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


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

vi .

4 Spoilage 4.1 Definitions 1.8 References 1.6 Pathogens: Growth and Survival 1. vii .3 Processing and its Effects on the Microflora 4.2 Initial Microflora 1.8 References MOLLUSCAN SHELLFISH 3.2 Definitions 2.5 Spoilage 2.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.4 Processing and its Effects on the Microflora 2.8 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 2.3 Initial Microflora 2.1 Introduction 2.8 Further Reading CRUSTACEAN SHELLFISH 4.1 Definitions 4.6 Pathogens: Growth and Survival 2.4 Spoilage 1.5 Factors Affecting Fresh Fish Spoilage 1.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.2 Initial Microflora 4.1 Definitions 3.7 References 3.6 Published Microbiological Criteria 4.7 References 4.7 Published Microbiological Criteria 1.3 Processing and its Effects on the Microflora 3.6 Published Microbiological Criteria 3. 4.2 Initial Microflora 3. CHILLED AND FROZEN RAW FISH 1.4 Spoilage 3.5 Pathogens: Growth and Survival 4. 3.7 Published Microbiological Criteria 2.5 Pathogens: Growth and Survival 3.3 Processing and its Effects on the Microflora 1.

1 Introduction 7. 2006 No. EU FOOD HYGIENE LEGISLATION 189 9.5 Regulation (EC) No.10 Further Reading 223 viii .5.10 Cyclic Imines 7.1 Definitions 5.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.8 Further Reading FERMENTED FISH 6.7 References 5.2 Definitions 8. S.3 Processing and its Effects on the Microflora 5.3 Processing and its Effects on the Microflora 6.8 Guidance 222 9.7 Further Reading FISH AND SHELLFISH TOXINS 7.2 Legislative Structure 190 9. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption 212 9.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.3 Regulation (EC) No.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.3 Stages of a HACCP Study 8. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.5 Pathogens: Growth and Survival 5. 9.4 Spoilage 6. CURED. 7.1 Introduction 8.9 Other Relevant Legislation 223 9.6 Published Microbiological Criteria 5.9 Azaspiracids 7.8 Ciguatoxins 7.5 Pathogens: Growth and Survival 6.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.4 Regulation (EC) No. SMOKED AND DRIED FISH 5.4 Spoilage 5.2 Initial Microflora 6.2 Initial Microflora 5.7 Food Hygiene (England) Regulations 2006.6 References 6.1 Introduction 189 9.1 Definitions 6.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.6 Lipophilic Shellfish Toxins (LST) 7.6 Regulation (EC) No.3 Tetrodotoxin 7.11 Conclusion 7. 14 (Hygiene requirements specific to the UK) 221 9. 852/2004 on the General Hygiene of Foodstuffs 191 9.4 Implementation and Review of the HACCP Plan 8.5 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.I. 8.

10. 10.5 Plesiomonas 10.7 Staphylococcus aureus 10.9 Vibrio parahaemolyticus 10.10 PATHOGEN PROFILES 10.1 Aeromonas spp.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .4 Listeria spp. 10.6 Salmonella spp.2 Clostridium botulinum 10.3 Clostridium perfringens 10.8 Vibrio cholerae 10.10 Vibrio vulnificus 10.


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

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

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

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

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

nitrogen (N2) and oxygen (O2). DMA is dimethyl amine. The proportion of each component is fixed when the mixture is introduced. colour. odour and texture. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. marbling. and texture of the fish. Fresh fish is raw fish that has not been processed. and flavour based on the maturity. transportation and sale. and maintained at or below 7 °C. This technique is primarily used for bulk storage. juiciness. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. Quality refers to palatability and organoleptic characteristics such as tenderness. hence. at -2 to -4 °C. usually carbon dioxide (CO2). transportation and sale. but not below 3 °C during storage. spoilage microflora describes an association of microorganisms that. Spoilage describes changes that render fish objectionable to consumers. Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases. Frozen fish is fish that has been cooled to. 2 . renders that fish objectionable to consumers. Organoleptic refers to qualities such as appearance. transportation and sale. 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. and maintained at temperatures just below the freezing point. colour. firmness.FISH AND SEAFOOD Chilled fish is fish that has been cooled to. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. through their development on fish. Superchilled fish is fish that has been cooled to. frozen or preserved. and maintained at or below -2 °C (normally below -12 °C) during storage. during storage. Evisceration is the removal of the viscera from a fish.

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

14 . Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. numbers of pathogenic bacteria in raw fish tend to be low. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . In addition.102 101 .26 °C 10 .37 °C Unknown Unknown TABLE 1. monocytogenes as part of the natural aquatic flora It is apparent from the above that there is potentially a very diverse range of organisms present on fish. and risk associated with the consumption of seafood is low (2.37 °C High for most species. 105 .1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. 4 . and mud Ubiquitous.19 µg/kg bodyweight from < 102 . sewage. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. vulnificus) 5 .1 . Escherichia coli Yersinia enterocolitica 101 .35 °C 8 .103 High (107 . stream water.106 cells/g. Toxin levels 0.>106 Salmonella spp. 4).106 cells/g (exception: V.109 cells/g) * Some sources considered L. However. birds.II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.0. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria. estuarine environments.FISH AND SEAFOOD TABLE 1. Plesiomonas shigelloides 3 .

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

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

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

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

Various methods for whole fish freshness evaluation by a trained panel have been proposed. and this pattern is the basis of freshness grading schemes. the gills from pink and shiny. 17. texture. and the flesh turns from bright and elastic to dull and soft (1). rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. so lower carbon dioxide concentrations may have better preservative effects. The eyes turn from convex and clear to concave and opaque. and metabolic activities of microorganisms present in the fish. 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. 1.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16.4.1 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). C (C=reject) Significantly. or for that matter evaluation of the taste of the fish. 18) and are probably the most widely used. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids).4 1. When working with fillets. A. Loss of freshness and spoilage cannot be separated as processes. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis.III. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). TABLE 1. to brown and slimy with an intense off-odour. as traditional spoilage patterns of the external organs are typically very distinct. Taste panel evaluation of cooked fish may be used to score or grade colour. reactions caused by activities of the fish’s own enzymes. with no smell. Degradation of whole fresh fish stored in ice generally follows a set pattern. smell and 9 .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. Over time. these are summarised in Table 1.CHILLED AND FROZEN RAW FISH off-odours. B.

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

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

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. a minority are often associated with the majority of the spoilage. The concept of specific spoilage organisms (SSOs) is not new. For example. careful temperature control during storage is not only important in terms of quality loss. by altering the gas atmosphere (extrinsic).FISH AND SEAFOOD 1.2 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). Fish spoil as a result of the chemical. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. 1. For fresh fish iced immediately after capture and continually stored in ice.and temperate-water fish spoilage rates may be many.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. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 .5 1. The factors are interactive and cannot be completely isolated. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. the results of altering one parameter may have far-reaching consequences on others. The reasons for the differences between tropical. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity.5. The sum of these changes may result in a different spoilage profile. the lower the temperature of storage. For fresh 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. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. 1. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. 5). biochemical and microbiological reactions taking place within and on the fish.5. the slower the spoilage processes proceed (within limits).5. 34). but also crucial for the assurance of consumer safety. Also. yoghurt spoilage by yeasts and clostridial spoilage of cheese (5) are examples where it has been recognised for many years that a particular minority of the microbial flora present in the product is responsible for its spoilage.

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

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

they do not produce significant product spoilage).3 °C (Group II) and these are non-proteolytic (i. C1. Adult disease is caused by the production of the botulinum toxin. 6). botulinum recorded up to 1963.1 Clostridium botulinum C.6 Pathogens: Growth and Survival Endogenous chemicals. but with proper cooking. presents a potential risk. B. 5) reports that fresh fish consumption has never been shown to cause human botulism. these risks are substantially reduced (1.e. algal toxins. thermosphacta in the spoilage of some MAP fish. previous exposure and access to supportive treatment.5. F and G) is well documented (44). 1. thermosphacta did not produce off-flavours until 2 . Fewer still are capable of growing on fresh fish.3. 5).6. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. types B. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. this is probably due to spoilage occurring before toxin elaboration. E and F include bacteria with minimum growth temperatures of approximately 3. E. botulinum. D. Growing evidence suggests a role for B. Of 404 intoxication outbreaks of type E C. especially type E botulinum intoxication. When inoculated into VP corned beef and sliced ham. Types A. a convenient but diverse species of bacterium with a number of different types. Acetate production has been reported as a good indicator of spoilage by this organism. Huss (1. E. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. Review of outbreak data suggests fresh fish to be safe. 1. age. human viruses. Of these. including antisera. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42).3 days after having reached 108 cfu/g (42). bacteria and higher parasites all present some risk associated with fish consumption (1. B.4 Brochothrix thermosphacta and lactic acid bacteria B. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. The heterogeneity of the types (A. C2. Some Ph. Recent studies have investigated the dominance of B. it is bacterial risks that increase after capture of the fish. 5. 1. lightly preserved fish products are associated with botulism. 75% (with a 35% mortality rate) occurred in Japan. However. where a particular 15 .CHILLED AND FROZEN RAW FISH present in fish into histamine. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. (and very rarely F and G) have been reported to cause human disease. B.

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

no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. in its habitat. is historically associated with faecally contaminated water. but proper cooking should substantially reduce risk of infection. are associated with human diarrhoeal illnesses. their potential to produce disease-causing exotoxin in food has not been fully examined (48). however. cytotoxic. Most cases have been sporadic rather than associated to large outbreaks. Despite this. monocytogenes incidence has been between 0. shigelloides is similar to Aeromonas spp.6. and especially Aeromonas hydrophila. They are aquatic organisms with an epidemiology that is yet to be fully understood. others are psychrotrophic mesophiles (49). and especially estuarine environment. Similar problems as those encountered for Aeromonas spp.CHILLED AND FROZEN RAW FISH disorders). Chill storage may not eliminate growth. Although the psychrophiles are unable to grow at body temperature. Since 2000. In the US. possibly due to under-reporting because of their likeness to E. Some isolates are true psychrophiles. infection. P. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. and proper cooking will substantially reduce risk. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis.55 cases per 100.4 Listeria monocytogenes L. sodium channel blocking and haemolytic activity (48). but the bacterium is known to survive and grow in the shallow marine. and fish and shellfish are probably the major reservoirs for the organism (50). although lack of data concerning infectious doses and the possibility that foodborne virulence is linked to the immune state of the host mean that full risk assessment is impossible. in assessing the risk of fishborne infection by P. V.20 °C. shigelloides are encountered. are present on fish and can grow to significant numbers during storage (1. proper chill storage will eliminate any risk associated with this organism. which is often fatal (>50%). 17). Again. 1. 1. coli on isolation media (50).3 Aeromonas Aeromonas spp. listeriosis cases have been reportable to the CDC. there is no risk from the consumption of properly handled and cooked fresh fish. 5.. The major virulence factor appears to be the production of toxins with enterotoxic.26 to 0. There is no risk associated with the consumption of properly chilled and cooked fresh fish. It is known that Aeromonas spp. with minimum growth temperatures around 0 °C and optima of 15 . It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. monocytogenes has been well documented as a foodborne human pathogen. L.000 17 .6. cholerae O1. the causative agent of cholera.

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

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

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

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

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

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

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

Hall G.. 25 . Jacobsen M. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish..H.eu/LexUriServ/LexUriServ.K. 59. Modified atmosphere packaging of fish and fish products. 322-6. 1. 479-88. 65-72. Gram L.. Klausen N. Blackie Academic & Professional. Liston J. 20023. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. London. Otero A. Huss H.europa.M.. in Quality Assurance in the Fish Industry. http://eur-lex.. Amsterdam. 1992.H.L.3 Pathogens Condon S. 1987.. 1997. Applied and Environmental Microbiology.9. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. 1993. 4. 5. Trolle G. Ed. Huss H. 137-46. 1. Sala F.. Garcia M. 2197-203.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products.J. 1987.CHILLED AND FROZEN RAW FISH Einarsson H. International Journal of Food Microbiology.9. Journal of Applied Bacteriology. in Fish processing technology. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Effect of culture age. Eds. 1992. International Journal of Food Microbiology. Huss H. Elsevier Science.H. 72. A rapid method for detection of histamine producing bacteria. Gram L.R..do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A.


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

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

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

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

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

cream. nuts and fruit Associated microflora Lactobacilli. cereus Bacillus licheniformis. protozoa and viruses B. Aspergillus. enteric viruses 2.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. however. and. yeasts Pathogenic microflora/toxins Salmonella.I provides an overview of common raw materials. moulds Bacillus Bacillus.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. streptococci. Shigella. cereus Staphylococcus aureus. histamine Cryptosporidium. and so are the potential microbial contaminants associated with them. Moulds Enterobacteriaceae Salmonella. Vibrio cholerae. moulds. Enterobacteriaceae. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter. moulds Bacillus. Penicillium. Clostridium Lactobacilli. Staphylococcus. and associated spoilage and pathogenic microorganisms.4.4 2. Bacillus subtilis B. Staph. yeasts Lactobacilli. intermediate processing. Escherichia coli. by the HACCP team or by a hazard and risk assessment programme. Clostridium perfringens. raw materials. anaerobic spore-formers. TABLE 2. Fusarium. 32 .FISH AND SEAFOOD 2. Clostridium botulinum. coliforms. B. mushrooms. be considered at an early stage during new product development. pseudomonads. staphylococcal toxin. In addition. Bacillus cereus. Listeria. moulds Bacillus. for chilled products. cereus B. Table 2.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. product assembly. coryneforms.3. spore-formers. shelf life determination. and milk Cheese Water/Ice Bacillus. 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. Pseudomonas. 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. Aeromonas. toxin.

The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. but will also need to ensure that the optimum shelf life can be safely obtained. this may be an incidental effect.4. or the use of microwave technology. but it also poses a risk of cross-contaminating other products undergoing tempering. defined holding time at a chill temperature (0 – 3 °C). 2. 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.4. Examples of tempering techniques include the use of heated air. 2. and contamination from equipment to clean fish muscle. depending on the size of the fillet 33 . line extensions and process reviews. For some components. It is recommended that the principles of HACCP are applied to novel processes.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes.3.3 Preparation of fish component of chilled and frozen recipe dishes 2. Band-sawn materials or fish dice are normally promptly further processed. there are a number of distinct methods used. Figures 2. Generic examples have been used to illustrate only the major points. to evenly raise the temperature of the entire frozen product from -20 °C or lower. 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.4. and potential opportunities for growth of foodpoisoning bacteria. up to -5 °C so as to improve the cutting and handling properties during processing.1 .CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. but it is assumed that any band sawing or dice process will introduce heat from blade friction.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. the core temperature. Where the raw materials are frozen. as a heat process is often required for functional or organoleptic reasons.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. Rapid controlled cooling of heat-processed components will help to minimise spoilage. Effects on the microflora are minimal during tempering (7). as the full complexity of the processes used for specific products cannot be covered in a general text.2. All have the same intention.

1 (2. 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. Raw material preparation stages for chilled or frozen coated fish products 34 .1. are placed into plastic boxes and covered in ice to preserve their condition. by mincing and mixing with a binding agent (as described in section 2. the scampi is washed in potable water to remove foreign bodies. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.4. Fig. The “blown” meat is graded. chilled or sold chilled on defrost with an expected shelf life of 6 . 2. On arrival at the processing plant.Scampi.2.4. 2. frozen in a nitrogen blast freezer and weighed into sacks for later processing.1.3.Shaped Fillets.3. drained and then refrozen for storage until required for further processing.1C . may not rise above 0 ºC during processing. with the coating heat-fixed. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. quickfrozen.1 Whole-fillet / scampi Scampi (Nephrops norvegicus).Fish Cakes). Frozen cores are usually batter-enrobed.7 days.FISH AND SEAFOOD processed.1. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. The processing of three examples types are described in Figure 2. after landing. 2. It is then size-graded.1B .2).1A .

Therefore. as the physical structure of the material has been destroyed. This is especially the case with block-sawn dust. chilled. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . spoil more rapidly than the dry ingredients. Fish cake cores may be batter-enrobed. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. that are to be reused for other foods. but.. material will need to be transferred swiftly to a chiller. from potato flake.1. quickfrozen. ii) Fish-filleting waste . Waste material produced from this type of processing is often reused in the third category (fish cakes). The mix is shaped and formed ready for packing or for enrobing. 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.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.g. more seriously. such as aerobic spore formers and Staphylococcus spp. additional spoilage organisms may be introduced. or promptly processed to avoid further quality loss.3. with an expected shelf life of 6 .7 days. because of its nature. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. iii) Fresh/fillet/minced fish block. 35 .block-sawn dust. are handled and stored as any other food product.2 Shaped products This type of product falls into two categories.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. It is for this reason that all by-products from primary processing. 2. The remaining dried ingredients are mixed with the fish. and water is added to rehydrate the components. or sold as a chilled product after thawing. fish fingers). or those where the trimmings.1.4. pathogens such as Salmonella and Listeria. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. those that use frozen blocks of whole fillets.3. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. unless it is poorly handled.off-cuts or belly flaps. once the dry components have been rehydrated. with the coating heat-fixed. The fish component of the cake will.4.

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

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

5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe. Figure 2. For example. The vegetables must withstand processing. A temperature no greater than 10 ºC for 12 hours cumulatively or 21. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7).4.4.g. There are two main categories. Flow diagram outline for recipe dish ingredients preparation 2. but they are supplied without a source of carbohydrate e. The protein source can be fish fillet or block. canned fish may be used in their preparation owing to it having wider availability and low wastage. 2. rice.5. and must not show evidence of disease. 38 . as this could introduce off-flavours and reduce shelf life.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal.2. Fig. to produce a specific product.3. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. pasta or potato.1 ºC for no more than 3 hours is recommended (1).

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

4. cereus.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. aureus toxin is well documented (10. but temperature-abused pasta can also spoil rapidly by mould growth. An incident involving Staph. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. This is a particular issue especially where products are heat treated to eliminate vegetative cells. Sauces can be multi-component preparations or more simple preparations. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings.) to the product (12.4. such as butter garnish with black pepper. 13).6. If the sauce contains modified starches for freeze/chill stability. cereus are well known and are still a common issue in the UK (9). Alternatively. especially in those products that have a vegetable component or rice accompaniments. Typically. 40 . 11). a process equivalent to at least a 5D Listeria kill will be necessary. rely on the use of microbiologically sound ingredients. On hydration. Products or dried ingredients contaminated with either spores of aerobic bacteria (e. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. For this reason all products.g. if subjected to temperatures between 20 and 40°C. where composite meals are heat-treated. and then cooled slowly allowing germination and re-growth. spore-formers can proliferate if adequate control of temperature is not maintained. a heat process in excess of 73 ºC will be required to rehydrate this component fully. Sauces or marinades are added to fish to broaden appeal or to accompany components. 2. with the range always expanding as new tastes and cuisines are developed. Placing such products in a standard chill store to cool down could result in condensation. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. and an increased risk of contamination from L. cereus). Potato toppings made from flake or powder will normally have little associated spoilage flora. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. B. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. can germinate and rapidly multiply. Appropriate handling facilities or equipment will be required to protect the product. Chilled pasta is becoming more popular than dried pasta as a component in many product lines.

that of a dairy-based sauce. etc.2. respectively.4 and 2.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. 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. 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. The format uses flavour compounds . suspended in an acidified sauce with an oil component. Without barriers. Figures 2. 41 . and handling of processed sauce can be less rigorous than. Materials prepared with regard to good hygienic practice need only be assembled in a tray or pouch before chilling or freezing in readiness for distribution. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. Marinade-type products are becoming more popular. spices.herbs. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit.5 illustrate the recipe dish ingredients preparation and assembly and packaging. 13).4. for instance.

7. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. 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.4.4. as it dissolves in the surface moisture to form a weak acid.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. These include: 2. Flow diagram outline assembly and packaging of recipe dish 2.5.FISH AND SEAFOOD Fig.7. which may restrict either microbial growth or enzyme activity.2. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions). botulinum to grow. the method replaces air in the sealed packaging with a defined mixture of gases. 42 .4. generally carbon dioxide and nitrogen. The oxygen-free conditions will however allow other anaerobic bacteria such as C. 2.2 Modified-atmosphere packaging Originally developed for the red meat industry.

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

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

This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage.. Incidents in such diverse materials as cream. botulinum when held in favourable conditions for growth (28) i. water. These are C. Additional hazards from marine bio-toxins. and Campylobacter are recorded in filter feeding molluscs such as mussels. Therefore. botulinum type E and Vibrio parahaemolyticus (25).6. soil. Counts of between 103 .6. have been linked with food poisoning and should also be considered. and many raw or processed foods (26). typhi. Germination of spores requires favourable conditions. Inappropriate holding times and temperatures will.1 Bacillus cereus This is a common contaminant of dust. permit rapid growth of B. cereus. along with previous discussion. A number of other Bacillus spp. 13. potato. histamine intoxication. 2. Cases quoting contamination of seafood with organisms such as S. low oxygen or anaerobic conditions. 2. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). and some effects related to product type and time to toxin production have been noted.104/ g should be treated with some concern (26). with pathogenic microorganisms or viruses. 2. may be of benefit within any risk assessment for chilled and frozen prepared fish products.6. Hazards attributable to the non-seafood components are listed below and this information. etc.3 Clostridium perfringens A study examining C. or a heat shock followed by holding at a temperature suitable for growth. agricultural run-off or sewage can contaminate in-shore species. perfringens contaminating seafood will depend on the source of raw material. in some cases.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. In addition. Vibrio spp. air. 45 . any risk of C. especially molluscan filter feeders. farmed or free-living crustacea (30).e. and this issue may be of greater consequence for warmwater. are dealt with elsewhere in this book. 27). spices and herbs and vegetable shoots have been reported (12.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs.

infected food handlers. These conditions will allow the production of the toxin. and this is believed to be due to a tolerance to brine used in the smoking process (32). at the present time.6.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. Sources of contamination are many because of the widespread distribution of this organism in the environment. These include inadequate refrigeration and preparing foods far in advance of planned service. will colonise factory environments in a similar fashion except for possibly Vibrio spp. indigenous or other. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33).6. aureus (35). 2. Shigella spp.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). All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. if any other potentially pathogenic organisms. 46 . L. monocytogenes as a contaminant in foods (31).FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. Rates of contamination with smoked seafood are much higher than those with unsmoked products. Chilling may help Shigella to survive. It is unclear. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32.6. growth may occur in foods (26). aureus enterotoxins in foods. and will therefore cause “food poisoning” even in a product that is cooked before consumption. An acceptance has now developed in the UK of the presence at low levels of L. and inadequate cooking or holding within the temperature growth range (27). but. 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). 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. 2.5 Shigella spp. when introduced or inoculated into sterile components of chilled prepared recipe dishes. the restrictions on growth rate seen with seafood will not apply. 2. but under the appropriate conditions. which is not affected by any subsequent cooking process. L. However.

aureus enterotoxins and B.7 Published Microbiological Criteria Due to the complexity of many processed food products. and are a recognised means of providing data on manufacturing standards. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems. For instance.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts. Microbial guidelines. the fresh product is often consumed before the results are obtained. The following tables (Table 2. and not to final products. but these may be eliminated by a subsequent heat-treatment stage. 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. and as such the monitoring is for verification purposes rather than control. such as batter mixes. cereus emetic toxin are often confused owing to similar symptoms. as published by for example. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40). 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. 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. 2. pasta. it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur. with respect to the number and types of ingredients used. are not mandatory. (36).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. unless the criteria is for a pathogenic organisms where only non detection is acceptable.III based on IFST Guidelines (39)) provide example guidelines for seafood products. although due to long analysis times required. Microbiological specifications do not necessarily relate to product safety.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. although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”. . and their association with similar materials. It should be remembered that 47 . 2. etc.II and 2.6.

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

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

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

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


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

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

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.

Different species are variously called quahog. Most clams are only a few centimetres in their maximum dimensions.1 Clam Clam is the common name of a large group of often edible. mostly marine bivalve molluscs. or siphons. Jacques Iceland scallop Bay scallop Weathervane scallop (also called Alaska scallop) Atlantic North East Atlantic North Atlantic West Atlantic Pacific – North America Pacific – North America Pacific – Japan Common scallop Sea scallop (also called giant smooth) Japan Atlantic – North America Pecten varius Pecten maximus Chlamys islandica Argopecten irradians Pecten caurinus Pecten aequisulcatus Pecten laquaetus Pecten yessoensis Penten magellanicus or Placopecten magellanicus Pecten meridionalis Pecten novaezealandiae Pecten jacobaeus Aequipecten gibbus Chlamys opercularis Chlamys varius Amusium balloti Mediterranean – South East Europe New Zealand Australia Mytilus galloprovincialis Mytilus canaliculus Mytilus planulatus Commercial scallop Australia New Zealand Great scallop Calico scallop Queen scallop Variegated scallop Scallop saucer Atlantic/Mediterranean Atlantic – North American Atlantic Atlantic/Mediterranean Australia 3. Buried clams leave tubes. A mantle tissue encloses the body of the clam. extended above the surface to maintain water currents needed for respiration and feeding. The muscular foot is used to burrow in mud or sand.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. hard-shell or soft-shell clam. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint. and other names. cherrystone. but 56 . laterally compressed. littleneck.1. geoduck.2.

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

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

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

Alcaligenes spp.3. Vibrios... have been observed microscopically in the digestive tract of Eastern and Pacific oysters.3. rakes. from which one or both shells have been removed.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp.1 Shellstock 3. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. dredges. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora.3.1. clams. In summary. but have never been cultured (14. Campylobacter coli Bacillus cereus Hepatitis non-A.1 Processing and its Effects on the Microflora Oysters. Water temperatures affect the concentrations of microflora in the water. mechanical harvesters. and yeast cells of Rhodotorula rubra and Trichosporon spp.2.. There are microorganisms. Moraxella spp.000 /g. 12. or by 60 . Temperature. whole or in part. nutrients. Other microorganisms found in lesser numbers are the Achromobacter spp. and shucked shellfish that is shellfish. observed microscopically. and Aeromonads. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. (9. 15). Acinetobacter spp.. responding to the environment in which they are harvested. At harvest. The largest groups found are the Pseudomonads.. Enterococci spp. 10. 13).. Flavobacterium spp.3 3. Pseudomonas spp. and cockles The processing of bivalves takes two forms.1. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp.. the initial flora composition and concentration of bivalves are highly variable... 3. spirochetes of the genus Cristispira and Saprospira. They are termed viable non-culturable microorganisms.1. that have never been cultured on laboratory media. mussels. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish.1 and 3. 3. For example. Bacillus spp. Micrococcus spp. shellstock (live) which is shellfish in the shell. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. 11.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. salinity.

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. by truck or boat. There are three primary reasons for wet storage: first. or in bay-approved shellfish-growing waters prior to sale or processing. usually in open-mesh containers and/or burlap sacks. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. The shellstock is transported dry. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. in containers from which they can be retrieved easily. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock. Thirdly.3. for transportation to the point of sale. Fig. or from other sources approved by the State Shellfish Control Agency (SSCA). or immediately placed in refrigerated trucks. Shellstock may be wet-stored at the processing facility in tanks of clean seawater. after harvesting.MOLLUSCAN SHELLFISH hand (Figure 3.1. Generic flow diagram of live shellfish from harvester to point of sale 61 . Water used for shellstock washing is from approved growing areas.1). Secondly. If by boat. Shellstock are placed in drycooler storage containers in the facility.

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

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

FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). 4. which may lead to contamination of product. There is a tendency to store such articles on the shucking benches or in packing rooms. Fig 3. 3. within one hour. Generic flow diagram of shucked shellfish from harvester to point of sale Shellfish are subject to contamination during the shucking and packing process when the following are not adhered to (19): 1. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. aprons. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7.2 ºC or less. 2. Separate rooms or lockers for storage of clothing.2. and gloves help prevent the contamination of shellfish. 64 . and sanitised before each filling. Shucking containers are rinsed clean with running water. Shucked meats are thoroughly drained. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker.

streptococci.47 to 6. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. 36). these include lactobacilli.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. Alcaligenes. shucking pails. Achromobacter and Flavobacterium levels decrease during storage. Pseudomonas. particularly oysters stored at 7 ºC. Flavobacterium. a carbohydrate. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. 33. 3. are an excellent medium for growth of bacteria. As described previously. thus producing acid and gas (38). These organisms may play a significant role in the spoilage processes. Micrococcus. glycogen levels range from 0. For example. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended.8% (37). or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. Controls that should be applied include using clean packing equipment.2 ºC or less. 31. Shucked shellfish are held and transported at temperatures of 7. In summary. where there is a breakdown of temperature control or prolonged storage of shellfish. Shucked shellfish are an excellent medium for the growth of bacteria. 34. rough. 35.2 ºC. 65 .4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. Much of the work has been done with oysters and clams. Glycogen. Proteus. and packing the shellfish in a timely manner. spoilage will occur. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. anywhere during the process of shellfish from harvesting to the point of sale. Temperature abuse will result in high bacterial counts. whereas bacterium such as Pseudomonas. is stored in the bivalve as a source of energy. The environment has an effect on the bacterial population found in shellfish. The spoilage process in shellfish may be divided into three stages: increase of acidity. blowers and other utensils that have cracked. showed that organisms such as Achromobacter. All equipment coming in contact with the shellfish such as colanders. skimmers. The initial flora found in shellfish is the major factor of spoilage in bivalves. Earlier studies. clean gloves. in oysters. Shellfish. Lactobacilli seem to be the predominant microflora in shellfish. This process changes the bacterial profile to microorganisms favouring low pH. abundant gas production. at the turn of the century. and yeasts. Glycogen is hydrolysed by the initial flora found in shellfish. and proteolysis (10). particularly shucked meats. 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. 32).

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

Inadequate refrigeration will accelerate the growth of V. cholerae non-01 is ubiquitous in the estuarine environment.000 population/year (8. Healthy individuals are susceptible to gastroenteritis. and vomiting. with a fatality rate of 50% for individuals with septicaemia. nausea. High-risk individuals. 49). vulnificus. vulnificus are fever. Control of oyster-associated 67 . 55. cholerae non-01 has been associated with septicaemia. 54. The bacterium is an estuarine species that causes wound infections.II (8. are particularly susceptible to primary septicaemia. however. Hazards from this bacterium can be controlled by thorough cooking of shellfish. Oysters are the most hazardous food associated with V. 56). cholerae non-01 (51). Primarily. in the range of 0.2 Vibrio vulnificus The symptoms of V. diabetes. 58).5. like V. and rapidly become inactive at low storage temperatures. V. All isolates are considered pathogenic.7 days. Onset of symptoms occurs in 48 hours and can be severe. 46.5. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. lasting 6 . 23 of 24 major USA-West Coast estuaries were positive for V. In one study. abdominal cramps.0. leukaemia. or primary septicaemia. parahaemolyticus growth in shellfish meats is temperature dependent.MOLLUSCAN SHELLFISH Control of V. cholerae. 53. and nausea within 24 . these individuals are advised not to eat raw shellfish. cirrhosis. chills. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. 3.1. It causes diarrhoea.48 hours of onset. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). Again. parahaemolyticus. The organism attaches to the intestine and release of a toxin is suspected. vulnificus is by rapid refrigeration during warm weather months. fever. 46. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness.4 . parahaemolyticus the estimated annual incidence rate is very low. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). the organism has been found in the Gulf of Mexico. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. V. or immunosuppression. It is one of the most severe foodborne infectious diseases. Control of V.8 case/100. those who have liver disease. 3. gastroenteritis. Death has been reported to occur within 36 hours from onset. 48). The occurrence of V.1. which have been implicated in shellfish-related disease outbreaks (31). 57. They are sensitive to cold. V.3 Vibrio cholerae non-01 Illness caused by V. and preventing cross contamination once the product is cooked.

(8. 3. fluvialis. cholerae non-01 illnesses is by cooking seafood thoroughly. 62. jejuni requires reduced levels of oxygen (3 . 3.48 hours with abdominal pain. salinity. nausea.S. Death may occur after severe fluid and electrolyte loss.5 Other Vibrios spp. cholerae 01. Illness can include abdominal cramps. V. respectively. 3. low pH.5. In addition. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. and illness lasts 7 . There are several other Vibrio spp. vulnificus.1. or cross-contaminated shellfish (57. The frequency of infection of these organisms is no different from other Vibrios. widespread in both polluted and unpolluted waters. acidity and heat. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). It is considered a fragile and sensitive organism to environmental stresses such as oxygen. cholerae 01 is transmitted by faecal contamination of food or water. associated with shellfish-borne illness outbreaks. They are common marine bacteria. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. 61. particularly in oysters. V. vomiting. 60). 68 . fever. including V. cholerae 01 establishing in the U. hollisae. these organisms cause gastroenteritis lasting 24 . The presence of these strains appears to be in low numbers and non-virulent (59). Onset of disease is 6 hours to 5 days. drying.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. abdominal pain.2 Campylobacter jejuni C. headache and fever. imported product from a country experiencing an epidemic. and V.5. cholerae 01 in molluscan shellfish. and southern Asia including India and Bangladesh.1. and shock. 63). Poor sanitation and contaminated water supplies will spread the disease. Gulf coast environment. In general.5%) and 2 . is the same as previously discussed for the non-01 strains. mimicus.5. Onset of illness is from 3 to 5 days after ingestion of contaminated food. preventing crosscontamination of cooked seafood. processors should know the product source. e. the pathogenic severity is lower than V.g. parahaemolyticus and V. It causes diarrhoea. The control of V. nausea. undercooked. dehydration. however. diarrhoea. Illnesses are associated with the warmer months. and nausea. In developing countries.FISH AND SEAFOOD V. and the transmission of the disease by consumption of raw.10% carbon dioxide for optimal growth conditions. V.10 days. Control of infection from these organisms is no different from the other Vibrios. There is evidence of strains of V.

MOLLUSCAN SHELLFISH TABLE 3. 69.5. jejuni does not multiply in shellstock at refrigerated temperatures. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. V. Human feeding studies suggest that about 400 . The seasonality of 69 . This finding is significant in that C. of Cases 4 362 0 1 0 a Includes V. Campylobacter appear to survive well in oysters during storage. however. 80) are ubiquitous in the marine environment. V. control of C. they survive well at refrigerated temperatures. To reduce the risk of shellfish-borne campylobacteriosis. 85). Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP).a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. 71). however. parahaemolyticus. 68. 82. from Pacific shellstock oysters purchased from a retail grocery store (74). 72. jejuni were recovered. V. mimicus.3. 76. 3. and V. C. C. They have been implicated in limited shellfishassociated gastroenteritis cases (8. although their role as a pathogen is highly controversial and circumstantial. vulnificus.3 Other organisms of concern 3. 81. 83. 84.II Molluscan shellfish associated outbreaks caused by Vibrio spp.5.500 cells/gram of Campylobacter may cause illness (64.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. cholerae 01 and non-01. whereas at 10 ºC Campylobacter survive only for 5 days (71). 70. have been isolated from shellfish beds throughout the world (67. 65. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. and their immediate proper refrigeration.4 to 114 cells. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. Recently. 66). the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. In one case. jejuni begins in the harvest of shellstock from approved waters. hydrophila group (75. 78) and P. V. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. ranging from 0. Campylobacter spp. fluviallis. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. 77. Although. of Incidents 4 279 0 1 0 No. hollisae Infective dose varies with host susceptibility and virulence of the strain. infection occurred among participants of a firemen's banquet. 73). shigelloides (79.

000 to 3. 70 . Pathogen monitoring of Selected High Risk Foods Domestic including: milk. Similar results were seen by other investigators (86. 2 Source (90).0001 negative for the presence3 Under review4 Source (89). suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. cholerae V. They are easily destroyed with mild cooking temperatures. FDA/ Administrative Guidelines. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. dried milk/whey drink powders.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. coli Enterotoxigenic E.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. prepared sandwiches.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. FDA/Administrative Guidelines 3 Source (91). soy products. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. aureus C. 87) of the outgrowth of A.000/gram (29). after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. vulnificus in molluscan shellfish. In one study. 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 <10.600. Test Standard Plate Counts E. and where indicated are intended to be used for official control purposes. Note: Although molluscan shellfish are not included in the selected high risk food.000 cfu/g1 <230/100 g2 <1000 per gram. being isolated more often during the warmer month. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. non-dairy frozen desserts. Control of these organisms in shellfish products is limited. coli Salmonella V. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. 3. hydrophila group stored at refrigeration temperatures. jejuni V. Caution should be exercised by individuals with haematological malignancies (88). They survive well in shellstock or shucked oyster meats held at refrigeration temperatures.

7.N. water and sediment from Galveston Bay. Effects of harvesting waters and storage conditions on yeast population in shellfish. in General Zoology. 1991. Cook D.E.W. Proceedings of the National Shellfisheries Association. Nauman R. Van Nostrand Reinhold.. Wiley-Blackwell. 33-6. 1940. Cook D. 231-51. The American Oyster. 1995. 71-6. 1994.J. 11. Walker W. Liston J. American Journal of Hygiene.K. U. 10.A. 44.C.S. 1984. Clinical Microbiology Review. Lovelace T. Microbiological characteristics of Dungeness crab (Cancer magister). Government Printing Office.M. 755-76. Journal of Food Protection.R. 2..C. Pfeifer D. 13. 1960. Thompson C.R. 245-7. Viability and release Salmonella charity and Escherichia coli from oyster faeces.. Fate of enteric bacteria in estuarine sediments and oyster faeces. 633-40. 29.D. 1981. Ray S. 544-8.L. Applied Microbiology. Veliger. A typhoid fever epidemic caused by carrier bootlegging oysters. 14. 30. American Journal of Public Health. 82-7. USA. Eliot C. Gill S. Hood M.R. 9. Galtsoff P.S. 71 . Jr..R.B. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas).. Biology of the oyster in relation to sanitation. Schmidt C. 19-34. Hackney C. American Journal of Public Health. Vanderzant C. Lee J. 46.W. Villee C. 64. Colwell R. 1970. Saunders Company. 30.K.R. Ward D. New York.. Jr. 1973. Microbiology of bivalve molluscan shellfish.. 15. 1983. Applied and Environmental Microbiology.. 58.7 1.MOLLUSCAN SHELLFISH 3.S. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). Infectious diseases associated with molluscan shellfish consumption. 12. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. Old H.E. References The Phylum Mollusca. Fish and Wildlife Service. Washington D. Applied and Environmental Microbiology. 1926. 1963. Applied Microbiology. in Fishery Bulletin 1964 No. Rippey S. 13.. Bacterial flora of the market oyster. 33643. Tall B. 1975. Colwell R. Fleet G.R. Eds. 4.. in Multilingual Dictionary of Fish and Fish Products.A. 1982.. 5.. Organisation for Economic Co-operation and Development.F.H. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. 6. Smith F. Rowse A. Journal of Milk and Food Technology. in Microbiology of Marine Food Products. 8. 42. Eds.A. 8. 419-25. 36.. 6. 17. 104-9. 26. 1936. W. 1968. 105-8. Tubiash H. 16. Ed.S. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. 72-8. Bernard F. 3. Philadelphia. 447-52. Galtsoff P. 7. Journal of the Mississippi Academy of Sciences. 1964.

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

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

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

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

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

in National Shellfish Sanitation Program Manual of Operations.. in U.. U. and HACCP: A Global Focus. Seafood Safety. Ed.E. Martin R. Food and Drug Administration. Food and Drug Administration Compliance Policy Guides.E. Ed. 1987. Program 7303.R. Food and Drug Administration.MOLLUSCAN SHELLFISH 88. fresh or frozenadulteration by bacteriological contamination Sec. Collette R. Food and Drug Administration. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. Baird-Parker T. mussels. Inc. Fish Inspection.030. Food and Drug Administration.. Ed.C. 1995. 7. 1995. Food and Drug Administration. The Microbiological Safety and Quality of Food.C. Food and Drug Administration. Technomic Publishing Co.W... Sherlock C. Aspen Publishers Inc. Food and Drug Administration.S.12).L.Pathogen monitoring of selected high risk foods (FY 88/89). Burdge D. 90. Lund B.S.S. Washington D.. 91.C. 1997. U. U. National Academy Press.. Pittsurgh. Gould G. Part I Sanitation of Shellfish Growing Areas. U. 3.600 (CPG 7119. Food and Drug Administration Compliance Program Guidance Manual.1987. Government Printing Office . U. Lancaster. 63-8. Food and Drug Administration.C.8 Further Reading Ahmed F. oysters. U. Chapter 5: Foods.. 1991.S. in U. Slavin J. Quality Control. Washington D.S.S. 89.S.M.A. Gaithersburg. 560. 77 .H.S. Washington D. 2000.. Clams. Smith J.W.


and Trachypenaeus spp. and available in 0. CRUSTACEAN SHELLFISH Linda Nicolaides.5 and 1. with shells and heads on or off are usually packed. packed and frozen in 0.g. the front pair ends in claws. e.. Species eaten from tropical waters include Penaeus spp. and are usually aquatic.5 and 1. crayfish and shrimp (1). Pandalus montagui and Crangon crangon (inshore shrimp).. Shrimps. Peeled and cooked shrimp are ready-to-eat after controlled thawing. peeled product. lobsters. with additional water to form a glaze. and frozen in 1 and 2.4. They possess a segmented body. Crayfish.5 kilo pack (boxes or bags). jointed limbs and a chitinous exoskeleton.. Such products are usually only partially cooked before freezing. M. Pandalus borealis (deep-water shrimp).0 kg polythene bags.0 kg polythene bags. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. Parapeneopsis spp. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). lobsters and crabs have ten legs. 79 . (deep-water shrimp). Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. Peeled ready-to-cook shrimp are raw. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda.Ph. crabs.

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

The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. In contrast. so they are either sold on the local market or are processed in local processing plants. gills and thorax. 4. When the shrimp arrive at the processing plant they are washed and then sorted into size. Hence. up to 16 days for tropical species compared with 8 . Vibrio spp.3 Processing and its Effects on the Microflora Crabs. graded according to size. corynebacteria. These animals can survive out of water if the surrounding environment is kept cool and damp. peeled and cooked. thus. the rate of deterioration of the crustacean shellfish by these organisms is reduced. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. Pseudomonas spp.. and Achromobacter spp.1). The rate of spoilage will be a function of time and temperature. Alcaligenes. from the time of capture and processing. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots. After capture and transport to the processing plant. 4. however. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected. In comparison shrimps die quickly after capture. The claws are usually bound together to prevent the animals fighting and causing damage to each other. with “head on” or “head off” whereby the edible tail is removed from the head.1 Prawns and shrimps An example of the process used for preparing frozen. They are then taken to market alive in either holding tanks. chilled 81 . they are washed. chilled sea water. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . bacterial contamination and spoilage are inevitable..1.3. or are found in brackish water around the shores. especially if they are stored in ice or iced water. A large proportion of shrimps are produced by aquaculture.for example. Shrimp will then be glazed and frozen in a raw state. or cooked. As with finfish. uncooked prawns and shrimp is presented in Figure 4. the genera associated with tropical species include Enterobacteriaceae. lobsters and crayfish are harvested from cages or pots containing bait. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. whether GHP are in place..CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. Shrimps can be processed whole. Shrimp might be further processed with the shell on. 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. Micrococcus spp. or transported directly to processing plants.10 days for temperate water species (3). or with the shell and central vein removed. glazed and frozen (see Figure 4.

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

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

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

monocytogenes has been attributed to crustaceans. the pathogen can grow at temperatures as low as 3 °C. is widely distributed in aquatic environments. 4. Researchers have demonstrated that inoculated shrimp supported toxin production by C. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. aureus on their hands. as part of the natural bacterial flora.5. 24.CRUSTACEAN SHELLFISH old. although contamination by types A and B may occur from the environment. pasteurisation and cooking. which can become colonised by V. aureus is not a member of the intrinsic flora associated with crustacean shellfish. 25). 22) as well as from undercooked crustacea when harvested from contaminated waters (23. Crustaceans have been linked with food intoxications attributed to Staph. 85 . The main source of contamination is the hands of processing personnel. especially in products that have been vacuum packaged. Vibrio cholerae. botulinum type E is commonly associated with fish and fishery products. so is able to survive heat treatments that eliminate vegetative cells. The pathogen has resistant spores. cholerae is divided into two groups. it is important that the cooking stage is controlled to eliminate the pathogen from the product and that procedures are in place to prevent any cross-contamination of the cooked product.5 Vibrio spp. aureus because of the ubiquity of this organism in the processing environment. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. 19).5.4 Clostridium botulinum C. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. therefore. the causative agent of cholera. Crabs are covered in a chitin exoskeleton. 4. To date. no incidence of foodborne illness caused by L. pregnant women and the immuno-compromised. cholerae. V. The former. Control should be achieved by promoting GHP and staff training programmes.g. 4. combined with the degree of handling of these types of product. botulinum type E at 10 °C but not at 4 °C (20).5. e. Once the competing microflora has been removed by a heat treatment. An epidemic that began in South America in 1991. about 50 .70% of healthy individuals carry toxin-producing strains of Staph. was associated with seafood consumption. cholerae non-O1. cholerae O1 and V. whilst boils and cuts are also common sources of the pathogen. However. particularly crabmeat (18.3 Staphylococcus aureus Staph. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. V.

parahaemolyticus.g. V. a marine halophile. but has the potential to cause fatal foodborne illness (26).5. A. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. although A. vulnificus is an invasive and lethal pathogen associated with wound infections. Hepatitis A virus presents the most serious hazard. Of these.5. 32). Caliciviruses. It is more commonly implicated in countries where crustacean shellfish are eaten raw.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. South America. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. the potential hazard exists as C. to date.5. however. 4.7 Aeromonas hydrophila and Aeromonas sobria A. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). Quantitative levels were below limits of detection in all cases (<0. e. 4.30 MPN/g) (27). hydrophila has been isolated from prawns (29). been implicated in a case of foodborne infection associated with crustaceans. However.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. occurs in crustaceans from warm environments.6 Campylobacter jejuni and Campylobacter coli C. in a survey carried out in the United States between 1977 and 86 . hydrophila and A.5. 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. jejuni has not. Astroviruses and non-A. 4. However. 4. particularly trematodes (6). whilst 5. coli. Parasitic infections are controlled by either cooking infected material prior to consumption. hydrophila is able to grow well at refrigeration temperatures (0 . Japan. 31. non-B Hepatitis viruses (31).8% of samples contained C.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. V. or freezing at a temperature of -23 °C for 7 days (30.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. Norovirus.

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. Provisions laid down concerning the use of additives would also apply to these types of products. 4. although it is one of the European Union (EU) legislated allergens. consistent with that in the stomach. The toxin causes amnesic shellfish poisoning (ASP) when consumed. only 0.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive.6. No foodborne incidents have been attributed to this source to date. They can survive low pH levels. 4. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. which results in a persistent and apparently permanent loss of short-term memory (36). from farm to fork (37. produced or imported into the European Union (EU). which could be transferred to the meat during unskilled removal from the shell (35). 87 . Norwalk viruses can also withstand heating to 60 °C.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. or chlorine concentrations similar to those found in drinking water. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). The risk of such hazards is far higher with fin fish and molluscan shellfish. 38.CRUSTACEAN SHELLFISH 1981. 39).1 EU Legislation Microbiological standards for cooked crustaceans and shellfish.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). 4. 4. A summary of microbiological criteria for crustacean shellfish is listed below.11 Chemical pollutants and biotoxins Researchers have demonstrated that crabs feeding in waters contaminated with algae do have potentially high levels of algal toxin domoic acid (DA) in their viscera.5.5. Selected parts from this regulation is presented below. are included in the Commission Regulation (EC) No. 2073/2005.

FISH AND SEAFOOD Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) Chapter 1 - Food safety criteria

L. monocytogenes Prepared crustacean products fall into the category described in the regulation as “Ready-to-eat foods available to support the growth of L. monocytogenes other than those intended for infants and for special medical purposes”. During capture and preparation there is a zero tolerance for the presence of L. monocytogenes on the product. However, to take into account the issues of natural habitats being contaminated with low levels of this pathogen there is a slight loosening of the regulation at the point of sale and for the duration of the products shelf life (refer to Table 9.I, Food Category 1.2). If the level of L. monocytogenes should be above this level then the consignment is considered to be unsatisfactory. Salmonella All cooked crustaceans and molluscan shellfish should be free from Salmonella at the point of sale and for the duration of their shelf life (refer to Table 9.I, Food Category 1.16). Should any sample test positive for Salmonella then the product is declared to be unsatisfactory. Chapter 2 - Process hygiene criteria

Process hygiene criteria for E. coli and coagulase-positive staphylococci in shellfish and shucked products of cooked crustaceans and molluscan shellfish are laid down (refer to Table 9.II, Food Category 2.4.1). 4.6.2 International requirements

International requirements are based upon having a risk-based, preventive management system in place at all stages of the supply chain. The requirements for this preventive approach is described as follows (37): • Fish and fishery products should be prepared in plants certified by the local competent authority. All certified plants should comply with the GHP (39); • The fisheries industry should take responsibility, implement and maintain safety management systems based upon HACCP; • The national competent authority is responsible for the certification of fish processing and manufacturing plants, verification of effective systems, and



issuing of certificates of compliance for export products. This includes auditing and inspection programmes; • National surveillance and monitoring programmes should be in place to demonstrate that all identified hazards are under control, e.g. biotoxins, and to identify potentially emerging hazards; Within this requirement is the flexibility for countries and trading blocks to set specific standards required to deliver an equivalent requirement with those at national level. Such standards need to be science-based, especially if they are more stringent that those recommended by the Codex Alimentarius Commission. Verification that GHP have been followed during the handling and preparation of crustacean shellfish for consumption is usually done by assessing the level of E. coli and/or coagulase-positive staphylococci in the products. Legal requirements generally require that these groups of indicator organisms are present in levels <100 cells/gram of product. Pathogenic bacteria such as Salmonella spp. should be absence is 25 grams multiplied by the number of subsamples tested. United States The FDA brought into force a Federal Mandate Seafood Rule in 1995 which forms the foundation for good practices required for the handling and processing of imported fish and fishery products into the U.S. The best practices include GHP and HACCP (37). All fish and fishery products produced in and imported into the U.S. after 17 December 1997 are required to demonstrate that HACCP principles were applied to their handling and processing. The USFDA has a zero tolerance for Salmonella in fish and fishery products; this includes farmed shrimp. The US FDA considers that the presence of pathogenic microorganisms in seafood is an indicator of adulteration (32). 4.7
1. 2.

Encarta 1997. Encarta 97 Encyclopedia World English Edition. Microsoft. The International Commission for Microbiological Specifications for Food. Fish and Fish Products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. The International Commission for Microbiological Specifications for Food. New York, Kluwer Academic/Plenum Publishers. 2005, 174-249. Cann D.C. Bacteriology of shellfish with reference to international trade, in Handling, Processing and Marketing of Tropical Fish. Ed. Tropical Products Institute. London, Ministry of Overseas Development. 1976, 377-94. D’Aoust J.Y., Gelinas R., Maishment C. Presence of indicator organisms and the recovery of Salmonella in fish and shellfish. Journal of Food Protection, 1980, 43, 679-82.




FISH AND SEAFOOD 5. 6. 7. 8. 9. 10. 11. Fraiser M.B., Koburger J.A. Incidence of salmonellae in clams, oysters, crabs and mullet. Journal of Food Protection, 47, 343-5. Reilly P.J.A., Twiddy D.R., Fuchs R.S. Review of the occurrence of Salmonella in tropical shrimp. FAO Fisheries Circular No. 851, 1992, (FIIU/C851). Reilly P.J.A. Salmonella and Vibrio cholerae in brackishwater cultured tropical prawns. International Journal of Food Microbiology, 1992, 16, 293-301. Twiddy D.R. Antibiotic-resistant human pathogens in integrated fish farms. ASEAN Food Journal, 1995, 10, 22-9. Olsufjew N.G., Petrow V.G. Detection of Erysipelothrix and Listeria in stream water. Zhurnal Mikrobiologii Epidemiologii i Immunobiologii, 1959, 30, 89-94. Anon. First Listeria finding in crabmeat confirmed by FDA. Food Chemistry News, 1987, 29 (14), 38. Ryser E.T., Marth E.H. Incidence and behaviour of Listeria monocytogenes in fish and seafood, in Listeria, Listeriosis and Food Safety. Eds. Ryser E.T., Marth E.H. New York, CRC Press. 2007, 617-54. Anon. Mexican crabmeat, Greek pasta automatically detained. Food Chemistry News, 29 (19), 43. Anon. FDA checking imported domestic shrimp, crabmeat for Listeria. Food Chemistry News, 1987, 29 (24) 15-7. Archer D.L. Review of the latest FDA information on the presence of Listeria in foods. WHO Working Group on Foodborne Listeriosis, Geneva, Switzerland. 1988, 15-9. Anon. FDA to sample shrimp for Salmonella, Listeria. Food Chemistry News, 1988, 30 (36), 9-10. Anon. Monitoring of changes to listeriosis problem urged. Food Chemistry News, 1989, 31 (20), 13-7. Richmond M. Report of the Committee on the Microbiological Safety of Food. London, HMSO. 1990, 133-7. Swartzentruber A., Schwab A.H., Duran A.P., Wentz B.A., Read R.B. Jr. Microbiological quality of frozen shrimp and lobster tail in the retail market. Journal of Food Protection, 1980, 40, 765-9. Wentz B.A., Duran A.P., Swaartzentruber A., Schwab A.H., McClure F.D., Archer D., Read R.B. Jr. Microbiological quality of crabmeat during processing. Journal of Food Protection, 1985, 48, 44-9. Garren D.M., Harrison M.A., Huang Y-W. Clostridium botulinum type E outgrowth and toxin production in vacuum-skin packaged shrimp. Food Microbiology, 1994, 11, 467-72. Molitoris E., Joseph S.E., Krichevsky M.I., Sindhuhardja W., Colwell R.R. Characterisation and distribution of Vibrio alginolyticus and Vibrio parahaemolyticus isolated from Indonesia. Applied and Environmental Microbiology, 1985, 50, 1388-94.

12. 13. 14.

15. 16. 17. 18.





CRUSTACEAN SHELLFISH 22. Varma P.R.G., Iver T.S.G., Joseph M.A., Zacharia S. Studies on the incidence of Vibrio cholera in fishery products. Journal of Food Science and Technology, 1989, 26, 341-2. Madden J.M., McCardell B.A., Morris J.G. Jr. Vibrio cholerae, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 525-42. Twedt R.M. Vibrio parahaemolyticus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 543-68. Oliver J.D. Vibrio vulnificus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 569-600. Pearson A.M., Dutson T.R. Quality Attributes and Their Measurement in Meat, Poultry and Fish Products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994. Reinhard R.G., McAdam T.J., Flick G.J., Croonenberghs R.E., Wittman R.F., Diallo A.A., Fernandes C. Analysis of Campylobacter jejuni, Campylobacter coli, Salmonella, Klebsiella pneumoniae and Escherichia coli 0157:H7 in fresh handpicked blue crab (Callinectes sapidus) meat. Journal of Food Protection, 1996, 59 (8), 803-7. Buchanan R.L., Palumbo S.A. Aeromonas hydrophila and Aeromonas sobria as potential food poisoning species: a review. Journal of Food Safety, 1985, 7, 15-29. Palumbo S.A., Maxino F. Williams A.C., Buchanan R.L., Thayer D.W. Starch ampicillin agar for the quantitive detection of Aeromonas hydrophila in foods. Applied and Environmental Microbiology, 1985, 50, 1027-30. Wekell M.M., Manger R., Colburn K., Adams A., Hill W. Microbiological quality of seafoods: viruses, bacteria and parasites, in Seafoods: Chemistry, Processing Technology and Quality. Eds. Shahidi F., Botta J.R. Glasgow, Blackie Academic and Professional. 1994, 196-219. Schantz J., McKerrow J.H. The dangers of eating raw fish. New England Journal of Medicine, 1989, 320, 1143-5. Ahmed F.E. Seafood Safety. Seafood Safety. Committee on Evaluation of the Safety of Fisheries Products, Food and Nutrition Board, Institute of Medicine. Washington D.C., National Academy Press. 1991. Blackwell J.H., Cliver D.O., Callis J.J., Heidelbaugh N.D., Larkin E.P., McKercher P.D., Tayer D.W. Foodborne viruses: their importance and need for research. Journal of Food Protection, 1985, 48, 717-23. Kapikian A.Z., Chanock R.M. Norwalk group of viruses, in Virology. Eds. Fields D.M., Knipe D.M. New York, Raven Press. 1990, 359-77. Horner R.A., Postel J.R. Toxic diatoms in western Eashington waters (US west coast). Hydrobiologia, 1993, 269/270, 197-205. Perl T.M., Bedard M.S.M., Kosatsky T., Hockin J.C., Todd E.C.D., Remis R.S. An outbreak of toxic encephalopathy caused by eating mussels contaminated with domoic acid. The New England Medical Journal, 1990, 322, 1775-80.

23. 24. 25. 26.


28. 29.


31. 32.


34. 35. 36.


FISH AND SEAFOOD 37. Ababouch L., Gandini G., Ryder J. Causes in Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. Australian Government. Australian Quarantine and Inspection Service Hazard Analysis and Critical Control Point (HACCP). A Guide to Compliance with Export Control (Fish and Fish Products) Orders 2005. FAO/WHO. General Hygiene Requirements. Rome, Food and Agriculture Organisation of the United Nations. 2003.



4.8 4.8.1

Further Reading Spoilage and pathogens

Gram L., Huss H.H. Fresh and Processed Fish and Shellfish, in Microbiological Safety and Quality of Food Volume 1. Eds. Lund B.M., Baird-Parker A.C., Gould G.W. Maryland, Aspen Publishers Inc. 2000, 472-506. Ababouch L., Gandini G., Ryder J. Causes of Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. International Commission on Microbiological Specifications for Foods. Fish and fish products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1998, 130-89. Sutcliffe P., Disney J. Handling, Processing and Marketing of Tropical Fish. London, Tropical Products Institute. 1977. Connell J.J. Control of Fish Quality. Oxford, Fishing News Books. 1995 Ward D.R., Hackney C. Microbiology of Marine Food Products. New York, Van Nostrand Reinhold AVI. 1991 Pearson A.M., Dutson T.R. Quality Attributes and their Measurement in Meat, Poultry and Fish products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994.

4.8.2 Pathogens - general
International Commission on Microbiological Specifications for Foods. Microorganisms in Foods 5. Characteristics of Microbial Pathogens. Eds. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1996. Varnam A.H, Evans M.G. Foodborne Pathogens - An Illustrated Text. London, Wolfe Publishing Ltd. 1991. Doyle M.P. Foodborne Bacterial Pathogens. New York, Marcel Dekker. 1989.


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

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

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

6 132. When a milder brine.4 52. % 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. only the most common processes can be described here.4% salt (Table 5. This should be distinguished from the salt content. Therefore. Thus. processes and producers’ private recipes. it is important to understand the concept of salt concentration. it is impossible to establish standardised processing stages in the curing of fish. it is important to state the temperature. Salt in the water phase links the two conditions of moisture and saltiness.0 . and salt content.FISH AND SEAFOOD 5.8 211. the fish is usually salted with a 70 . is employed.80º brine (184. A 100º brine solution means that the solution is saturated and no more salt can be dissolved.I Brine strength at 18 °C (after Bannerman (2)) Brineometer degrees 10 20 30 40 50 60 70 80 90 100 96 Weight of salt (g/litre of brine) 26. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.8 211. about 50º (132.6 264. a 90º or above brine (237.4 184. TABLE 5. Salt concentration means the percentage of salt in the water contained in the fish. When a 100º brine is used.I).0 g of salt per litre of brine).2 237. a 0º brine saturation means pure water.6 g of salt or more per litre of brine) may cause salt burn in this way. The dissolution of salt in water is dependent on the temperature. or salt in the water phase. it causes swelling because the fish absorbs water from the salt solution (1). sometimes the salt crystallises onto the surface of the fish. The strength of the solution is described as degrees of saturation (brineometer degrees). smoking or marinating processes. etc.2 105.3 Processing and its Effects on the Microflora Owing to the wide variety of products.0 158. When working with salting. For smoking salmon. Different brine strengths are required according to the curing processes and/or the type of product. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. all using salting steps.8 79. At 18 °C a 100º brine is produced when the solution contains 26.2 g of salt per litre of brine).

outgrowth of halophilic bacteria may occur. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). which consequently inhibits the diffusion of the water from the bulk of the product to the surface. 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. which include calcium and magnesium. However. Salting can be done in a number of different ways. producing characteristic pink patches. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. The most common fish used in the northern hemisphere is cod (Gadus morhua). and a ‘putty’ fish product is also obtained (1). and the salt has penetrated throughout the fish (3). The fish is then stacked in piles of about 2 metres and the water is allowed to drain away.0. however. 5.1. When pure salt is used. See Figure 5.15% calcium and magnesium. low in calcium and magnesium. and such delays. Also.1 Dry salting / Kench curing Usually. mainly in tropical conditions. when the air humidity is high.35% and 0. Usually it is followed by other preservation techniques such as drying and/or smoking. respectively (1). It takes about 3 weeks until proper salt penetration and moisture loss are achieved. respectively. and essentially free from iron and copper (3). can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage.3.1 ppm and 10 ppm. are recommended to help make the product whiter. if very pure sodium chloride is used for curing purposes.1 Salting Salting of fish for preservation purposes has been used since pre-history. but the concentration of copper and iron must be kept under 0. This is because fatty fish turn rancid quickly. ‘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. are likely to cause hydroscopicity of the product and. and salt can speed this oxidation process. it tends to lead to a slight yellowing and softening of the product. and for a heavy salting a ratio of 1:3 salt:fish is used (1). lean white fish is used for this purpose. the fish is mixed with one part of salt per eight parts fish. caught in North Atlantic waters. for which there is a high demand in Mediterranean countries. This condition is known as ‘putty fish’ (4).15 . having a short section at the end of the tail. The salt used should be of food-grade quality.1. addition of levels of 0. The calcium and magnesium impurities. ‘Green cured’ split cod with about 60% of the backbone removed. SMOKED AND DRIED FISH 5.3.CURED. On the other hand. the fish is restacked with fresh salt after about a week. 97 . can be dried further to produce ‘klipfish’ or ‘bacalão’. For a light salting. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts.

For large fish it is advisable to split it before salting. such as Staphylococcus aureus could survive and grow at such water activities in the product. Only small fish can be salted whole. it appears that water activities much less than 0. Processing of dried salted cod 5. Fig.2 Wet salting or brining As referred to above.1.90 are rarely achieved. Where fish are salted in their whole form. although toxin production would be unlikely (4).1.3. such as 98 . The greatest barriers to salt penetration are the skin and scales. so that the risk of spoilage is high. Food-poisoning organisms.5. eviscerated and/or split).FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. when a light brine is used. the process is mainly for flavouring purposes prior to the use of other preservation processes.

03 2.80 5.11 4.06 4.90 (1). if split fish is dry-salted. the liquid that comes out of the fish forms a brine. See Table 5. Histamine formation is of importance when using scombroid and some clupeidean species.98 5.16 7.85 3. can be cured in a saturated brine solution for 6 days and. The main objective of the brining method of salting fish is to limit air penetration during processing.3 days. but as it is placed in containers.88 5.45 3.1.23 2. For preservation purposes.74 9.78 2.23 8.25 5.II.98 6. the fish is dry-salted.0 2.3 Dry / wet salting In this process.0 4.64 4.26 2.51 3.41 3.19 3. The fish are kept immersed in their own brine near saturation point for several days.0 3. they can be stored at 28 °C. a saturated brine is used.3.17 4.76 4.84 1. Fresh sardines.5 10.76 4. sardines.60 2.49 4. 5.26 7.70 3.03 4. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.96 1.46 4. TABLE 5.5 3.26 3.65 2.5 4. with a shelf life of about 4 weeks (water activity (aw) about 0.17 5.76 99 .26 1.CURED. herrings and other fatty fish can be used.78 6. the brine is then replaced over the fish.61 3.47 6.73 1.23 3. SMOKED AND DRIED FISH smoking.30 2.51 5.66 5.05 7. and a red burgundy colour will develop in the product.23 2.44 2. Here.10 1.85 3.86 2. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.00 3.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. after being drained and pressed at 465 kg/m2 for 15 hours.91 2.98 6.78) (1).11 9.61 3. There are several variations of this process.95 3.25 5.66 5.11 3. For example.5 3. thus.76 4. the fish enzymes play an important role by maturing or ripening the product. headed and gutted.99 2. This is why it is the preferred method for fatty fish.41 4. Histamineforming bacteria do not generally grow at water activities below 0. but mackerel. Anchovies are the main fish used for these purposes.56 6. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 . and can be collected and replaced again over the fish.44 2.26 4.04 5.35 4.09 8. the product obtained is called gaspé-cured (5).54 5. limiting the amount of rancidity that can develop.41 5.09 8.33 5.70 6. with pressure being applied to the top of the container to help the brine to drain away.41 6.76 4.22 6.76 4.

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

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

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

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

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

and their sensitivity to salt (13). thus allowing any psychrotrophic C. Hot-smoked products require a longer cooling stage. the products are cooled. A typical temperature set is as follows: firstly. secondly. typically mesophilic.3. a final cooking period at 80 °C (12). See Figure 5.5. if a 105 . These products should be vacuum packed at room temperature. thirdly. stored at 4 °C and packed. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). but the proteolytic types A and B. refrigerated storage and vacuum packaging still allow its growth (12). are of concern when smoking fish from tropical waters (13). Vacuum packing is recommended only for coldsmoked salmon. Smoked salmon fillets are usually vacuum packed whole or after slicing. and hotsmoked salmon (kippered) (3).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. Afterwards. condensation will occur inside the pack.CURED. If the refrigeration chain is maintained. Length of the smoking process at each stage will depend on the species and the type of product. This temperature should be maintained during storage and distribution (3). botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. a partial cooking period at 50 °C. botulinum type E present an even longer time than before to grow and produce toxin (14). On the other hand. The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. if present. 5. a drying period at 30 °C. Furthermore. The exception is vacuum packed fish. can grow with salt levels of 10% in the water phase. which underlines the danger of storage at abuse temperatures of >10 °C (13). During smoking. further cooled to 3 °C within 12 hours and then packed. Smoked products can be also packed under controlled-atmosphere packaging. SMOKED AND DRIED FISH During cold-smoking. 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. in order to produce toxin. and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). monocytogenes. as. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). Although other human pathogens that can grow at refrigeration temperature. there is no point in the process that can fully assure the absence of L.2. gravad salmon (also called gravad lax or gravlax). can be isolated from seafood. Pathogenic vibrios. but they should be cooled to a temperature of 10 °C within 3 hours. Neither the smoking temperature nor the salt content is enough to kill the organism. such as Yersinia enterocolitica. if they are taken from a chill room at 3 °C and packed. To avoid growth of non-proteolytic C.

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

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

and less than 0. consistency of flavour and colour. 5. nutritional neutrality and antioxidant activity (27). 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.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil.4 Clear smoke treatments The “Clear” or “Tasteless” smoking process is a processing technique whereby smoke is generated as in a normal food smoking process but is filtered to remove the characteristic smoke flavours before application on to otherwise untreated (uncured) tuna. 5. In experiments comparing traditionally smoked fish with liquid smoke. When liquid smoke was used.03 μg/kg of benzopyrene. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. both of which are considered potential carcinogens. It has been shown that liquid smoke can reduce the formation of PAHs (3).3.6 to 2.8% and still provide protection for 7 days (25). 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).06 μg/kg of benzoanthracene in the final foodstuff (28). drying. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25).5.4 Spoilage Brining. so as to impart an unnatural red colouration to the flesh. or applying it as an atomised spray within a modern automatic smoke kiln. 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. Fresh fish spoilage may be measured by 108 . injecting it into the fish flesh. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. smoking. the salt level could effectively be reduced from 4. The European Scientific Commitee for Food. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. applying it as a dip after brining. Liquid smoke was considered to provide advantages of safety. According to some recent research. 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. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. or smoke extracts in organic solvents.

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

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

flavus. 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. The heat during hot-smoking will kill vegetative microorganisms but not all spores. Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. Marine vibrios were the dominant flora in smoked salmon that was drysalted. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). able to grow on dried salted fish. where a high internal temperature is reached. Aspergillus spp. SMOKED AND DRIED FISH infrequently at low levels. Aspergillus clavatus and Aspergillus niger.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. Refrigeration is necessary. botulinum type E. will have a different residual post-smoking flora from 'hot-smoked' fish. as already stated. which does not create the salty conditions required for spoilage by those halophilic microorganisms.CURED. For more information on spoilage see Table 5. 36).V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). Vacuum packing limits chemical oxidation. with a lower aw limit for growth of 0. not the growth of spoilage organisms. it was. but this will not prevent the growth of spoilage bacteria. Table 5. 33. 111 . including A. so the bacteria spoiling these products will be psychrotrophic or psychrophilic. but. It has been reported that the type of salting (dry. The brining to 3 . Cold-smoked fish will contain microbial flora representative of that of the raw fish. however. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations. around the water activity of saturated sodium chloride. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. Pseudomonadaceae. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. has been isolated from dried fish (35). LAB. In Vietnam. and microbial spoilage is more likely. Smoked fish products tend to have a light brine treatment. The preservative effect of the smoke will inhibit some spoilage microorganisms. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32.5% sodium chloride in the water phase will inhibit some bacteria.. The salt in the water phase must be high enough to inhibit the growth of C. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. Fish processed by cold-smoking.IV.747.

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

97 0. The concentration required to prevent growth at room temperature can vary from as low as 3.5-10 4. pressure on the traditional methods of curing. However. under optimal conditions. could not grow and produce toxin. and as botulism is a serious hazard in foods.5 0-11 3.75 0. the fermented products being the most dangerous group (7).5 5. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. 9 and 8 of the outbreaks.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13).5.5-8.CURED.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. botulinum Staph. The presence of salt in cured products has a great effect on the growth of the bacteria. the importance of cold-chain integrity must be emphasised. respectively.0-9. botulinum type E Staph. sodium 113 . popular demand for healthy foods has led to a change in the tastes preferred by the public and. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C.8 0. and C.5-10.8 1. smoked and dried. In the United States. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39).V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. Before. the non-proteolytic type E strains can grow in up to 5% sodium chloride. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). although the combination of the two factors. when present in the raw material.5% to 5%. Since neither the smoking nor the drying parts of this curing process are particularly severe. botulinum. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. with this change.9 0.5 5. Examination of 165 outbreaks of botulism caused by fish products showed that smoked.97 0. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. SMOKED AND DRIED FISH TABLE 5. salted and pickled products accounted for 10. but if the fish products are stored at temperatures <10 °C.86 0. types A and B are not affected by 3 . aureus Yeasts Moulds Lactic acid bacteria pH 5. the products were heavily salted.61 Organisms Most Pseudomonadaceae C. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13).5-8. botulinum type E. botulinum type E and production of toxin.5 4. It has been shown that.

5. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . coli all occur on fish products as a result of contamination from animal/human reservoirs. the risk of infection can be eliminated by proper cooking. if the person handling the product is a carrier of Staph. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene.3 Enterobacteriaceae The pathogens Salmonella. and the minimum pH for growth is 4. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin.4. 5. when the bacterial load of the product is lower. the organism can survive most of the light curing processes. 5. growth and toxin formation may occur.30% of the population positive for enterotoxin-producing strains (41).86 and 10% sodium chloride. the human nose. they do not grow well in the raw material. as they require more handling.5. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13).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. As with other organisms. if recontamination occurs and storage conditions are favourable. if storage temperatures are over 10 °C.5% salt in the water phase. An outbreak of Shigella in an Italian cruise ship. has been reported (42). 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.or cold-smoked fish should have at least 3. The organism is mesophilic with a minimum growth temperature of 7 °C. Shigella and E. are particularly susceptible to contamination and. throat and skin.94 (7). Usually. but. Infection by Shigella sonnei has increased in the UK. gills and guts as well as in several lightly preserved fish 114 . aureus.5.5. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. the organism may contaminate the product. it should contain not less than 3.0 . The FDA established that hot. they will grow rapidly (7). after processing.0% salt in the water phase (30). Thus.4 Listeria monocytogenes The psychrotrophic L.5. but. As they are poor competitors. but requires higher temperatures for toxin production (>10 °C). related to the consumption of a raw smoked fish meal. aureus growth. Once the product is contaminated. more importantly. if it contains sodium nitrite ≥100 ppm. In foodstuffs. Sliced smoked fish products.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. However. It is a halotolerant bacterium able to grow at water activities as low as 0.

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

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

where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. * Staph.M. 37(revised). 2007. Doré I. Food spoilage fungi. Fernandes R. 82 (revised). Chapman & Hall. Hot Smoking of Fish.. Smoked. in Micro-Facts: The Working Companion for Food Microbiologists. 4. Preservation of fish by curing (drying. Horner W. Assurance of Seafood Quality. Eds.W.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. Martin R. 334. Marinades. in Fish Processing Technology.. 1990. Wareing P. McLay R. New York. 56.F. botulinum * Limit of tolerance 1x 103 ETEC/g. or presence of toxin 0. monocytogenes Salmonella spp. Regenstein C.E. Hall G. 2.7 1. Urner Barry Publications Inc. parahaemolyticus Vibrio vulnificus C. London. 1991. Van Nostrand Reinhold. 7. 120-38. Flick G. References Regenstein J. New York. coli (ETEC) L.. Van Nostrand Reinhold. except in the viscera of dungeness crab. FAO Fisheries Technical Paper. Catching. 1984. Leatherhead. The Smoked and Cured Seafood Guide.M. Leatherhead Publishing. SMOKED AND DRIED FISH TABLE 5. 8. Introduction to Fish Technology. 1982. 6.F. Fernandes R. Ed. 1994. aureus * Vibrio cholerae V. Handling and Processing Eels. 9. Birnie K. 3273. in The Seafood Industry. cured and dried fish. 381-406.CURED.A. 1997. 254-82 5. salting and smoking).. 117 . Huss H. Wareing P.. LT or ST positive Presence of the organism Presence of the organism Positive for enterotoxin or ≥104 (Most Probable Number) Presence of toxigenic 01 or non-01 ≥ 1 x104 (Kanagawa positive or negative) Presence of the organism Presence of viable spores or vegetative cells in products that will support their growth. 1993. Eds.. 1979. Torry Advisory Note No. Horne J. 3. Bannerman A. Torry Advisory Note No. USA. Torry Advisory Note No. Moody M. Flick G.H.8 ppm (saxitoxin equivalent) 20 ppm domoic acid.

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

Fungi and Food Spoilage. Hocking A. London. 197-208.M. CRC. SMOKED AND DRIED FISH 26.. 1997.V.. Pitt J. Adebajo L. Pitt J. Gill T. Shewan J. 1991. 1941. Maga J. The microbiology of sea-water fish. in Fish as Food. Food Microbiology. Sirvas S.D. Wheeler K.D.A. 88-90.. Letters in Applied Microbiology. 1995.E.I. Commission of the European Communities.. Smoke flavourings. 107-11. 742. 1993. Journal of Food Technology. Importance of autolysis and microbiological activity on quality of cold-smoked salmon. Luxembourg. Food Research International. 1979. Journal of the Fisheries Research Board of Canada. New York. Journal of Tropical Medicine and Hygiene. Fungi associated with Indonesian dried fish. 1986. CEC.S. Studies on salt fish. 1994. 1996. Huss H. Jay J. Lucas F. Hocking A. Sprouse R. Ed. 30. Antimicrobial properties of smoke.D.. Modern Food Microbiology.E. Ahmed F. in Smoke in Food Processing. 38.. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. Fungal flora of South Vietnamese fish and rice. Hocking A. Committee on Evaluation of the Safety of Fishery Products.H. 119 . Pszczola D. 49 (1). Journal of Applied Bacteriology. Taylor L. Røntved S. 35. Pitt J. 43. 74. in Reports of the Scientific Committee for Food (34th Series).M. Cann D. 42. 33.. Rockville. Simini B. Interactions among xerophilic fungi associated with dried salted fish. 40. 29. 1971. in Fish and Fishery Products Hazards and Controls Guide. Nahrung. 29 (2).R. An outbreak of Shigella. 123-9. 218-25. 27.. New species of fungi from Indonesian dried fish.M.. Lancet. Hansen L. Smoked and smoke-flavoured fishery products. Seafood Safety.Y. Frank M. ceviche. 1961.. Anggawati A. 39. 28. Cox J. Fuchs R. 1996.A. 348.K.F. Ed.. Springer Science.T. US Food and Drug Administration.. 32. 311-7. 5. Press Inc. Townsend J. 181-8. Academic Press. Food Technology. 31. 22.O. Hess E. Blackie. Incidence of Listeria monocytogenes in an acidified product.A.A. The control of botulism in hot-smoked trout and mackerel. 37.I. Diyaolu S. Washington DC. 38 (3). 34. Hocking A. 41.F.C. Scientific Committee for Food. 27686. Oxford. V. 2005. Mycotaxon.. 3 (4).. 70-2. Food and Drug Administration. Ed. Wheeler K.D. 98100.J.. 74.. 74 (2). Studies on Sporendonema epizoum from ‘dun’ salt fish.CURED. 36. Institute of Medicine.. 1995. Borgstrøm G. 1991. Tour highlights production and uses of smoke-based flavours. Ed. Food and Drug Administration. 351-7. 12.1988.I. 14. Boca Raton. 1994.. Loessner M. National Academic Press. 164-9.. 1985.B. Golden D.

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

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


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

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

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

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

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. in many cases. When this is a simple carbohydrate source such as sugar. 4). and the anaerobic conditions that prevail in the bulk of the material. 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. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. or boiled or roasted rice. Other halophilic species have been identified from fish/salt products. In another microbiological study of four fish sauces. however. In some cases. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. angkak (Philippines and Thailand) and look-pang (Thailand) are used. which included nam-pla and patis.1 – 6. and studies 127 . traditional starter culture preparations such as koji (Japan). 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. These preparations contain a mixed microflora of moulds and yeasts.3. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp.FERMENTED FISH Where other ingredients are used. Hansenula. mean that microorganisms have been generally held to play little or no role in the process that follows. It also demonstrated their significant proteolytic activity during the first month of fermentation. 6. Endomycopsis and Saccharomyces . Bacillus spp. the Philippine fish sauce. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). were found to be the predominant isolates. Generally. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). (3. principally species of the genera Rhizopus. either directly or as the partially saccharified rice they produce. after one month.3 6.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. however. This. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. 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. sufficient to saturate the water phase present and produce an aw of 0.74 or below. Similar results were seen with patis. they can contribute to the initial microflora.4. More recently.

1. Belacan (Condiment) Fig.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 .2. 6. Patis (Condiment) 128 . 6.12). Fig.

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

which is eventually separated from the residual fish tissues. 6.V and Figures 6.5). particularly glutamate. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. 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 . 6.3. for example. volatile fatty acids and nucleotides. Hoi-Malangpu-Dong (side dish.6 – 6.5. In contrast to free-swimming fish. filtered and bottled. shellfish such as mussels often have appreciable levels of carbohydrate in their tissues and in these cases addition of carbohydrate is not necessary for a lactic fermentation to occur (see. Figure 6.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids.8. Some examples of products and their formulation are presented in Table 6. Fig.

south of Thailand.6.5-1. C NE. NE.4 are also produced using shrimps.12 months Shelf life Use 1 . C NE.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .6.FERMENTED FISH TABLE 6. N S 5 . N.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. north. In these cases.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . 3:1:0. central. NE Formulation Fish (whole or pieces): salt: roasted rice.12 months Main dish 2 Pla-jao C. N. the prefix 'pla' is replaced by 'kung'.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.7 days 5 .20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. side dish) 131 . S.03 Preparation time 6 . Fig. Kung-Chom (staple food. north-east. C.12 days 3 . 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0.2-4 Fish (pieces): salt:khaomark (fermented rice). Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. NE 10 . N.

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

3 years 1 . 133 .FERMENTED FISH (from Arroyo et al.6.12 months 1 .1 4.0 Shelf life 3 weeks 6 .5.9.5 . 2.0 .7 .8 pH 4.3 5.9 4.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.8 2. Philippines Journal of Food Science and Technology.5 .9 .6.1 .8 .0 . .3 .5..4.6 3. 106) Fig.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).0 .4 . White balao-balao TABLE 6.2 7.5 3.9 4.

FISH AND SEAFOOD 6. also serves as a valuable source of the more readily fermented carbohydrate.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.5) despite other factors. also lower the unit cost of the product and this is probably a significant factor in many cases. 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. For example. 19). although the final population of LAB may be similar.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi.3. being similar (18.4. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. of course.VII). alternatively. Where they are used. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. Garlic. When added as boiled or roasted rice alone.1 . 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). a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. TABLE 6. the rice may be added in a partially saccharified form such as kao-mark (Thailand). such as salt content. Rice is the most common carbohydrate source used. Addition of carbohydrate will. In several products.0 . 21).3.9) compared with the white product (pH 4. Although the ability to ferment starch is not widespread in the LAB. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. a common ingredient in these products.2. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. 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 . This results in a more active lactic fermentation and a lower pH. inulin.

3 days. will not reliably exclude air. presumably correlating with the salt content. 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). Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage.5. There is little published information on the factors that limit the shelf life of the low-salt products. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. The mixture is allowed to ferment for 2 .5 in the first 2 . the Korean fermented fish product. is something of an exception since millet is used along with garlic as the carbohydrate source. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). Excessive growth and acid production by the LAB could render products too sour for acceptability. 135 . allowing other spoilage organisms to grow. Results of a product survey were consistent with this. Clearly. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products.7 to below 4. around 12%. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. reporting the product’s pH to range from 4. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others.2. the pH drops from 6. but they are likely to behave similarly to lactic fermented products produced from other commodities.7 up to 6.2 at the end of fermentation. lead to an increase in pH. which ranged from 7. The growth of yeasts and moulds could.5:9:3. In a successful fermentation. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. 6. thereafter declining to about 4. 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.8 .17.3 weeks at around 20 °C.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). The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. such as wrapping in banana leaves. Traditional packing techniques used with lactic fermented fish products. in turn. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. Generally.9%. and there is a marked regional variation in the preferred type. the level of salt employed is on the high side for fish/salt/carbohydrate products. markedly extending the shelf life (25).FERMENTED FISH Sikhae.

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

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

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

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

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

and a second updated edition dealing with the pharmacology.C.M. 5). Nevertheless. Ph. It remains to be seen if this approach gains momentum in the future. a number of new toxins and toxin families have been identified. While some of these toxins only occur in seafood from a restricted geographical region. Although most of these antibody-based test kits have yet to receive international regulatory approval. physiology and detection of marine and freshwater toxins has also appeared (8)..D. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. Increasingly sophisticated instrumentation (e. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. The manual on harmful marine algae and their products. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. Wright.3) and some practical applications for detecting shellfish toxins have been reported (4. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. Scientific and Cultural Organisation) has been updated (7). 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. FISH AND SHELLFISH TOXINS Jeffrey L. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins. known and unknown. C.7.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. several scientific reports have described their efficacy and potential. including the well known marine toxins 141 . and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. most notably the azaspiracids and the cyclic imines as well as some additional polyethers. Another book dealing with the clinical effects of a variety of poisons and toxins.. Since the original chapter in this series was published (6).g.

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

Human symptoms begin with tingling and numbness of the lips.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. and humans seem to be particularly susceptible. and fingers within 5 . Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. 143 . 7. 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). this is never sufficient to render the seafood safe to consume. tongue. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins.1.2. However.30 minutes of consumption. 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. 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. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water.

3. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). 15.8 µg/g) of shellfish or seafood tissue. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). a criticism has been that such methods are time consuming. a pre-column method has received Association of Analytical Communities (AOAC) approval (18).3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. China. 24) (Figure 7. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). Two approaches have been taken here. and islands of the South Pacific. Since its discovery. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. The latter test is accepted by the United States Food and Drug Administration (US FDA) for use in the United States National Shellfish Sanitation Program. and while simple and relatively inexpensive to operate. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. as well as other parts. 23. 7. specifically 144 . There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. 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®). and has also been evaluated by a number of EU laboratories.2. Thailand. 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. After some development and various trials. While the chemical detection methods are most promising for the future.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. Taiwan. However it remains as the monitoring method of choice in many countries. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. and over ten analogs are known (22. Most intoxication events have occurred in South East Asia. TTX most commonly occurs in many species of puffer fish.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. The mouse bioassay has been in place for decades and the method has been standardised.2). other derivatives have been reported. 17). including Japan.FISH AND SEAFOOD 7.

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

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

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

are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. although in the latter study the probes for Pseudonitzschia pungens. but just over 15 years ago a third toxin. and this is typically the method of choice. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). Dinophysis spp.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. The benthic Prorocentrum spp. To further complicate matters. and these methods have been reviewed (40. shellfish tissue can be efficiently extracted with aqueous methanol (1:1).FISH AND SEAFOOD 7. 41). dinophysistoxin-1 (DTX-1). 48. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC.. The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. 49). 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). 7. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. and dinophysistoxin-2 (DTX-2) (Figure 7.4).4. produce a variety of other 148 . Alternatively. However. the heterotrophic dinoflagellates Dinophysis spp. Fortunately DA is conveniently detected by a variety of LC-linked analytical methods and this is the recommended approach for DA monitoring in EU countries. Since then. though this method is most commonly used to analyse plankton and seawater (42). In the context of this chapter. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. australis. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). as can LC/MS. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. DA can be detected by the mouse bioassay but the regulatory limit for shellfish is set below this level at 20 µg/g (20 mg/kg) shellfish tissue. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. in Monterey Bay. and consequently were often overlooked as the real source of the toxins. DTX-2 was discovered in contaminated Irish shellfish (46). and an immunosensor method has also been reported (4).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.. California. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative.

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

3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. it does not provide information on the DSP toxin profile.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). 7.5. various European countries.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. A regulatory level of 80 . At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. and a detection limit of 100 µg/kg has been reported (12). The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . and while this provides critical seafood safety information. it was agreed that the established mouse bioassay (52) with an observation time of 24 hours is currently the preferred method for the detection of the acute toxicity of acetone soluble DSP toxins. most progress in DSP toxin detection has been using new or improved LC/MS methods and a detection limit of 10 µg/kg has been reported. 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. DSP episodes have been reported from Japan. 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). Since the late 1970s.3 days. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. In recent years. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. New Zealand and Chile. vomiting. or indeed on the presence of other associated LSTs that can occur with DSP toxins and which result in a positive mouse bioassay result (vide infra). and DTX-2. DSP-Check is a commercially 150 . 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. Due to the lipophilic nature of the toxins. even with the minimal of clean up steps (48). Human symptoms include diarrhoea.5. nausea. DSP episodes have been reported worldwide and are included in many monitoring programs (47). 7. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. and abdominal pain.

and another rapid antibodybased test kit for DSP toxins has also been reported (53). with good agreement with the mouse bioassay and LC–based methods (54.5 and 7. Indeed.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. with a claimed detection limit of 20 µg/kg. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). 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. 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. In yet another approach.5. 7. 58).6. Within this group. Some representative examples of both groups of toxins are shown in Figures 7.6 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). Fig. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. 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. 55). fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. for many years these compounds were included in the DSP toxin complex. However. 7.

Japan. 58). 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. 7. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. 57. Structure of the two parent yessotoxins.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). YTX and homo-YTX 7.FISH AND SEAFOOD Fig. which may co-occur in blooms with other organisms. 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). 7. the toxin content and profile may vary among different strains. eastern Canada. many parts of Europe and the Mediterranean. While 152 . New Zealand and South America (49. As is typical for many algal species.6.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum.6.6.

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

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

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

The diversity of benthic phytoplankton that exist on reefs further complicates the picture. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7.1 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.FISH AND SEAFOOD transferred through the food chain to the larger carnivores. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. lipid soluble polyethers. a group of closely related polyethers that are considered the source of ciguatoxins. Distribution through the food chain begins with 156 . Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red. but more are suspected to exist. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. 7. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins. Fig.8. These include species belonging to the genera Ostreopsis and Coolia. CTXs are large.8). green and brown seaweeds and also occur in sediments and coral debris. A detailed comprehensive review of ciguatoxins has recently been published (74). complex. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. and only a few structures have been identified.

and temperature sensation reversal.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. Like the brevetoxins.76). and although the gastronomical problems may dissipate within a day or so. and it is estimated that ingestion of as little as 0. all of which can be linked to the effect of CTX on excitable membranes. which can be confused with chronic fatigue syndrome. only trace amounts of CTXs can cause illness following consumption of contaminated fish. 78). and there are suggestions that such incidents are increasing and spreading worldwide (74). the neurological symptoms can persist for days. Local island populations attempt to avoid poisoning by catching only smaller fish. where cold objects feel hot and vice versa. Globally. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. Patients have reported symptoms lasting weeks and even months. sharks. and moray eels. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms.8. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. although not all CTXs are equipotent. These include paresthesias (numbness and tingling of the extremities).1 µg of ciguatoxin can result in human illness (74. which can return following the consumption of alcohol and certain foods such as oily fish (74. myalgia. 75). and the sensation of loose teeth (77). 7. groupers. which in turn are consumed by carnivores such as snappers. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. 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. pruritus. Another symptom that can develop over time is extreme fatigue. there are neither standards nor an official testing program. barracudas. Even when suspect samples are available. Other symptoms include a metallic taste. 75. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. 7.3 Detection methods As a consequence of their potency. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . weeks or even months (74 . The onset of symptoms can occur rapidly within 24 hours. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. and in fact CTX poisoning is often only identified after consumption of contaminated fish. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue.8.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. arthralgia.

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

7. Portugal.9. AZA toxins do 159 . In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. which are ingested by P. 90). Following the original episode.). Ensis siliqua. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. these molluscs were never considered to be the biogenetic source. There has been considerable debate over the source of azaspiracids. These symptoms are not unlike those experienced following the consumption of DSP toxins. scallops (Pecten maximus). Mytilus galloprovincialis). Donax spp. 89). oysters (Crassostrea gigas. cockles (Cerastoderma edule). 7. and off the coast of Morocco (83. The bivalves affected include mussels (Mytilus edulis. cassipes produces AZAs in culture. it has been impossible to establish that P. vomiting. 84.9. cassipes (83).2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. Structure of the three principal azaspiracids found in shellfish 7. clams (Tapes philipinarum. which can persist for 2 . However. While this result is not necessarily in debate. Scandinavia. and to date this has been the only source from which these toxins have been found at toxic levels.3 days. Ostrea edulis).FISH AND SHELLFISH TOXINS Fig. as in other cases of shellfish toxins.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. and to date no long-term effects have been recorded (83. AZA occurrence has been reported by several European countries including France. but current data indicates that unlike the DSP compounds. and some time after the original episode in Ireland.9. and most recently in a crustacean (Cancer pagurus). it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). diarrhoea and stomach cramps. 88.

However. and erosion and bleeding in the stomach. 7.12). Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. The mouse bioassay has been used to monitor shellfish extracts. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. 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 closely related ptierotoxins (Figures 7. it is recommended that the entire shellfish be used for AZA analysis since it has been shown that these toxins can migrate to other tissues. AZAs also display cytotoxicity towards various mammalian cell lines (83. 85). but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. Once again. the spirolides. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. and ASP. parenchymal cell pyknosis in the pancreas. 7.10 Cyclic imines These toxins are represented by several distinct structural types.9. though the possibility exists that they may inhibit other phosphatases. 92).10 – 7. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. 91. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). Importantly these pathological outcomes were reported as being different from those caused by DSP. the pinnatoxins. Based on IP minimum lethal dose data. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . interferences can arise when analysing a matrix as complex as a crude shellfish extract and it is recommended that a simple solid phase extraction (SPE) step be introduced prior to LC/MS analysis (93). and necrosis of epithelial cells on the microvilli (91). Using the same approach. abnormalities in the thymus and spleen. but all contain a spirocyclic or cyclic imine function within their structure. PSP. 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. which are particularly amenable to detection by positive ion electrospray mass spectrometry.

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. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. 161 . Cyclic imines are often referred to as fast acting toxins due to their “all or nothing” effect in the mouse bioassay over a very narrow dose range. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis).1 Origins and distribution The original observation of toxicity in oysters led to a wider survey of other shellfish species in New Zealand when it was discovered that the toxin was present in a variety of bivalve molluscs including mussels. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin. 7. cockles.10.10. Fig.. surf clam and New Zealand abalone.FISH AND SHELLFISH TOXINS health. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . but they are included here for completeness. Subsequent taxonomic studies revised this again to a new dinoflagellate K. two additional congeners. 95). scallops. Structure of the cyclic imine gymnodimine 7. selliformis (97).10.1. it was proposed that a dinoflagellate identified as Gymnodinium spp.10). In only two cases (gymnodimines and spirolides) has a planktonic source been identified. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. Some time later. Pacific oyster. 7. At the time of the initial observation of gymnodimine. they are sometimes referred to as emerging toxins of uncertain human health concern. 95). Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94.520 Da (94.

However. Reduction of the imine group eradicated the toxicity.FISH AND SEAFOOD 7.1. and lack of responsiveness to stimuli. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. 7. 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. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection.10. There is a rapid onset of symptoms including respiratory distress.100). Whether isolated from dinoflagellate cells directly or from shellfish tissue. Based on this diminished oral toxicity.2 Toxicology A review of the toxicology of all cyclic imines has been published (98).10. The time lapse between ingestion and death is between 5 and 15 minutes. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . and so spectroscopic detection by UV methods is not favoured. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. almost a dozen derivatives in total have been identified to date (94. together with a lack of evidence of human intoxication. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine.11).900 µg/kg). Since their original discovery which resulted in the characterisation of spirolides A-F (99. 95) (Figure 7. 50%) of 96 µg/kg.3 Detection methods The characteristic “fast-acting. Oral administration of the toxin to mice by gavage resulted in about an 8. They were originally discovered in extracts of mussels (M. abdominal breathing. finishing with 100% methanol.to10-fold reduction of toxicity (LD50 600 .1. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. 95). In particular.10. paralysis of the hind quarters and eventually complete immobility. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. it has been stated that regulation against gymnodimine is not required (98). 7. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. A later study using purified toxin established an LD50 dose (Lethal Dose. and this is the detection method of choice. Most cyclic imines lack an extended chromophore.

10.11.2. All data reported here are for desmethyl spirolide C. fasted mice).166 µg/kg. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia.707 µg/kg. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). in fjords in Denmark and Norway. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. fasted mice). southern Ireland. Interestingly. 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. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). Structure of four spirolide toxins A-D 7. those spirolides possessing the vicinal dimethyl grouping (see Figure 7. or by oral ingestion (381 .8 µg/kg). the gulf of Maine. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. 7. 7. While oral toxicity is considerably less than by IP injection.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra).11.2. by gavage (87 . Fig.2 Toxicology The spirolides are the most studied group of the cyclic imines. 95). these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . the western coast of Scotland. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 .10.

2.FISH AND SEAFOOD (100). 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. 95.12). Three members of this group have been reported.B contain a spiro imine moiety similar to that found in the spirolides. 102). the mouse bioassay can be used to detect these fast acting toxins. 7. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. four members of the group have been identified – two of them being the C-34 diastereomers. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry.3 Detection methods As discussed for gymnodimine. However. While the mechanism of action remains to be clearly defined. 7. Fig. 7. consistent with modulation of acetyl cholinergic receptors (103). which includes two diastereomers. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C.12. To date.3 Pinnatoxins and pteriatoxins Pinnatoxins A .10.10. A number of methods following this procedure have been described (94. and in fact these compounds share a number of other structural features.

new toxin groups such as the azaspiracids and cyclic imines have also been reported.2 Toxicology There is little information describing the toxicology of these compounds (98). However. The LD99 (Lethal Dose.10. Only the acute toxicity in mice following IP injection has been reported. This is generally a result of increased monitoring and continually improving detection technologies and methods. 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.100 µg/kg). while pinnatoxin D was essentially inactive.3.3. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. like the gymnodimines and spirolides. Coupled with this. Despite this progress.3 Detection methods No formal detection methods for these compounds have been reported.most likely a dinoflagellate.1 Origins and distribution Shellfish of the genus Pinna.FISH AND SHELLFISH TOXINS 7. 99%) values for pinnatoxins A-C are in the range (22 . 7. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). there is no compelling evidence to suggest that they have resulted in human toxicity (98). While this is recognised as a real possibility for a wide range of toxins. have long been suspected of causing human illness after ingestion. 7.11 Conclusion There has been considerable progress in the detection of known marine toxins. 7. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. which in many cases are now found to be more widespread than previously believed. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. There is strong circumstantial evidence to suspect that these toxins. to date action or 165 .10. With increasing knowledge of the chemistry of toxins. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). but this remains to be conclusively established (106). once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. arise from an algal source . which occupy tropical waters around Okinawa and the south east coast of China. and more advanced means of detection. The LD99 dose for the pteriatoxins is reported to be in the range (8 .3.180 µg/kg).10.

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

Shoji Y. Noguchi T. Ed. Determination of marine biotoxins for regulations: from the mouse bioassay to coupled LC-MS methods. Risk assessment of marine toxins. AOAC International. 18. prechromatographic oxidation and liquid chromatography with fluorescence detection.. Analytical Biochemistry.M.. 2008. Interspecies distribution and biogenetic origin of tetrodotoxin and its derivatives. 21.. Neurotoxins from marine dinoflagellates: A brief review. 177-96.R. Villar-Gonzalez A. Marine Drugs.L. Gaithersburg. 2006. in Methods of Analysis of the Association of Official Analytical Chemists. CRC Press.. Journal of Shellfish Research. Eds. Paris. 20. in Manual on Harmful Marine Microalgae. Stobo L.FISH AND SHELLFISH TOXINS 11.. Boca Raton. Pure and Applied Chemistry. Barrett R. 2002.V. 283-317.A.. Marine Biotoxins. 455-60. Electrospray ionisation mass spectrometry of tetrodotoxin and its analogs: Liquid chromatography/mass spectrometry. 5-52.E. Tetrodotoxin – distribution and accumulation in aquatic organisms and cases of human intoxication. in Seafood and Freshwater Toxins. Cembella C. Van Amerongen A. Naoki H. Wang D-Z. Wageningen Academic Publishers.. 1989. 2005. 1995. Lacaze J-P. Paralytic shellfish poisoning toxins in shellfish. 6. Turrell E. Fukuyo Y. Murata M. Larsen J. 61. Wageningen..A.M.H. in parallel with the regulatory AOAC mousse bioassay.A. 117-34. Taxonomy of harmful dinoflagellates. a new rapid test. Evaluation of MIST Alert rapid test kits for the detection of paralytic shellfish poisoning toxins in shellfish..... in Rapid Methods for Biological and Chemical Contaminants in Food and Feed. Toxicon. Christian B.. Eds.. 635-48. Smith E. Quilliam M. Rapid detection methods for marine toxins: needs and limits. Yotsu-Yamashita M. 24. Yotsu M.. Krys S. CRC Press. Miyazawa T. Anderson D. Chemical analysis. Aune T. 3-20. 14.. Food and Agriculture Organisation. Luckas B. method 2005. Analytical and Bioanalytical Chemistry. Scott A. 219-27.. Association of Official Analytical Chemists. in FAO Food and Nutrition Paper 80. 230-42. Toxicon.. Barug D. Jellet J. Roberts R.J.L. 349-71. 22. tandem mass spectrometry. 13. Botana L. 2004. Food and Agriculture Organisation of the United Nations. Food and Agriculture Organisation of the United Nations... 51.A. Marine Drugs. 505-8. 16. Yasumoto T. 290.M. Ben-Gigirey B. Lauwaars A. Laycock M. 21. 2008. 10-7. Detection of paralytic shellfish poisoning (PSP) toxins in shellfish tissue using MIST Alert. Arakawa O. 1407-25. Yasumoto T. 6.F. Botana L.C. Surveillance of algal toxins in shellfish from Scottish waters.. Hallegraeff G.C. Ed. 12. UNESCO.. Rome. in Seafood and Freshwater Toxins. 2008.. 2002. 17. 391 (1). 19. 167 .. 2001. Ed. Endo A. Boca Raton.06.. Paralytic Shellfish Poisoning. 15. Mackintosh F. Petrie J. Taylor F. 2008. 2008.M. and liquid chromatography/tandem mass spectrometry. 2008. 23.M. 40.

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

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

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

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

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

80. 81.








89. 90.



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


94. 95.





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

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


Rhea Fernandes and Dr. Peter Wareing Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 8.1 Introduction

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


853/2004 require a fully operational, and maintained HACCP system, according to Codex, to be in place. 8.2 Definitions

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


Establish the corrective action to be taken when monitoring shows that a CCP is not under control. 5. The team should include members familiar with all aspects of the production process as well as specialists with expertise in particular areas such as production. Determine the CCPs.The application of methods.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. 4. Verification . Each stage is discussed below in detail. The team should also include 177 . procedures. 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. 2.Obtaining evidence that the elements of the HACCP plan are effective. 8. ingredient and packaging buyers. Figure 8.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . food chemistry or engineering. quality assurance or control.1 Assemble HACCP team HACCP requires management commitment of resources to the process. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. in addition to monitoring to determine compliance with the HACCP plan. 7. food microbiology. 3.A point. Establish the critical limit(s). from primary production to final consumption. tests and other evaluations. Validation . hygiene managers. procedure. Establish procedures for verification to confirm that the HACCP system is working effectively.1 is a flow diagram illustrating this 12-stage logic sequence. Conduct a hazard analysis. Establish documentation concerning all procedures and records appropriate to these principles and their application. 8. 6.3. Establish a system to monitor control of the CCP. operation or stage in the food chain including raw materials.

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

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

It may include details of the factory and equipment layout. cleaning and hygiene procedures and storage conditions. For example.6 List all potential hazards associated with each step.3. conduct a hazard analysis. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. 8.5 On-site confirmation of the flow diagram The HACCP team should confirm that the flow diagram matches the process that is actually being carried out.3. it is essential that the details of each step are fully appreciated and recorded. However. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards.3. Ideally. 8. or circumstances of any reclaim or rework activity. 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. and therefore a detailed description of the procedure applied must be available. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. it should also include details of CCP steps. 8. once determined.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters.FISH AND SEAFOOD product. the qualitative and/or quantitative evaluation of the presence of hazards. The team should then conduct a hazard analysis to identify which hazards are of such a nature that their elimination or reduction to an acceptable level is essential to the production of safe food. time/temperature data.4. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. 180 ● ● . survival or multiplication of pathogenic microorganisms. The operation should be observed at all stages. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. because the hazard analysis and decisions regarding CCPs are based on these data. features of equipment design. ingredient specifications. It is essential that the diagram is accurate. If HACCP studies are applied to proposed new process lines or products.

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

3.1 Fish and seafood Again. More than one critical limit may be defined for a single step. application of the decision tree should be flexible.7. For example. and its use may not always be appropriate. given the enormous variety of fish products and processes in use.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.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. Criteria used to set critical limits must be measurable and may include physical. it can be said that effective control measures are likely to include the following: • • • • • 8. then the process must be modified to include a control measure.FISH AND SEAFOOD 8. If a significant hazard has been identified at a step where control is required for safety. it is necessary to specify both time and temperature for a thermal process.3. and the reader is referred to the appropriate product chapter. biological or sensory parameters. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. Where possible. 8. but for which no control exists at that step or any other. However.8. Some examples relevant to fish products are: • • • Time and temperature limits for pasteurised and cooked products Temperature limits for chilled fish Brine concentration in cured fish 182 . 8. However. chemical. structured approach to decision making.2) to provide a logical.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. it is not possible to generalise on likely CCPs. critical limits should be specified and validated for each CCP.


• • • 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.


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

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

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.9. European Communities L 268. 7. 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.B. 41). 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption Regulation (EC) No. packaging.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. Subsequently. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. and sets out what those enforcing the provisions have to do. J. 9. 24. manufacturing. N. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. p. transportation.1).1991. handling and offering for sale or supply to the consumer. 190 • • .2 Legislative Structure From 1 January 2006. p.7. as last amended by Regulation (EC) No 806/2003.15). Regulation 854/2004 relates to the organisation of official controls on products of animal origin.9. 24. distribution. storing. 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. processing. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin.1991. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. as last amended by Regulation (EC) No 806/2003. A number of more detailed implementing and transitional measures have been adopted at EC level. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No.1992. p.

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



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:


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

EY. a new mark must be applied to the product.2 Fishery product requirements 9. please refer to Regulation (EC) 853/2004. (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 food business operator may apply the same identification mark to both types of food. Section I of Regulation (EC) 853/2004.4. ES.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. and the characters easily decipherable and be clearly displayed.2. the mark must show the approval number of the establishment. EG.2 Form of the identification mark (i) The mark must be legible and indelible.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. EÜ.4. the mark must be oval shaped and include the abbreviation CE. In this case. EF. the code would be UK. EK. EK. (iv) If the identification mark is applied in an establishment within the Community. for other codes. Extracts of the requirements of Annex II. 198 . EO.4. Section I of Regulation (EC) 853/2004 are given as follows: 9.2. when a product's packaging and/or wrapping is removed or is further processed in another establishment.4. (ii) The mark must indicate the name of the country in which the establishment is located and written out in full or shown as a two-letter code in accordance with the relevant ISO standard.FISH AND SEAFOOD 9. For the UK.1. (ii) However. (iii) Additionally.2. 9. EB or WE. 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. EC.1. the new mark must show the approval number of the establishment where this occurred.

the approval number of the establishment and the abbreviation for the community where appropriate. and fishery products carried in bulk. 9. Parts A. echinoderms.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. and are intended for further handling. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. Extracts of the requirements of Annex III.LEGISLATION 9. for full requirements. Therefore. C and D.1. it applies to those animals when not placed on the market live. Chapter IV. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . Part A and Chapter V apply to retail. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels.2. it is sufficient to apply the mark to the exterior of that package only. This section does not apply to bivalve molluscs. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. a) requirements for establishments.2. the wrapping or the packaging. the mark may be applied to the outer surface of the container or packaging.e. With the exception of Chapters I and II. they would need to be obtained following Section VII on live bivalve molluscs.4. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. Chapter III. its wrapping or the packaging. Section VIII of Regulation 853/2004. 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. (v) When the mark is applied directly to products of animal origin. specifically relating to fishery products are given below. reference should be made to the actual Regulation. as amended. processing. (ii) For products of animal origin that are placed in transport containers or large packages. granulate and powdered products of animal origin carried in bulk. i. or be printed on a label affixed to the product. including vessels.4.3 Method of marking (i) The mark is to be applied directly to the product. the colourings used must be EC authorised colours. wrapping or packaging in another establishment.

from the place of production to the first establishment of destination. There is a derogation from point a. Chapter VII of Regulation 852/2004.. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. removing fins.2. Structural and equipment requirements refer to the following points 1. or to handle or process them after harvesting. i. clean seawater may be used for the handling and washing of fishery products. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. (a) primary production covers the farming.1 Chapter I .Requirements for vessels Food business operators are required to ensure that they meet the structural and equipment requirements as well as hygiene requirements required for vessels used to harvest fishery products from their natural environment. 2. refrigeration and wrapping. 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. fishing and collection of live fishery products with a view to their being placed on the market. it must be placed somewhere that will not contaminate the water supply. bleeding. 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. heading. A. if a vessel has water coming in that is used with fishery products. gutting. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material.e.) 9. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter.4. this supplements the requirements of Annex II. in that record keeping requirements stated by point 7 in Part A of Annex I of Regulation 852/2004 do not apply to operators involved in small-scale coastal fishing (as defined through Regulation (EC) 1198/2006) if the activity is under 24 hours.2.

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

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

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

they must be cooled rapidly. (iii) A document from the manufacturer. D. (iv) (wild) Atlantic and Pacific salmon. (b) fishery products from the following species.4. except when supplied to the final consumer. Once produced. (ii) mackerel. and whole fish must be gutted and washed beforehand. (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. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. (iii) sprat. 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.2.2. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . 9. and (c) marinated and/or salted fishery products. if the processing is insufficient to destroy nematode larvae.Requirements for certain processed fishery products A.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. must accompany fishery products referred to above when placed on the market. cooling must continue until a temperature 204 . or a stabilising treatment. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. stating the type of process they have undergone. raw materials must have no guts.4 Chapter IV .FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. If no other method of preservation is used.

B. C. Shelling/shucking is to be done hygienically. 2073/2005).Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. there is an exception: food business operator may refrain from chilling the fishery products when whole fishery products are used directly in the preparation of fish oil for human consumption. Total volatile nitrogen Unprocessed fishery products cannot be placed on the market if chemical tests reveal that TVB-N or TMA-N limits are exceeded (limits are set in Regulation (EC) No. pressing. However.LEGISLATION approaching that of melting ice is reached.2. 2074/2005. 2073/2005. refining and purification steps before being placed on the market for the final consumer. centrifugation. the following conditions need to be met: A. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. 205 . raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. 2074/2005). and the raw material is processed within 36 hours after loading. and after this.4.5 Chapter V . separation. 9. 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). 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. (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.2. B. processing. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport.

(ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. Molidae. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. The scientific name must accompany the common name on the label. 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.2. Diodontidae and Canthigasteridae. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. Fresh.2. 206 . 9.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. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption.2.7 Chapter VII . and when fishery products are wrapped on board fishing vessels. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. prepared and processed fishery products belonging to the family Gempylidae.Storage of fishery products (i) Fresh fishery products. Fishery products contaminated with parasites may not be placed on the market for human consumption. 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. it should be kept at no more than -9 °C.6 Chapter VI .FISH AND SEAFOOD D. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice.4. 9. E. thawed unprocessed fishery products.2. In the case of whole frozen fish in brine intended for the manufacture of canned food.4.

(b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. (ii) If fishery products are kept under ice.3 Other information: Rules relating to visual inspections for the detection of parasites in fishery products are stated in Annex II (obligations on the competent authorities) of Regulation (EC) No. (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 .).4. species belonging to the Merlucciidae family.2. Sebastichthys capensis. The temperature of frozen fishery products must not fluctuate above -15°C.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. 9. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar.Transport of fishery products The above points for Chapter VII apply in relation to transport.8 Chapter VIII . 2074/2005. 9.2. provided that the journey is short and the competent authority allows this. melt water must not remain in contact with the products. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. Helicolenus dactylopterus. 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. species belonging to the Gadidae family. The following in relation to TVB-N limits is extracted from the regulation: Unprocessed fishery products shall be regarded as unfit for human consumption where organoleptic assessment has raised doubts as to their freshness and chemical checks reveal that the following TVB-N limits are exceeded: (a) 25 mg of nitrogen/100 g of flesh for the species: Sebastes spp.4. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated.2.

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

209 . Requirements for production areas (i) The competent authority classes (classed A. they need to be sent to a processing establishment and undergo specified treatments such as sterilisation in hermetically sealed containers or specific heat treatments such as immersion in boiled water so the internal temperature of the mollusc flesh reaches at least 90 ºC and this temperature is maintained for at least 90 seconds.3. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. C.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority.4. 9. processing or relaying.Hygiene requirements for the production and harvesting of live bivalve molluscs A. 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. poles or other fixed means are to identify the boundaries of the sites.2 Chapter II . (ii) Provided the requirements of chapter V are met. collection from: .Class B means treatment in a purification centre or after relaying is done before it can be placed on the market .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.Class C means relaying over a long time before it can be placed on the market For bivalve molluscs not subject to Class B or C treatments. B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with. B. Transport needs to allow sufficient drainage for best survival conditions.

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

160 micrograms of azaspiracid equivalents per kilogram.4.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.4.7 Chapter VII .8 Chapter VIII .3. dispatch centre or processing establishment. they cannot be re-immersed in or sprayed with water.6 Chapter VI .Other requirements (i) The temperature at which live bivalve molluscs are stored/ transported in should be at one that does not adversely affect food safety or their viability. including the identification mark. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.3. the species of bivalve mollusc and date of packaging (at least the date and month) are required. 9.3.9 Chapter IX .4. 9. the label.Identification marking and labelling In addition to general identification marking (contained in 9. 211 . the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.LEGISLATION (v) azaspiracids. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre.2.4. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.4. Finally.1). 9. Registration document requirements also apply. must be waterproof.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards. Also. Requirements of Chapters III and IV are to be complied with. (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.3. by way of derogation from directive 2000/13/EC.

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

5. A summary is given as follows: 9.2 This states official controls for the production and placing on the market of live bivalve molluscs.5. live bivalve molluscs must not exceed 46. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. Also. in 90% of the samples. live echinoderms. live tunicates and live marine gastropods. 4. Classification of production and relaying areas: This section reinforces that specified in Annex III. - - - - B. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. Live bivalve molluscs from Class C areas must not exceed 46. coli per 100 g of flesh and intravalvular liquid. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. three dilutions MPN test specified in ISO 16649-3.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. Live bivalve molluscs from Class B areas must not exceed. coli per 100 g of flesh and intravalvular liquid.Official controls concerning live bivalve molluscs from classified production areas A. chapter V is reinforced. The reference method for this analysis is the five-tube. as previously stated.1 Chapter II . In the remaining 10 % of samples. coli per 100 g of flesh and intravalvular liquid. The reference method for this analysis is the five-tube. Annex II 9.600 E. 213 .2. 000 E. that microbiological criteria and chemical contaminants amongst other requirements are met. Sampling plans and frequency are required. Regulation (EC) 854/2004.000 E. section VII.

Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions. which has applied since 1 January 2006. E. boundaries or class of a production area or its closure. establishes microbiological criteria for a range of foods.5.2 Chapter III . 9. 2073/2005. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met.6 Regulation (EC) No. Decisions after monitoring There are specific requirements whereby if sampling results indicate that health standards for molluscs are exceeded or there is a risk to human health.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. 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. 214 .5. Procedures for re-opening a closed production area are stated D. opening or closing a production area. Recording and exchange of information Requirements are stated for competent authorities in relation to establishing and maintaining a current list of approved production and relaying areas and their role to inform interested parties of any changes of location. the competent authority is to close the production area. 9. Additional monitoring requirements This concerns the competent authority in relation to monitoring of classified production areas where bivalve mollusc harvesting is forbidden and of relaying and production zones to ensure there is no malpractice etc. F. 9.FISH AND SEAFOOD C.2. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying.

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

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. other than those intended for infants and for special medical purposes(4) (8) L. monocytogenes. monocytogenes.3 Ready-to-eat foods unable to support the growth of L. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life . monocytogenes L. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.TABLE 9.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 1.2 Ready-to-eat foods able to support the growth of L. monocytogenes 1.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L.

tunicates and gastropods 1(16) 0 230 MPN/100 g of flesh and intravalvular liquid 100 mg/kg 200 mg/kg HPLC (19) ISO TS 16649-3 Salmonella 1.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1.25 Live bivalve molluscs and live echinoderms.27 Fishery products which have undergone enzyme maturation treatment in brine. manufactured from fish species associated with a high amount of histidine (17) Histamine . tunicates and gastropods 9 (18) 2 E. coli (15) 1.Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.17 Live bivalve molluscs and live echinoderms.

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

if all the values observed indicate the absence of the bacterium. — unsatisfactory. 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. monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory. if any of the values are > the limit. excluding live bivalve molluscs and live echinoderms. coli. if the mean value observed exceeds m or more than c/n values are between m and M or one or more of the values observed are > M. — unsatisfactory. 1097-101. if all the values observed indicate the absence of the bacterium. a maximum of c/n values observed are between m and M 3. Interpretation of the test results relating to Table 9. where the limit refers to a pooled sample. L. — unsatisfactory. if the presence of the bacterium is detected in any of the sample units. 82. no values observed exceed the limit of M. — unsatisfactory.I The limits given refer to each sample unit tested. L. (1) 219 . 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. — unsatisfactory. L. monocytogenes in other ready-to-eat foods: — satisfactory. monocytogenes in ready-to-eat foods able to support the growth of L. 1999. J. if the following requirements are fulfilled: 1. The test results demonstrate the microbiological quality of the batch tested(1).LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. if the presence of the bacterium is detected in any of the sample units. Salmonella in different food categories: — satisfactory. if all the values observed are ≤ the limit. The test results may be used also for demonstrating the effectiveness of the hazard analysis and critical control point principles or good hygiene procedure of the process. AOAC Internat. if all the values observed indicate the absence of the bacterium. tunicates and gastropods in relation to testing E. the mean value observed is ≤ m 2.

1 Shelled and E.4. coli 5 shucked products of cooked crustaceans and molluscan shellfish 2 100 cfu/g 1000 cfu/g EN/ISO 6888-1 or 2 FISH AND SEAFOOD Coagulasepositive staphylococci 5 Improvements in production hygiene 220 .TABLE 9.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.

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

For other food. and was kept at above 8 °C or the recommended temperature. 3).food.I.S. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment.8 Guidance In the U. 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. 2006 No.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 .FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours.I. S. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions.K. for a limited period consistent with food safety. 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. Alternatively.gov. was kept at above 8 °C or the recommended temperature for an unavoidable reason. 2006 No. such as that below. or. the Food Standards Agency has published guidance notes on the requirements of the EU hygiene and microbiological criteria regulations which can be accessed at the following link: http://www. or at a temperature above 63 °C and to reheat food to a temperature of at least 82 °C (The Food Hygiene (Scotland) Regulations 2006. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. 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. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S. 9. including: Shelf life: where the minimum durability or 'use by' indication is required according to Regulation 20 or 21 of the Food Labelling Regulations 1996 (form of indication of minimum durability and form of indication of ‘use-by date’) the period up to and including that date.

Regulation (EC) No. 2074/2005.2004. Regulation (EC) No.2004. 223 . L226. 25. 25. 2076/2005. Commission Regulation (EC) No. L139. 2076/2005.2005. Regulation (EC) No. 2074/2005. 854/2004 is now set out in a Corrigendum (OJ No. L338. 852/2004 is now set out in a Corrigendum (OJ No. The revised text of Regulation (EC) No. 2074/2005 and Regulation (EC) No.4.2004. 1). 22) as amended by Regulation (EC) Nos.2004. 206). as amended.LEGISLATION 9.10. 853/2004 is now set out in a Corrigendum (OJ No. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. as read with the corrigenda at OJ No. L226.4.4. Regulation (EC) No.6. 55).2004. Regulation (EEC) 2136/89. 62) as amended by Regulation (EC) No. laying down common marketing standards for preserved sardines. 14. namely. L139. 1663/2006. 1688/2005.9 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.2006.6. 1. these are not be discussed here. 3) as amended by Regulation (EC) No. 2076/2005. 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. L139. 25. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 30. 9. 2073/2005 on microbiological criteria for foodstuffs (OJ No. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. 1662/2006. 2074/2005. 1441/2007. L226. 22. 30. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No.2004. 1019/2008 and as read with Regulation 2073/2005.6. 83) as amended by Regulation Nos 882/2004. 30.12.10 Further Reading Regulation (EC) No. 2075/2005 and Regulation (EC) No. Regulation (EC) No. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. L283. 2076/2005. The revised text of Regulation (EC) No. Regulation(EC) No. The revised text of Regulation (EC) No.

12. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.2005. 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. 21.10. (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 338.2000. 22.FISH AND SEAFOOD Commission Regulation (EC) No 2074/2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC)No 882/2004 of the European Parliament and of the Council. derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004.1.2005.12. 22. 23.2001.

1.5 µm 10.0 x 1. and D-values at 45 and 51 °C have been reported as 29. no growth is observed (1).4 Heat resistance Aeromonads are sensitive to moderately high temperatures.5 minutes. Some death will occur at this pH if the temperature is 4 °C. At pH 4.3 . PATHOGEN PROFILES 10.10 (3). under optimal incubation temperatures.1 10. Aeromonas has the potential to grow over a pH range of ca 4 . 10. 0.1.5. 225 . .1.3 minutes. The optimum growth temperature is 28 °C (2). The minimum pH value for growth is thought to be pH 5.1.2 Oxygen requirements Facultative anaerobe.5 pH Under otherwise optimal conditions.45 °C (1).5. 10. respectively (1). At 5 °C. a 10 to 1. Morphology Gram-negative rods.1.000-fold increase in number is observed after one week in food systems (1). and 2.1 Aeromonas spp.3 Temperature Aeromonas grows at temperatures of 2 . 10.

10. which may be inactivated by heating at 80 ºC for 20 . spore-forming rods.3 Temperature All strains of C. being capable of slow growth and toxin production at low temperatures .2. but the spores of this organism are more so. botulinum are known.0 µm. B. and is sensitive to > 4. All C.30 minutes. Group I will not grow at temperatures of 10 ºC or less. types B and F can be either. All type A strains are proteolytic and type E strains are usually non-proteolytic.2. to a lesser extent.2.5% NaCl (4). F are the types that are responsible for most cases of human botulism (5.4 Heat resistance The vegetative cells of C.2 Oxygen requirements Although C. botulinum types produce heat labile toxins. 0.2. at 85 ºC for 5 minutes. forming at least seven different toxins.even as low as 3 ºC (7).4 x 1. E and. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C.22.1. 10. 10.1 Morphology Gram-positive.5 . 226 .6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . 6). botulinum is an obligate anaerobe. There are four main groupings of the organism. 10. botulinum is prevented.2. botulinum grow reasonably well in the temperature range of 20 45 ºC.2 Clostridium botulinum Seven different types of C. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. Thus an aerobically packed product may not support the growth of the organism on the surface.7 . 10. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. and groups I and II are those responsible for the botulism cases. Types A. or at 90 ºC for a few seconds. A to G. botulinum are not particularly heat resistant.2% sodium chloride (NaCl) (1).FISH AND SEAFOOD 10. but Group II strains are psychrotrophic. but the interior of the food may do so.

Note also that if humectants other than salt are used. However. parcooked meals with pH value > 5. safety margins must be introduced to allow for process variability. It will not. botulinum depends on solute. the minimum aw for growth may be lower. perfringens .94 for Group I and 0.3. These concentrations correspond to limiting aw of 0.9 x 2. 10.97) (10.7 Characteristics of C.is an obligate anaerobe. In commercial situations. pH and temperature. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.2. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0.PATHOGEN PROFILES 10. These values have been established under carefully controlled laboratory conditions.0 µm. 11)). 9).0. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. therefore.2. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains.2.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. 10.1. The organism will grow well in the centre of meat or poultry dishes. where oxygen levels are reduced.21 minutes) (9).0 and aw > 0. grow on the surface of foods unless they are vacuum or gas-packed. 0.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. 10.0 .3.1 Clostridium perfringens Morphology Gram-positive spore-forming rods.97 for Group II (5). vacuum packed. respectively (8.2 Oxygen requirements C. 10.3 .10.6 and pH 5. particularly by cooking.g. botulinum spores The most heat resistant spores of Group I C. 227 .3 10.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. they may survive mild heat treatments (70 .like other clostridia .

10. As in the case of other factors limiting the growth/survival of this organism. pH. perfringens.5 pH C. Its optimum temperature for growth is 43 . etc. (It is unusual for spores to be formed in foods after the growth of this organism (14)). type of solute. 10. perfringens appears to be 0.93 and 0.3. Salt concentrations of 6 .3. 16).3.10 ºC. Under otherwise ideal conditions. will survive greater extremes of pH (and aw) (12. although C.it is destroyed by heating at 60 ºC for 10 minutes (14.3 Temperature The most significant characteristic of C.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. lower concentrations may be effective in combination with other factors. The D-value for C. 13).FISH AND SEAFOOD 10. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12.3.3. vegetative cells die rapidly (12). perfringens in roast beef at 60 ºC is 14. the limits for aw are affected by temperature. 10. perfringens has the potential ability to grow within the temperature range 15 .97 (glycerol and sucrose respectively) depending on the solute used to control aw (13.50 ºC. however. 18). although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. The lowest aw recorded to support the growth of C.6 Aw/Sodium chloride C. perfringens can vary quite considerably in their heat resistance.7 Characteristics of C. It grows best at pH values between 6 and 7 (the same pH as most meats). 10. perfringens is not a tolerant organism with respect to pH.8% inhibit growth of most C. perfringens in vacuum packed beef (18). depending on strain and other conditions. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. perfringens spores The spores of C. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. Recorded heat 228 . In addition. While some growth can occur at 50 ºC.3. the enterotoxin is not heat resistant . Spores. Some studies indicate that the presence of 3% NaCl delays growth of C.5 minutes (17). 0 . 15. At cold temperatures. perfringens is not tolerant of low water activities.45 ºC. perfringens strains. 13). and their heat resistance is also affected by the heating substrate.

4 ºC is probably a more likely minimum for growth in foods (19). however.0. 10. growth at low temperatures can be very slow. so can be destroyed by pasteurisation. and where acetic acid is used as an acidulant. monocytogenes is not a particularly heat resistant organism.8 minutes for heat sensitive spores (12). pH < 5. requiring days or weeks to reach maximum numbers.4 .PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. 10. 0.at refrigeration temperatures down to. D-values for L. is between 30 and 37 ºC.especially at refrigeration temperatures.4 10. 10.98 and 0.4 Heat resistance L. it is not a sporeformer.4.albeit slowly . monocytogenes in crawfish tail meat at 55.2. or even below 0 ºC. In foods. Its optimum growth temperature. However. monocytogenes is reported to be 45 ºC (20). and temperature. Morphology Gram-positive short rods.19 minutes.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic.3 . however.2 Oxygen requirements Aerobe or microaerophilic.1 Listeria spp. Under ideal conditions. the lowest limit for growth is likely to be considerably higher .3 is the lowest value where growth has been recorded.2.0 minutes for heat resistant spores to 1. 1.5 . the organism is able to grow at pH values well below pH 5 (pH 4. being potentially capable of growing . The upper temperature limit for the growth of L. using hydrochloric acid as an acidulant).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.6 .4. 10. -0.4. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens. respectively (21).23.0 µm.4. 10.5 x 0. 60 and 65 ºC were reported as 10.64.2 has been suggested as the lowest working limit (22). 229 .4.

10.5.5 .12% NaCl or more).5.5.5 . 10.5 pH Plesiomonas has a pH range for growth of pH 4. 10.90. 10. 0.5. The bacterium may survive for long periods at aw values as low as 0. It is likely to survive.97.7 x 1.FISH AND SEAFOOD 10.4. 25). peritrichous flagella. It grows best at aw of ≥ 0.83 (20).6. 25). but has been shown to be able to grow at an aw level of 0.1.2 Oxygen requirements Facultative anaerobe.3.0 µm.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.3 µm.8.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).5.0.6 10.6 Aw/Sodium chloride L.5 10. or even grow. the optimum being 30 ºC (23). 10. at salt levels found in foods (10 .1 Plesiomonas Morphology Gram-negative.0 x 1 . 10. 10. monocytogenes is quite tolerant of high NaCl/low water activities. 0.3 Temperature Plesiomonas can grow in the temperature range of 8 .0 . Morphology Gram-negative short rods.8 .1 Salmonella spp. 230 .45 ºC.5 (24.5.

Salmonellae are quite resistant to drying. and appropriate heat treatments must be determined experimentally for low-aw foods. temperature and pH.0.3 Temperature Salmonellae can grow in the temperature range of 7 . substrate. 10.7. 10. Furthermore. However.6. under otherwise ideal conditions.6 Aw/Sodium chloride Where all other conditions are favourable. therefore.6. Salmonellae are quite resistant to freezing. a heat resistant organism. depending on serotype.6.4 Heat resistance Salmonella is not a spore-forming organism. D60 ºC values normally range from about 1 to 10 minutes. although salt tolerance increases with increasing temperature (28). and with an appropriate acid. The growth of Salmonella is generally inhibited by the presence of 3 .PATHOGEN PROFILES 10. However.20 times more heat resistant than the average strain of Salmonella at high aw (27).9. with a z-value of 4 .6. 10.6.5 ºC. 231 .945.8 . It is not.4% NaCl. The optimal pH for Salmonella growth is between 6.37 ºC. Salmonella senftenberg 775W is about 10 .5.2 Oxygen requirements Facultative anaerobe. Salmonella has the potential to grow at aw levels as low as 0. depending on the type of acid and temperature. strains vary in their ability to withstand heating. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments.5.93 (as reported in dried meat and dehydrated soup). some strains are able to grow at temperatures as low as 4 °C (26). Growth is slow at temperatures below about 10 ºC. pasteurisation and equivalent treatments will destroy the organism under normal circumstances. or possibly 0.48 ºC. 10. Some death will occur at pH values of less than about 4. the optimum being 35 .5 pH Salmonella has a pH range for growth of pH 3.5 .

The limits for toxin production are also narrower than for growth. aureus than citric acid).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. fat content. aw and temperature (sensitivity to acid increases with temperature). the D60 ºC value is 2 .4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH.5).7 .2 to 9.37 ºC (29).0 .9 µm diameter.7.5 ºC. stressed cells can also be less tolerant of heating. 10. Most strains of staphylococci can grow within the pH range 4. As is the case with other bacteria. It can survive well at low temperatures.7. 10.20 minutes. depending on aw. aureus can grow within the temperature range 7 – 48. aw. Under most circumstances.7 10. 31). aureus viability. 10. but may cause some cell damage (30). with an optimum of 35 . the organism is heat sensitive and will be destroyed by pasteurisation.7.1 Staphylococcus aureus Morphology Gram-positive cocci.0.). however. NB: The growth of Staph. under otherwise ideal conditions (29.3 (optimum 7.3 Temperature Under otherwise ideal conditions.2 Oxygen requirements Facultative anaerobe.FISH AND SEAFOOD 10. aureus is more limited under anaerobic than aerobic conditions. 0. Staph.7.7. etc.7. The following relate to limits for growth only. Freezing and thawing have little effect on Staph. 232 . 10. In meat macerate.

6 µm 10. 10.0 . It can grow over the aw range 0. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant. 31) between 0.2 .0. However.>0.99 aerobically under otherwise optimal conditions. aureus can grow in 7 .86 is the generally recognised minimum in foods (32).7. strain.8.3 Temperature The optimum temperature for growth has been reported as 30 ºC. and solute. * dependent on temperature.5 . with a temperature range for growth of 10 .8.1 Vibrio cholerae Morphology Gram-negative curved rods.9.2 Oxygen requirements Facultative anaerobe.99 little or no toxin production in anaerobically packed foods.0 (optimum 7. 233 .PATHOGEN PROFILES 10.83 .7. 0. 10.43 ºC (35). 10.8 10.2.8. 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. 34).4 . but certain strains can grow in 20% NaCl. In general. Staph. atmosphere. Staphylococci are more resistant to salt present in foods than other organisms.7.8 x 1.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5.5) (29.7 Limits permitting toxin production 10 .87 and > 0. In general.6 Aw/Sodium chloride Staph. an aw of 0.10% NaCl. pH.

8.6 (35.97 . under otherwise favourable conditions. cholerae can grow over the aw range Heat resistance V.FISH AND SEAFOOD Also. 10.984 (35).9. V. cholerae in crab meat homogenate at 60 ºC is 2. 10.5 ºC). with an optimum of 0.5 pH This organism is tolerant of high pH values. The D-value for V. there is evidence that V. 37. It can grow in the presence of 0.998. 10.6. 10. 10. but not acid conditions.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. but only moderately sensitive to freezing (40).3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.9. with optimum at 7. 234 .2 Oxygen requirements Facultative anaerobe.8.9. parahaemolyticus is easily destroyed by drying. are 5 and 43 ºC. 10.0. cholerae may persist on frozen fish for >180 days (36).9 10.1 . respectively. cholerae is sensitive to heat at temperatures of >45 ºC.0% NaCl (35). declining in number during storage. it has a pH range for growth of pH 5 . 38). and it is sensitive to refrigeration temperatures (0 .6 Aw/Sodium chloride V. with an optimum of 37 ºC (39).8.65 minutes (35).

The optimum concentration of NaCl equates to aw of approximately 0.29 minutes.5 minutes. depending on solute. 10.10 Vibrio vulnificus 10. respectively (39). 10. 10. vulnificus is heat sensitive.9. V.5 pH The pH range for growth is 4.PATHOGEN PROFILES 10. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0. but the optimum concentration is 2 . 10. 41).8 .4% (39.10.3 Temperature The optimum temperature for growth has been reported at 37 ºC. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43). 235 .43 ºC (42).4 Heat resistance V.10.922. and are readily destroyed by cooking.11.8. 41). 10.02 . with a temperature range for growth of 8 . parahaemolyticus will grow at aw levels as low as 0. with an optimum pH of 7.0.10. parahaemolyticus is unable to grow in the absence of salt.5% NaCl.6 .4 Heat resistance V.9.9. parahaemolyticus are not heat resistant.6 (41). 10. The D-values at 55 ºC for clam and crab homogenate are 0.10.1 Morphology Gram-negative curved rods. respectively.2 Oxygen requirements Facultative anaerobe.6 Aw/Sodium chloride V.98 (35. and 2. The organism can survive for long periods at freezer temperatures (42).

D..H. 1989. Doyle M.. 2003. Diseases Caused by Bacteria. 236 . 1994. Eds. in Clostridium botulinum: Ecology and Control in Foods.M. Marcel Dekker.5% (42). Montville T. 383-408. Smith J. Stelma G.J. 1989. Clostridium botulinum.O. Dodds K. 1994. in Microorganisms in Foods. 2001.H.D. London. 1993. Fratamico P. New York.K. Great Britain. 2.6 Aw/Sodium chloride V.P.N.W.E. Stelma G.L.. 9.. Kim J.11 1. Eds. Murrell K.R. with an optimum pH of 7. Dodds K.M. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.W.5 . 6.D. 10. Eds.10. Clostridium botulinum. Potential hazards associated with REPFEDS.L. Austin J. Microbiology and Molecular Biology. Clostridium botulinum.. 1-19.W. Washington D.FISH AND SEAFOOD 10.O.10. 7.A.H.5.P. 329-49.W. ASM Press. Gaze J... New York.P.R. in Foodborne Pathogens. Aeromonas hydrophila.. 4. New York. in International Handbook of Foodborne Pathogens. Marcel Dekker.. Volume 1.. Eds. Betts G. 10.5 pH The pH range for growth is 5 .M. Marcel Dekker Inc. 5.H.L. 97-131.. vulnificus grows over the range 0. Marcel Dekker.. Caister Academic Press. 5-19.W. Ed. in Food Microbiology: Fundamentals and Frontiers.R. with an optimum of 2. Eds.L. International Commission on Microbiological Specifications for Foods.N. Beuchat L. Hauschild A. 111-89. Foegeding P. References International Commission on Microbiological Specifications for Foods.8 (42). Marcel Dekker. Blackie. 6 (1). Eds.H. Juneja V. Volume 1: Diseases Caused by Bacteria. Hauschild A. Miliotis M. Peck M. Ed. Novak J. 10. 3. in Foodborne Disease Handbook.. Murrell K.. Palumbo S. 357-67. Doyle M. Hui Y. 1993.. New York. Bier J. Food Control. Clostridium botulinum and Clostridium perfringens. Johnson E. in Foodborne Bacterial Pathogens.. Gorham J. Faris A. in Clostridium botulinum: Ecology and Control in Foods.. 279-303. 57-63. New York. Hui Y. Ed. 1995.. Abeyta C.10. Eds. 1-27. Lund B. Aeromonas hydrophila group. Marcel Dekker. Principles of control.0% NaCl. Cliver D. 11. New York.D.C. Marcel Dekker.. Dodds K. in Foodborne Bacterial Pathogens. Cliver D. Volume 5: Microbiological Specifications of Food Pathogens. Aeromonas.H. Aeromonas species. 2005. Bhunia A.. New York. Notermans S. 1996. 8. Doyle M.... 121-76. Kovacek K. in Foodborne Disease Handbook. Gorman J. Hauschild A.

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

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

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


uk Institute of Fisheries Management (IFM) 22 Rushworth Avenue West Bridgford Nottingham NG2 7LF United Kingdom Tel: + 44 (0)115 9822317 Fax: + 44 (0)115 9826150 Email: info@ifm.ifm.uk Web site: www.uk/Topics/Fisheries Seafish Industry Authority 18 Logie Mill Logie Green Road Edinburgh EH7 4HS United Kingdom Tel: + 44 (0)131 5583331 Fax: + 44 (0)131 5581442 Web site: www.org.gsi.gov.org/ .defra.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 Prepared Fish Products Association (PFPA) Federation House 6 Catherine Street London WC2B 5JJ United Kingdom Tel: + 44 (0) 208 78362460 The Scottish Government Fisheries Group Environment and Rural Affairs Department Pentland House 47 Robb's Loan Edinburgh EH14 1TY United Kingdom Tel: + 44 (0) 8457 741741 or + 44 (0)131 5568400 Fax: + 44 (0)1397 795001 Email: ceu@scotland.seafish.gov.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.scotland.holt@ifm.gsi.gov.org.gov.co.uk Web site: www.uk or v.uk Web site: www.org.

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hpa.gov.org Web site: www.uk/en/index.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.co.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.gov.gsi.au 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.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.dh.bsi-global.seafood.co.csiro.ifst.fdf.uk Institute of Food Research (IFR) Norwich Research Park Colney lane Norwich NR4 7UA United Kingdom Tel: + 44 (0) 160 3255000 Fax: + 44 (0) 160 3507723 Web site: www.com/ 243 .uk 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.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.ifr.com 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.cmar.org.nz Web site: www.ac.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: generalenquiries@fdf.org.uk Web site: www.

org.scot.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.int Web site: http://www.who.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.who. jsp Food Safety 10 .nhs.uk Web site: www.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.who.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: hpsenquiries@hps.sik.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.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.euro.nhs.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.net/web/indexen.int.sofht.int Web site: www.co. Web site: www.euro.uk/index.se Web site: www.int/ SIK .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.scot.de 244 .uk Web site: www.hps.co.uk/scieh.sfam.int/foodsafety World Health Organization (WHO) Headquarters CH .org Web site: www.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: foodsafety@euro.codexalimentarius.

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


INDEX Achromobacter spp. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. growth and survival in crustacean shellfish 86 Aeromonas sobria. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. pathogenic microorganisms in molluscan shellfish 59-60 as contaminant in enrobing process 37 as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45 growth in vegetable and rice 247 . 69-70 Aeromonas hydrophila. source of PSP toxins 142 Alexandrium ostenfeldii. causing spoilage of dried fish 110. 102 Bacillus spp. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. planktonic source of spirolides in shellfish 163 Allergens. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. in crustacean shellfish 86-7 ATP. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. preserved by dry/wet salting 99 Anisakis spp. to test microbiological criteria 19 Aeromonadaceae. in crustacean shellfish 87 Alteromonas spp. 111 Astroviruses. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. 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. 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.

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

cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. of fish – different processes 94 Dun. as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45-6 pathogen profile 227-9 Coated fish products. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). to determine fresh fish spoilage 11 Eels. 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. 149-50. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. as contaminants in scampi 36. 106 Contacts 241-5 Conway method. source of azaspiracids 159 source of DSP toxins 148-9. smoked and dried fish 114 Enzymic spoilage. as autochthonous bacterial flora of fish 3 Crabs. of prepared fish products 44 Escherichia coli. 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. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. processing 334 Cockles. for detection of DA 148 ELISA kit (DSP-Check). hot-smoking 107 ELISA assay. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. to detect NSP toxins 155 ELISA kit (Biosense). to determine fresh fish spoilage 11 Corynebacteria. growth and survival in crabmeat 84 growth and survival in cured. 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. adverse condition on poorly preserved fish 110 EC grading scheme. 152 Diphyllobothrium.INDEX pathogen profile 226-7 Clostridium perfringens. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 .

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

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

contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P. smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. 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. shigelloides. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay. reduction in smoking process 108 Norcardiopsis dassonvillei. of crustacean shellfish – effects of processing 81-3 of cured. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 .FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. as TTX producer 145 Norovirus. 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. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. caused by lipid oxidation 11 Oysters. for detection of azaspiracids 160 for detection of ciguatoxins 157 for detection of DA 148 for detection of DSP toxins 150. 153 for detection of LST toxins 151 for detection of NSP toxins 155 for detection of pinnatoxins and pteriatoxins 165 for detection of PSP toxins 144 for detection of spirolides 164 for detection of tetrotoxin 145 Mussels. in crustacean shellfish 867 Norwalk virus. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini.

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

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

for evaluation of fish quality 9 Torrymeter. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru. processing and effects on microflora 63-4 Skin resistance. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 . as indicator of fresh fish spoilage 11 Toxins. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. as contaminant in scampi 36 as contaminant in enrobing process 37 as hazard in dry salting of fish 89 growth and survival in chilled and frozen prepared fish products 46-7 growth and survival in crustacean shellfish 84 growth and survival in cured. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO. of crustacean shellfish 80.INDEX Shigella spp. in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. as measure of fish degradation 10 Sliming. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. contact details 241-2 Trematode infections foodborne 137 Trichosporon. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. concerning crustacean shellfish 89 Vacuum packaging. fish and shellfish 141-74 Trade associations. 101 Storage. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. 14 Torry Scale. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. of fish after capture 5 Surstrømming. grading of 31 hot-smoking 107 US legislation. in molluscan shellfish 60 Tuna. role in fish degradation 10. fish sauce 123 Shrimps. smoked and dried fish 114 pathogen profile 232-3 pathogenic microorganisms in molluscan shellfish 59-60 potential for growth in pasta component of seafood ready meals 40 Stockfish. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. 13. fermented fish product 126 Taste panel evaluation.

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

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