant C. New York. Pfeifer D. Journal of Milk and Food Technology. 36. 19-34.. 231-51. 7. in Multilingual Dictionary of Fish and Fish Products. 15. 544-8. Ed. Applied Microbiology. Microbiology of bivalve molluscan shellfish. Schmidt C. Lovelace T. 9. Ray S. 16. Colwell R. Applied Microbiology. Bacterial flora of the market oyster. Eds. USA.W. 82-7. in Microbiology of Marine Food Products. References The Phylum Mollusca.MOLLUSCAN SHELLFISH 3. Liston J. Van Nostrand Reinhold. Fleet G.E. 12. Ward D. Hood M. Journal of the Mississippi Academy of Sciences.K. American Journal of Hygiene. Jr. 30.. 1940. Lee J. Rippey S. Eliot C. 6. Journal of Food Protection. Applied and Environmental Microbiology. Nauman R. 447-52.. 8. The American Oyster. 17. Viability and release Salmonella charity and Escherichia coli from oyster faeces. 1964. 3. Wiley-Blackwell.A. 7. 1973.F. Washington D. 33-6.. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. Organisation for Economic Co-operation and Development. 1936. Villee C. Proceedings of the National Shellfisheries Association.A. in Fishery Bulletin 1964 No. Smith F. 1975.M. Veliger.R. 105-8. Tall B. Government Printing Office. 33643. Colwell R.. Biology of the oyster in relation to sanitation.R. Galtsoff P. 13. 633-40. 1968. 46. 5. Gill S. 419-25. 71 . Eds. Old H.L.E. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. 26. 1995. 1991....R. Thompson C.J. 1926. 10. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica).C. Saunders Company. Effects of harvesting waters and storage conditions on yeast population in shellfish. W. 11.7 1. Infectious diseases associated with molluscan shellfish consumption. Fate of enteric bacteria in estuarine sediments and oyster faeces. 4. American Journal of Public Health. 72-8.S. Cook D.K. 44. Clinical Microbiology Review. 104-9. 58. Jr. Hackney C.S.. in General Zoology. 1984. 14. 8. Microbiological characteristics of Dungeness crab (Cancer magister). Rowse A. 1963. 71-6. 30. 245-7. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas).B.. 64.R. 13. Philadelphia.R. 29. Tubiash H.C.

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

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

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

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

Ed. 1991.C. Government Printing Office . Washington D.MOLLUSCAN SHELLFISH 88. mussels. 90. Food and Drug Administration. 560.C. Martin R. in U. Food and Drug Administration. Lancaster. Clams. Aspen Publishers Inc. 77 ..S. Gaithersburg. Sherlock C.S. 7. 91. Slavin J. Seafood Safety. 89.600 (CPG 7119.H. Ed.M. Pittsurgh.W. U. Food and Drug Administration. National Academy Press. U.S.E. Inc.. Technomic Publishing Co. Collette R. Smith J.12). U. Fish Inspection.S.E. Chapter 5: Foods. in National Shellfish Sanitation Program Manual of Operations. Burdge D..030. Ed. Food and Drug Administration.C. Food and Drug Administration. Part I Sanitation of Shellfish Growing Areas. and HACCP: A Global Focus.L.S.. 3.Pathogen monitoring of selected high risk foods (FY 88/89). Food and Drug Administration Compliance Policy Guides. 2000. oysters.. Baird-Parker T. 1987.. Washington D. Lund B. in U. Program 7303.C. Quality Control.S.W. The Microbiological Safety and Quality of Food.A.S. Food and Drug Administration. U. 1997.. U. 1995. 63-8. Gould G. Food and Drug Administration Compliance Program Guidance Manual. Washington D.1987.S. U.. fresh or frozenadulteration by bacteriological contamination Sec. 1995. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease.8 Further Reading Ahmed F.. Food and Drug Administration. Food and Drug Administration.R.


(deep-water shrimp). 79 . lobsters and crabs have ten legs. and are usually aquatic. packed and frozen in 0.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. Such products are usually only partially cooked before freezing. with additional water to form a glaze. Peeled ready-to-cook shrimp are raw. Pandalus montagui and Crangon crangon (inshore shrimp). and frozen in 1 and 2. jointed limbs and a chitinous exoskeleton. e.Ph. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). CRUSTACEAN SHELLFISH Linda Nicolaides. M.g. They possess a segmented body. Pandalus borealis (deep-water shrimp). Shrimps.0 kg polythene bags. peeled product.0 kg polythene bags.5 and 1... crabs. crayfish and shrimp (1). FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. and Trachypenaeus spp. lobsters. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. Species eaten from tropical waters include Penaeus spp. and available in 0.5 kilo pack (boxes or bags).4.5 and 1. with shells and heads on or off are usually packed. Parapeneopsis spp. Crayfish. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. the front pair ends in claws.. Peeled and cooked shrimp are ready-to-eat after controlled thawing.

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

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

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

An offensive odour is also produced. and have a sweetish. the meat is either repacked and sold in the shells. the joint between the tail and carapace is open and the limbs hang loosely. 4. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. The shell will be soft. there is no indication of life. since crabs and lobsters are traditionally consumed immediately after cooking. The shells have a crisp. Once removed and cleaned. and a strong ammoniacal odour is produced. immediately prior to boiling. or in hermatically sealed cans that have undergone an appropriate heat treatment. for crabs.CRUSTACEAN SHELLFISH 4. In comparison.2 Cooked crabs and lobsters As indicated earlier.4. with the growing demand for ready-to-eat products from crab and lobster meat. contaminating microflora may include enteric bacteria. With spoiled crabs and lobsters. bright and dry appearance. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. or packaged and processed.4.1 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 a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. whilst boiled crustaceans have a pleasant slightly sweet smell. indicative of protein degradation and the growth of the principal spoilage bacterium Sh.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. bacteria of public health significance. If the tail drops.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. Heat is produced if the shrimps decompose in a confined space. slightly iodine smell. 4. 4. 83 .3. If hygienic procedures are not followed. If boiled. spoiling prawns and shrimps have shells that are soft. there is no associated spoilage problem. dull and sticky. viruses or parasites. However. they include Sh. putrefaciens and Pseudomonas spp. wet and ‘soapy’ or ‘jammy’ to the touch. putrefaciens.

Although foodborne illness had not been associated with the product. Following this. The above results prompted the FDA to develop a compliance programme for the pathogen. An import alert was put in place in June 1987. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . in 24.2 Listeria spp. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). shrimp and shellfish (11).5. 16). later surveys in 1990 recovered the pathogen from retail raw. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). However. fish fingers.cooked prior to consumption. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. cooked and smoked fish. shrimps and cockles sold (17). The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . and usually for both. Moreover. 4. monocytogenes in cooked ready-to-eat foods.5 4. animal manure. even if GMP is followed.5. 6.FISH AND SEAFOOD 4. as well as for Salmonella. The presence of L. monocytogenes was absent from 40 samples of cooked prawns. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. in domestic/imported shrimps (15). Further surveys identified Listeria spp. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments.5% of frozen raw shrimps from different countries (13). although efficient cooking will quickly eliminate it (6). Aquatic birds and the hands of workers have also been identified as sources of the pathogen. Hence. a common pond fertiliser used in aquaculture. Salmonella cannot be completely removed from raw shrimp during processing. 7). Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. 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. nearly 30 years elapsed before the potential hazard of L. 5.1 Pathogens: Growth and Survival Salmonella spp.

the pathogen can grow at temperatures as low as 3 °C. therefore. 25). V. 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. the causative agent of cholera. aureus is not a member of the intrinsic flora associated with crustacean shellfish. which can become colonised by V.g. monocytogenes has been attributed to crustaceans. was associated with seafood consumption. is widely distributed in aquatic environments. To date. combined with the degree of handling of these types of product. Crustaceans have been linked with food intoxications attributed to Staph. whilst boils and cuts are also common sources of the pathogen. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. pasteurisation and cooking. 4. cholerae. about 50 . cholerae non-O1. 4. V. no incidence of foodborne illness caused by L. 19). botulinum type E is commonly associated with fish and fishery products. 24. e. although contamination by types A and B may occur from the environment. as part of the natural bacterial flora. Once the competing microflora has been removed by a heat treatment. pregnant women and the immuno-compromised. The main source of contamination is the hands of processing personnel.70% of healthy individuals carry toxin-producing strains of Staph. especially in products that have been vacuum packaged. The pathogen has resistant spores. aureus on their hands. However. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. Vibrio cholerae. cholerae is divided into two groups. 22) as well as from undercooked crustacea when harvested from contaminated waters (23.CRUSTACEAN SHELLFISH old.5.5. Control should be achieved by promoting GHP and staff training programmes. 4. particularly crabmeat (18. An epidemic that began in South America in 1991. Crabs are covered in a chitin exoskeleton. aureus because of the ubiquity of this organism in the processing environment. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. The former. 85 .4 Clostridium botulinum C.5 Vibrio spp.5. cholerae O1 and V. botulinum type E at 10 °C but not at 4 °C (20). although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. so is able to survive heat treatments that eliminate vegetative cells.3 Staphylococcus aureus Staph. Researchers have demonstrated that inoculated shrimp supported toxin production by C.

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

risk-based system based upon the seven principles of Hazard Analysis and Critical Control Point (HACCP) supported by pre-requisite or good practices programmes at all stages of the food chain. 4.11 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. 4. 38. No foodborne incidents have been attributed to this source to date. consistent with that in the stomach. The risk of such hazards is far higher with fin fish and molluscan shellfish. 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. 87 . 4. which results in a persistent and apparently permanent loss of short-term memory (36). 39). They can survive low pH levels.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). A summary of microbiological criteria for crustacean shellfish is listed below. from farm to fork (37. Norwalk viruses can also withstand heating to 60 °C. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34).6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive.5. which could be transferred to the meat during unskilled removal from the shell (35). produced or imported into the European Union (EU). only 0. although it is one of the European Union (EU) legislated allergens.6. 2073/2005.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. or chlorine concentrations similar to those found in drinking water. Selected parts from this regulation is presented below. The toxin causes amnesic shellfish poisoning (ASP) when consumed. Provisions laid down concerning the use of additives would also apply to these types of products.CRUSTACEAN SHELLFISH 1981.5.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. 4.

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.


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

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

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

only the most common processes can be described here.80º brine (184.6 g of salt or more per litre of brine) may cause salt burn in this way. etc. For smoking salmon.8 211. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. Therefore. it is important to understand the concept of salt concentration. This should be distinguished from the salt content. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. or salt in the water phase. When working with salting. it is impossible to establish standardised processing stages in the curing of fish. Salt in the water phase links the two conditions of moisture and saltiness. When a milder brine. the fish is usually salted with a 70 . a 0º brine saturation means pure water.FISH AND SEAFOOD 5.4 184. At 18 °C a 100º brine is produced when the solution contains 26.0 158.4 52.8 79. TABLE 5. it is important to state the temperature. about 50º (132. processes and producers’ private recipes.0 . sometimes the salt crystallises onto the surface of the fish.4% salt (Table 5. and salt content.6 264. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.I Brine strength at 18 °C (after Bannerman (2)) Brineometer degrees 10 20 30 40 50 60 70 80 90 100 96 Weight of salt (g/litre of brine) 26.I). a 90º or above brine (237.3 Processing and its Effects on the Microflora Owing to the wide variety of products. Thus. is employed. The dissolution of salt in water is dependent on the temperature. it causes swelling because the fish absorbs water from the salt solution (1).2 237.2 105.6 132. Different brine strengths are required according to the curing processes and/or the type of product.2 g of salt per litre of brine). all using salting steps. % 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. smoking or marinating processes. The strength of the solution is described as degrees of saturation (brineometer degrees). Salt concentration means the percentage of salt in the water contained in the fish. When a 100º brine is used.8 211.0 g of salt per litre of brine).

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

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

17 5. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.51 5. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .51 3.61 3.30 2. the liquid that comes out of the fish forms a brine. TABLE 5. the fish enzymes play an important role by maturing or ripening the product.95 3.84 1. See Table 5. the fish is dry-salted.5 4.41 6. sardines.23 3.66 5. There are several variations of this process. they can be stored at 28 °C.64 4. herrings and other fatty fish can be used. The fish are kept immersed in their own brine near saturation point for several days.76 4.73 1.61 3. Fresh sardines.76 4.0 3.70 3.04 5. This is why it is the preferred method for fatty fish. Anchovies are the main fish used for these purposes.85 3. if split fish is dry-salted.05 7.60 2.78 6.44 2.54 5.96 1. Here.11 3. but as it is placed in containers.1. with a shelf life of about 4 weeks (water activity (aw) about 0.26 4.35 4.00 3. the product obtained is called gaspé-cured (5). For preservation purposes.3 Dry / wet salting In this process.23 2.41 5. headed and gutted. but mackerel.65 2. the brine is then replaced over the fish.91 2.26 2.11 4.98 6.76 4.98 6.5 3.33 5.66 5.86 2. can be cured in a saturated brine solution for 6 days and. with pressure being applied to the top of the container to help the brine to drain away.II.06 4.5 10. after being drained and pressed at 465 kg/m2 for 15 hours.26 3.0 2.23 2.25 5. 5.3.76 4.10 1.76 4.41 4.99 2. and a red burgundy colour will develop in the product.09 8.90 (1).09 8. thus. Histamineforming bacteria do not generally grow at water activities below 0. SMOKED AND DRIED FISH smoking.26 1. a saturated brine is used.19 3.3 days.46 4.70 6.85 3.41 3. and can be collected and replaced again over the fish.45 3.44 2.76 99 .0 4.98 5.23 8. limiting the amount of rancidity that can develop.03 2.26 7.74 9.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) 2.88 5.80 5.78) (1). The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.03 4.78 2.49 4.56 6. For example.11 9.16 7.5 3.CURED.25 5. Histamine formation is of importance when using scombroid and some clupeidean species.17 4.22 6. The main objective of the brining method of salting fish is to limit air penetration during processing.47 6.

then packed in barrels with a 70º brine. but it has further smoking. Afterwards. has a shelf life of several years. Usually. Ceviche. However. drying and salt-boiling steps until a hard translucent block is obtained (4). Another salt-boiled Indonesian product. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten.6 days without refrigeration (1). Once cooled. ‘ikan kaju’. is made of different varieties of fish. usually flavoured with onions and hot chillies. usually requiring refrigeration below 10 °C for longer-term storage.2% acetic acid and 2 . Another variation uses sour cream instead of the second brine step.2 months.3. underlines the need for careful control (4).30 minutes. salt.3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. Herring can also be cured for several days in an 80 to 90º brine with 2. The lime juice plays the role of the vinegar. or a pH <5 and they are stabilised for a limited period. a Peruvian product. or can be boiled by being placed in a basket. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. they can be cut and cured for 3 days in a solution with 3% white distilled vinegar.5% 120grain distilled vinegar.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. even if the product is hermetically sealed. until the flesh is soft. and packed (3). All these products are ‘semi-preserves’.4% of salt) for distribution and sale (1). which is dipped into a boiling brine for 10 . because of the salt content of the product. up to 1 . Other preservation techniques can be used for extending the shelf life of these products. and lasts for 3 . herring or other fatty species are used. and 6% salt. and even death. The fish can be soaked in a high concentration of vinegar (about 4 . the occasional implication of pindang in cases of foodborne illness. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. As referred to above. giving a cooked appearance to the product. only the nonspore-forming organisms are eliminated in the process and. containing 1 kg of salt for every 25 kg of fish. it can be repacked in milder brine (1 . and spices) is added to the brine.7%) for as long as 3 weeks. A much firmer product is obtained when the fish is salted prior to acidification. They will have a salt content >6% (w/w) in the water phase. placed in a marinade of lime juice. they are packed for consumption within 2 weeks at chill temperature (6). whereas the latter method preserves the fish for only a few days. Traditional jellied eels are a product where pieces of fish are cooked in boiling water. There is epidemiological evidence that these products have been the cause of some 100 . outgrowth of bacterial spores after processing may occur (4). In a second step.FISH AND SEAFOOD 5. flavouring (sugar. The first process can give a longer shelf life. 5.3. After this.

5. Matjes are made of fat young herring. They are an important dried product. Provided the product is kept in dry and fairly cool conditions. gutted. Gravad fish is a Scandinavian product made of salmon fillets. still made without prior salting on a large scale. Foodborne pathogens. Some bacteria and enzymes are still active in the presence of acid and salt. etc. It is important to assure the quality of the raw material. are generally air-dried at sea.3. marinated products will keep for several months at a temperature of 4 °C. bacterial toxins and parasites (7). it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. and the flesh will break down. In Scandinavian countries. At pH levels below 4. It takes about 2 days to marinate completely. depending on the thickness. In the first case. sea cucumbers and seaweeds. such as squid. Usually. 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). SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins.CURED. jellyfish. air flows naturally. 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. split and dried gadoid species) are traditionally made in cold climates. are likely to survive in these mild-processed products. by using either solar heaters or mechanical dryers. 101 . Other products. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. the fish is dried naturally in a relatively cold environment. some fish species such as moray eel and varieties of dog fish. In some coastal areas of Portugal. used as an appetiser. South America and the Caribbean.There are several products named marinades (matjes. Stockfish is extensively used in Portugal. Stockfish products have a golden brown colour and are very hard. but sometimes it is dried without prior salting.) where no acid is added to the process. the temperature and the amount of pressure applied (5). the fish is dried after salting or brining. and the stacks are placed in the refrigerator. a very fine delicacy for Oriental food lovers. sugar and spices. Stockfish (dressed. mixed with salt. it will last for many years (5). which are placed in a mixture of salt. Italy and Spain (5). are sundried and. Dried shrimps are widely used in Asia. The most common method used for fish products is the air-drying process.4 Air drying This is probably the oldest method of fish preservation. such as Listeria monocytogenes and Clostridium botulinum. Shark pectoral fins. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). Dried octopus is also a very common dish in the interior of the country. 5.0) (7). while in the latter case there is a provision for fans or other means of mechanically moving the air (1). under a light pressure. are commonly dried in Asian countries. gravad fish. Fresh dill is placed between the layers of fish. after frying. and a fine product is obtained.

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

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

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

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

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

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

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

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

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

Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. with a lower aw limit for growth of 0. including A. Refrigeration is necessary. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. In Vietnam. able to grow on dried salted fish. 36). The brining to 3 . The salt in the water phase must be high enough to inhibit the growth of C. and microbial spoilage is more likely.CURED. The heat during hot-smoking will kill vegetative microorganisms but not all spores. It has been reported that the type of salting (dry.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. 33. flavus. Fish processed by cold-smoking. but. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations. so the bacteria spoiling these products will be psychrotrophic or psychrophilic. Pseudomonadaceae. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32.. Aspergillus clavatus and Aspergillus niger. around the water activity of saturated sodium chloride. SMOKED AND DRIED FISH infrequently at low levels. The preservative effect of the smoke will inhibit some spoilage microorganisms. however. Vacuum packing limits chemical oxidation. Marine vibrios were the dominant flora in smoked salmon that was drysalted. Cold-smoked fish will contain microbial flora representative of that of the raw fish. For more information on spoilage see Table 5. it was. has been isolated from dried fish (35).747.IV. Smoked fish products tend to have a light brine treatment. not the growth of spoilage organisms. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. Aspergillus spp. as already stated. but this will not prevent the growth of spoilage bacteria. 111 .V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). will have a different residual post-smoking flora from 'hot-smoked' fish. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). LAB. which does not create the salty conditions required for spoilage by those halophilic microorganisms. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life. Table 5.5% sodium chloride in the water phase will inhibit some bacteria. where a high internal temperature is reached. botulinum type E.

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

and as botulism is a serious hazard in foods. and C. botulinum. Before.CURED.8 0. when present in the raw material.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C.75 0. The presence of salt in cured products has a great effect on the growth of the bacteria. The concentration required to prevent growth at room temperature can vary from as low as 3. botulinum type E Staph. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13).9 0. under optimal conditions. botulinum Staph. aureus Yeasts Moulds Lactic acid bacteria pH 5.97 0.97 0. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C.5-8. Examination of 165 outbreaks of botulism caused by fish products showed that smoked.5 5.0-9. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. with this change. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C.5 4.86 0. sodium 113 . but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39).5-10 4. In the United States. popular demand for healthy foods has led to a change in the tastes preferred by the public and. botulinum type E. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. 9 and 8 of the outbreaks. salted and pickled products accounted for 10. smoked and dried. pressure on the traditional methods of curing. the products were heavily salted.5 0-11 3.5-8. Since neither the smoking nor the drying parts of this curing process are particularly severe. the fermented products being the most dangerous group (7). SMOKED AND DRIED FISH TABLE 5.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. However. botulinum type E and production of toxin.8 1.5 5. but if the fish products are stored at temperatures <10 °C.61 Organisms Most Pseudomonadaceae C. could not grow and produce toxin. types A and B are not affected by 3 . respectively.5. the non-proteolytic type E strains can grow in up to 5% sodium chloride. although the combination of the two factors. It has been shown that.5-10.5% to 5%. the importance of cold-chain integrity must be emphasised.

Thus.5. are particularly susceptible to contamination and. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). However.94 (7). but requires higher temperatures for toxin production (>10 °C). when the bacterial load of the product is lower. the risk of infection can be eliminated by proper cooking. Once the product is contaminated. it should contain not less than 3. 5. as they require more handling. and the minimum pH for growth is 4. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. the organism may contaminate the product. growth and toxin formation may occur.5% salt in the water phase. but. 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. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. 5. As they are poor competitors. The organism is mesophilic with a minimum growth temperature of 7 °C. aureus growth. Usually. aureus.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. if recontamination occurs and storage conditions are favourable.86 and 10% sodium chloride.5.or cold-smoked fish should have at least 3. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . if the person handling the product is a carrier of Staph. coli all occur on fish products as a result of contamination from animal/human reservoirs. throat and skin. As with other organisms.0 .3 Enterobacteriaceae The pathogens Salmonella. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. more importantly. It is a halotolerant bacterium able to grow at water activities as low as 0. An outbreak of Shigella in an Italian cruise ship. the organism can survive most of the light curing processes. In foodstuffs. if it contains sodium nitrite ≥100 ppm.30% of the population positive for enterotoxin-producing strains (41).0% salt in the water phase (30). the human nose. Infection by Shigella sonnei has increased in the UK. after processing. 5.5.4 Listeria monocytogenes The psychrotrophic L.5. but. related to the consumption of a raw smoked fish meal.4.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. they do not grow well in the raw material. Sliced smoked fish products. Shigella and E. The FDA established that hot. they will grow rapidly (7). has been reported (42). if storage temperatures are over 10 °C. gills and guts as well as in several lightly preserved fish 114 .

in the EU Fisheries R&D-FLAIR program. a number of fish products. lightly salted caviar. which include intermediate hosts.45 °C. Some LAB.95 (45). such as ‘lightly preserved’ fish products.8 . are considered unsafe. 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 CT 3162-FAIR program. most of them are of little concern to public health. monocytogenes pathogenic for only specific segments at risk. there were no reported outbreaks related to cured fish. Its limiting conditions for growth are 0 .514.12%. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. SMOKED AND DRIED FISH products such as cold-smoked fish. marinades (ceviche) (43) and gravad fish (44). a round worm.CURED. do not survive 1 min at 60 °C. which. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. the only concern to public health is when eating raw or uncooked fish products.2. They have complicated life cycles.9. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). This includes matjes herring. and the tape worm Diphyllobothrium spp. As they are heat-sensitive. a short period of freezing either of the raw material or of 115 . which means that the process must conform to a level of ‘no detectable L.6 and water activities below 0. from freshwater fish. salt levels of 8 . from seawater fish.5. or nematodes. and in CT95-1207FAIR). the USA established ‘zero tolerance’ for the organism.0. in combination with curing agents and refrigerated storage. pH 4. widely used as starter cultures in food processes. gravad fish.92 . Thus. cold-smoked fish. both being detected in cold-smoked salmon. The round worms.the round worm Anisakis spp. The use of LAB in fish preservation is a topic that has been studied in projects (UP. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. Of microbiological concern in the northern hemisphere are two parasites . favours growth of psychrotrophic LAB. have been shown to have an anti-listerial activity in many food products. Until 1997. Two outbreaks in the United States have been reported to be linked to salmon (10). monocytogenes’ in 25 g of the finished product. and other local traditional products (7).5 Parasites Despite the presence of parasites in fish being very common. 5. which considered L. has also been detected in cod (7). As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. Pseudoterranova dicipiens. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. Unlike some European countries. monocytogenes in levels at or above 102 per gram (46).

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

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

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

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

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

25 (5). 58 (5). Beuchat L. Davies A. 1995. 844-6.P... Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. Letters in Applied Microbiology. 11. 94-8.S. 1994.. 1994. Tran T. 1995. Contamination pattern of Listeria monocytogenes and other Listeria spp. 1987.R. New York.K. 293301.178-80.A. 1075-80. Bayliss A.. Poysky F. Eds. Jemmi T. 309-16. International Journal of Food Microbiology. 1991. Dillon R.J.E.R. Kramer D. Hitchins A. International Journal of Food Microbiology. 14. Ben Embarek P. 27 (6). Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. 1973.R. Patel T. Ratnam S. and temperature on the growth of Vibrio parahaemolyticus.. 1988. 61 (12).. Beuchat L.. solute.A. International Journal of Food Microbiology. Yndestad M.N. Dallas H. Journal of Food Protection. 4389-95. Prevalence of Listeria in smoked fish. 23 (1). 42. Fuchs R. 1992. Taylor S.C. 25. 502-8. 55 (11). pH and sodium chloride. Paranjpye R.. Rørvik L. Journal of Food Safety.. Letters in Applied Microbiology. detection and growth of Listeria monocytogenes in seafoods: a review. Incidence of Listeria in hot and cold-smoked fish. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. Interacting effects of pH..CURED. Hudson J. Marine toxins of microbial origin. Keusch A. 1734. Food Technology. and salt concentration on growth and survival of Vibrio parahaemolyticus. Applied Microbiology. Eklund M. 309-18. 121 . in a salmon slaughterhouse and smoked salmon processing plant. Caugant D. Nicolaides L.A. 1995.T.. Histamine production by psychrophilic flora.. van Spreckens K. Combined effects of water activity. Presence. Liston J.. 866-70. Journal of Food Protection. 1994. Poindexter C. 217-36. SMOKED AND DRIED FISH Sutherland J. Letters in Applied Microbiology.. Pelroy G. 1994. in Seafood Quality Determination. 21 (3). Peterson M.. 19.. 1992. 11. 19-27. Roberts T.. Elsevier. 394-6. Lashbrook L. Applied Microbiology.W. Food Microbiology.. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. 1974. Slade A. Competition of food bacteria with Listeria monocytogenes during enrichment culture. temperature.


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

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

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

and the initial microflora is likely to be considerably larger in these cases. states that sugar is often included with the salt. Penaeus indicua Many of the fish used in fermentation are uneviscerated.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. it was found that several different sources were used for the fish . or buried salmon. and old descriptions of their production often refer to the addition of whey. For example. in a survey of fermented fish products in north-east Thailand. 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.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. 126 . some fermented fish products remain popular. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. especially in fish/salt products. Table 6.2 Initial Microflora Composition of the initial microflora of fish is described in some detail elsewhere in this Handbook and will not be reiterated here. and their microflora tends to reflect their local environment more than that of marine species. made from herring. relatively mild-tasting product. 6. TABLE 6. freshwater fish are the usual raw material. A modern account of their production. malt or flour (1). rakefisk or surfisk. which employ lower levels of salt than Asian products. which has been increasing in popularity over the last 50 years. These are often described as fish/salt products.FISH AND SEAFOOD In Northern Europe. the most popular varieties of which are rakørret. and a large local freshwater reservoir (2). is a traditional. and counts ranging from below 104 /g to in excess of 107 have been reported. Norwegian gravlaks. The diversity of fish used in fermented products (see. in Sweden. fermented trout. for example. There are also more heavily fermented products.the flooded rice fields. Here. however. paddy ponds beside rice fields used for collecting fish when the field has dried up. in Norway and surstrømming.

Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. principally species of the genera Rhizopus. in many cases.4.74 or below. either directly or as the partially saccharified rice they produce. Hansenula. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. mean that microorganisms have been generally held to play little or no role in the process that follows. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). angkak (Philippines and Thailand) and look-pang (Thailand) are used. When this is a simple carbohydrate source such as sugar.3 6. It also demonstrated their significant proteolytic activity during the first month of fermentation. however. Bacillus spp. they can contribute to the initial microflora. More recently. which included nam-pla and patis. 4). traditional starter culture preparations such as koji (Japan). Mucor. 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. Endomycopsis and Saccharomyces . In another microbiological study of four fish sauces. In some cases.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. This. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). after one month.FERMENTED FISH Where other ingredients are used. however. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6).3. and studies 127 . Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. and the anaerobic conditions that prevail in the bulk of the material. 6. the Philippine fish sauce. sufficient to saturate the water phase present and produce an aw of 0. Generally. Other halophilic species have been identified from fish/salt products. These preparations contain a mixed microflora of moulds and yeasts. were found to be the predominant isolates. or boiled or roasted rice. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp.1 – 6. Similar results were seen with patis. 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.

6.12). Belacan (Condiment) Fig.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 . 6. Patis (Condiment) 128 .1.2. Fig.

however. In fish sauce production. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are. 6.4. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. the 129 .6. Ika-Shiokara (side dish) Fig. eviscerated fish are sometimes used to slow the autolytic process.FERMENTED FISH Fig. 14). This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. thought to be largely the result of autolytic breakdown of the fish tissues. To produce semi-solid fish pastes as opposed to fish sauces.3.

6. which is eventually separated from the residual fish tissues. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. snack) 130 .V and Figures 6. In contrast to free-swimming fish. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. 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.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. volatile fatty acids and nucleotides. Figure 6.8. filtered and bottled.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. Fig.3. Hoi-Malangpu-Dong (side dish. for example. Some examples of products and their formulation are presented in Table 6.6 – 6. 6.5.5). particularly glutamate.

3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 .7 days 5 . south of Thailand. Kung-Chom (staple food.5-1. C. C NE. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus.12 days 3 . side dish) 131 . N.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. the prefix 'pla' is replaced by 'kung'. NE. NE 10 .6. N. S.12 months Main dish 2 Pla-jao C. north.2-4 Fish (pieces): salt:khaomark (fermented rice). NE Formulation Fish (whole or pieces): salt: roasted rice. central. In these cases.03 Preparation time 6 . 3:1:0. Fig. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. N S 5 . N.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.FERMENTED FISH TABLE 6. C NE.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .6.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE.12 months Shelf life Use 1 .4 are also produced using shrimps.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. north-east.

typically in the range 2 ..FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. Phillipines Journal of Food Science and Technology.2. but there is considerable variation in this. This is readily explained by the susceptibility of LAB to elevated salt concentrations.7. 106) Fig. Red burong isda 132 . 2.1 The use of higher salt levels results in a longer production phase but a better keeping quality product.salt and lactic acid. If acidity is a desired sensory property of the product. This can be clearly seen from the data on reported shelf lives of a number of fish salt carbohydrate products and their respective pH ranges and salt contents (Table 6. It also indicates that. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6.3.6.VI). while they are able to grow at high salt levels. giving a reduced rate of acid generation. The LAB isolated from these products are those with more marked salt tolerance but. their growth and metabolism are nonetheless slowed.10%. (from Arroyo et al. then this will take longer to develop than if lower salt levels are used. the salt is of far greater importance in determining the product’s long-term stability. of the two factors contributing to shelf life . Generally. these products are salted at lower levels than the fish sauces and pastes.

9 4.4.2 7. White balao-balao TABLE 6.8.5 .7 .0 .FERMENTED FISH (from Arroyo et al.5.8 pH 4.1 .4 .8 .4.5 . 2.0 Shelf life 3 weeks 6 .6. Philippines Journal of Food Science and Technology.3 5.6.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.9. 106) Fig.1 ..6 3.9.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).2 4.17.2 4. 133 .0 .12 months 1 .8 2.5.5 3.9 4.8 .0 .5.5.9 .3 years 1 .

FISH AND SEAFOOD 6. Garlic. alternatively. This results in a more active lactic fermentation and a lower pH. 19). When added as boiled or roasted rice alone. inulin. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. being similar (18. Where they are used. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. In several products. also serves as a valuable source of the more readily fermented carbohydrate.1 . 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 .0 . of course.3. also lower the unit cost of the product and this is probably a significant factor in many cases. For example.4.3. Addition of carbohydrate will.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. a common ingredient in these products. Although the ability to ferment starch is not widespread in the LAB. TABLE 6. 21).VII). a number of amylolytic species have been described and an amylolytic strain of Lactobacillus plantarum has been isolated from burong bangus in the Philippines (17). In som-fak 9% of LAB isolated were able to ferment rice whereas 19% could ferment garlic and omission of garlic from the formulation resulted in lower acid production during fermentation (20.5) despite other factors. Rice is the most common carbohydrate source used.2. they increase the amount of soluble sugars produced and thus the range of LAB that can grow.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. the rice may be added in a partially saccharified form such as kao-mark (Thailand). efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. although the final population of LAB may be similar.9) compared with the white product (pH 4. such as salt content.

3 days. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. 6. the level of salt employed is on the high side for fish/salt/carbohydrate products. Excessive growth and acid production by the LAB could render products too sour for acceptability. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases.4 Spoilage The ASCA handbook on fermented foods in the region ascribes shelf lives to various fermented fish products ranging from 2 weeks in the case of some of the low-salt lactic fermented products to 5 years for some of the high salt sauces and pastes (16).3 weeks at around 20 °C. Generally.5 in the first 2 . Results of a product survey were consistent with this. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. The mixture is allowed to ferment for 2 . will not reliably exclude air. the pH drops from 6.2 at the end of fermentation. is something of an exception since millet is used along with garlic as the carbohydrate source. There is little published information on the factors that limit the shelf life of the low-salt products.17. such as wrapping in banana leaves.7 to below 4. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. The growth of yeasts and moulds could. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. around 12%. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. the Korean fermented fish product. which ranged from 7. Traditional packing techniques used with lactic fermented fish products. 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). but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. allowing other spoilage organisms to grow. in turn. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23).5.8 . In a successful fermentation. and there is a marked regional variation in the preferred type. markedly extending the shelf life (25). The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products.FERMENTED FISH Sikhae. thereafter declining to about 4.9%. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. 135 .5:9:3. reporting the product’s pH to range from 4. but they are likely to behave similarly to lactic fermented products produced from other commodities. Clearly.7 up to 6. lead to an increase in pH.2. presumably correlating with the salt content. 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.

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

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

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

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

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

While some of these toxins only occur in seafood from a restricted geographical region. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. FISH AND SHELLFISH TOXINS Jeffrey L. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. Ph. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins.. Another book dealing with the clinical effects of a variety of poisons and toxins. 5). Increasingly sophisticated instrumentation (e. most notably the azaspiracids and the cyclic imines as well as some additional polyethers.. including the well known marine toxins 141 . known and unknown.g. a number of new toxins and toxin families have been identified. The manual on harmful marine algae and their products.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. C.D. several scientific reports have described their efficacy and potential. Wright.3) and some practical applications for detecting shellfish toxins have been reported (4. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. Scientific and Cultural Organisation) has been updated (7). Nevertheless. Since the original chapter in this series was published (6).M. It remains to be seen if this approach gains momentum in the future. An emerging technology is the use of immunosensors (1 . FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7.7.C. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins. physiology and detection of marine and freshwater toxins has also appeared (8). Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins. Although most of these antibody-based test kits have yet to receive international regulatory approval. and a second updated edition dealing with the pharmacology.

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

Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. this is never sufficient to render the seafood safe to consume. 143 . However.1. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis.2 Toxicology PSP toxins affect the central nervous system of mammals. 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). tongue. 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.30 minutes of consumption.2. and fingers within 5 . Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water.FISH AND SHELLFISH TOXINS STX GTX2 GTX3 GTX5 (B1) C1 C2 NEO GTX1 GTX4 GTX6 (B2) C3 C4 R1 H H H H H H OH OH OH OH OH OH R2 H H OSO3 H H OSO3 H H OSO3 H H OSO3 - R3 H OSO3 H H OSO3 H H OSO3 H H OSO3 H R4 H H H SO3 SO3 SO3 H H H SO3 SO3 SO3 - Fig. Human symptoms begin with tingling and numbness of the lips. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. 7. and humans seem to be particularly susceptible.

and over ten analogs are known (22. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). a pre-column method has received Association of Analytical Communities (AOAC) approval (18). 17). 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. 15. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. 7. Alternative HPLC methods have subsequently reduced this to a single HPLC run. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). specifically 144 . or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. Most intoxication events have occurred in South East Asia. 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. While the chemical detection methods are most promising for the future. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. 7. as well as other parts.8 µg/g) of shellfish or seafood tissue.2. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). other derivatives have been reported. and has also been evaluated by a number of EU laboratories. The mouse bioassay has been in place for decades and the method has been standardised.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. Thailand. and islands of the South Pacific. However it remains as the monitoring method of choice in many countries. Two approaches have been taken here. 23. including Japan.3. Since its discovery. and while simple and relatively inexpensive to operate.2). There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. 24) (Figure 7. TTX most commonly occurs in many species of puffer fish.FISH AND SEAFOOD 7. Taiwan. a criticism has been that such methods are time consuming. After some development and various trials.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. China.

27). the blue-ringed octopus. The onset of symptoms usually occurs rapidly. and in mild cases results in distal muscle weakness. preventing depolarisation and propagation of action potentials in nerve cells. Severe poisoning causes paralysis and respiratory failure. Since then. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. and once again the mouse bioassay has been used to detect the presence of the toxin. where puffer fish of the genus Fugu are a delicacy. In Japan.2 mg of purified toxin can be lethal. Typically this has been accomplished using HPLC followed by post 145 . 7. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. All of the observed toxicity is secondary to the action potential blockade.3.2. 7. Fig. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. respiratory failure and coma. nausea and vomiting. and more recently by an actinomycete Norcardiopsis dassonvillei (26). and even newts. 23. molluscs. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. raising speculation that TTX is produced by a bacterial source. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. Structure of tetrodotoxin 7. gonads and roe (24). horseshoe crabs.2 Toxicology Like the PSP toxins. it is served in speciality restaurants with qualified chefs to properly prepare the fish. 26). 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. A review of TTX detection methods has been published (12. An oral dose of 1 . 25. hypotension. and blocks diffusion of sodium through the sodium channel.FISH AND SHELLFISH TOXINS in the liver.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions.3.

Currently 12 isomers of DA have been reported (see Figure 7. Structure of domoic acid and three naturally occurring photoisomers 7. and while promising. but it is known that such congeners are less toxic than DA itself (30). 7. a polar glutamate agonist. However this method has been criticised for being complex.4.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28). but 146 . 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. Fig. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. and more rapid LC-MS methods [(M+H)+ m/z 320] have been reported for the detection and identification of TTX and its various derivatives (27).3.4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA).FISH AND SEAFOOD column oxidation to the fluorescent derivatives. 7.3 for examples). but all of them are considerably less toxic than DA itself (29). Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. need further development (24).

and this has led to ASP toxicity in birds. as does impaired renal function which increases serum concentration and residence time of the toxin. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). 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). 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. 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. nausea. Another factor is the movement of the toxin through the food chain. To a great extent. Okinawa and more northerly parts of Japan. Recently. cormorants and pelicans (34). found in the Pacific around Vietnam. and clams. Penetration of the blood brain barrier is poor in healthy subjects. For example.10% of the administered dose. which has been well documented in studies along the west coast of the US. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. Pseudonitzschia australis. the Mediterranean. 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. but any impairment of the blood brain barrier results in additional risk. southern and western portions of Ireland. disorientation and coma. including persistent short term memory deficits (36 .39). 7. Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. seizures. 33).4. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. The oral absorption of DA is around 5 . and in locations around New Zealand (32.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. 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 is mainly distributed in the blood. and sea animals such as sea lions and even whales (35). the Philippines. multiseries. another diatom species Nitzschia navisvaringica. 147 . has been reported as a medium-level producer of DA (33).FISH AND SHELLFISH TOXINS only three strains (P. Victims display a variety of symptoms including headache. scallops. Levels of DA in the other strains are generally <1 pg/cell.

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

analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. which include the pectenotoxins and the yessotoxins. Dinophysis acuta.4. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. DTX-5. Dinophysis fortii. Fig.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. 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. In this case Dinophysis was suspected as the culprit organism. Today over ten Dinophysis spp. although occasionally fatty acyl esters linked through the 7-OH group (referred to as DTX-3 compounds) are also found and are likely a result of bioconversion in shellfish tissue. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). such as Prorocentrum lima. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. The parent acids are the only DSP congeners to be found in shellfish tissue. 149 . 49). are also recognised as a threat to human health.5. are recognised as DSP toxin producers. Lately these compounds. Dinophysis caudata. Prorocentrum maculosum. and Dinophysis norvegica among others (47. and include Dinophysis acuminata..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. This was probably not the first DSP episode. DSP toxins produced by benthic Prorocentrum spp. In comparison. and Prorocentrum belizeanum. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. While these esters are not toxic in vitro. Structure of okadaic acid and the two related DSP toxins 7. 7.

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

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. Indeed. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . However. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. In yet another approach.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).5 and 7. Within this group. 58). Some representative examples of both groups of toxins are shown in Figures 7. and another rapid antibodybased test kit for DSP toxins has also been reported (53). Fig. Some doubt remains as to the significance of these compounds as marine or shellfish toxins with respect to human health since the PTXs for example are co-extracted with the DSP toxins. with a claimed detection limit of 20 µg/kg. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. 55).5.6. for many years these compounds were included in the DSP toxin complex. 7. but with the increased availability of pure toxins permitting detailed toxicology studies it is now recognised that both these groups of compounds possess a different toxic profile from the DSP toxins. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). with good agreement with the mouse bioassay and LC–based methods (54. 7. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49).

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

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

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

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

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

These include paresthesias (numbness and tingling of the extremities). though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. 7. the neurological symptoms can persist for days. 75). although not all CTXs are equipotent. The onset of symptoms can occur rapidly within 24 hours. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. 78). and temperature sensation reversal. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. barracudas.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. 7. 75. which in turn are consumed by carnivores such as snappers. weeks or even months (74 . The recommended method to extract fish tissue has been described in detail (74) and the initial warm acetone extract is partitioned against hexane and diethyl ether where the CTXs are concentrated.8.8. and it is estimated that ingestion of as little as 0. and there are suggestions that such incidents are increasing and spreading worldwide (74). pruritus. Other symptoms include a metallic taste.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. which can return following the consumption of alcohol and certain foods such as oily fish (74. Patients have reported symptoms lasting weeks and even months. Even when suspect samples are available. and although the gastronomical problems may dissipate within a day or so. Globally. and in fact CTX poisoning is often only identified after consumption of contaminated fish. Local island populations attempt to avoid poisoning by catching only smaller fish. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel.3 Detection methods As a consequence of their potency. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. and moray eels. Like the brevetoxins.1 µg of ciguatoxin can result in human illness (74. myalgia. there are neither standards nor an official testing program. only trace amounts of CTXs can cause illness following consumption of contaminated fish. Another symptom that can develop over time is extreme fatigue. sharks. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. which can be confused with chronic fatigue syndrome. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. where cold objects feel hot and vice versa. and the sensation of loose teeth (77). they bind to voltage-dependant sodium channels in the cell membrane causing them to open. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 .76). arthralgia. groupers. all of which can be linked to the effect of CTX on excitable membranes.

9).25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. which are based on an LC-MS/MS approach. Recent reviews of this new group of toxins have appeared (83.9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. and another dealing with the chemistry and detection methods has been published (85). 7. The original structure proposed revealed an entirely new class of shellfish toxin. followed by clean-up on silica and aminopropyl cartridges. but this only has application in a clinical setting (81). which features characteristic fragment ions corresponding to sequential losses of three molecules of water. Since the original discovery and identification of the toxin. another in vitro sodium channel-based assay. and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. Recently. Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. high-throughput screen (74). and the corrected structure was published in 2004 (86) (Figure 7. The majority of these have been proposed based on LC/MS analytical data.30%) and AZA3 (5 .9). minor revisions of a few stereocenters were made. Immunoassays may be an alternative detection method (79).80%). 158 . This is then subjected to LCMS/MS analysis. has been developed. In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). almost 20 additional derivatives have been reported (83 . Acetone extracts of fish tissue are subjected to solvent partitioning. with AZA2 (10 . specific for CTX activity. A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine. Usually the major AZA derivatives found in shellfish are AZA1 (50 . although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). and only a few structures (AZA 1 . at C-3 and C-23). only a few laboratories are able to undertake such analyses.FISH AND SEAFOOD Lethal Dose (MLD) 0. and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. Chemical detection methods are severely compromised by the lack of standards.g. Consequently. and following synthesis of the molecule some time later.20%) present in lesser amounts (Figure 7. following consumption of contaminated mussels Mytelius edulis (82). 84).85).

vomiting. Scandinavia. The bivalves affected include mussels (Mytilus edulis. 88.3 days. 84. which can persist for 2 . oysters (Crassostrea gigas.9. While this result is not necessarily in debate. Following the original episode. diarrhoea and stomach cramps. 7. it has been impossible to establish that P. scallops (Pecten maximus). 90).). Ensis siliqua. which are ingested by P. Ostrea edulis). and some time after the original episode in Ireland. but current data indicates that unlike the DSP compounds.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. Structure of the three principal azaspiracids found in shellfish 7. and to date this has been the only source from which these toxins have been found at toxic levels. cockles (Cerastoderma edule). AZA occurrence has been reported by several European countries including France.9. these molluscs were never considered to be the biogenetic source. 89). Donax spp.9. as in other cases of shellfish toxins. There has been considerable debate over the source of azaspiracids. These symptoms are not unlike those experienced following the consumption of DSP toxins. and off the coast of Morocco (83. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. Portugal. and most recently in a crustacean (Cancer pagurus). Mytilus galloprovincialis). cassipes (83).FISH AND SHELLFISH TOXINS Fig. AZA toxins do 159 . 7. and to date no long-term effects have been recorded (83. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). clams (Tapes philipinarum. However. cassipes produces AZAs in culture.

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

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

This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. and so spectroscopic detection by UV methods is not favoured. The time lapse between ingestion and death is between 5 and 15 minutes.100). The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. together with a lack of evidence of human intoxication. and this is the detection method of choice.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). 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. Reduction of the imine group eradicated the toxicity.to10-fold reduction of toxicity (LD50 600 .2 Spirolides The spirolides comprise the largest group of cyclic imine compounds.1. 50%) of 96 µg/kg. In particular. Most cyclic imines lack an extended chromophore. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. paralysis of the hind quarters and eventually complete immobility. Since their original discovery which resulted in the characterisation of spirolides A-F (99.1. it has been stated that regulation against gymnodimine is not required (98). Whether isolated from dinoflagellate cells directly or from shellfish tissue.3 Detection methods The characteristic “fast-acting. Oral administration of the toxin to mice by gavage resulted in about an 8. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. 7. and lack of responsiveness to stimuli.10. finishing with 100% methanol. Based on this diminished oral toxicity.900 µg/kg). edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. abdominal breathing. They were originally discovered in extracts of mussels (M. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. However. 95) (Figure 7. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. There is a rapid onset of symptoms including respiratory distress. 95).10.FISH AND SEAFOOD 7. A later study using purified toxin established an LD50 dose (Lethal Dose. 7. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine.11). The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . almost a dozen derivatives in total have been identified to date (94.10.

95). fasted mice). 7. fasted mice).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. in fjords in Denmark and Norway. 7.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. 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.2.10. Structure of four spirolide toxins A-D 7.11. southern Ireland. All data reported here are for desmethyl spirolide C.2. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101).707 µg/kg. by gavage (87 .2 Toxicology The spirolides are the most studied group of the cyclic imines. While oral toxicity is considerably less than by IP injection. the western coast of Scotland.166 µg/kg. those spirolides possessing the vicinal dimethyl grouping (see Figure 7. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . or by oral ingestion (381 . Interestingly.11. Fig. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98).10. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. the gulf of Maine.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . 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.

2. four members of the group have been identified – two of them being the C-34 diastereomers.FISH AND SEAFOOD (100). The compounds are zwitterionic and therefore somewhat more polar than the spirolides. 102).B contain a spiro imine moiety similar to that found in the spirolides. To date. 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. and in fact these compounds share a number of other structural features. While the mechanism of action remains to be clearly defined. the mouse bioassay can be used to detect these fast acting toxins. Three members of this group have been reported. 95. However. 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 .3 Pinnatoxins and pteriatoxins Pinnatoxins A . A number of methods following this procedure have been described (94. Fig. consistent with modulation of acetyl cholinergic receptors (103).3 Detection methods As discussed for gymnodimine. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7.12. which includes two diastereomers. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. 7. 7.

3. but this remains to be conclusively established (106).FISH AND SHELLFISH TOXINS 7. and more advanced means of detection. Only the acute toxicity in mice following IP injection has been reported. The LD99 dose for the pteriatoxins is reported to be in the range (8 . There is strong circumstantial evidence to suspect that these toxins. to date action or 165 . 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. there is no compelling evidence to suggest that they have resulted in human toxicity (98).180 µg/kg). 99%) values for pinnatoxins A-C are in the range (22 . Coupled with this. new toxin groups such as the azaspiracids and cyclic imines have also been reported.1 Origins and distribution Shellfish of the genus Pinna. while pinnatoxin D was essentially inactive. 7.3 Detection methods No formal detection methods for these compounds have been reported.11 Conclusion There has been considerable progress in the detection of known marine toxins.100 µg/kg).10.most likely a dinoflagellate. The LD99 (Lethal Dose. arise from an algal source . Despite this progress. However. With increasing knowledge of the chemistry of toxins.2 Toxicology There is little information describing the toxicology of these compounds (98). This is generally a result of increased monitoring and continually improving detection technologies and methods. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins.3. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). which occupy tropical waters around Okinawa and the south east coast of China. While this is recognised as a real possibility for a wide range of toxins.3. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. 7. 7.10. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). like the gymnodimines and spirolides.10. which in many cases are now found to be more widespread than previously believed. have long been suspected of causing human illness after ingestion. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other.

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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.


3.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . Validation . 4.1 Assemble HACCP team HACCP requires management commitment of resources to the process. 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. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available.A point. 7. 5. 8. Establish procedures for verification to confirm that the HACCP system is working effectively.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. food microbiology. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. procedures. 6. tests and other evaluations. Figure 8. in addition to monitoring to determine compliance with the HACCP plan. It is recommended by the Codex Alimentarius that the practical application of the HACCP principles be approached by breaking the seven principles down into a 12-stage logic sequence. ingredient and packaging buyers. food chemistry or engineering. Determine the CCPs. Verification .1 is a flow diagram illustrating this 12-stage logic sequence. quality assurance or control. operation or stage in the food chain including raw materials. procedure.Obtaining evidence that the elements of the HACCP plan are effective. 3. Establish documentation concerning all procedures and records appropriate to these principles and their application. hygiene managers. Each stage is discussed below in detail. The team should also include 177 . 8. from primary production to final consumption. Establish the critical limit(s).The application of methods. Establish a system to monitor control of the CCP. Conduct a hazard analysis. 2.

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

3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. 8.3. aquaculture drug residues and potentially pathogenic Vibrio spp.3. may need to be considered specifically. marine toxins associated with particular fish and shellfish species. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. freezing. contaminated fish and shellfish should not be used. there may be species-specific hazards related to certain fish and shellfish. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. etc. pH.3. composition.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. Expertise on such inherent hazards. physical and chemical structure (including water activity (aw). It is also important to identify the consumer target groups. is a cooking process required?).3. However. shelf life. naturally present on fish from warm waters (> 15 °C). fermentation. processing conditions (e.g. 8. packaging.3. including practices onboard fishing vessels. In addition.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. If the processing does not include a kill step. storage and distribution conditions and instructions for use. or in aquaculture operations. such as enteric viruses in shellfish. 8. These factors are likely to have a significant effect on some potential hazards.). Vulnerable groups of the population. which should be described in detail. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments.).1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora. etc.1. It should provide an accurate representation of all steps in the production process from raw materials to the end179 .3. such as children or the elderly. 8.2 Describe the product It is important to have a complete understanding of the product.g. heat treatment. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation.4 Construct a flow diagram The flow diagram should be constructed by the HACCP team and should contain sufficient technical data for the study to progress. The description should include information such as the product name.

time/temperature data.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. survival or multiplication of pathogenic microorganisms. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. features of equipment design. 8.3. and therefore a detailed description of the procedure applied must be available.6 List all potential hazards associated with each step. Ideally. ingredient specifications. or circumstances of any reclaim or rework activity. For example. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. If HACCP studies are applied to proposed new process lines or products. The operation should be observed at all stages. 8. it is essential that the details of each step are fully appreciated and recorded.4.5 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. However. It may include details of the factory and equipment layout. 8. because the hazard analysis and decisions regarding CCPs are based on these data. the qualitative and/or quantitative evaluation of the presence of hazards.FISH AND SEAFOOD product. conduct a hazard analysis. 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. It is essential that the diagram is accurate. cleaning and hygiene procedures and storage conditions. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. and identify any measures to control identified hazards The HACCP team should list all hazards that may reasonably be expected to occur at each step in the production process.3. 180 ● ● .3. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. it should also include details of CCP steps. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. once determined. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised.

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

2) to provide a logical. However.1 Fish and seafood Again.3. 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 . biological or sensory parameters. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. then the process must be modified to include a control measure. and its use may not always be appropriate. it is not possible to generalise on likely CCPs. Where possible. More than one critical limit may be defined for a single step. If a significant hazard has been identified at a step where control is required for safety.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.3. but for which no control exists at that step or any other. However. For example.FISH AND SEAFOOD 8. 8. critical limits should be specified and validated for each CCP.7.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. Criteria used to set critical limits must be measurable and may include physical. application of the decision tree should be flexible.3. 8. given the enormous variety of fish products and processes in use. it can be said that effective control measures are likely to include the following: • • • • • 8. structured approach to decision making.3. 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. chemical.8.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. it is necessary to specify both time and temperature for a thermal process. and the reader is referred to the appropriate product chapter.


• • • 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.


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

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

the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. p.9. as last amended by Regulation (EC) No 806/2003. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. distribution. European Communities L 268. 7. Council Directive 92/48/EEC of 16 June 1992 laying down the minimum hygiene rules applicable to fishery products caught on board certain vessels in accordance with Article 3(1)(a)(i) of Directive 91/493/EEC (OJ L 187. storing. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. as last amended by Regulation (EC) No 806/2003.B. manufacturing. packaging. Subsequently.1991.2 Legislative Structure From 1 January 2006. 24. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption Regulation (EC) No. transportation.9. 190 • • .1). J. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. N. and sets out what those enforcing the provisions have to do. 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. handling and offering for sale or supply to the consumer. processing. 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.7. 41).1992. p. 9. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No.1991.15). 24. A number of more detailed implementing and transitional measures have been adopted at EC level. p.

LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. penalties and powers of entry. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. 2073/2005. the legislation is directly applicable law.I. Wales and Northern Ireland. The regulation lays down general hygiene provisions for which food business operators carrying out primary production must comply with as laid down in Part A of Annex 1. national legislation in the form of a Statutory Instrument (S. 852/2004. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland.I. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. 9. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. and of hunting and fishing. e. As regulations. which will be covered later in this chapter. domestic preparation or handling.g.7. The Food Hygiene (England) Regulations 2006 (S. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. setting offences.3 Regulation (EC) No. There are some exemptions.14. is required to give effect to the EU regulations. etc. and equivalent legislation in Scotland.11) The EU hygiene regulations apply to all stages of food production including primary production. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. for example. Primary products are defined as products of primary production including products of the soil. 2006 No. food storage that is for private or domestic consumption. Wales and Northern Ireland).1993. 1 . of stock farming. p. Likewise Regulation 852/2004 will not apply to collection centres and tanneries meeting the definition of food business because they handle raw material for the production of gelatine or collagen. Additionally the requirements of EC Regulation 853/2004 must be complied with. right through to the implementation of procedures based on HACCP principles. Although the regulations have the force of law. 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. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. with primary production. 191 . 19. revocation of existing implementing legislation.) in England. and binding in its entirety on all member states from the date of entry into force.



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:


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

4. (iv) If the identification mark is applied in an establishment within the Community. In this case.4. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. EÜ. the new mark must show the approval number of the establishment where this occurred. Section I of Regulation (EC) 853/2004. for other codes.4. please refer to Regulation (EC) 853/2004. EC. ES.2.2. EK. Section I of Regulation (EC) 853/2004 are given as follows: 9. EB or WE. and the characters easily decipherable and be clearly displayed. EF. (ii) However. the mark must be oval shaped and include the abbreviation CE.1. EO. the food business operator may apply the same identification mark to both types of food.4. when a product's packaging and/or wrapping is removed or is further processed in another establishment. Extracts of the requirements of Annex II. EG. a new mark must be applied to the product.FISH AND SEAFOOD 9. (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.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. For the UK. the code would be UK.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. 9.1. EK. 198 .2. (iii) Additionally. EY. (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. the mark must show the approval number of the establishment.2 Form of the identification mark (i) The mark must be legible and indelible. 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.2 Fishery product requirements 9.

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

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

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

chilled and wrapped. 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.FISH AND SEAFOOD (vi) A water tank needs to be placed where there is no contamination of the water supply and hand washing taps should be designed to prevent contamination. The brine must not be a source of contamination.2.Requirements during and after landing 9. provided no damage occurs to the flesh. other than those kept alive. (ii) In the case of food business operators responsible for auction and wholesale markets etc. c) Fishery products. Chapter II . the product must be kept at no more than -9 °C. there need to be lockable facilities to refrigerate detained fishery products and separate lockable facilities to store fishery products that are unfit for human 202 . where fishery products are displayed for sale. e) In cases where whole fish intended for canning is frozen in brine. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. B. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. landed as soon as possible. Hygiene requirements Hygiene requirements refer to the following: a) Parts of vessels used for storage of fishery products must be cleaned and maintained in a good state with no contamination. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above.2. and products washed thoroughly. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. must be chilled or. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen.4. if this is not possible. Handlers may use spiked instruments to move large fish or fish that might injure them. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. b) Factory vessels on board which crustaceans and molluscs are cooked.

(ii) Heading and gutting must be carried out hygienically. Food business operators need to co-operate with the relevant competent authorities so as to allow them to carry out official controls as laid down in Regulation (EC) No. Additionally. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. part I. points 1 and 2.2. including vessels. 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. handling fishery products A. They should be wrapped. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 .LEGISLATION consumption. Requirements for frozen products An establishment on land that freezes fishery products needs to have equipment that meets that for freezer vessels as stated in Section VIII. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. B. they must be stored under ice in appropriate facilities. C.Requirements for establishments. packaged and chilled as quickly as possible after preparation.3 Chapter III . C. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. (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.2. prepared or processed immediately after arrival at an establishment on land. Fillets and slices must then be removed from work tables as soon as possible after preparation. dispatched. and where necessary. 9. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. Chapter I. 854/2004.4.

(ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . (iii) A document from the manufacturer. 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. must accompany fishery products referred to above when placed on the market. (iii) sprat.4. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. (iv) (wild) Atlantic and Pacific salmon.2. (ii) mackerel. (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. and whole fish must be gutted and washed beforehand. Once produced. If no other method of preservation is used.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. stating the type of process they have undergone.2. and (c) marinated and/or salted fishery products. D. cooling must continue until a temperature 204 . if the processing is insufficient to destroy nematode larvae. 9.4 Chapter IV . except when supplied to the final consumer.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. they must be cooled rapidly. or a stabilising treatment. raw materials must have no guts. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing.Requirements for certain processed fishery products A.

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. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. 9. (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.LEGISLATION approaching that of melting ice is reached.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. However. B. separation. processing. 2073/2005). 2074/2005). 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. centrifugation.2.4.5 Chapter V . 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. B. Shelling/shucking is to be done hygienically. 2073/2005. 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. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. the following conditions need to be met: A. refining and purification steps before being placed on the market for the final consumer. pressing. 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). C. 205 .2. 2074/2005. and the raw material is processed within 36 hours after loading.

Molidae. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption.Storage of fishery products (i) Fresh fishery products. Fresh.7 Chapter VII .2. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market.2. In the case of whole frozen fish in brine intended for the manufacture of canned food.4.4. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. The scientific name must accompany the common name on the label. it should be kept at no more than -9 °C. and when fishery products are wrapped on board fishing vessels. may only be placed on the market in wrapped/packaged form and must be appropriately labeled to provide information to the consumer on preparation/cooking methods and on the risk related to the presence of substances with adverse gastrointestinal effects. Fishery products contaminated with parasites may not be placed on the market for human consumption.6 Chapter VI .2. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. 206 . in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. Diodontidae and Canthigasteridae. 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. E.FISH AND SEAFOOD D.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. prepared and processed fishery products belonging to the family Gempylidae. thawed unprocessed fishery products. 9. 9.2.

This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. (ii) If fishery products are kept under ice. 9. Helicolenus dactylopterus. melt water must not remain in contact with the products.2. 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.8 Chapter VIII . species belonging to the Merlucciidae family.2. 9.4. Sebastichthys capensis.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability.). species belonging to the Gadidae family. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated.4. (d) 60 mg of nitrogen/100 g of whole fishery products used directly for the preparation of fish oil for human consumption as referred to in the second sub-paragraph of Part B(1) of Chapter IV of Section VIII of Annex III to 207 . The temperature of frozen fishery products must not fluctuate above -15°C.2. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. provided that the journey is short and the competent authority allows this. 2074/2005.Transport of fishery products The above points for Chapter VII apply in relation to transport.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. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. 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.

The reference method to be used for checking the TVB-N limits involves distilling an extract deproteinised by perchloric acid as set out in Chapter III For distillation apparatus and routine methods which may be used to check the TVB-N limits.4. tunicates and marine gastropods. they supplement the requirements of Annex II to that Regulation.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. where the raw material complies with points (a). Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. reference should be made to the actual regulation.3. Extracts of the requirements of Annex III. VI. as amended. The information required in the registration document is given in point 4 and includes the health status of the production area. Chapters V. 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. and point 3 of Chapter VII applies to retail. however. this Section also applies to live echinoderms. The sample must consist of about 100 g of flesh. Live bivalve mollusc requirements - 9. The copy of 208 . specifically relating to bivalve molluscs are given below. they supplement the requirements of Annex I to that Regulation.FISH AND SEAFOOD Regulation (EC) No 853/2004.4. 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. taken from at least three different points and mixed together by grinding.3 The regulation details specific hygiene requirements for bivalve molluscs. please refer directly to the regulation. (b) In the case of other operations. Apart from the provisions on purification. Section VII of Regulation 853/2004. (b) and (c) of Part B(1) of that Chapter. Chapter IX applies to pectinidae harvested outside those areas. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. for full requirements. VIII and IX. 9.1 Chapter I . shellfish species etc.

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

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

must be waterproof. 160 micrograms of azaspiracid equivalents per kilogram. the species of bivalve mollusc and date of packaging (at least the date and month) are required. 9. 211 .Identification marking and labelling In addition to general identification marking (contained in 9. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.7 Chapter VII . Registration document requirements also apply.3. by way of derogation from directive 2000/13/EC.9 Chapter IX .4.2. (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.Other requirements (i) The temperature at which live bivalve molluscs are stored/ transported in should be at one that does not adversely affect food safety or their viability.3. the label.6 Chapter VI . 9.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.8 Chapter VIII .4. Also. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.1).4. dispatch centre or processing establishment.3.LEGISLATION (v) azaspiracids. Requirements of Chapters III and IV are to be complied with.4.4. including the identification mark.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. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. Finally. 9. they cannot be re-immersed in or sprayed with water.3.

The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004.FISH AND SEAFOOD 9. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given. echinoderms. 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. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. 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. In this regulation. or - - 212 . This refers to official controls or production and placing on the market of fishery products. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. Therefore the health standards for fishery products as previously stated in Annex III.5. they derive from: (i) poisonous fish. (ii) fishery products not complying with the requirement concerning biotoxins. Fishery products in particular must comply with the requirements of Annex III of the regulation. It is a requirement that food business operators give assistance to ensure that official controls carried out by the competent authority can be done properly. Section VIII.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. Chapter V of Regulation 853/2004 apply. chemical. or (iii) bivalve molluscs. 9.5 Regulation (EC) No.

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

Recording and exchange of information Requirements are stated for competent authorities in relation to establishing and maintaining a current list of approved production and relaying areas and their role to inform interested parties of any changes of location. 214 .2 Chapter III . dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met.FISH AND SEAFOOD C. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. establishes microbiological criteria for a range of foods.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. 9. the competent authority is to close the production area.2. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. E. which has applied since 1 January 2006.5. 9.5. Procedures for re-opening a closed production area are stated D. boundaries or class of a production area or its closure. 9. 2073/2005.6 Regulation (EC) No. F. 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. 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. 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.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. opening or closing a production area.

presence or number of microorganisms.6. and the legislation applies to all businesses involved in food production and handling. Two kinds of criteria have been established: food safety criteria. 9.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. ensuring that foods being placed on the market do not pose a risk to human health.4 referring to fishery products. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. 9. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. volume.II.6.LEGISLATION The aim of this legislation is to complement food hygiene requirements. 215 . per unit(s) of mass. and/or on the quantity of their toxins or metabolites. The criteria are outlined in Table 9. The relevant criteria are outlined in Table 9.1 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. area or batch. with chapter 2.I. a batch of foodstuffs or a process. based on the absence.

TABLE 9. other than those intended for infants and for special medical purposes(4) (8) L. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1. 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.2 Ready-to-eat foods able to support the growth of L. monocytogenes. monocytogenes L. monocytogenes.I Food safety criteria Sampling plan(1) n 10 5 Absence in 25 g EN/ISO 11290-1 c m M Products placed on the market during their shelf life Limit(2) Analytical reference method(3) Stage where the criterion applies Food category Microorganisms 1. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life .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.3 Ready-to-eat foods unable to support the growth of L. monocytogenes 1.

26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1. coli (15) 1. manufactured from fish species associated with a high amount of histidine (17) Histamine .Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1. tunicates and gastropods 1(16) 0 230 MPN/100 g of flesh and intravalvular liquid 100 mg/kg 200 mg/kg HPLC (19) ISO TS 16649-3 Salmonella 1.27 Fishery products which have undergone enzyme maturation treatment in brine. tunicates and gastropods 9 (18) 2 E.25 Live bivalve molluscs and live echinoderms.17 Live bivalve molluscs and live echinoderms.

1996.25. 2. according to which the whole batch is to be deemed unsafe. 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. uncut and unprocessed vegetables and fruits. — fresh. when he is not able to demonstrate. Other categories of products can also belong to this category. Valle M. Single samples may be taken at retail level. including cocoa and chocolate products. to the satisfaction of the competent authority. — — bottled or packed waters. This criterion shall apply to products before they have left the immediate control of the producing food business operator. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. beer.4 or aw ≤ 0. biscuits and similar products. m = M. when recontamination is not possible after this treatment (for example. Engraulidae. Pomatomidae. products heat treated in their final package).94. sugar. that the product will not exceed the limit 100 cfu/g throughout the shelf life. 79.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample.0 and aw ≤ 0. References: 1. 5 This criterion shall apply if the manufacturer is able to demonstrate. shall not apply. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. Clupeidae. The operator may fix intermediate limits during the process that must be low enough to guarantee that the limit of 100 cfu/g is not exceeded at the end of shelf life. Dervin C. products with pH ≤ 5. — bread.. The most recent edition of the standard shall be used.1 . monocytogenes. products with a shelf-life of less than five days shall be automatically considered to belong to this category... Assay of biogenic amines involved in fish decomposition. to the satisfaction of the competent authority. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . cider.92. Malle P. wine. excluding sprouted seeds. coli is used here as an indicator of faecal contamination. E. Scombresosidae. live bivalve molluscs.. c = number of sample units giving values between m and M For point 1. AOAC Internat. Products with pH ≤ 4.. Duflos G. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. honey and confectionery. soft drinks. A pooled sample comprising a minimum of 10 individual animals. Bouquelet S. Coryfenidae. spirits and similar products. J. 43-9. Particularly fish species of the families: Scombridae. Bouquelet S.1. subject to scientific justification. Malle P.

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

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 2.1 Shelled and E.TABLE 9.4.

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

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

L338. The revised text of Regulation (EC) No. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. 852/2004 is now set out in a Corrigendum (OJ No. Regulation (EEC) 2136/89. 1662/2006. 1663/2006. 22) as amended by Regulation (EC) Nos. Regulation (EC) No. laying down common marketing standards for preserved sardines. 2076/2005.10. 9. The revised text of Regulation (EC) No. Regulation (EC) No.2005. 22. L139. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito.2004.2004.4. 854/2004 is now set out in a Corrigendum (OJ No. Regulation (EC) No. 30. L139. 2074/2005. 14. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 25. 55).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.2004.6. Regulation (EC) No. as amended. namely. L139. 3) as amended by Regulation (EC) No. 25. 83) as amended by Regulation Nos 882/2004. 25.10 Further Reading Regulation (EC) No. 2074/2005. 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. L283. L226.2004.12. 1. L226. 2076/2005. 2075/2005 and Regulation (EC) No. 1). L226.6.2006.LEGISLATION 9. 2073/2005 on microbiological criteria for foodstuffs (OJ No. 30. 2076/2005. 2074/2005 and Regulation (EC) No. 2074/2005. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. Commission Regulation (EC) No. 1019/2008 and as read with Regulation 2073/2005.4. these are not be discussed here. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 206).2004. 1441/2007. 2076/2005.2004. Regulation(EC) No. 30.4. The revised text of Regulation (EC) No. 62) as amended by Regulation (EC) No. 1688/2005.6. as read with the corrigenda at OJ No. 223 . 853/2004 is now set out in a Corrigendum (OJ No. Regulation (EC) No.

2000.2005.1. 23.2005.FISH AND SEAFOOD Commission Regulation (EC) No 2074/2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC)No 882/2004 of the European Parliament and of the Council. (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. 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 854/2004.2001.12. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. 22. 6) 224 . 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. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17. 21. 22. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.12.10.

3 minutes. 0.10.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.1 10.1.0 . under optimal incubation temperatures.10 (3). Aeromonas has the potential to grow over a pH range of ca 4 .5 pH Under otherwise optimal conditions. The minimum pH value for growth is thought to be pH Oxygen requirements Facultative anaerobe.3. At 5 °C.1.5 µm 10.5 minutes. At pH 4.1 Aeromonas spp.000-fold increase in number is observed after one week in food systems (1).0 x 1. no growth is observed (1).3 . 10. The optimum growth temperature is 28 °C (2). respectively (1). 10. Some death will occur at this pH if the temperature is 4 °C.1.3 Temperature Aeromonas grows at temperatures of 2 . 225 .5. and 2. a 10 to 1. PATHOGEN PROFILES 10.45 °C (1). 10. Morphology Gram-negative rods.5.1. and D-values at 45 and 51 °C have been reported as 29.

and is sensitive to > 4. spore-forming rods. There are four main groupings of the organism. 10. but the interior of the food may do so.5% NaCl (4). 10. forming at least seven different toxins. E and.2. botulinum types produce heat labile toxins.2.30 minutes. Thus an aerobically packed product may not support the growth of the organism on the surface. All C.2. 226 . B. botulinum are not particularly heat resistant. 10.FISH AND SEAFOOD 10.1 Morphology Gram-positive. Group I will not grow at temperatures of 10 ºC or less. which may be inactivated by heating at 80 ºC for 20 .2.4 x 1.even as low as 3 ºC (7). 10. 10. or at 90 ºC for a few seconds.3 Temperature All strains of C. 6).2% sodium chloride (NaCl) (1).2. and groups I and II are those responsible for the botulism cases. types B and F can be either. but the spores of this organism are more so. botulinum is prevented.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . to a lesser extent.4 Heat resistance The vegetative cells of C. botulinum grow reasonably well in the temperature range of 20 45 ºC.22. being capable of slow growth and toxin production at low temperatures . but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different.2 Clostridium botulinum Seven different types of C. All type A strains are proteolytic and type E strains are usually non-proteolytic. but Group II strains are psychrotrophic.2 Oxygen requirements Although C. at 85 ºC for 5 minutes. botulinum are known.1. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C.7 . Types A.5 . 0.0 µm. botulinum is an obligate anaerobe. A to G. F are the types that are responsible for most cases of human botulism (5. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth.

particularly by cooking.2 Oxygen requirements C.1 Clostridium perfringens Morphology Gram-positive spore-forming rods.0.3 .85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e.21 minutes) (9). The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. botulinum spores The most heat resistant spores of Group I C.10.PATHOGEN PROFILES 10. 9). they may survive mild heat treatments (70 .1. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. grow on the surface of foods unless they are vacuum or gas-packed.7 Characteristics of C. Note also that if humectants other than salt are used.2.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. where oxygen levels are reduced. 10. 10.9 x 2.6 and pH 5. parcooked meals with pH value > 5. 227 .5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4.0 µm.0 and aw > 0. botulinum depends on solute.3.0 . The organism will grow well in the centre of meat or poultry dishes. the minimum aw for growth may be lower. 10.3 10. safety margins must be introduced to allow for process variability. perfringens . It will not.is an obligate anaerobe. 10. 11)). However.2. pH and temperature. These concentrations correspond to limiting aw of 0.97 for Group II (5).97) (10.3. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. therefore. In commercial situations. 0.like other clostridia .g. vacuum packed.2. respectively (8. These values have been established under carefully controlled laboratory conditions.94 for Group I and 0. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.

Salt concentrations of 6 . As in the case of other factors limiting the growth/survival of this organism. In addition. 15. At cold temperatures. however.5 minutes (17). the enterotoxin is not heat resistant . etc. although C. 10.3.50 ºC. perfringens appears to be 0. While some growth can occur at 50 ºC.6 Aw/Sodium chloride C. 13). perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures.3. Its optimum temperature for growth is 43 . perfringens. and their heat resistance is also affected by the heating substrate.7 Characteristics of C. type of solute. Spores. Recorded heat 228 . vegetative cells die rapidly (12). (It is unusual for spores to be formed in foods after the growth of this organism (14)). pH. 10.3.it is destroyed by heating at 60 ºC for 10 minutes (14.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. perfringens in vacuum packed beef (18). perfringens can vary quite considerably in their heat resistance.45 ºC. the limits for aw are affected by temperature. 10. The lowest aw recorded to support the growth of C. Under otherwise ideal conditions. 18). perfringens strains.3. 16).8% inhibit growth of most C. will survive greater extremes of pH (and aw) (12.5 pH C. perfringens spores The spores of C. 10. depending on strain and other conditions.3. 0 .4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. It grows best at pH values between 6 and 7 (the same pH as most meats). although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. 13).3. Some studies indicate that the presence of 3% NaCl delays growth of C. perfringens in roast beef at 60 ºC is 14.3 Temperature The most significant characteristic of C. perfringens is not a tolerant organism with respect to pH. perfringens is not tolerant of low water activities.10 ºC. The D-value for C. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12.FISH AND SEAFOOD 10.93 and 0. lower concentrations may be effective in combination with other factors. perfringens has the potential ability to grow within the temperature range 15 . very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8.

19 minutes. 229 .4. D-values for L.1 Listeria spp. monocytogenes is not a particularly heat resistant organism.8 minutes for heat sensitive spores (12).98 and 0.2 Oxygen requirements Aerobe or microaerophilic. 10.4. -0. However. however. and where acetic acid is used as an acidulant. 1. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.23.4 Heat resistance L. the lowest limit for growth is likely to be considerably higher . 60 and 65 ºC were reported as 10.5 . using hydrochloric acid as an acidulant). is between 30 and 37 ºC. pH < 5.3 . the organism is able to grow at pH values well below pH 5 (pH 4. Its optimum growth temperature.6 .5 x 0. 10. monocytogenes is reported to be 45 ºC (20).4.especially at refrigeration temperatures. monocytogenes in crawfish tail meat at 55. or even below 0 ºC. it is not a sporeformer.4. 10. In foods.0 µm. requiring days or weeks to reach maximum numbers.2.4 ºC is probably a more likely minimum for growth in foods (19).2. so can be destroyed by pasteurisation.4 . and temperature. The upper temperature limit for the growth of L.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. Under ideal conditions.4 10. growth at low temperatures can be very slow.at refrigeration temperatures down to.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. however.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. Morphology Gram-positive short rods. being potentially capable of growing .0 minutes for heat resistant spores to 1. 10. 0.0.2 has been suggested as the lowest working limit (22). 10. respectively (21).4.3 is the lowest value where growth has been recorded.albeit slowly .64.

3. 10.8. It grows best at aw of ≥ 0.7 x 1. The bacterium may survive for long periods at aw values as low as 0.1. 10.5 pH Plesiomonas has a pH range for growth of pH 4.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.2 Oxygen requirements Facultative anaerobe. 10. 0.FISH AND SEAFOOD 10.4. at salt levels found in foods (10 .1 Plesiomonas Morphology Gram-negative.6. It is likely to survive.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).5 (24. monocytogenes is quite tolerant of high NaCl/low water activities. 25).6 Aw/Sodium chloride L. 0.8 . but has been shown to be able to grow at an aw level of 0.5 . 10.0. 10.1 Salmonella spp.3 µm.6 10.0 µm. peritrichous flagella.5.97.0 x 1 . 10.5 10. 230 .5.0 .3 Temperature Plesiomonas can grow in the temperature range of 8 .90.5.45 ºC.83 (20).12% NaCl or more). 10. or even grow.5 .5. Morphology Gram-negative short rods.5. 25). the optimum being 30 ºC (23).5.

under otherwise ideal conditions. The growth of Salmonella is generally inhibited by the presence of 3 .6 Aw/Sodium chloride Where all other conditions are favourable. therefore. Some death will occur at pH values of less than about 4. substrate. However. with a z-value of 4 . the optimum being 35 .5 .6. and appropriate heat treatments must be determined experimentally for low-aw foods.3 Temperature Salmonellae can grow in the temperature range of 7 . depending on the type of acid and temperature.7. Furthermore. 10. temperature and pH.20 times more heat resistant than the average strain of Salmonella at high aw (27).5 pH Salmonella has a pH range for growth of pH 3. some strains are able to grow at temperatures as low as 4 °C (26).6.93 (as reported in dried meat and dehydrated soup).0. 231 .8 . D60 ºC values normally range from about 1 to 10 minutes. Salmonella has the potential to grow at aw levels as low as 0. Salmonellae are quite resistant to freezing. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. Growth is slow at temperatures below about 10 ºC. although salt tolerance increases with increasing temperature (28). Salmonella senftenberg 775W is about 10 . depending on serotype. 10.4 Heat resistance Salmonella is not a spore-forming organism. 10.2 Oxygen requirements Facultative anaerobe.5. or possibly 0. The optimal pH for Salmonella growth is between 6.5 ºC.6. a heat resistant organism.6. Salmonellae are quite resistant to drying. strains vary in their ability to withstand heating.945.PATHOGEN PROFILES 10.48 ºC. 10. It is not.9. pasteurisation and equivalent treatments will destroy the organism under normal circumstances.5.6.37 ºC. However.4% NaCl. and with an appropriate acid.

31). stressed cells can also be less tolerant of heating. NB: The growth of Staph. aureus viability.7. 0. In meat macerate. with an optimum of 35 . etc.3 Temperature Under otherwise ideal conditions.1 Staphylococcus aureus Morphology Gram-positive cocci.7. aureus can grow within the temperature range 7 – 48. 10.7 . the D60 ºC value is 2 . Staph. 10. The limits for toxin production are also narrower than for growth.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH.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.2 to 9.FISH AND SEAFOOD 10. It can survive well at low temperatures.0. aw. aw and temperature (sensitivity to acid increases with temperature).9 µm diameter.0 . The following relate to limits for growth only.7. under otherwise ideal conditions (29. fat content.20 minutes.7 10. depending on aw. 232 .7.5 ºC. aureus is more limited under anaerobic than aerobic conditions.3 (optimum 7. aureus than citric acid). the organism is heat sensitive and will be destroyed by pasteurisation.2 Oxygen requirements Facultative anaerobe.5).7.). but may cause some cell damage (30). Most strains of staphylococci can grow within the pH range 4. As is the case with other bacteria. however.37 ºC (29). 10. Under most circumstances.7. 10. Freezing and thawing have little effect on Staph.

10. * dependent on temperature. 0. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.86 is the generally recognised minimum in foods (32).2 Oxygen requirements Facultative anaerobe.9.3 Temperature The optimum temperature for growth has been reported as 30 ºC.7. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.83 . but certain strains can grow in 20% NaCl.2.87 and > 0. Staph.0. atmosphere. 10. an aw of 0.10% NaCl.0 . 10.8 10. It can grow over the aw range 0. strain. and solute.99 aerobically under otherwise optimal conditions.5) (29.PATHOGEN PROFILES 10.8.8 x 1.5 .7 Limits permitting toxin production 10 .>0. pH. with a temperature range for growth of 10 . However.1 Vibrio cholerae Morphology Gram-negative curved rods.2 .7. aureus can grow in 7 .6 µm 10. In general. An effect of increasing salt concentration is to raise the minimum pH of growth. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31.0 (optimum 7.8. In general.4 .6 Aw/Sodium chloride Staph. 34). Staphylococci are more resistant to salt present in foods than other organisms.8. 233 .7.99 little or no toxin production in anaerobically packed foods.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. 31) between 0.43 ºC (35).

10.5 pH This organism is tolerant of high pH values.9.8. under otherwise favourable conditions. and it is sensitive to refrigeration temperatures (0 .8. 37.9. parahaemolyticus is easily destroyed by drying.6 Aw/Sodium chloride V. 10. with an optimum of 0. 10.1 .0% NaCl (35). with an optimum of 37 ºC (39). 234 .6.9.FISH AND SEAFOOD Also.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.998.8. 10. V.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods.4. with optimum at 7. declining in number during storage. there is evidence that V.65 minutes (35).2 Oxygen requirements Facultative anaerobe.97 .6 (35.9. it has a pH range for growth of pH 5 .0. 38). 10. cholerae in crab meat homogenate at 60 ºC is 2. 10. but only moderately sensitive to freezing (40).5 ºC).9 10. are 5 and 43 ºC. It can grow in the presence of 0. cholerae may persist on frozen fish for >180 days (36). but not acid conditions. respectively. The D-value for V.984 (35).4 Heat resistance V. cholerae can grow over the aw range 0. cholerae is sensitive to heat at temperatures of >45 ºC.

10. respectively (39).02 . it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43).98 (35. parahaemolyticus are not heat resistant. V.8.29 minutes.9.6 . The organism can survive for long periods at freezer temperatures (42). 10.5 minutes.10.3 Temperature The optimum temperature for growth has been reported at 37 ºC. and are readily destroyed by cooking. and 2.4% (39. with an optimum pH of 7.9.1 Morphology Gram-negative curved rods. 10. but the optimum concentration is 2 .4 Heat resistance V. 10.0. vulnificus is heat sensitive. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0. with a temperature range for growth of 8 .6 (41).2 Oxygen requirements Facultative anaerobe.10. respectively.9.6 Aw/Sodium chloride V. depending on solute. 10. 10.PATHOGEN PROFILES 10.5% NaCl. The D-values at 55 ºC for clam and crab homogenate are 0. 10. parahaemolyticus is unable to grow in the absence of salt.10. The optimum concentration of NaCl equates to aw of approximately 0. 41). 235 .43 ºC (42).4 Heat resistance V.8 . 41). parahaemolyticus will grow at aw levels as low as 0.11.922.5 pH The pH range for growth is 4.10 Vibrio vulnificus 10.

. 279-303. Ed.5. Novak J. Montville T. ASM Press. New York. Murrell K. London. Aeromonas species. 111-89. 383-408.. Bier J. 10.. Clostridium botulinum.. Palumbo S.. International Commission on Microbiological Specifications for Foods. Murrell K. Gaze J. Marcel Dekker.K.H. Hui Y.FISH AND SEAFOOD 10. Cliver D.. New York.W. in Clostridium botulinum: Ecology and Control in Foods. 1995. in Foodborne Bacterial Pathogens. 1994. 121-76.. 9. in Clostridium botulinum: Ecology and Control in Foods. Food Control.W. 2003.. Marcel Dekker. 4.. Bhunia A. Marcel Dekker. vulnificus grows over the range 0. Smith J. Eds.M. Gorman J. Hauschild A. Eds. Blackie. 97-131. 6 (1).10.L.D. Marcel Dekker Inc. Volume 1: Diseases Caused by Bacteria.. Marcel Dekker.O.R. Hauschild A.L. Lund B. Potential hazards associated with REPFEDS.. Volume 5: Microbiological Specifications of Food Pathogens. in Foodborne Bacterial Pathogens. Dodds K. Eds.H. Dodds K.10. Eds..M. 10. Doyle M. Aeromonas hydrophila group. Eds.6 Aw/Sodium chloride V.P. 2005.11 1. Gorham J. Hui Y.H. with an optimum pH of 7... Diseases Caused by Bacteria.D. New York. 5. 6. in Microorganisms in Foods. Clostridium botulinum and Clostridium perfringens. Miliotis M. Beuchat L.C.L. 1989. Abeyta C. 10. Caister Academic Press.W. Aeromonas.N.H. 1989. Doyle M.. 7.H. 57-63. Cliver D.E. New York.5 pH The pH range for growth is 5 .. in Foodborne Pathogens. 329-49. Principles of control. Austin J. 2001. in Foodborne Disease Handbook. Doyle M. 2. New York.R. 1-19..P.. in International Handbook of Foodborne Pathogens. Marcel Dekker. 1993. Ed. in Foodborne Disease Handbook. Kovacek K.D. New York.R.. Clostridium botulinum. Eds.10. New York. in Food Microbiology: Fundamentals and Frontiers. Fratamico P. Johnson E. Notermans S. Stelma G. with an optimum of 2. 8.J. Eds. Clostridium botulinum.L.D.P. 5-19. Hauschild A.. 1-27. Microbiology and Molecular Biology.W.M.. 1993.W. 1994.5% (42).O. Ed. 236 . Aeromonas hydrophila. Stelma G.N. 357-67.5 .0% NaCl. Faris A. Kim J. Juneja V. Marcel Dekker. 3. 1996. Peck M. Washington D. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.A.. Foegeding P. References International Commission on Microbiological Specifications for Foods.8 (42). Betts G. Dodds K..H. 11. Volume 1. Great Britain.

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

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

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


org.gsi.uk Prepared Fish Products Association (PFPA) Federation House 6 Catherine Street London WC2B 5JJ United Kingdom Tel: + 44 (0) 208 78362460 The Scottish Government Fisheries Group Environment and Rural Affairs Department Pentland House 47 Robb's Loan Edinburgh EH14 1TY United Kingdom Tel: + 44 (0) 8457 741741 or + 44 (0)131 5568400 Fax: + 44 (0)1397 795001 Email: ceu@scotland.ifm.uk Web site: www.scotland.gov.org.gov.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.defra.gov.holt@ifm.gsi.co.uk Web site: www.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: cawoods@denholmseafoods.uk Web site: www.org/ .uk/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: helpline@defra.org.uk or v.seafish.gov.uk Institute of Fisheries Management (IFM) 22 Rushworth Avenue West Bridgford Nottingham NG2 7LF United Kingdom Tel: + 44 (0)115 9822317 Fax: + 44 (0)115 9826150 Email: info@ifm.

uk Web site: www.suite. Northern Ireland Fisheries and Rural Policy Division 4th Floor North Dundonald House Stormont Estate Belfast BT4 3SB United Kingdom Tel: + 44 (0) 28 90524999/ 4991 Fax: + 44 (0) 28 90378323 Email: dardhelpline@dardni.dk Web site: www.C.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.gov.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.org Web site: www.is/ 242 .no Web site: www.ie/templates/homepage.bim.gov.fao.dk Danmarks Fiskehandlere H. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.com Web site: www.uk/index/fisheriesfarming-and-food.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/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .org/fishery/ European Association of Fish Producers’ Organisations H.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: matis@matis.dk Web site: www.ifpea@eircom.fiskehandlerne.eapo.is Web site: www. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.fiskeriforskning.FISH AND SEAFOOD Department of Agriculture and Rural Development. Andersens Boulevard 37.matis.difta.dardni.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: post@fiskeriforskning.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.org.nz/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.org 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.ifst.uk/en/index.ac.au Web site: www.CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: contact@nfi.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.org.uk Web site: www.hpa.fdf.ifr.gsi.bsi-global.nz Web site: www.gov.dh.com/ 243 .uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.com Web site: www.co.uk Web site: www.org Other Sources of Information British Standards Institution 389 Chiswick High Road London W4 4AL United Kingdom Tel: + 44 (0) 208 9969001 Fax: + 44 (0) 208 9967001 Email: cservices@bsigroup.org.csiro.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.co.cmar.gov.au New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: info@seafood.seafood.

jsp Food Safety 10 .sofht.who.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.co.uk Web site: www.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.se Web site: www.uk Web site: www.scot.php Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: admin@sofht. Web site: www.scot.nhs.who.uk/index.sfam.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: foodsafety@euro.se/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: fao-who-cc@bgvv.int.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.int Web site: http://www.uk/scieh.sik.euro.nhs.codexalimentarius.who.hps.int/foodsafety World Health Organization (WHO) Headquarters CH .co.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.who.int/ 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.org.euro.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: hpsenquiries@hps.org Web site: www.de 244 .net/web/indexen.

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


111 Astroviruses. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. 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. planktonic source of spirolides in shellfish 163 Allergens. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. preserved by dry/wet salting 99 Anisakis spp. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. source of PSP toxins 142 Alexandrium ostenfeldii. 69-70 Aeromonas hydrophila. 102 Bacillus spp. to test microbiological criteria 19 Aeromonadaceae. in crustacean shellfish 87 Alteromonas spp. as autochthonous bacterial flora of fish 3 Aeromonas spp. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. growth and survival in crustacean shellfish 86 Aeromonas sobria. causing spoilage of dried fish 110. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. in crustacean shellfish 86-7 ATP. pathogenic microorganisms in molluscan shellfish 59-60 as contaminant in enrobing process 37 as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45 growth in vegetable and rice 247 . as 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.INDEX Achromobacter spp. 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.

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

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

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

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

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

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

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

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

FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. 152-3 256 . of shellstock 61. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 67 pathogen profile 235-6 Vibrionaceae. processing for seafood recipe meal centres 39 Vibrio spp. 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 molluscan shellfish 67-8 Vibrio parahaemolyticus. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. 68 Vibrio cholerae non-01. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. 62 Yersinia spp. 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.

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