Leatherhead Food International Leatherhead Food International

Edited by
Rhea Fernandes

This edition first published 2009 by Leatherhead Publishing, a division of Leatherhead Food International Ltd Randalls Road, Leatherhead, Surrey KT22 7RY, UK URL: http://www.leatherheadfood.com and Royal Society of Chemistry Thomas Graham House, Science Park, Milton Road, Cambridge, CB4 0WF, UK URL: http://www.rsc.org Regstered Charity No. 207890

ISBN: 978-1-905224-76-0

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Associate Prof. Covadonga Arias Microbial Genomics Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America Dr. Simon Derrick Special Projects The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom Carlos Abeyta, Jr. Supervisory Microbiologist Pacific Regional Laboratory Northwest U.S. Food and Drug Administration 22201 23rd Dr. S.E. P. O. Box 3012 Bothell WA 98021-4421 United States of America Linda Nicolaides, M.Ph., FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom

Prof. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom Jeffrey L.C. Wright C.M., Ph.D., FCIC Carl B. Brown Distinguished Professor of Marine Science UNCW Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America Rhea Fernandes and Dr. Peter Wareing Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom Eugenia Choi and Dr. Jenny Pfleger Regulatory Advisors Leatherhead Food International Randall Road Leatherhead Surrey KT22 7RY United Kingdom


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

vi .

2 Initial Microflora 4.2 Definitions 2.6 Published Microbiological Criteria 3.4 Spoilage 4.7 Published Microbiological Criteria 2.8 References MOLLUSCAN SHELLFISH 3. 3.2 Initial Microflora 1.7 References 3.5 Factors Affecting Fresh Fish Spoilage 1.5 Pathogens: Growth and Survival 3.3 Processing and its Effects on the Microflora 4.3 Processing and its Effects on the Microflora 1.6 Published Microbiological Criteria 4.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.5 Spoilage 2.1 Definitions 3.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1. CHILLED AND FROZEN RAW FISH 1.4 Processing and its Effects on the Microflora 2.7 Published Microbiological Criteria 1.7 References 4. vii .1 Definitions 4.6 Pathogens: Growth and Survival 2.5 Pathogens: Growth and Survival 4.4 Spoilage 3.6 Pathogens: Growth and Survival 1.3 Processing and its Effects on the Microflora 3.2 Initial Microflora 3.4 Spoilage 1.3 Initial Microflora 2.8 Further Reading CRUSTACEAN SHELLFISH 4.1 Introduction 2. 4.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.8 References 1.1 Definitions 1.

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 6.6 Lipophilic Shellfish Toxins (LST) 7.3 Regulation (EC) No.2 Initial Microflora 5. SMOKED AND DRIED FISH 5.2 Initial Microflora 6. 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 9.10 Further Reading 223 viii .1 Definitions 5.6 Published Microbiological Criteria 5.4 Spoilage 6.1 Definitions 6. EU FOOD HYGIENE LEGISLATION 189 9.5 Pathogens: Growth and Survival 6.8 Further Reading FERMENTED FISH 6.8 Ciguatoxins 7.10 Cyclic Imines 7.7 Further Reading FISH AND SHELLFISH TOXINS 7.9 Azaspiracids 7.7 References 5.2 Definitions 8.1 Introduction 8. CURED.4 Regulation (EC) No. 7.3 Processing and its Effects on the Microflora 6.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.I.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.2 Legislative Structure 190 9.1 Introduction 7. 14 (Hygiene requirements specific to the UK) 221 9. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.6 References 6.1 Introduction 189 9.5 Pathogens: Growth and Survival 5.4 Amnesic Shellfish Poisoning (ASP) Toxins 7. 9.9 Other Relevant Legislation 223 9. 8.3 Stages of a HACCP Study 8.5.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.7 Food Hygiene (England) Regulations 2006. S.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.11 Conclusion 7. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.3 Tetrodotoxin 7. 852/2004 on the General Hygiene of Foodstuffs 191 9.3 Processing and its Effects on the Microflora 5.4 Spoilage 5. 2006 No.4 Implementation and Review of the HACCP Plan 8.5 Regulation (EC) No.6 Regulation (EC) No.8 Guidance 222 9.

10.9 Vibrio parahaemolyticus 10.3 Clostridium perfringens 10.4 Listeria spp.10 Vibrio vulnificus 10.10 PATHOGEN PROFILES 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .8 Vibrio cholerae 10.5 Plesiomonas 10.7 Staphylococcus aureus 10.2 Clostridium botulinum 10.6 Salmonella spp. 10.1 Aeromonas spp. 10.


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

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

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

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

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

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

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

Toxin levels 0. and risk associated with the consumption of seafood is low (2. numbers of pathogenic bacteria in raw fish tend to be low.37 °C High for most species.FISH AND SEAFOOD TABLE 1.14 .106 cells/g. birds.0. 4 .26 °C 10 . sewage.1 .109 cells/g) * Some sources considered L. and mud Ubiquitous. However.>106 Salmonella spp. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. 105 .102 101 . Plesiomonas shigelloides 3 .1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.35 °C 8 . during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria. In addition. Escherichia coli Yersinia enterocolitica 101 .103 High (107 . 4). vulnificus) 5 .106 cells/g (exception: V.37 °C Unknown Unknown TABLE 1. stream water.19 µg/kg bodyweight from < 102 . Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00.II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil. 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. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . estuarine environments.

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

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

the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). Gases used in MAP of fish most commonly include carbon dioxide and nitrogen. phosphates. The microflora of meat is not the same as that of whole. Many additives have been tried in conjunction with changed atmosphere. Replacement of oxygen with nitrogen. does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. discolour pigmented fish (11). VP is one of the oldest forms of altering the interior gaseous environment of a pack. botulinum. including salt. and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). the pathogens that may be able to grow before spoilage occurs. and the promotion of a predominantly Gram-positive. High concentrations of carbon dioxide have the most pronounced microbial effect. as they are able to grow at temperatures as low as 3.3 °C and produce toxins.CHILLED AND FROZEN RAW FISH surrounding gas. an inert and odourless gas. gutted or filleted fish. nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. Micrococcus and Moraxella). although less information on their effectiveness is available. and increase in-pack drip (12). This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. The products. but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). slower-growing flora (11). the inhibition of common 'spoilage' bacteria (Pseudomonas. but can dissolve into fish liquids and deform packages. may not be considered ‘fresh’ fish. Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). which are able to grow rapidly at low temperature. and the problems of muscle structure damage by the modified-atmosphere (11). a modified-atmosphere may contain any gas necessary for the desired effect. but only in recent years has it seen a marked expansion in use and market share. Sulphur dioxide. In the United States. after additive application. the higher pH and reduction potential (Eh) of the fish muscle. The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). Flavobacterium. without overt signs of spoilage. MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). Modified-atmosphere preservation of fish was first reported in the 1930s. Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains.

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

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

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

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

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).5. a minority are often associated with the majority of the spoilage. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks.and temperate-water fish spoilage rates may be many. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH.5. The reasons for the differences between tropical. by altering the gas atmosphere (extrinsic). For example.3 Recognised specific spoilage organisms (SSOs) The degree of spoilage leading to sensory rejection of fish is partly dependent on the perception of the consumer. 1. the results of altering one parameter may have far-reaching consequences on others.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. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. 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. For fresh fish. the slower the spoilage processes proceed (within limits). Also. 34). but also crucial for the assurance of consumer safety.5 1. The sum of these changes may result in a different spoilage profile. For fresh fish iced immediately after capture and continually stored in ice. 1. 5). biochemical and microbiological reactions taking place within and on the fish. the lower the temperature of storage. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. Fish spoil as a result of the chemical. The factors are interactive and cannot be completely isolated. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 .FISH AND SEAFOOD 1. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. The concept of specific spoilage organisms (SSOs) is not new. careful temperature control during storage is not only important in terms of quality loss.5.

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

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

Of 404 intoxication outbreaks of type E C.CHILLED AND FROZEN RAW FISH present in fish into histamine. However.3 °C (Group II) and these are non-proteolytic (i. botulinum recorded up to 1963. 1. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose.e. botulinum.3. 1. Acetate production has been reported as a good indicator of spoilage by this organism. D. (and very rarely F and G) have been reported to cause human disease. a convenient but diverse species of bacterium with a number of different types. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. algal toxins. Recent studies have investigated the dominance of B. where a particular 15 . Some Ph. Growing evidence suggests a role for B. but with proper cooking. especially type E botulinum intoxication. Fewer still are capable of growing on fresh fish. When inoculated into VP corned beef and sliced ham. lightly preserved fish products are associated with botulism. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). including antisera. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. Adult disease is caused by the production of the botulinum toxin. C2. Huss (1. B. previous exposure and access to supportive treatment. 75% (with a 35% mortality rate) occurred in Japan.6. 5) reports that fresh fish consumption has never been shown to cause human botulism. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). The heterogeneity of the types (A. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. E. presents a potential risk. thermosphacta did not produce off-flavours until 2 . E and F include bacteria with minimum growth temperatures of approximately 3. age. thermosphacta in the spoilage of some MAP fish. it is bacterial risks that increase after capture of the fish. 5). they do not produce significant product spoilage).5. B.3 days after having reached 108 cfu/g (42).4 Brochothrix thermosphacta and lactic acid bacteria B. 6). types B. bacteria and higher parasites all present some risk associated with fish consumption (1. this is probably due to spoilage occurring before toxin elaboration. E.1 Clostridium botulinum C. human viruses. 5.6 Pathogens: Growth and Survival Endogenous chemicals. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. B. Review of outbreak data suggests fresh fish to be safe. Of these. 1. Types A. these risks are substantially reduced (1. F and G) is well documented (44). C1.

Generally.2 Vibrio parahaemolyticus and other vibrios V. the majority are associated with products not cooked immediately before consumption. whether stored aerobically or in modified-atmospheres. storage and preparation (11).g. 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. France. V. 11). salted or pickled) fish have also been associated with botulism. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). which was probably contaminated from an uncooked food source or an excreting foodhandler. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. Despite the ubiquitous nature of type E C. The first report of the organism being a foodborne agent was with shirasu. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. 1978 . Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. which is believed to be required for virulence. parahaemolyticus cases have increased across the world.g. The bacterium causes primary septicaemia. A unique clone of V. the majority of outbreaks are caused by consumption of raw fish products (e. again. Japan. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile.6. and proper cooking (boiling for 1 minute. I-sushi. a Japanese boiled and semi-dried sardine dish. there is no specific guideline that describes a minimum level of V. and North America. were associated with finfish between 1978 and 1998 (46). due to the cultural preference for raw fish dishes. Proper chilling. Currently.2005). Since 1996. 1. Russia. including epidemics in India. Proper temperature control (<3. and when in contact with contaminated water (no CDC reported cases from fish. Only about 1% of marine isolates produce a thermostable haemolysin.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. In Japan. which can cause human gastroenteritis. is small. parahaemolyticus is a marine organism. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . This strain has been responsible for 50 to 80% of all V. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. vulnificus is associated with warm marine and estuarine waters. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. botulinum in the marine environment (isolated from around 90% of marine and environmental samples from northern European waters and its consequent presence in seafood (a surveyed incidence of as high as 65%) (2. parahaemolyticus outbreaks.3 °C) will eliminate all risk. the risk posed to the consumer of fresh fish. smoked. V. sushi) (5). parahaemolyticus O3:K6 is responsible for many of the recent V. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). 5). USA. Other lightly or semi-preserved (e. accounted for the majority of cases (1. Southeast Asia.

possibly due to under-reporting because of their likeness to E.CHILLED AND FROZEN RAW FISH disorders). They are aquatic organisms with an epidemiology that is yet to be fully understood.20 °C. and especially estuarine environment. coli on isolation media (50). L.. with minimum growth temperatures around 0 °C and optima of 15 . Although the psychrophiles are unable to grow at body temperature.6. There is no risk associated with the consumption of properly chilled and cooked fresh fish. are present on fish and can grow to significant numbers during storage (1. Most cases have been sporadic rather than associated to large outbreaks. however. sodium channel blocking and haemolytic activity (48). Chill storage may not eliminate growth. others are psychrotrophic mesophiles (49). 1. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. but proper cooking should substantially reduce risk of infection. and proper cooking will substantially reduce risk. in its habitat. cytotoxic. P. Again.4 Listeria monocytogenes L. shigelloides are encountered.26 to 0.3 Aeromonas Aeromonas spp. shigelloides is similar to Aeromonas spp. listeriosis cases have been reportable to the CDC. there is no risk from the consumption of properly handled and cooked fresh fish. in assessing the risk of fishborne infection by P. is historically associated with faecally contaminated water. In the US. infection. 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. and fish and shellfish are probably the major reservoirs for the organism (50). though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. are associated with human diarrhoeal illnesses. their potential to produce disease-causing exotoxin in food has not been fully examined (48). which is often fatal (>50%). cholerae O1. 17). the causative agent of cholera. and especially Aeromonas hydrophila. 1. It is known that Aeromonas spp. V. but the bacterium is known to survive and grow in the shallow marine. Similar problems as those encountered for Aeromonas spp. Since 2000. 5. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. Despite this. Some isolates are true psychrophiles.000 17 . proper chill storage will eliminate any risk associated with this organism. monocytogenes has been well documented as a foodborne human pathogen.6.55 cases per 100. monocytogenes incidence has been between 0. The major virulence factor appears to be the production of toxins with enterotoxic.

6. there is a possibility for growth of the organism on chill-stored fish. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses... foetuses.. Lightly preserved products. some Vibrio spp.6. Photobacterium spp. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). 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. 1.48 minutes in salmon have been reported (53). It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. 1. D60 of 1. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . alcoholics. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. Being a psychrotroph. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). leads to the production of histamine.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. neonates.2002 (55)) and is also common worldwide. but more especially by Morganella morganii. especially pickled fish. which has a maximum permissible level (for fish products from fish species associated with high amount of histidine) of 200 mg/kg fish in the EC.FISH AND SEAFOOD persons. but cooking will significantly reduce the risk of infection.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. Like other toxins. 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. Klebsiella pneumoniae and Hafnia alvei.pregnant women. it is not apparent to the consumer and it cannot be destroyed by cooking. full risk analysis is not possible (52). Infections are mostly associated with the consumption of raw or 18 . It is regularly isolated from seafoods (51). at risk groups . 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. Clostridium spp.98 minutes in cod and D60 of 4. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. If fish are not properly refrigerated. Fortunately. but proper cooking will reduce any risk. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. and Lactobacillus spp. The organism poses a serious risk in chilled products not cooked before consumption.

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

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

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

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

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

A. 1984. Torry Research Station of the Ministry of Agriculture. Labuza T. Florida. Huss H. 317-21.. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging.C.. Trolle G. Dalgaard P. 1986. International Journal of Food Microbiology.M. The Microbiology of Seawater Fish.9. Changes in microflora and chemical compounds in mackerel stored by partial freezing. 317 (Suppl. Borgstrom G.H. Kitts D. Torry Research Station of the Ministry of Agriculture. Akiba M.. Ed. Magno-Orejana F. in FAO Fisheries Report No. Olley J.. Cann D. 1992. Ross T. 1984.. International Journal of Food Microbiology. Cho Y. Rome. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Huss H.1983. 6. Aberdeen. McMeekin T. 1. Fisheries and Food. 24 .P.G. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. Picard G. Ogden I. Estimation of the bacteriological quality of fish by automated conductance measurements.M. Torry Research Station. Hobbs G.9. Smith G. International Journal of Food Microbiology. 4.). Microbiological quality and shelf life prediction of chilled fish. Food and Agriculture Organisation.. Journal of Food Protection.L. Ed. 51. in Seafood Science and Technology. Application of predictive microbiology to assure the quality and safety of fish and fish products. Technomic Publishers. Ravn Jørgensen B.E. 1992. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. 295-307. 1961.1 Further Reading Processing Anon.H. 741-50. 271-85. Gibson D. Eds. Bulletin of the Faculty of Fisheries (Hokkaido University).D. Villemure G.. Gram L. Academic Press.. in Fish as Food.. 35. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches. 1. Shinano H. Fisheries and Food.2 Spoilage Shewan J. Houston N...S. Gibson D.. 55. 1. 1985. International Journal of Food Microbiology. in Torry Advisory Note No 93. Journal of Food Science. 15.. Aberdeen..M. 65-72. 1988..9 1. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. Sumner J. Jones Y.H. 62-70.FISH AND SEAFOOD 1.. 127-34. Lancaster.. 1989. 13-32. Shahidi F. Taoukis P.. Handling and processing of scad. Food and Agriculture Organisation of the United Nations. Huss H. 1997.D. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. 1989. 73-89... Simard R.

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


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

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

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

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

other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. and pathogenic flora associated with seafood. it may be used for other products or markets depending on the product specification. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). Only after death.3 2. A primary role in fish spoilage has been assigned to this organism. After capture and during any subsequent processing. see Table 2. In summary. which may become a focus of contamination deep within the muscle. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. Recent public opinion has favoured the use of dolphin. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. Grading is based on size of fish (>25 kg).e. will the saprophytic flora start to invade the surface flesh (1. For a summary of background information on the hazards associated with fisheries products and ingredients.3. Dalgaard et al. 6). Tuna that does not meet the highest of grades based on colour. and have emphasised the origin and sustainability of the fish as key marketing points. handling or preparation stage. storage. fat content. The edible tissue of some species of fish are more susceptible to gaping than others. and ingredients added to the final product. as a result there is an increase in spoilage rate thereby reducing the product shelf life. skin. i. 2). or visible damage from gaff marks (4).I. 31 .CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. sensory assessment of freshness etc. and processing conditions such as dehydration or poor filleting practices. the microbial flora of seafood is associated with the gills. remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. searing or bar marking). product handlers. Contamination will. including seasonal variation. and the potentially toxic by-products of histamine-producing bacteria (5. on the whole. and intestines of the fish. whilst gaping is also dependent on a range of other factors that effect the muscle composition. species (Bluefin>Bigeye>Yellowfin>Albacore). Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. is not used for sushi or other high risk (and high value) products.and turtlefriendly long lining to the less discriminatory netting.

staphylococcal toxin. Vibrio cholerae. the effects of processing and combination or use of ingredients on identified microorganisms should also be considered with respect to both the safety and quality/shelf life of the product. Aeromonas. shelf life determination. cereus B. moulds Bacillus Bacillus. Fusarium. spore-formers. protozoa and viruses B. cereus Bacillus licheniformis. 32 . Staph. TABLE 2. for chilled products. Listeria. Clostridium Lactobacilli. and so are the potential microbial contaminants associated with them. by the HACCP team or by a hazard and risk assessment programme. yeasts Pathogenic microflora/toxins Salmonella. anaerobic spore-formers. and associated spoilage and pathogenic microorganisms. however. coryneforms. nuts and fruit Associated microflora Lactobacilli. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter.FISH AND SEAFOOD 2. product assembly. 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. Clostridium perfringens. Staphylococcus.I provides an overview of common raw materials. B. intermediate processing. cereus Staphylococcus aureus. Aspergillus. Pseudomonas. Clostridium botulinum. moulds. cream. enteric viruses 2. moulds Bacillus. toxin. Shigella. and. mushrooms. coliforms. Bacillus cereus. Enterobacteriaceae. raw materials.4 2. Moulds Enterobacteriaceae Salmonella.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. pseudomonads. Escherichia coli. Table 2.4. Bacillus subtilis B. In addition. be considered at an early stage during new product development. and milk Cheese Water/Ice Bacillus.3. streptococci. yeasts Lactobacilli.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. histamine Cryptosporidium. moulds Bacillus. Penicillium.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse.

Examples of tempering techniques include the use of heated air. Figures 2.4.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. Band-sawn materials or fish dice are normally promptly further processed. this may be an incidental effect. Where the raw materials are frozen.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. line extensions and process reviews.2. but it is assumed that any band sawing or dice process will introduce heat from blade friction. 2. the core temperature. It is critical that the time and product temperature during tempering be controlled to avoid elevated temperatures (>5 °C) on the product surface allowing microbial growth or the onset of spoilage in raw material. and contamination from equipment to clean fish muscle. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. Generic examples have been used to illustrate only the major points.3 Preparation of fish component of chilled and frozen recipe dishes 2.4. or the use of microwave technology. but it also poses a risk of cross-contaminating other products undergoing tempering. All have the same intention. as a heat process is often required for functional or organoleptic reasons. up to -5 °C so as to improve the cutting and handling properties during processing. For some components. but will also need to ensure that the optimum shelf life can be safely obtained. It is recommended that the principles of HACCP are applied to novel processes. Effects on the microflora are minimal during tempering (7).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. depending on the size of the fillet 33 .3. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface.4.1 . as the full complexity of the processes used for specific products cannot be covered in a general text.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. and potential opportunities for growth of foodpoisoning bacteria. there are a number of distinct methods used. to evenly raise the temperature of the entire frozen product from -20 °C or lower. defined holding time at a chill temperature (0 – 3 °C). 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. Rapid controlled cooling of heat-processed components will help to minimise spoilage. 2.

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

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

perfringens could. or sold as a chilled product after thawing. Breaded or battered fish fillets are fully heat-processed before consumption. vibrios and other fishassociated spoilage organisms. e. Natural fillets d. If a crumb coating is required. egg.g. e. and possibly Staphylococcus spp. rusk or crumb. is the addition of a pre-dust.g. and may include milk powder. absorb any excess moisture on the surface of the fish portion. Fish products. such as Listeria and Vibrio spp. Bacillus spp. and if required. The function of this is to improve the adhesion of subsequent coatings. The first stage. flour. Products are preformed and presented to the process (either three. any excess is removed with an air knife. A large variety of product formats and types can be batter-enrobed. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. Tempered band-sawn materials b. with an expected shelf life of 6 . chilled. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. with the coating heat-fixed.FISH AND SEAFOOD In all of the above examples. quickfrozen.. Over time. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. spices. milk powder and spices. and to carry flavourings where required.e.4 Battering/enrobing process Figure 2. fish cakes Portions or fillets may be batter-enrobed. because of the intensive handling required during processing. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process. as well as ensuring that there is minimal contamination from human processors. Batter contains water. Initially. IQF/pre-formed fillets c. a. 2.4.2 illustrates the typical stages of enrobing or breading. which typically consists of flour. starch. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. which can occasionally be isolated from protein-handling processes. Spore-forming anaerobes such as C. 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. a characteristic flora will develop from the batter components. lactobacilli from the milk powder and flour. and Enterobacteriaceae from flour. which is product-dependent and not always required. i. potentially. rapidly multiply if strict temperature control was not maintained.7 days. raising agents. some of this initial flora 36 . from flour.or fivestage) frozen or chilled.

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

Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). to produce a specific product. There are two main categories. canned fish may be used in their preparation owing to it having wider availability and low wastage. and must not show evidence of disease. Figure 2. as this could introduce off-flavours and reduce shelf life. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products. The vegetables must withstand processing.2.g.1 ºC for no more than 3 hours is recommended (1).3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes. pasta or potato.4. 2. Flow diagram outline for recipe dish ingredients preparation 2. Fig. but they are supplied without a source of carbohydrate e.4. The protein source can be fish fillet or block. 38 .5.3.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal. For example. rice. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. A temperature no greater than 10 ºC for 12 hours cumulatively or 21.

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

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

Without barriers. 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. respectively. spices. 13). that of a dairy-based sauce. The format uses flavour compounds . and handling of processed sauce can be less rigorous than. flour products.5 illustrate the recipe dish ingredients preparation and assembly and packaging. 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. Figures 2.4 and 2.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. for instance. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. Materials prepared with regard to good hygienic practice need only be assembled in a tray or pouch before chilling or freezing in readiness for distribution. Marinade-type products are becoming more popular.2. etc. suspended in an acidified sauce with an oil component. 41 .herbs.4.

the method replaces air in the sealed packaging with a defined mixture of gases. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. which may restrict either microbial growth or enzyme activity. generally carbon dioxide and nitrogen.2 Modified-atmosphere packaging Originally developed for the red meat industry. 42 . 2.4.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).7. botulinum to grow. as it dissolves in the surface moisture to form a weak acid. The oxygen-free conditions will however allow other anaerobic bacteria such as C.4.2. and tuna loins are packed by placing the product in a plastic pouch and removing all the air.7. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth.4. Flow diagram outline assembly and packaging of recipe dish 2.5. unless the temperature is strictly controlled to below 5 °C throughout the supply chain.FISH AND SEAFOOD Fig. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. These include: 2.

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

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

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

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

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

L.II IFST Guidelines for part-cooked foods – e. Maximum= Levels at end of stated shelf life 48 . coated fish. cereus Staph. pasta. monocytogenes B.g. aureus V. coli GMP* <102 Maximum 104 TABLE 2.fish products.III IFST Guidelines for processed foods . parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E. TABLE 2. 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.FISH AND SEAFOOD the microbial criteria for other products (rice. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp. fish cakes Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Pathogens GMP* Maximum Criteria for absence not generally applicable Indicators and spoilage organisms Organism E. batter etc) should also be consulted when establishing the specifications for raw materials and final product. fish fingers.

Letters in Applied Microbiology. Latt T. 2.G.H. Bacterial histamine production as a function of temperature and time of incubation.. Slavin J. Washington D. 7. Mejholm O.. New York. 13.W. 7 (24).. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. Hoshino T. Martin R. Murrel H.. Journal of Food Science. 663-87. Communicable Disease Report.M... Powers E. Fish: microbiological spoilage and safety. 38 (11). 1997. Aureli P. Wienke A.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). 5 (3)... 1962. Journal of Milk Food Technology. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. Importance of Photobacterium phosphoreum in relation to spoilage.A. Tu A. 15.R. 1997. 8. 6. Hackney C.T. References US Food and Drug Administration..L. 10.. 5.R.. 3.. Masuoka Y. 1991. IBH Publishing Co. Food and Drug Administration.L.R. 49 4. 373-8. International Commission on Microbiological Specifications for Foods. Letters in Applied Microbiology. Pvt. Ishizaki K.L. London. in Fish Inspection Quality Control and HACCP: A Global Focus..K. Kumeta H. 1993. 1976. 1991. Ltd. Van Nostrand Reinhold. Eds.. 96. Kluwer Academic. Huss H. 1311-7. Matsuyama H. Hobbs G. Food Science and Technology Today. 14. Journal of Hygiene. 233-7. 25 (1). 1. Gilbert R. Cambridge. 204-16. 67-73. Pathogenic microorganisms associated with fresh produce. Bartlett C. Incidence and levels of Bacillus cereus in processed spices.. . Microbiology of processed spices. Sakamoto T. 1976. Ward D. Taylor S.C.M. 1982.C. Recent Advances in Food Science. Christiansen T. Yumoto I. Oxford. Beuchat L. Lawyer R. Shewan J. 59 (2). 215.. 166-73. Collette R.R. Behling A.. 39 (10). 668 -70. Taylor and Francis. Dalgaard P. Powers E. Fish and Fisheries Products: Hazards and Controls Guidance.J. Journal of Milk Food Technology. 11. Ohgiya S. Toxin Related Diseases: Poisons Originating from Plants. Outbreaks of foodborne illness in humans. 2.C. Communicable Disease Report. 47.. 24 (5).. England and Wales Quarterly Report. The bacteriology of fresh and spoiling fish and some related chemical changes. Proper processing and handling of raw tuna. London. 12. 1997. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature.J. Brown T. Animals and Spoilage. 1997. Bartram P. 2005. 70-2.8 1. 167-93.M. 1986.E..R. 9. Woolaway M. Journal of Food Protection. Microbiology of Marine Food Products. 2001...

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

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


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

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

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

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

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

The adductor muscle is the only edible part of the animal. or filaments. gutted. vinegar solution. are united. with the posterior ends of the shells (valves) broad and rounded. The familiar marine mussel is asymmetrical.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. The posterior halves of the valves contain a large adductor muscle that holds the valves closed. or packed with jelly. 58 . in mussel liquor. Bait: used live extensively for baiting fish hooks for line fishing. mussel in butter sauce. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). The shells are fluted and round with a broad. sauces. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves.) Dried: peeled. scallops can swim or leap by a series of rapid closings and openings of the valves.2. flattened ear or wing at the hinge area. Internally. Live: whole with shells. mussel paste. In some species. and the anterior end smaller and pointed. Salted: cooked meats bottled in brine.1.2.1. Fresh: shelled meats (often only the adductor muscle and the roe are eaten.3 Mussel Mussels are marine bivalve molluscs that live either partially buried in the sea bottom or attached to rocky surfaces by means of byssus threads.FISH AND SEAFOOD 3. The common scallop is about 5 cm long. scallops possess a single large adductor muscle. boiled. Smoked: meats may be canned in edible oil. brine. also with other ingredients. Unlike other sessile bivalve molluscs. Semi-preserves: cooked meats with vinegar-acidified brine and spices. and held apart. cooked meats packed in dry salt for transport. 3. They have filibranch gills. Fresh: cooked meats. by interlocking tufts of hair-like cilia. Canned: cooked meats. smouldered and afterwards dried (Japan). the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. in which the individual branches.

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

rakes.000 /g. 10. shellstock (live) which is shellfish in the shell. (9. 3. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. dredges. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. and cockles The processing of bivalves takes two forms. Acinetobacter spp. 12. Pseudomonas spp. mussels. spirochetes of the genus Cristispira and Saprospira. 13).1 and 3.. but have never been cultured (14... Vibrios. Water temperatures affect the concentrations of microflora in the water. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. nutrients..3. that have never been cultured on laboratory media.3. observed microscopically. In summary. For example. from which one or both shells have been removed. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. 15)..1 Processing and its Effects on the Microflora Oysters. whole or in part. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. Bacillus spp.1. Moraxella spp. Campylobacter coli Bacillus cereus Hepatitis non-A. and shucked shellfish that is shellfish. Other microorganisms found in lesser numbers are the Achromobacter spp..1. 11. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. or by 60 .3 3.. Enterococci spp. There are microorganisms. At harvest. responding to the environment in which they are harvested.. They are termed viable non-culturable microorganisms.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. mechanical harvesters. Temperature. Micrococcus spp.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs.2. Flavobacterium spp. clams. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish.3. 3.1 Shellstock 3. the initial flora composition and concentration of bivalves are highly variable. and Aeromonads. and yeast cells of Rhodotorula rubra and Trichosporon spp. The largest groups found are the Pseudomonads. salinity. Alcaligenes spp..1.

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

16. At this temperature the multiplication of faecal coliforms (E. The most important factor in controlling bacterial growth in shellstock is temperature. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. Harvest conditions are another factor that influences the growth of bacteria in shellstock. The washing of sediment and detritus soon after harvesting will prevent further contamination. Over a period. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. however. to avoid possible contamination of the shellstock. handling.FISH AND SEAFOOD 3. the bacterial counts in the meats will increase. 17). and Citrobacter species) and Vibrios. At harvest.3. Excessive sediment on the shells and dead shellfish may increase bacterial 62 .1. for a considerable length of time. shellstock are placed in refrigerated trucks for transport or placed in wet storage. coli. Generally. vulnificus. 21. shellstock are not refrigerated on the harvest boat. Furthermore. there is evidence that in the absence of adequate refrigeration. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21.2 ºC (45 ºF) or below (19). some pathogenic organisms will survive and multiply in shellfish (20. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). are prevented. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. Water used for shellstock washing should be of good sanitary quality. hydrophila (18). Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). 25). 24. If long delays occur before reaching the processing facility and/or refrigeration. and thus contaminating the shucked meat at the point of sale. with the exception of A. and the sanitary practices applied during harvesting and shipping.1. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. Enterobacter. without adequate refrigeration. including V. 23. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. An outbreak of infectious hepatitis in Sweden in 1956. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. 22). 26). Klebsiella. Generally. when it reaches the processing facility.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. and temperature storage of shellstock to point of sale. synthetic seawater for the purpose of de-sanding and salt uptake. bivalves tightly close their shells thus trapping water and microflora. with prolonged storage the microbiological quality of the bivalve decreases.

thus contributing to high bacterial counts in the final product (28). Containers are placed in packing boxes for shipping at refrigerated temperatures.1. and packing shellfish into containers (Figure 3. are applied to the shucking operation of shellfish. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. 3.3. washing and culling shellfish prior to storage is essential. Thus. for the effects of microflora during the processing of shellstock. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. shellstock is subjected to contamination when stored on the floor in standing water.3. or subjected to splash from foot traffic.2 Shucked shellfish 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. lobsters. washing. Shuckers routinely use either cotton or rubber gloves during the shucking operation. In the blower-washer. 3. Under these conditions if the shuckers enter the packing room. the finished product may become contaminated. If shellfish are not reasonably clean at the time of processing. and fish. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported.2 Effects of microflora during processing The same considerations as previously discussed. through which the washed meats are discharged for rinsing and then packed in containers. Studies have shown the use of rubber gloves reduces the contamination level. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. whereas cotton become wet and 63 . To remove the shellfish attached to the remaining valve. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations.2). removal of shellfish (shucking).3.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage.1. After washing of meats.2. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station. clothing becomes very soiled. the tank is opened through a drain gate and chute. In addition. 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). Due to the nature of the shucking operation.1.2.1 Processing description The processing of shucked shellfish is limited to washing of shellstock.

FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). within one hour. There is a tendency to store such articles on the shucking benches or in packing rooms. 64 . 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): 1.2. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. Shucked meats are thoroughly drained.2 ºC or less. Shucking containers are rinsed clean with running water. Fig 3. which may lead to contamination of product. 4. and gloves help prevent the contamination of shellfish. aprons. 2. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. 3. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. Separate rooms or lockers for storage of clothing. and sanitised before each filling.

spoilage will occur. 3. is stored in the bivalve as a source of energy. and proteolysis (10). blowers and other utensils that have cracked. In summary. are an excellent medium for growth of bacteria. All equipment coming in contact with the shellfish such as colanders. whereas bacterium such as Pseudomonas. showed that organisms such as Achromobacter. abundant gas production. Temperature abuse will result in high bacterial counts. Controls that should be applied include using clean packing equipment. 34.47 to 6. 35. For example. a carbohydrate. Shucked shellfish are an excellent medium for the growth of bacteria. 31. where there is a breakdown of temperature control or prolonged storage of shellfish. clean gloves. thus producing acid and gas (38). particularly oysters stored at 7 ºC. Shellfish. This process changes the bacterial profile to microorganisms favouring low pH. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. Alcaligenes. The spoilage process in shellfish may be divided into three stages: increase of acidity. and yeasts. Lactobacilli seem to be the predominant microflora in shellfish. rough. Flavobacterium. 65 . and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. particularly shucked meats. skimmers.2 ºC or less. and packing the shellfish in a timely manner. Glycogen is hydrolysed by the initial flora found in shellfish. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. Shucked shellfish are held and transported at temperatures of 7. These organisms may play a significant role in the spoilage processes. 36). The environment has an effect on the bacterial population found in shellfish. anywhere during the process of shellfish from harvesting to the point of sale. Achromobacter and Flavobacterium levels decrease during storage. these include lactobacilli. at the turn of the century. The initial flora found in shellfish is the major factor of spoilage in bivalves. Micrococcus. Glycogen.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. Earlier studies. 33.2 ºC. 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. streptococci. glycogen levels range from 0. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. shucking pails. in oysters. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. Proteus. 32).4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish.8% (37). Pseudomonas. As described previously. Much of the work has been done with oysters and clams. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended.

000 population/year (46). The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. ranging from epidemic cholera to sporadic cases of diarrhoea. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. Major outbreaks are associated with the warmer months. diarrhoea.FISH AND SEAFOOD 3. parahaemolyticus causes gastroenteritis lasting 24 . from harvesting to finished product (i. These organisms can be present in the shellfish from the point of harvesting. parahaemolyticus.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. of V. V. It is a common marine bacterium. coastal areas). 66 . lot and retail outlet. Between May and September 1997. parahaemolyticus from shellfish is low. 3. only a small percentage of strains in the marine environment are potentially pathogenic (45). nausea. Fortunately. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. primarily in brackish or saltwater. where outbreaks occur regularly. This bacterium is present in all the major processing steps of a shellfish plant.5. Diseases caused by Vibrios are enteric in nature. prevention of cross-contamination. One of 250 cases reported involved shucked products with low levels. and very common in Japan. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). 41).5. however. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). headache and fever.e. They can be introduced to the shellfish from the environment or added during processing.1 Vibrio parahaemolyticus V. during processing. Products involved were from the same processor.48 hours with abdominal pain. shellstock and packing of shellfish in jars) (29). widespread in both polluted and unpolluted waters. and estimates based from a hospital study show less than 0.5 cases/100. although non-hemolytic strains have been associated with illness (43. Sporadic cases are frequent along the coast of the United States.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. and to the point of sale. parahaemolyticus possibly producing illnesses (40). The incidence of V. and harvesting of shellfish in the cooler months. approximately 23 cfu/g. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. 3.1. 44). there is a recent report of low levels of V. V. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration.

Inadequate refrigeration will accelerate the growth of V.000 population/year (8. High-risk individuals. Death has been reported to occur within 36 hours from onset. V.7 days. The organism attaches to the intestine and release of a toxin is suspected. are particularly susceptible to primary septicaemia. Primarily.II (8.2 Vibrio vulnificus The symptoms of V. 54.0. V. cirrhosis.1. vulnificus are fever. 46. 49).1. with a fatality rate of 50% for individuals with septicaemia.8 case/100. They are sensitive to cold. Healthy individuals are susceptible to gastroenteritis. chills. 55. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. Control of V. It causes diarrhoea. 3. Onset of symptoms occurs in 48 hours and can be severe. 48). The bacterium is an estuarine species that causes wound infections. cholerae non-01 is ubiquitous in the estuarine environment.48 hours of onset. 23 of 24 major USA-West Coast estuaries were positive for V. The occurrence of V. parahaemolyticus. diabetes. fever. those who have liver disease. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). 3. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. 56). In one study. nausea. vulnificus. leukaemia. lasting 6 . these individuals are advised not to eat raw shellfish. All isolates are considered pathogenic. and nausea within 24 . Oysters are the most hazardous food associated with V. parahaemolyticus the estimated annual incidence rate is very low. like V.5. in the range of 0. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. gastroenteritis. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. Hazards from this bacterium can be controlled by thorough cooking of shellfish. the organism has been found in the Gulf of Mexico. cholerae non-01 has been associated with septicaemia. 57. and rapidly become inactive at low storage temperatures. V. Again. Control of oyster-associated 67 . cholerae non-01 is generally a less severe gastroenteritis than that caused by V.4 . or immunosuppression.3 Vibrio cholerae non-01 Illness caused by V.MOLLUSCAN SHELLFISH Control of V. 46. abdominal cramps. parahaemolyticus growth in shellfish meats is temperature dependent. or primary septicaemia. which have been implicated in shellfish-related disease outbreaks (31). It is one of the most severe foodborne infectious diseases. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). cholerae.5. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. however. cholerae non-01 (51). and preventing cross contamination once the product is cooked. 58). and vomiting. 53. vulnificus is by rapid refrigeration during warm weather months.

1. (8. Onset of disease is 6 hours to 5 days. nausea. In addition. cholerae non-01 illnesses is by cooking seafood thoroughly. parahaemolyticus and V. Onset of illness is from 3 to 5 days after ingestion of contaminated food. vomiting. They are common marine bacteria. imported product from a country experiencing an epidemic.48 hours with abdominal pain. diarrhoea. hollisae. 61. particularly in oysters. jejuni requires reduced levels of oxygen (3 . including V. drying. dehydration.5.10% carbon dioxide for optimal growth conditions. respectively. processors should know the product source. cholerae 01 is transmitted by faecal contamination of food or water. undercooked. 3. vulnificus. abdominal pain. however. cholerae 01. cholerae 01 in molluscan shellfish. 3. fluvialis. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. Poor sanitation and contaminated water supplies will spread the disease. V. or cross-contaminated shellfish (57. widespread in both polluted and unpolluted waters.1. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. 68 . The frequency of infection of these organisms is no different from other Vibrios. The presence of these strains appears to be in low numbers and non-virulent (59).g. nausea.10 days. In developing countries. 60). and nausea. There is evidence of strains of V. headache and fever. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. Gulf coast environment. 3.5. is the same as previously discussed for the non-01 strains. preventing crosscontamination of cooked seafood. V. associated with shellfish-borne illness outbreaks. The control of V. and shock.5 Other Vibrios spp. 63).5. Illness can include abdominal cramps. Illnesses are associated with the warmer months. and southern Asia including India and Bangladesh. Death may occur after severe fluid and electrolyte loss. There are several other Vibrio spp. e. these organisms cause gastroenteritis lasting 24 .5%) and 2 .S. and illness lasts 7 . salinity. and the transmission of the disease by consumption of raw. 62.2 Campylobacter jejuni C. acidity and heat. In general. It causes diarrhoea. fever. V. mimicus.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools.FISH AND SEAFOOD V. cholerae 01 establishing in the U. and V. low pH. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). Control of infection from these organisms is no different from the other Vibrios. the pathogenic severity is lower than V.

Although. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. Human feeding studies suggest that about 400 .5. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. hydrophila group (75.500 cells/gram of Campylobacter may cause illness (64. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. V. 73). 70. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). cholerae 01 and non-01.MOLLUSCAN SHELLFISH TABLE 3. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. of Incidents 4 279 0 1 0 No. V. mimicus. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. jejuni begins in the harvest of shellstock from approved waters. V. ranging from 0.5. jejuni does not multiply in shellstock at refrigerated temperatures. infection occurred among participants of a firemen's banquet. 77. The seasonality of 69 . fluviallis. Campylobacter spp.3 Other organisms of concern 3. 80) are ubiquitous in the marine environment. 84. 66). V. Campylobacter appear to survive well in oysters during storage. 71). C. 68. This finding is significant in that C. 76. 83. they survive well at refrigerated temperatures. C. 72. 78) and P. Recently.4 to 114 cells. 3. 81. however. 82. from Pacific shellstock oysters purchased from a retail grocery store (74). however. shigelloides (79. have been isolated from shellfish beds throughout the world (67. jejuni were recovered. 69. control of C. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. They have been implicated in limited shellfishassociated gastroenteritis cases (8. hollisae Infective dose varies with host susceptibility and virulence of the strain. 65.3. whereas at 10 ºC Campylobacter survive only for 5 days (71). To reduce the risk of shellfish-borne campylobacteriosis. parahaemolyticus. vulnificus. although their role as a pathogen is highly controversial and circumstantial. of Cases 4 362 0 1 0 a Includes V.II Molluscan shellfish associated outbreaks caused by Vibrio spp. and their immediate proper refrigeration.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. 85).1 Aeromonas hydrophila group and Plesiomonas shigelloides A. In one case. and V.

87) of the outgrowth of A. They are easily destroyed with mild cooking temperatures. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. parahaemolyticus Staph. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. soy products. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. In one study.000/gram (29). Test Standard Plate Counts E. non-dairy frozen desserts. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. vulnificus in molluscan shellfish. Caution should be exercised by individuals with haematological malignancies (88). Similar results were seen by other investigators (86. jejuni V. coli Enterotoxigenic E. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. 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. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500.000 to 3. prepared sandwiches. FDA/ Administrative Guidelines. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. Control of these organisms in shellfish products is limited. FDA/Administrative Guidelines 3 Source (91). cholerae V.600. 2 Source (90). 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. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. 70 .6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. hydrophila group stored at refrigeration temperatures. coli Salmonella V. dried milk/whey drink powders.0001 negative for the presence3 Under review4 Source (89). aureus C. 3. being isolated more often during the warmer month.

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

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

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

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

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

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

M.S. Burdge D. U. U.MOLLUSCAN SHELLFISH 88.L. 3.S.S.R.. The Microbiological Safety and Quality of Food.W. Martin R.. Aspen Publishers Inc. 89. Clams.C. Food and Drug Administration. Slavin J..030. Inc. National Academy Press.C. Government Printing Office . Food and Drug Administration. U.A. Seafood Safety.8 Further Reading Ahmed F.Pathogen monitoring of selected high risk foods (FY 88/89).S. 63-8. U. Smith J. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. Food and Drug Administration. Washington D. Washington D.E. oysters.C.S. 1987.S. Food and Drug Administration Compliance Policy Guides. mussels. Food and Drug Administration. Part I Sanitation of Shellfish Growing Areas. 560. Sherlock C. and HACCP: A Global Focus. Chapter 5: Foods. Food and Drug Administration. 2000. fresh or frozenadulteration by bacteriological contamination Sec. Ed. Food and Drug Administration Compliance Program Guidance Manual. Ed. 91. 7..W. in U.S. Food and Drug Administration. Food and Drug Administration.S. Fish Inspection.H. Baird-Parker T.. Gould G. Technomic Publishing Co..C. U. Quality Control. Lancaster. Food and Drug Administration. 1991. 90.. U. 1995.E.12). in U. 1995.600 (CPG 7119.1987. Program 7303. Lund B. Washington D... Gaithersburg. 77 . 1997. Ed. in National Shellfish Sanitation Program Manual of Operations. Pittsurgh. Collette R.


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

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

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

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

the joint between the tail and carapace is open and the limbs hang loosely. since crabs and lobsters are traditionally consumed immediately after cooking. spoiling prawns and shrimps have shells that are soft. However. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. If the tail drops. 4. there is no indication of life.4.4 Spoilage The main genera of Gram-negative bacteria responsible for the spoilage of crustacean shellfish are the same as those that cause spoilage in finfish. wet and ‘soapy’ or ‘jammy’ to the touch. Heat is produced if the shrimps decompose in a confined space. the meat is either repacked and sold in the shells. immediately prior to boiling. and have a sweetish.4. with the growing demand for ready-to-eat products from crab and lobster meat.3.CRUSTACEAN SHELLFISH 4. dull and sticky. or packaged and processed. The shells have a crisp. and a strong ammoniacal odour is produced.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. 83 . In comparison. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. If boiled. With spoiled crabs and lobsters. contaminating microflora may include enteric bacteria. putrefaciens. slightly iodine smell. The shell will be soft. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. or in hermatically sealed cans that have undergone an appropriate heat treatment. Once removed and cleaned. 4. putrefaciens and Pseudomonas spp.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. viruses or parasites. they include Sh. An offensive odour is also produced. bright and dry appearance. whilst boiled crustaceans have a pleasant slightly sweet smell. If hygienic procedures are not followed.2 Cooked crabs and lobsters As indicated earlier. there is no associated spoilage problem. bacteria of public health significance. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. 4. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. for crabs.

shrimp and shellfish (11). the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). in 24. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. monocytogenes in cooked ready-to-eat foods. monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). However. 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.cooked prior to consumption. 16). monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 .5% of frozen raw shrimps from different countries (13). even if GMP is followed. 6. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. An import alert was put in place in June 1987. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12).1 Pathogens: Growth and Survival Salmonella spp.5. in domestic/imported shrimps (15). and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14.2 Listeria spp. as well as for Salmonella. Further surveys identified Listeria spp. shrimps and cockles sold (17). The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . although efficient cooking will quickly eliminate it (6). Hence. a common pond fertiliser used in aquaculture. The above results prompted the FDA to develop a compliance programme for the pathogen. and usually for both. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. 4. animal manure. Moreover. later surveys in 1990 recovered the pathogen from retail raw.FISH AND SEAFOOD 4. Although foodborne illness had not been associated with the product. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9).5. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. 7). often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. Salmonella cannot be completely removed from raw shrimp during processing. fish fingers. Following this. monocytogenes was absent from 40 samples of cooked prawns. cooked and smoked fish. nearly 30 years elapsed before the potential hazard of L.5 4. 5. The presence of L.

22) as well as from undercooked crustacea when harvested from contaminated waters (23.5. cholerae. The main source of contamination is the hands of processing personnel. The pathogen has resistant spores. 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. V. 4. V.5 Vibrio spp. whilst boils and cuts are also common sources of the pathogen.70% of healthy individuals carry toxin-producing strains of Staph. However.3 Staphylococcus aureus Staph. botulinum type E is commonly associated with fish and fishery products.5. Control should be achieved by promoting GHP and staff training programmes. monocytogenes has been attributed to crustaceans. Once the competing microflora has been removed by a heat treatment. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. An epidemic that began in South America in 1991. so is able to survive heat treatments that eliminate vegetative cells. the pathogen can grow at temperatures as low as 3 °C. the causative agent of cholera. especially in products that have been vacuum packaged. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. as part of the natural bacterial flora. 4. cholerae is divided into two groups. 25). 24. aureus because of the ubiquity of this organism in the processing environment. which can become colonised by V.5. was associated with seafood consumption. is widely distributed in aquatic environments. pregnant women and the immuno-compromised. 19). therefore. aureus is not a member of the intrinsic flora associated with crustacean shellfish. Vibrio cholerae. 85 . combined with the degree of handling of these types of product. Crustaceans have been linked with food intoxications attributed to Staph. To date. pasteurisation and cooking.4 Clostridium botulinum C. Crabs are covered in a chitin exoskeleton. 4. although contamination by types A and B may occur from the environment. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. no incidence of foodborne illness caused by L.CRUSTACEAN SHELLFISH old.g. cholerae non-O1. aureus on their hands. The former. about 50 . e. Researchers have demonstrated that inoculated shrimp supported toxin production by C. particularly crabmeat (18. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. cholerae O1 and V. botulinum type E at 10 °C but not at 4 °C (20).

4. vulnificus is an invasive and lethal pathogen associated with wound infections. hydrophila and A. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. V. Quantitative levels were below limits of detection in all cases (<0. Hepatitis A virus presents the most serious hazard.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. or freezing at a temperature of -23 °C for 7 days (30. 32).30 MPN/g) (27). whilst 5. Norovirus.7 Aeromonas hydrophila and Aeromonas sobria A. 4.2 °C) so that GHP coupled with controlled low temperature storage is essential to prevent the pathogen from reaching levels that may present a hazard.6 Campylobacter jejuni and Campylobacter coli C. been implicated in a case of foodborne infection associated with crustaceans. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture.g. in a survey carried out in the United States between 1977 and 86 . to date. V. however.5. parahaemolyticus. e. Of these.5.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. non-B Hepatitis viruses (31). jejuni has not.5. hydrophila has been isolated from prawns (29). Astroviruses and non-A. coli.8% of samples contained C. 4. Caliciviruses. However. Contamination by this organism can be controlled by rapid chilling of crustaceans after harvesting and avoidance of time/temperature abuse during distribution and processing chains.5. It is more commonly implicated in countries where crustacean shellfish are eaten raw. 4. a marine halophile.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. but has the potential to cause fatal foodborne illness (26). although A. South America. occurs in crustaceans from warm environments. Parasitic infections are controlled by either cooking infected material prior to consumption. hydrophila is able to grow well at refrigeration temperatures (0 . However. 31. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. Japan. A. particularly trematodes (6). they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). the potential hazard exists as C.

The risk of such hazards is far higher with fin fish and molluscan shellfish. only 0.CRUSTACEAN SHELLFISH 1981. 4.11 Chemical pollutants and biotoxins Researchers have demonstrated that crabs feeding in waters contaminated with algae do have potentially high levels of algal toxin domoic acid (DA) in their viscera. 4. 38. which results in a persistent and apparently permanent loss of short-term memory (36). Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). from farm to fork (37. Norwalk viruses can also withstand heating to 60 °C.6. 87 . Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control.5. They can survive low pH levels. or chlorine concentrations similar to those found in drinking water.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. consistent with that in the stomach. 39). are included in the Commission Regulation (EC) No. produced or imported into the European Union (EU). 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. A summary of microbiological criteria for crustacean shellfish is listed below.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33).1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. 4. 4. 2073/2005.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive.5. No foodborne incidents have been attributed to this source to date. although it is one of the European Union (EU) legislated allergens. Selected parts from this regulation is presented below. Provisions laid down concerning the use of additives would also apply to these types of products. The toxin causes amnesic shellfish poisoning (ASP) when consumed. which could be transferred to the meat during unskilled removal from the shell (35).

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

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

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

8 79. smoking or marinating processes.2 105. TABLE 5. about 50º (132.FISH AND SEAFOOD 5. Different brine strengths are required according to the curing processes and/or the type of product. it is important to understand the concept of salt concentration. For smoking salmon.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) 26.8 211.80º brine (184.3 Processing and its Effects on the Microflora Owing to the wide variety of products. all using salting steps. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. the fish is usually salted with a 70 . Thus.4% salt (Table 5.2 g of salt per litre of brine).2 237.8 211.I).0 g of salt per litre of brine). The strength of the solution is described as degrees of saturation (brineometer degrees). sometimes the salt crystallises onto the surface of the fish.0 158. it causes swelling because the fish absorbs water from the salt solution (1). etc. only the most common processes can be described here.4 184. % Salt in the water phase = % salt content % salt content + % moisture x 100 Another important point to remember when wet salting is used is the concentration or strength of a brine. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. or salt in the water phase.6 264. Therefore.6 132. Salt in the water phase links the two conditions of moisture and saltiness. When working with salting. a 0º brine saturation means pure water. Salt concentration means the percentage of salt in the water contained in the fish. it is impossible to establish standardised processing stages in the curing of fish. and salt content. When a milder brine. a 90º or above brine (237. processes and producers’ private recipes.6 g of salt or more per litre of brine) may cause salt burn in this way. At 18 °C a 100º brine is produced when the solution contains 26. The dissolution of salt in water is dependent on the temperature. it is important to state the temperature. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat.0 . When a 100º brine is used. is employed. This should be distinguished from the salt content.4 52.

It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. Usually it is followed by other preservation techniques such as drying and/or smoking. but the concentration of copper and iron must be kept under 0. Also. This condition is known as ‘putty fish’ (4). are likely to cause hydroscopicity of the product and. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. For a light salting.15% calcium and magnesium.1.1 Salting Salting of fish for preservation purposes has been used since pre-history. it tends to lead to a slight yellowing and softening of the product.1 ppm and 10 ppm. and salt can speed this oxidation process. are recommended to help make the product whiter. respectively (1).0. caught in North Atlantic waters. the fish is mixed with one part of salt per eight parts fish. low in calcium and magnesium. and the salt has penetrated throughout the fish (3). The salt used should be of food-grade quality. 97 . for which there is a high demand in Mediterranean countries. See Figure 5. The calcium and magnesium impurities. 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. and such delays. and essentially free from iron and copper (3).35% and 0. can be dried further to produce ‘klipfish’ or ‘bacalão’. addition of levels of 0. outgrowth of halophilic bacteria may occur. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. mainly in tropical conditions.3. the fish is restacked with fresh salt after about a week. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). However. The most common fish used in the northern hemisphere is cod (Gadus morhua).CURED. when the air humidity is high. SMOKED AND DRIED FISH 5. producing characteristic pink patches. This is because fatty fish turn rancid quickly.15 . ‘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. if very pure sodium chloride is used for curing purposes. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. respectively. and for a heavy salting a ratio of 1:3 salt:fish is used (1).1. ‘Green cured’ split cod with about 60% of the backbone removed. 5. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. however. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. lean white fish is used for this purpose. having a short section at the end of the tail. When pure salt is used. On the other hand. Salting can be done in a number of different ways. which include calcium and magnesium.1 Dry salting / Kench curing Usually. and a ‘putty’ fish product is also obtained (1).3.

such as 98 . Where fish are salted in their whole form. although toxin production would be unlikely (4).90 are rarely achieved. Only small fish can be salted whole. The greatest barriers to salt penetration are the skin and scales. Fig. such as Staphylococcus aureus could survive and grow at such water activities in the product. For large fish it is advisable to split it before salting.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat.2 Wet salting or brining As referred to above. eviscerated and/or split).3. when a light brine is used.5. Food-poisoning organisms. Processing of dried salted cod 5.1. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. it appears that water activities much less than 0. the process is mainly for flavouring purposes prior to the use of other preservation processes. so that the risk of spoilage is high.1.

96 1.64 4.17 4.80 5.49 4.10 1.70 6. The main objective of the brining method of salting fish is to limit air penetration during processing.35 4.76 4.26 2.23 2.23 3. the liquid that comes out of the fish forms a brine.65 2.85 3.03 2.98 5. the fish enzymes play an important role by maturing or ripening the product. the product obtained is called gaspé-cured (5).00 3. The fish are kept immersed in their own brine near saturation point for several days. For example. but as it is placed in containers. Histamine formation is of importance when using scombroid and some clupeidean species.3 Dry / wet salting In this process.19 3.41 5.46 4.85 3.11 4.54 5.33 5.76 4.76 4.98 6.51 5.44 2.73 1.95 3.3. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.41 3. Histamineforming bacteria do not generally grow at water activities below 0.5 10. See Table 5. with pressure being applied to the top of the container to help the brine to drain away. can be cured in a saturated brine solution for 6 days and.26 7.11 9.0 2.09 8. the fish is dry-salted.47 6.06 4.51 3.76 4.03 4.66 5.CURED.1.26 3. with a shelf life of about 4 weeks (water activity (aw) about 0.66 5.II.04 5.23 2.26 1.25 5.45 3.91 2.26 4.99 2. the brine is then replaced over the fish.78 2.II Salt in the water phase (after Ian Doré (5)) Moisture (%) 40 45 50 55 60 65 70 75 80 85 90 Percentage salt in the flesh (by weight) 2. 5.17 5.5 4. herrings and other fatty fish can be used.61 3.90 (1).76 99 . TABLE 5.78) (1). Fresh sardines.86 2.5 3.76 4.16 7.5 3. if split fish is dry-salted. and a red burgundy colour will develop in the product.0 4. after being drained and pressed at 465 kg/m2 for 15 hours.0 3. they can be stored at 28 °C.22 6.74 9. headed and gutted.84 1.09 8. limiting the amount of rancidity that can develop.88 5.25 5.05 7. There are several variations of this process. Anchovies are the main fish used for these purposes. For preservation purposes. This is why it is the preferred method for fatty fish.78 6.41 4.98 6.70 3. and can be collected and replaced again over the fish. Here. sardines. thus.30 2.41 6. but mackerel. SMOKED AND DRIED FISH smoking.3 days.61 3.11 3.56 6. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .60 2.23 8. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.44 2. a saturated brine is used.

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

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

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

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

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

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

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

Hot-smoked products made from white fish will usually keep better than those from fatty fish. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). 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). a very expensive and fashionable smoked product in the Netherlands. break down of fats and the moderation of smoke flavour. Growth of these moulds on the tuna results in the extraction of moisture. and many closely related Aspergilli (Aspergillus ochraceus. When refrigerated at 3 °C. are used. Katsuobushi. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). Boiled or steamed clams. then hot smoking and finally sun drying it. chilled or freeze-thawed. Natural smoke extracts. are usually 80º brined for 5 . The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. rewetted and remoulded to give the desired shape and consistency.3½ hours using the typical temperature set for hot-smoking referred to above (12). 18). 17. 5. Aspergillus oryzae. Aspergillus ostianus. but mechanical drying is used instead of sun drying. Aspergillus tamarii) and several Penicillium spp. synthetic smoke flavours. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. and substances unconnected with smoke but with a smoky flavour or smell. including Eurotium repens and Eurotium rubrum. 21. Mackerel. are usually hot-smoked after being immersed in 80º brine or dry-salted (6).3. Vacuum packaging retards the onset of rancidity when fatty fish is used. (Penicillum cyclopium and Penicillium puterellii) (19). fatty fish will remain in good condition for 6 days. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. In terms of safety. They will keep well frozen. The product is removed from the drying cycle. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. They are generally not cooked prior to consumption. 23). white fish about 8 days. but seems. similar to masmeen. 22. apart from the improved appearance. resulting in a characteristic flavour (20. cooking in brine. such as yeast derivatives. 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). The production method is similar to masmeen.5. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. to have no further advantage over other packing methods for other fish (12). moulding it into long thick sections.CURED. The classical liquid smoke is 107 . 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’ (16.6 hours and smoked for 3 . Eels.

it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (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). applying it as a dip after brining. injecting it into the fish flesh. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. When liquid smoke was used. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0.06 μg/kg of benzoanthracene in the final foodstuff (28). or smoke extracts in organic solvents.03 μg/kg of benzopyrene. consistency of flavour and colour. The process relies on the generation of the smoke to produce elevated levels of carbon monoxide (CO) which interact with haemoglobin in the muscle to form the very stable bright pink/red carboxymyoglobin complex. and less than 0.5. both of which are considered potential carcinogens. The European Scientific Commitee for Food.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. spraying is preferable because it is easiest to control flavour and acceptable with this method (3).6 to 2. It has been shown that liquid smoke can reduce the formation of PAHs (3). nutritional neutrality and antioxidant activity (27). 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.8% and still provide protection for 7 days (25). Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. so as to impart an unnatural red colouration to the flesh. Liquid smoke in combination with sodium chloride in a hot-process smoke flavoured fish was found to be effective in preventing growth and toxin production associated with the growth of C. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines.4 Spoilage Brining. drying. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). According to some recent research. smoking. Liquid smoke was considered to provide advantages of safety. the salt level could effectively be reduced from 4. or applying it as an atomised spray within a modern automatic smoke kiln. 5. In experiments comparing traditionally smoked fish with liquid smoke. Fresh fish spoilage may be measured by 108 . pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. 5.3.4 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.

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

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

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

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

SMOKED AND DRIED FISH TABLE 5.86 0. The presence of salt in cured products has a great effect on the growth of the bacteria. the fermented products being the most dangerous group (7).61 Organisms Most Pseudomonadaceae C.8 0.5 5. aureus Yeasts Moulds Lactic acid bacteria pH 5. The concentration required to prevent growth at room temperature can vary from as low as 3. However. types A and B are not affected by 3 .5. although the combination of the two factors. the importance of cold-chain integrity must be emphasised.CURED.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. botulinum type E Staph.97 0.5 5. popular demand for healthy foods has led to a change in the tastes preferred by the public and. botulinum type E and production of toxin. botulinum type E. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C.5-8.5-10 4. Since neither the smoking nor the drying parts of this curing process are particularly severe. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). 9 and 8 of the outbreaks. It has been shown that. Before.8 1.5-8.5 0-11 3.97 0. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. could not grow and produce toxin. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39).1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). In the United States. pressure on the traditional methods of curing. respectively. botulinum. when present in the raw material. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. under optimal conditions. and C.75 0. but if the fish products are stored at temperatures <10 °C.5 4.5% to 5%.5-10. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. and as botulism is a serious hazard in foods. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. the products were heavily salted. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). the non-proteolytic type E strains can grow in up to 5% sodium chloride.9 0. botulinum Staph. with this change.0-9. sodium 113 . salted and pickled products accounted for 10. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. smoked and dried.

The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). However. but requires higher temperatures for toxin production (>10 °C). The organism is mesophilic with a minimum growth temperature of 7 °C.4.5.94 (7). related to the consumption of a raw smoked fish meal. Thus. the risk of infection can be eliminated by proper cooking. coli all occur on fish products as a result of contamination from animal/human reservoirs. has been reported (42). Sliced smoked fish products. throat and skin.5. 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.4 Listeria monocytogenes The psychrotrophic L. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0.3 Enterobacteriaceae The pathogens Salmonella. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . are particularly susceptible to contamination and. 5.or cold-smoked fish should have at least 3. As they are poor competitors. 5. It is a halotolerant bacterium able to grow at water activities as low as 0. The FDA established that hot.5. gills and guts as well as in several lightly preserved fish 114 . but.0 . growth and toxin formation may occur. Usually. 5. they do not grow well in the raw material.5% salt in the water phase. after processing. the organism can survive most of the light curing processes.86 and 10% sodium chloride. As with other organisms. Shigella and E.5. when the bacterial load of the product is lower. it should contain not less than 3. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. more importantly. An outbreak of Shigella in an Italian cruise ship. if storage temperatures are over 10 °C. aureus. they will grow rapidly (7).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. and the minimum pH for growth is 4. the human nose.0% salt in the water phase (30).2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. if recontamination occurs and storage conditions are favourable.30% of the population positive for enterotoxin-producing strains (41). aureus growth. Once the product is contaminated. In foodstuffs. if it contains sodium nitrite ≥100 ppm. as they require more handling. but. Infection by Shigella sonnei has increased in the UK. the organism may contaminate the product. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. if the person handling the product is a carrier of Staph.

The use of LAB in fish preservation is a topic that has been studied in projects (UP.the round worm Anisakis spp. favours growth of psychrotrophic LAB. Until 1997. do not survive 1 min at 60 °C.514. Of microbiological concern in the northern hemisphere are two parasites .5 Parasites Despite the presence of parasites in fish being very common.9. They have complicated life cycles. which. which include intermediate hosts.12%. a number of fish products. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). researchers are looking for other ways of preventing the pathogen’s occurrence or growth. a short period of freezing either of the raw material or of 115 . the only concern to public health is when eating raw or uncooked fish products. and other local traditional products (7). Its limiting conditions for growth are 0 . and in CT95-1207FAIR).CURED. there were no reported outbreaks related to cured fish.92 . the USA established ‘zero tolerance’ for the organism. which considered L. 5.95 (45). salt levels of 8 . such as ‘lightly preserved’ fish products. Unlike some European countries. cold-smoked fish. monocytogenes pathogenic for only specific segments at risk. have been shown to have an anti-listerial activity in many food products.6 and water activities below 0. or nematodes. the CT 3162-FAIR program.0. Thus.8 . from freshwater fish. in the EU Fisheries R&D-FLAIR program. Pseudoterranova dicipiens.2. and the tape worm Diphyllobothrium spp. Two outbreaks in the United States have been reported to be linked to salmon (10). As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. monocytogenes in levels at or above 102 per gram (46). Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. has also been detected in cod (7). gravad fish. most of them are of little concern to public health. 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. in combination with curing agents and refrigerated storage. a round worm. lightly salted caviar.5. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). As they are heat-sensitive. monocytogenes’ in 25 g of the finished product. widely used as starter cultures in food processes. This includes matjes herring. both being detected in cold-smoked salmon. Some LAB. are considered unsafe. which means that the process must conform to a level of ‘no detectable L. SMOKED AND DRIED FISH products such as cold-smoked fish. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. pH 4. The round worms.45 °C. marinades (ceviche) (43) and gravad fish (44). from seawater fish. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere.

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

Birnie K. Wareing P. Martin R. 1994. * Staph. Flick G. Torry Advisory Note No. References Regenstein J. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5.M. Bannerman A. Fernandes R.E. Torry Advisory Note No. Doré I. 1993. Preservation of fish by curing (drying. McLay R. SMOKED AND DRIED FISH TABLE 5. New York. 1997. Eds. 82 (revised). Moody M.M. Leatherhead Publishing. Handling and Processing Eels. 1984. Marinades.. USA. 9. cured and dried fish. Catching..CURED. salting and smoking). Assurance of Seafood Quality. 254-82 5. or presence of toxin 0. 3. 120-38. except in the viscera of dungeness crab. New York. monocytogenes Salmonella spp.W. Urner Barry Publications Inc. Van Nostrand Reinhold.. The Smoked and Cured Seafood Guide. Huss H. 37(revised). Smoked.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. Regenstein C. 381-406. 2. FAO Fisheries Technical Paper. botulinum * Limit of tolerance 1x 103 ETEC/g. 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. 56. London.F.. Leatherhead. 1979. 2007. Ed. 117 . Food spoilage fungi. Chapman & Hall. Van Nostrand Reinhold. Horne J. 1991. Introduction to Fish Technology. 1990. Horner 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. in Fish Processing Technology.. 1982. coli (ETEC) L. 3273. Eds.A. aureus * Vibrio cholerae V. Hot Smoking of Fish. 4.7 1. in Micro-Facts: The Working Companion for Food Microbiologists.H. 7. Flick G. Fernandes R..F. Wareing P. 8. 334. Hall G. parahaemolyticus Vibrio vulnificus C. 6. Torry Advisory Note No. in The Seafood Industry.

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

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

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

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


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

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

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

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

Similar results were seen with patis. however. In another microbiological study of four fish sauces. These preparations contain a mixed microflora of moulds and yeasts. More recently. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). Other halophilic species have been identified from fish/salt products. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. Endomycopsis and Saccharomyces . were found to be the predominant isolates. however. and the anaerobic conditions that prevail in the bulk of the material. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. When this is a simple carbohydrate source such as sugar. Mucor. Hansenula. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. 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. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). the Philippine fish sauce.1 – 6. Bacillus spp. mean that microorganisms have been generally held to play little or no role in the process that follows. in many cases. after one month. In some cases. and studies 127 .organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. It also demonstrated their significant proteolytic activity during the first month of fermentation. (3. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). or boiled or roasted rice. Generally.3.74 or below. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that.FERMENTED FISH Where other ingredients are used. they can contribute to the initial microflora.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. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). 6. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns.3 6. angkak (Philippines and Thailand) and look-pang (Thailand) are used. This. traditional starter culture preparations such as koji (Japan). 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. which included nam-pla and patis. either directly or as the partially saccharified rice they produce. principally species of the genera Rhizopus. 4).4. sufficient to saturate the water phase present and produce an aw of 0.

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

however. 6. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. In fish sauce production. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues.FERMENTED FISH Fig. eviscerated fish are sometimes used to slow the autolytic process. To produce semi-solid fish pastes as opposed to fish sauces. 14). Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are.3. thought to be largely the result of autolytic breakdown of the fish tissues.4.6. Ika-Shiokara (side dish) Fig. the 129 .

3. In contrast to free-swimming fish. Fig.5).6 – 6. snack) 130 . Hoi-Malangpu-Dong (side dish.8.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. Figure 6. which is eventually separated from the residual fish tissues. 6.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. for example.5. Some examples of products and their formulation are presented in Table 6. particularly glutamate. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. volatile fatty acids and nucleotides.V and Figures 6. 6. shellfish such as mussels often have appreciable levels of carbohydrate in their tissues and in these cases addition of carbohydrate is not necessary for a lactic fermentation to occur (see. filtered and bottled. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur.

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

but there is considerable variation in this. giving a reduced rate of acid generation. these products are salted at lower levels than the fish sauces and pastes. Phillipines Journal of Food Science and Technology.. their growth and metabolism are nonetheless slowed. typically in the range 2 . while they are able to grow at high salt levels. then this will take longer to develop than if lower salt levels are used. 106) Fig. of the two factors contributing to shelf life .3. This can be clearly seen from the data on reported shelf lives of a number of fish salt carbohydrate products and their respective pH ranges and salt contents (Table 6.6. It also indicates that. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6.7.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. Generally. the salt is of far greater importance in determining the product’s long-term stability. (from Arroyo et al.2.salt and lactic acid.1 The use of higher salt levels results in a longer production phase but a better keeping quality product. 2. Red burong isda 132 . If acidity is a desired sensory property of the product.VI). The LAB isolated from these products are those with more marked salt tolerance but. This is readily explained by the susceptibility of LAB to elevated salt concentrations.10%.

3 years 1 .0 Shelf life 3 weeks 6 .3 5.4 .9 . 106) Fig.2 7.6.7 .4.8 .5 .FERMENTED FISH (from Arroyo et al.6.0 .12 months 1 .6 3.8 .2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).5.9 4.2 4.1 .8 2.5 .9.5 3. . Philippines Journal of Food Science and Technology.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.0 .17. White balao-balao TABLE 6. 133 .3 . 4.2 4.4..8 pH 4.

VII). Addition of carbohydrate will.3.3. When added as boiled or roasted rice alone.5) despite other factors.1 . TABLE 6. Garlic. also serves as a valuable source of the more readily fermented carbohydrate. Where they are used. This results in a more active lactic fermentation and a lower pH.9) compared with the white product (pH 4. a common ingredient in these products.FISH AND SEAFOOD 6.2 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.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. Rice is the most common carbohydrate source used. such as salt content. a number of amylolytic species have been described and an amylolytic strain of Lactobacillus plantarum has been isolated from burong bangus in the Philippines (17). a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. In 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. the rice may be added in a partially saccharified form such as kao-mark (Thailand). Although the ability to ferment starch is not widespread in the LAB. For example.4. 21). nare sushi pla-chao pla-paeng-daeng pa-chao Country Philippines Japan Thailand Thailand Laos Saccharified rice kao-mark Kha-namak Starter culture ang-kak koji ang-kak - 134 .0 . a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. being similar (18. inulin. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. although the final population of LAB may be similar. of course. alternatively. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. In several products. 19). also lower the unit cost of the product and this is probably a significant factor in many cases.2.

red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. The growth of yeasts and moulds could.7 up to 6. and there is a marked regional variation in the preferred type. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. There is little published information on the factors that limit the shelf life of the low-salt products. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. such as wrapping in banana leaves. The same product can range from a brown turbid liquid where the fish has been subject to extensive autolysis to brown partly dried pieces of fish. Excessive growth and acid production by the LAB could render products too sour for acceptability. Clearly. 135 . allowing other spoilage organisms to grow. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. lead to an increase in pH. reporting the product’s pH to range from 4. The mixture is allowed to ferment for 2 . which ranged from 7.FERMENTED FISH Sikhae.7 to below 4.5 in the first 2 . but they are likely to behave similarly to lactic fermented products produced from other commodities. presumably correlating with the salt content. Results of a product survey were consistent with this. Traditional packing techniques used with lactic fermented fish products. Generally.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). A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. 6.3 days. in turn. the Korean fermented fish product. In a successful fermentation.9%.17.2 at the end of fermentation. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products.2.8 .5. 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). and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). will not reliably exclude air. is something of an exception since millet is used along with garlic as the carbohydrate source.5:9:3. markedly extending the shelf life (25). the level of salt employed is on the high side for fish/salt/carbohydrate products. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage.3 weeks at around 20 °C. thereafter declining to about 4. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. the pH drops from 6. around 12%.

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

A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). Foodborne trematode infections are emerging as a major public health problem. which is generally consumed raw (29). Puwastien P. and warm weather can result in rapid growth of C. botulinum and the production of toxin. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). particularly in south-east Asia and the western Pacific regions (30). Institute of Nutrition.FERMENTED FISH botulism have become less frequent in recent years. Rakefisk and Herring in the Norwegian Tradition.. and these are often used in the production of fermented fish products. Salting levels are often insufficiently low to prevent growth of C.M. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada. giving an alkaline fermentation. 137 . though 27% survival was still recorded (33). With this particular organism.O. Lutefisk. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. walrus nose. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. although the absence of any source of fermentable carbohydrate means that the process is putrefactive. Thailand. Saisithi P. beaver tail and paw. 2. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31). References Riddervold A. Oslo. One study with the trematode Haplorchis taichui. Dhanamitta S. Journal of Food Science. Microbiology and chemistry of fermented fish. Dollar A. Greenland and Alaska. they could act as an important vehicle of transmission. while increasing the acetic acid content of the formulation had little or no effect (32).. 105-10... whale meat. Fermented Fish Products Study (Thailand) Phase 1. Many of these products are described as fermented. 1989. Baseline Study (1987-1988). 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). 3. Kasemsarn B. Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. affecting an estimated 40 million people. botulinum. 1990. 31. and salmon heads and eggs. 1966. 6.6 1.. Yhoung-Aree J. Liston J. Mahidol University. salt content was found to have the greatest impact on the viability of larvae. Raw materials subject to this process include seal flipper. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. Novus Press. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. which will persist into the product. Where these products are consumed without final cooking.

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

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

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

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

and sub-tropical waters and habitats (15). dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. some are the result of biotransformation processes by the filter-feeding host. Gymnodinium catenatum. 7.2.1).g. The Ncarbamoyl sulfates are readily hydrolysed to STX. all of which contain a tetrahydropurine skeleton (Figure 7. 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. Many of these species are extremely common and are found throughout the world in a variety of cold.FISH AND SEAFOOD has been produced (9).1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium.2 Paralytic Shellfish Poisoning (PSP) Toxins The PSP toxins are arguably the most recognised group of marine toxins and continue to be regarded among the most dangerous. and a tropical species Pyrodinium bahamense (16). Consequently. a great variety of shellfish from all latitudes. and a review on the risk assessment of marine toxins has also been produced (14). 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. PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. All PSPs degrade rapidly in alkaline conditions. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). and another. Saxitoxin Group instead of the Paralytic Shellfish Poisons). In addition. Around twenty PSP toxins are known. temperate. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). 7. neoSTX and Gonyautoxins (GTX) 1-4. with intoxications occurring in many countries even today (15). even crustaceans such as lobsters. 142 . Not all the toxins found in shellfish extracts are microalgal products. can become contaminated with these toxins. The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13).

Human symptoms begin with tingling and numbness of the lips. and humans seem to be particularly susceptible. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. and fingers within 5 . Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water.2 Toxicology PSP toxins affect the central nervous system of mammals.2. 7.30 minutes of consumption. tongue. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. this is never sufficient to render the seafood safe to consume. 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.1. However. Neurophysiological studies have shown that PSP toxins all possess the same mode of action though some derivatives such as STX and neoSTX are considerably more potent than the C-toxins (15). 143 . leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis.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.

8 µg/g) of shellfish or seafood tissue. 15.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. a criticism has been that such methods are time consuming. 7. China. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. and over ten analogs are known (22.2. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. including Japan. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19).2).1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes.3. 17).3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. Alternative HPLC methods have subsequently reduced this to a single HPLC run. 24) (Figure 7. However it remains as the monitoring method of choice in many countries. Most intoxication events have occurred in South East Asia. and has also been evaluated by a number of EU laboratories. specifically 144 . Two approaches have been taken here. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. a pre-column method has received Association of Analytical Communities (AOAC) approval (18). An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®).FISH AND SEAFOOD 7. 7. and while simple and relatively inexpensive to operate. 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. other derivatives have been reported. Since its discovery. as well as other parts. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). The mouse bioassay has been in place for decades and the method has been standardised. TTX most commonly occurs in many species of puffer fish. Taiwan. While the chemical detection methods are most promising for the future. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. 23. After some development and various trials. Thailand. and islands of the South Pacific. 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.

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

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

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

and dinophysistoxin-2 (DTX-2) (Figure 7. However. as can LC/MS. and consequently were often overlooked as the real source of the toxins.3 Detection methods DA is water soluble and can be extracted from shellfish tissue by the acidic PSP extraction method although this may result in some decomposition. The benthic Prorocentrum spp. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). In the context of this chapter. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. and these methods have been reviewed (40. 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.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47.FISH AND SEAFOOD 7. DTX-2 was discovered in contaminated Irish shellfish (46). though this method is most commonly used to analyse plankton and seawater (42).4.. Since then. 48. Dinophysis spp. California.. and this is typically the method of choice.4). are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. but just over 15 years ago a third toxin. 49). Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. dinophysistoxin-1 (DTX-1). are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). 7. australis. and an immunosensor method has also been reported (4). 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. the heterotrophic dinoflagellates Dinophysis spp. in Monterey Bay. Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. although in the latter study the probes for Pseudonitzschia pungens. 41). To further complicate matters. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . produce a variety of other 148 . Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). 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). Alternatively.

This was probably not the first DSP episode. are also recognised as a threat to human health.4. While these esters are not toxic in vitro. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. In this case Dinophysis was suspected as the culprit organism. Dinophysis fortii. and Prorocentrum belizeanum. which include the pectenotoxins and the yessotoxins. The parent acids are the only DSP congeners to be found in shellfish tissue. In comparison. and include Dinophysis acuminata.1 Origins and distribution The first member of the group was identified in contaminated Japanese shellfish in 1985 and DSP toxins continue to be a threat in those areas. Dinophysis caudata. Today over ten Dinophysis spp. Prorocentrum maculosum. Dinophysis acuta. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish..5. DSP toxins produced by benthic Prorocentrum spp. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. 149 . Fig. DTX-5. such as Prorocentrum lima. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48).FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. Lately these compounds. 7. 49). and Dinophysis norvegica among others (47. 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. 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. are recognised as DSP toxin producers. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. Structure of okadaic acid and the two related DSP toxins 7.

Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. even with the minimal of clean up steps (48). A regulatory level of 80 . it does not provide information on the DSP toxin profile.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. New Zealand and Chile. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. DSP episodes have been reported from Japan. DSP-Check is a commercially 150 . The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 .3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. and abdominal pain. DSP episodes have been reported worldwide and are included in many monitoring programs (47). 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.5. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. Human symptoms include diarrhoea.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. various European countries. both critical enzymes involved in many key metabolic pathways in mammalian cells (50) and consequently affect a host of other biological processes in a cell including tumor promotion. 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). nausea.5. Since the late 1970s.3 days.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. which can lead to an underestimation of the total DSP content in a sample. vomiting. 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). 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. and DTX-2. 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. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). and while this provides critical seafood safety information. and a detection limit of 100 µg/kg has been reported (12). Due to the lipophilic nature of the toxins. 7. 7. In recent years. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996.

FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). 58). 7. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . Fig. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. for many years these compounds were included in the DSP toxin complex.5. and another rapid antibodybased test kit for DSP toxins has also been reported (53).5 and 7.6. but with the increased availability of pure toxins permitting detailed toxicology studies it is now recognised that both these groups of compounds possess a different toxic profile from the DSP toxins. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. 7. with good agreement with the mouse bioassay and LC–based methods (54. with a claimed detection limit of 20 µg/kg. Indeed. In yet another approach. Some representative examples of both groups of toxins are shown in Figures 7. 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. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57.6 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). 55). Within this group. However. 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.

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

7). Their toxicology and mechanism of action remain uncertain. the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present. Some of the PTXs are potent cytotoxins in vitro (60). For many years it was believed that 153 .FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. which aids detection by LC/UV. but various congeners are acutely toxic in the mouse bioassay following IP injection. it may be compromised by other biological materials in the matrix. some of which are found as glycosides and sulfates. but less so following oral dosing (61).6. Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. While LC/UV detection is possible for PTXs. Although there is no evidence that yessotoxins are responsible for human illness. chemical analysis is best carried out by LC/MS and several approaches have been described (57.3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue. Because of the number and complexity of the YTX group. The structures and chemical data including the molecular weight of each member have been collated (56). 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 YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. 64). and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. though its cell effects may be linked to modulation of calcium channels (62). which is further compounded by differences in the experimental design. 58). 7. a regulatory level of 1 mg/kg shellfish tissue has been set. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7. However.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. Many YTX toxins contain a conjugated diene system. The YTX group has not been reported to cause human illness. Estimates of acute toxicity of the parent compound following IP injection vary considerably. the PTXs show considerably less toxicity when administered orally. but a fluorescent derivative has been prepared for others. 7. There are no clear cases of human illness resulting from ingestion of PTXs (59). The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds. The crude extract may be used directly in the mouse bioassay.

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

pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. The mouse bioassay is used as the primary detection method. The mean time to onset of symptoms is around 3 hours. 69). vertigo. 7. In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. nausea. with a mean duration of symptoms of around 1 day. the competitive ELISA method correlated most closely with the mouse bioassay results. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. malaise. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65.7. The brevetoxin metabolites are reported to be less toxic than the parent compound (70).7. which can result in a cascade of associated events in a cell (66).FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). and mammalian body fluids without pretreatment or dilution (72). 7. with hot spots in the coral reef areas of the Caribbean and French Polynesia. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. and diarrhoea.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). and an action level has been set at 800 µg/kg (or 80 µg PbTx-2 equivalents/100 g or 20 MU/100 g or 4 µg/mouse) of shellfish tissue to trigger regulatory action. The human health implications following exposure to NSPs have been reviewed (69). 66. While the mouse bioassay continues to be used. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . 7. and in severe cases seizure. Ingestion of brevetoxins results in a variety of symptoms including parathesia. In this application. shellfish homogenates. this family of toxins has proven to be a challenge both to identify and monitor.

1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins. These include species belonging to the genera Ostreopsis and Coolia. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7. but more are suspected to exist. Fig. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red.8).FISH AND SEAFOOD transferred through the food chain to the larger carnivores. a group of closely related polyethers that are considered the source of ciguatoxins. green and brown seaweeds and also occur in sediments and coral debris. CTXs are large. 7. lipid soluble polyethers. complex. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin.8.8. and only a few structures have been identified. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. The diversity of benthic phytoplankton that exist on reefs further complicates the picture. A detailed comprehensive review of ciguatoxins has recently been published (74). although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins. Distribution through the food chain begins with 156 .

Nevertheless CTX poisoning continues to be a threat to local and visiting populations. which in turn are consumed by carnivores such as snappers. although not all CTXs are equipotent. sharks. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. weeks or even months (74 . arthralgia. and in fact CTX poisoning is often only identified after consumption of contaminated fish. and it is estimated that ingestion of as little as 0. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. and the sensation of loose teeth (77). and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue.8. Like the brevetoxins. 7. The onset of symptoms can occur rapidly within 24 hours.8. where cold objects feel hot and vice versa. and temperature sensation reversal. Another symptom that can develop over time is extreme fatigue. Even when suspect samples are available.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. myalgia. which can be confused with chronic fatigue syndrome. Patients have reported symptoms lasting weeks and even months.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. barracudas. 78). the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. 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. Other symptoms include a metallic taste. and moray eels. there are neither standards nor an official testing program. Local island populations attempt to avoid poisoning by catching only smaller fish. which can return following the consumption of alcohol and certain foods such as oily fish (74. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . 7. groupers. and there are suggestions that such incidents are increasing and spreading worldwide (74).1 µg of ciguatoxin can result in human illness (74. all of which can be linked to the effect of CTX on excitable membranes. and although the gastronomical problems may dissipate within a day or so. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. the neurological symptoms can persist for days. pruritus. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. Globally. only trace amounts of CTXs can cause illness following consumption of contaminated fish.76). they bind to voltage-dependant sodium channels in the cell membrane causing them to open. These include paresthesias (numbness and tingling of the extremities).3 Detection methods As a consequence of their potency. 75. 75).

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

9. AZA occurrence has been reported by several European countries including France. 88. cassipes (83). However. 89). and off the coast of Morocco (83. and to date this has been the only source from which these toxins have been found at toxic levels. 90).3 days.9. The bivalves affected include mussels (Mytilus edulis.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. and some time after the original episode in Ireland. these molluscs were never considered to be the biogenetic source. Mytilus galloprovincialis). and most recently in a crustacean (Cancer pagurus). Portugal. scallops (Pecten maximus). There has been considerable debate over the source of azaspiracids. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). which are ingested by P.FISH AND SHELLFISH TOXINS Fig.9. Scandinavia. but current data indicates that unlike the DSP compounds.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. Donax spp. as in other cases of shellfish toxins. which can persist for 2 . While this result is not necessarily in debate. clams (Tapes philipinarum. 7. cassipes produces AZAs in culture. Structure of the three principal azaspiracids found in shellfish 7. cockles (Cerastoderma edule). vomiting.). Ostrea edulis). and to date no long-term effects have been recorded (83. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. diarrhoea and stomach cramps. In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. Ensis siliqua. 7. AZA toxins do 159 . oysters (Crassostrea gigas. 84. These symptoms are not unlike those experienced following the consumption of DSP toxins. it has been impossible to establish that P. Following the original episode.

Using the same approach. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. 92). Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. but all contain a spirocyclic or cyclic imine function within their structure. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. it is recommended that the entire shellfish be used for AZA analysis since it has been shown that these toxins can migrate to other tissues. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. abnormalities in the thymus and spleen. 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. and erosion and bleeding in the stomach. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). parenchymal cell pyknosis in the pancreas. though the possibility exists that they may inhibit other phosphatases. However. Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83.12). Based on IP minimum lethal dose data. 7. and closely related ptierotoxins (Figures 7. Importantly these pathological outcomes were reported as being different from those caused by DSP. and ASP. the pinnatoxins.9. which are particularly amenable to detection by positive ion electrospray mass spectrometry. 7. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 .10 Cyclic imines These toxins are represented by several distinct structural types. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. AZAs also display cytotoxicity towards various mammalian cell lines (83. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. the spirolides. 91. Once again.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. 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). The mouse bioassay has been used to monitor shellfish extracts. PSP. 85). 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.10 – 7.

two additional congeners. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94.FISH AND SHELLFISH TOXINS health. Subsequent taxonomic studies revised this again to a new dinoflagellate K. cockles. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure.10). they are sometimes referred to as emerging toxins of uncertain human health concern. Some time later.10. selliformis (97). 95). Structure of the cyclic imine gymnodimine 7.1 Origins and distribution The original observation of toxicity in oysters led to a wider survey of other shellfish species in New Zealand when it was discovered that the toxin was present in a variety of bivalve molluscs including mussels. 95).520 Da (94.10. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7.. In only two cases (gymnodimines and spirolides) has a planktonic source been identified.1.1 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. Fig. 161 . 7. it was proposed that a dinoflagellate identified as Gymnodinium spp. scallops. surf clam and New Zealand abalone. At the time of the initial observation of gymnodimine. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin. 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). The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 .10. 7. but they are included here for completeness. Pacific oyster.

FISH AND SEAFOOD 7.900 µg/kg). cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. In particular. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. it has been stated that regulation against gymnodimine is not required (98).to10-fold reduction of toxicity (LD50 600 .100). Oral administration of the toxin to mice by gavage resulted in about an 8. finishing with 100% methanol. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . Reduction of the imine group eradicated the toxicity. together with a lack of evidence of human intoxication. and this is the detection method of choice. Most cyclic imines lack an extended chromophore. Since their original discovery which resulted in the characterisation of spirolides A-F (99.1. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection.3 Detection methods The characteristic “fast-acting. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine.10. 95) (Figure 7. abdominal breathing.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. 7. Whether isolated from dinoflagellate cells directly or from shellfish tissue. and lack of responsiveness to stimuli. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach.10. the first step in the process is typically an initial clean-up step using a short C18 cartridge and elution with water containing increasing amounts of methanol. The time lapse between ingestion and death is between 5 and 15 minutes. 7. 50%) of 96 µg/kg.1.2 Toxicology A review of the toxicology of all cyclic imines has been published (98).10. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. Based on this diminished oral toxicity. There is a rapid onset of symptoms including respiratory distress. They were originally discovered in extracts of mussels (M. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. 95).11). almost a dozen derivatives in total have been identified to date (94. and so spectroscopic detection by UV methods is not favoured. A later study using purified toxin established an LD50 dose (Lethal Dose. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. paralysis of the hind quarters and eventually complete immobility. However.

the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). the gulf of Maine. in fjords in Denmark and Norway. by gavage (87 . 95).2. Interestingly. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). southern Ireland.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. or by oral ingestion (381 . They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94.2. Structure of four spirolide toxins A-D 7. All data reported here are for desmethyl spirolide C. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. 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.8 µg/kg).10.11. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 .1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. 7. While oral toxicity is considerably less than by IP injection. fasted mice).11. 7. the western coast of Scotland.166 µg/kg. those spirolides possessing the vicinal dimethyl grouping (see Figure 7. Fig. fasted mice).707 µg/kg.2 Toxicology The spirolides are the most studied group of the cyclic imines. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . R1 = CH3) of the cyclic imine ring are the most toxic of the group (98).10.

To date. and in fact these compounds share a number of other structural features. four members of the group have been identified – two of them being the C-34 diastereomers. consistent with modulation of acetyl cholinergic receptors (103). Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. The compounds are zwitterionic and therefore somewhat more polar than the spirolides.2. A number of methods following this procedure have been described (94. the mouse bioassay can be used to detect these fast acting toxins. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C. 7. 95.12). The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. Fig. Three members of this group have been reported.B contain a spiro imine moiety similar to that found in the spirolides. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . 7. However.3 Pinnatoxins and pteriatoxins Pinnatoxins A . which includes two diastereomers. 7.10. While the mechanism of action remains to be clearly defined.12.3 Detection methods As discussed for gymnodimine.10. more useful information on the amount and distribution of toxins present in a shellfish extract is obtained by LC-MS analysis usually following a simple clean-up step.FISH AND SEAFOOD (100). 102).

The LD99 (Lethal Dose. which in many cases are now found to be more widespread than previously believed. However. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. like the gymnodimines and spirolides. new toxin groups such as the azaspiracids and cyclic imines have also been reported. Despite this progress. but this remains to be conclusively established (106).FISH AND SHELLFISH TOXINS 7. to date action or 165 .10. While this is recognised as a real possibility for a wide range of toxins. and more advanced means of detection. which occupy tropical waters around Okinawa and the south east coast of China. 7.100 µg/kg). 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. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). There is strong circumstantial evidence to suspect that these toxins.most likely a dinoflagellate.3. 99%) values for pinnatoxins A-C are in the range (22 . With increasing knowledge of the chemistry of toxins. have long been suspected of causing human illness after ingestion.10. The LD99 dose for the pteriatoxins is reported to be in the range (8 . 7. while pinnatoxin D was essentially inactive. Coupled with this. arise from an algal source . 7. This is generally a result of increased monitoring and continually improving detection technologies and methods. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other.2 Toxicology There is little information describing the toxicology of these compounds (98).3. there is no compelling evidence to suggest that they have resulted in human toxicity (98). Only the acute toxicity in mice following IP injection has been reported. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins.3.180 µg/kg).11 Conclusion There has been considerable progress in the detection of known marine toxins.10.1 Origins and distribution Shellfish of the genus Pinna.3 Detection methods No formal detection methods for these compounds have been reported. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures.

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

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

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


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

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


procedures. 6. The team should also include 177 . Figure 8. Validation . Establish procedures for verification to confirm that the HACCP system is working effectively. operation or stage in the food chain including raw materials.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . 4. Verification .A point. food microbiology. 7. Establish a system to monitor control of the CCP. Determine the CCPs. food chemistry or engineering.Obtaining evidence that the elements of the HACCP plan are effective. Establish the corrective action to be taken when monitoring shows that a CCP is not under control.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. 3. procedure. tests and other evaluations. quality assurance or control. 8. 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. Establish documentation concerning all procedures and records appropriate to these principles and their application. from primary production to final consumption.1 Assemble HACCP team HACCP requires management commitment of resources to the process. ingredient and packaging buyers. 2. 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. Establish the critical limit(s). in addition to monitoring to determine compliance with the HACCP plan.The application of methods. Conduct a hazard analysis.3. Each stage is discussed below in detail. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. 8. hygiene managers. 5.1 is a flow diagram illustrating this 12-stage logic sequence.

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

etc. or in aquaculture operations. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . there may be species-specific hazards related to certain fish and shellfish. heat treatment. It is also important to identify the consumer target groups. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team.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. 8. freezing. contaminated fish and shellfish should not be used. In addition. Expertise on such inherent hazards. physical and chemical structure (including water activity (aw).). The description should include information such as the product name.g. marine toxins associated with particular fish and shellfish species.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. However. may need to be considered specifically. naturally present on fish from warm waters (> 15 °C).3.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora.2 Describe the product It is important to have a complete understanding of the product. Vulnerable groups of the population. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation. which should be described in detail.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.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.1. etc. storage and distribution conditions and instructions for use.3. is a cooking process required?). processing conditions (e.3. 8. packaging.3. such as enteric viruses in shellfish. fermentation. including practices onboard fishing vessels. 8. aquaculture drug residues and potentially pathogenic Vibrio spp.3. If the processing does not include a kill step. These factors are likely to have a significant effect on some potential hazards.3. pH. shelf life. such as children or the elderly. composition. 8. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments.g.

It is also important to include observation of production outside normal working hours such as shift patterns and weekend working.3. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. time/temperature data. ingredient specifications. 8. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. 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. However. it is essential that the details of each step are fully appreciated and recorded. it should also include details of CCP steps. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised.3.5 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. survival or multiplication of pathogenic microorganisms. the qualitative and/or quantitative evaluation of the presence of hazards. cleaning and hygiene procedures and storage conditions. 8. It may include details of the factory and equipment layout. once determined. 8. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. features of equipment design.FISH AND SEAFOOD product.3. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects.4. 180 ● ● .1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. If HACCP studies are applied to proposed new process lines or products. because the hazard analysis and decisions regarding CCPs are based on these data. 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. For example. conduct a hazard analysis. and therefore a detailed description of the procedure applied must be available. Ideally.6 List all potential hazards associated with each step. It is essential that the diagram is accurate. or circumstances of any reclaim or rework activity. The operation should be observed at all stages.

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

7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. but for which no control exists at that step or any other.3. it is not possible to generalise on likely CCPs. Criteria used to set critical limits must be measurable and may include physical. 8.8. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively.7. 8. More than one critical limit may be defined for a single step. 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 . chemical.1 Fish and seafood Again. However.2) to provide a logical. Where possible.3. it is necessary to specify both time and temperature for a thermal process. critical limits should be specified and validated for each CCP.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. and the reader is referred to the appropriate product chapter.3. If a significant hazard has been identified at a step where control is required for safety.3. application of the decision tree should be flexible. and its use may not always be appropriate. However.FISH AND SEAFOOD 8. then the process must be modified to include a control measure. 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. given the enormous variety of fish products and processes in use. structured approach to decision making. For example. it can be said that effective control measures are likely to include the following: • • • • • 8. biological or sensory parameters.8 Careful selection of sources for raw materials Adequate temperature control Effective sanitation Food handler hygiene Prevention of cross-contamination Establish critical limits for each CCP Critical limits separate acceptable from unacceptable products.


• • • 8.3.9

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.


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

EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr. to sale or supply to the final consumer.9. A number of standards are provided in food legislation. and one that plays an important role in most countries’ food legislation. 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. transport and delivery vehicles requirements. The new legislation was designed to establish conditions. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. Hygiene is a general concept that covers a wide subject area. including microbiological criteria. eliminate or acceptably control pathogen contamination of food. from structural conditions in the factory or process facility. under which food is produced to optimise public health and to prevent. final product specifications. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . have been replaced by several linked measures on food safety rules and associated animal health controls. and conditions of raw materials. however. 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. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9.1 Introduction Hygiene is an important aspect of ensuring food safety. the existence of microbiological standards alone cannot protect consumer health. to personnel requirements. with the intention of introducing consistency and clarity throughout the food production chain from primary production.

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. N. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No.15). 190 • • . Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No.1991. distribution. transportation.2 Legislative Structure From 1 January 2006. 41). Subsequently. as last amended by Regulation (EC) No 806/2003. processing. as last amended by Regulation (EC) No 806/2003. p. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. p.9.7. and sets out what those enforcing the provisions have to do. Council Directive 91/493/EEC of 22 July 1991 laying down the health conditions for the production and the placing on the market of fishery products (OJ L 268. 9. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption Regulation (EC) No. 24. p.B. J.1991.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. 24. European Communities L 268. 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. handling and offering for sale or supply to the consumer.1). packaging. A number of more detailed implementing and transitional measures have been adopted at EC level. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. manufacturing. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. storing. 7.9.1992.

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



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:


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

1 Identification marking An identification mark needs to be applied following the requirements of Annex II.4. EC. when a product's packaging and/or wrapping is removed or is further processed in another establishment. EG. (iv) If the identification mark is applied in an establishment within the Community. (iii) Additionally. ES. please refer to Regulation (EC) 853/2004.2. EK. the code would be UK. EÜ. EY. the food business operator may apply the same identification mark to both types of food. For the UK.4. the mark must show the approval number of the establishment. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not.2 Fishery product requirements 9. In this case.2. the new mark must show the approval number of the establishment where this occurred. EO. EF. 198 .1. (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. EK.1. (ii) However.FISH AND SEAFOOD 9. Section I of Regulation (EC) 853/2004 are given as follows: 9. for other codes.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.4. Section I of Regulation (EC) 853/2004. 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.2. and the characters easily decipherable and be clearly displayed.2 Form of the identification mark (i) The mark must be legible and indelible. a new mark must be applied to the product. 9. the mark must be oval shaped and include the abbreviation CE.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. Extracts of the requirements of Annex II.

i. for full requirements. 9. and fishery products carried in bulk.2. Chapter IV. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. Therefore. Chapter III. (v) When the mark is applied directly to products of animal origin. the colourings used must be EC authorised colours. With the exception of Chapters I and II.2. it applies to those animals when not placed on the market live. Parts A. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 .3 Method of marking (i) The mark is to be applied directly to the product. and are intended for further handling. the approval number of the establishment and the abbreviation for the community where appropriate. including vessels. The requirements of this section accompany those in Regulation (EC) 852/2004.4.1. it is sufficient to apply the mark to the exterior of that package only. they would need to be obtained following Section VII on live bivalve molluscs. a) requirements for establishments. granulate and powdered products of animal origin carried in bulk. its wrapping or the packaging. (iii) In the case of liquid. specifically relating to fishery products are given below.LEGISLATION 9. (ii) For products of animal origin that are placed in transport containers or large packages.4. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. This section does not apply to bivalve molluscs. as amended. tunicates and marine gastropods when placed on the market live. Section VIII of Regulation 853/2004. or be printed on a label affixed to the product. processing. wrapping or packaging in another establishment. C and D. reference should be made to the actual Regulation. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. the mark may be applied to the outer surface of the container or packaging. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. echinoderms.e. Extracts of the requirements of Annex III. Part A and Chapter V apply to retail. the wrapping or the packaging.

There is a derogation from point a. bleeding. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products. i.) 9. fishing and collection of live fishery products with a view to their being placed on the market. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. if a vessel has water coming in that is used with fishery products. The definition of fishery products covers primary production as well as its associated operations.1 Chapter I . it must be placed somewhere that will not contaminate the water supply.. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 . the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking. from the place of production to the first establishment of destination. gutting. (a) primary production covers the farming. refrigeration and wrapping. A. 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. removing fins.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. or to handle or process them after harvesting. clean seawater may be used for the handling and washing of fishery products.2. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material. Structural and equipment requirements refer to the following points 1.FISH AND SEAFOOD c) for the water supply. Chapter VII of Regulation 852/2004.2. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter.e. this supplements the requirements of Annex II. heading. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products.4.

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

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

(iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. handling fishery products A. Chapter I. 9.2. 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. packaged and chilled as quickly as possible after preparation. dispatched. (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. including vessels. prepared or processed immediately after arrival at an establishment on land. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed.3 Chapter III . there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. They should be wrapped. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. 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 establishments. and where necessary. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice.LEGISLATION consumption. they must be stored under ice in appropriate facilities.2. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. B. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . part I. C. 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. Additionally. C. (ii) Heading and gutting must be carried out hygienically. points 1 and 2.4. 854/2004. Fillets and slices must then be removed from work tables as soon as possible after preparation.

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

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

Fresh. E. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing.2. In the case of whole frozen fish in brine intended for the manufacture of canned food. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption. food business operators must ensure that wrapping material is not contaminated and is stored in such a way that it is not exposed to contamination and is easy to clean where necessary.Wrapping and packaging of fishery products (i) Containers in which fresh fishery products are kept under ice must be waterresistant and ensure that melt water does not remain in contact with the products. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum.6 Chapter VI .7 Chapter VII . (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product.4. 9.2.FISH AND SEAFOOD D. Fishery products contaminated with parasites may not be placed on the market for human consumption. Diodontidae and Canthigasteridae. 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. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae.4. it should be kept at no more than -9 °C.2. and when fishery products are wrapped on board fishing vessels.2. prepared and processed fishery products belonging to the family Gempylidae. 9. thawed unprocessed fishery products. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. Molidae. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. The scientific name must accompany the common name on the label. 206 .Storage of fishery products (i) Fresh fishery products.

LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. (ii) If fishery products are kept under ice. species belonging to the Gadidae family. (d) 60 mg of nitrogen/100 g of whole fishery products used directly for the preparation of fish oil for human consumption as referred to in the second sub-paragraph of Part B(1) of Chapter IV of Section VIII of Annex III to 207 .2. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar.2.4. 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. 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. provided that the journey is short and the competent authority allows this. melt water must not remain in contact with the products.2.). This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. 9. The temperature of frozen fishery products must not fluctuate above -15°C. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. Helicolenus dactylopterus.Transport of fishery products The above points for Chapter VII apply in relation to transport. 9. Sebastichthys capensis. 2074/2005.3 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. species belonging to the Merlucciidae family.8 Chapter VIII .

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

C. 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. 209 .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. processing or relaying. poles or other fixed means are to identify the boundaries of the sites.Hygiene requirements for the production and harvesting of live bivalve molluscs A.Class A means they can be placed directly on the market . 9.2 Chapter II . Requirements for production areas (i) The competent authority classes (classed 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. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified.4. B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with. 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. Transport needs to allow sufficient drainage for best survival conditions.3.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market . collection from: . (ii) Provided the requirements of chapter V are met.

3 Chapter III . requirements if there are many batches of live bivalve molluscs.FISH AND SEAFOOD 9. shells of live bivalve molluscs are to be washed throughout with clean water and the molluscs are to come from one of the following: (i) Class A production area (ii) Relaying area (iii) Purification centre (iv) Another dispatch centre 9.4.3. the purification system to allow them to resume /maintain filter-feeding activity. Before dispatch. 210 . 20 milligrams of domoic acid per kilogram (iii) okadaic acid. 1 milligram of yessotoxin equivalent per kilogram.3. (iv) yessotoxins. containers and a label to identify purification. dinophysistoxins and pectenotoxins together.Hygiene requirements for purification and dispatch centres A.4 Chapter IV . B. Requirements for dispatch centres Handling should not result in contamination to any degree.4. 800 micrograms per kilogram. for: (i) paralytic shellfish poison (PSP).Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc.5 Chapter V . 9. the levels of marine biotoxins should not exceed the following limits. (ii) amnesic shellfish poison (ASP). and there are requirements for tanks and water storage containers. Requirements for purification centres There are requirements for live bivalve molluscs to be washed.4.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements.3. 160 micrograms of okadaic acid equivalents per kilogram.

The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction. 9. 9.3.9 Chapter IX . must be waterproof.7 Chapter VII .LEGISLATION (v) azaspiracids. dispatch centre or processing establishment.4. including the identification mark. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. 9. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.1). Registration document requirements also apply. by way of derogation from directive 2000/13/EC.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.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards. 211 .2. 160 micrograms of azaspiracid equivalents per kilogram. the label.8 Chapter VIII . Requirements of Chapters III and IV are to be complied with. Finally.4.Identification marking and labelling In addition to general identification marking (contained in 9.4. (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.4.6 Chapter VI . Also. the species of bivalve mollusc and date of packaging (at least the date and month) are required.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. they cannot be re-immersed in or sprayed with water.

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

5. In the remaining 10 % of samples. 213 . three dilutions MPN test specified in ISO 16649-3.2 This states official controls for the production and placing on the market of live bivalve molluscs. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. live tunicates and live marine gastropods. coli per 100 g of flesh and intravalvular liquid. The reference method for this analysis is the five-tube. Monitoring of classified relaying and production areas This is to ensure there is no malpractice.LEGISLATION - the competent authority considers that they may constitute a risk to public or animal health or are for any other reason not suitable for human consumption. coli per 100 g of flesh and intravalvular liquid. The reference method for this analysis is the five-tube.Official controls concerning live bivalve molluscs from classified production areas A. Sampling plans and frequency are required.1 Chapter II . coli per 100 g of flesh and intravalvular liquid. section VII. Classification of production and relaying areas: This section reinforces that specified in Annex III. chapter V is reinforced. Also.5. - - - - B. in 90% of the samples. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated.2. Live bivalve molluscs from Class C areas must not exceed 46.000 E. Regulation (EC) 854/2004. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. live bivalve molluscs must not exceed 46. as previously stated. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. live echinoderms. that microbiological criteria and chemical contaminants amongst other requirements are met. 4.600 E. Annex II 9. Live bivalve molluscs from Class B areas must not exceed. 000 E. A summary is given as follows: 9.

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

4 referring to fishery products. with chapter 2. a batch of foodstuffs or a process. and the legislation applies to all businesses involved in food production and handling.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.II. area or batch. Two kinds of criteria have been established: food safety criteria. 9. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process.6. presence or number of microorganisms. The criteria are outlined in Table 9. per unit(s) of mass. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product.I. The relevant criteria are outlined in Table 9.6. 9. based on the absence.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. 215 . volume.LEGISLATION The aim of this legislation is to complement food hygiene requirements. and/or on the quantity of their toxins or metabolites. ensuring that foods being placed on the market do not pose a risk to human health.

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

25 Live bivalve molluscs and live echinoderms. coli (15) 1.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1.27 Fishery products which have undergone enzyme maturation treatment in brine. tunicates and gastropods 9 (18) 2 E. 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 1.17 Live bivalve molluscs and live echinoderms. manufactured from fish species associated with a high amount of histidine (17) Histamine .Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.

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

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

coli 5 shucked products of cooked crustaceans and molluscan shellfish 2 100 cfu/g 1000 cfu/g EN/ISO 6888-1 or 2 FISH AND SEAFOOD Coagulasepositive staphylococci 5 Improvements in production hygiene 220 .II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2.4.1 Shelled and E.TABLE 9.

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

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

9. 2073/2005 on microbiological criteria for foodstuffs (OJ No.2004. 30.10. 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. 30. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. 55). L226. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. L338.6. as amended. 2076/2005. 2075/2005 and Regulation (EC) No.2004.6. 25. 2076/2005. 1).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. L139. namely. 2074/2005. 1441/2007. 2074/2005. L283. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. L226. L139. The revised text of Regulation (EC) No. 852/2004 is now set out in a Corrigendum (OJ No. 854/2004 is now set out in a Corrigendum (OJ No.4. Regulation (EC) No.2004. Regulation (EEC) 2136/89. as read with the corrigenda at OJ No. these are not be discussed here. laying down common marketing standards for preserved sardines.2004. 22) as amended by Regulation (EC) Nos. The revised text of Regulation (EC) No.2004. 1663/2006. 2076/2005. 223 . 83) as amended by Regulation Nos 882/2004. L226.10 Further Reading Regulation (EC) No. 2074/2005.4.2006. Regulation (EC) No.LEGISLATION 9. 1688/2005.6. 206). Regulation(EC) No. 14.2005. 2076/2005.4. 1662/2006. Regulation (EC) No. 22. Commission Regulation (EC) No. 3) as amended by Regulation (EC) No. The revised text of Regulation (EC) No. Regulation (EC) No. 1.12. 853/2004 is now set out in a Corrigendum (OJ No. 25. 62) as amended by Regulation (EC) No. 30. 1019/2008 and as read with Regulation 2073/2005. Regulation (EC) No. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. L139. 2074/2005 and Regulation (EC) No. 25. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito.2004.

as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.2005. (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004 ( OJ L 338. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. 6) 224 . 23.10.2001. 22. 21. 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 854/2004.12.1.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. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17. 22.2005.2000. 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 278.12.

0 . The optimum growth temperature is 28 °C (2). 225 .3 . Aeromonas has the potential to grow over a pH range of ca 4 .1.5. a 10 to 1.10. Morphology Gram-negative rods.1 Aeromonas spp.1.3 minutes.5 µm 10.1. under optimal incubation temperatures.3 Temperature Aeromonas grows at temperatures of 2 .1.1.000-fold increase in number is observed after one week in food systems (1).3.10 (3).5.2 Oxygen requirements Facultative anaerobe.5 minutes. 10. Some death will occur at this pH if the temperature is 4 °C. no growth is observed (1).4 Heat resistance Aeromonads are sensitive to moderately high temperatures.1. and 2.5 pH Under otherwise optimal conditions. 0. At 5 °C.0 x 1. 10. 10. The minimum pH value for growth is thought to be pH 5. respectively (1).1 10.45 °C (1). PATHOGEN PROFILES 10. and D-values at 45 and 51 °C have been reported as 29. At pH 4.

which may be inactivated by heating at 80 ºC for 20 .2. E and. Types A. and groups I and II are those responsible for the botulism cases. All C.7 . 10.3 Temperature All strains of C.30 minutes. 0. All type A strains are proteolytic and type E strains are usually non-proteolytic. Group I will not grow at temperatures of 10 ºC or less. A to G. but the interior of the food may do so. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C. spore-forming rods. botulinum grow reasonably well in the temperature range of 20 45 ºC. 10. but Group II strains are psychrotrophic. 10. forming at least seven different toxins.2.22. 226 . and is sensitive to > 4. Thus an aerobically packed product may not support the growth of the organism on the surface. B.5% NaCl (4).2 Oxygen requirements Although C.2% sodium chloride (NaCl) (1). 10.even as low as 3 ºC (7). F are the types that are responsible for most cases of human botulism (5.1 Morphology Gram-positive.4 Heat resistance The vegetative cells of C. to a lesser extent. 6).5 . 10.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . botulinum types produce heat labile toxins. There are four main groupings of the organism. or at 90 ºC for a few seconds. types B and F can be either. being capable of slow growth and toxin production at low temperatures .2 Clostridium botulinum Seven different types of C.2.FISH AND SEAFOOD 10.2. botulinum are known. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth.1. botulinum is an obligate anaerobe. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different.4 x 1.0 µm. at 85 ºC for 5 minutes.2. but the spores of this organism are more so. botulinum is prevented. botulinum are not particularly heat resistant.

g. perfringens . where oxygen levels are reduced. pH and temperature. therefore. they may survive mild heat treatments (70 . grow on the surface of foods unless they are vacuum or gas-packed.0 . botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. parcooked meals with pH value > 5.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. botulinum spores The most heat resistant spores of Group I C.3 10. However.7 Characteristics of C.0. 9).is an obligate anaerobe. In commercial situations. 10. particularly by cooking. These concentrations correspond to limiting aw of 0. botulinum depends on solute.2 Oxygen requirements C.97 for Group II (5). vacuum packed. Note also that if humectants other than salt are used.97) (10.94 for Group I and 0. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. safety margins must be introduced to allow for process variability.2. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains.6 and pH µm. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.PATHOGEN PROFILES 10.3 . It will not.1.9 x 2.2.21 minutes) (9).1 Clostridium perfringens Morphology Gram-positive spore-forming rods. 227 . 10.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. 10.like other clostridia . These values have been established under carefully controlled laboratory conditions. the minimum aw for growth may be lower. 11)). 0.10. 10. respectively (8.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. The organism will grow well in the centre of meat or poultry dishes.2.0 and aw > 0.

depending on strain and other conditions.3.3. (It is unusual for spores to be formed in foods after the growth of this organism (14)).50 ºC. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures.93 and 0.10 ºC. perfringens is not tolerant of low water activities. 13). perfringens in vacuum packed beef (18).3 Temperature The most significant characteristic of C. 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. 10. perfringens in roast beef at 60 ºC is 14.3. and their heat resistance is also affected by the heating substrate. perfringens has the potential ability to grow within the temperature range 15 .4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. 16). perfringens is not a tolerant organism with respect to pH. perfringens spores The spores of C. pH.3. While some growth can occur at 50 ºC. Spores. 15. As in the case of other factors limiting the growth/survival of this organism. perfringens appears to be 0. Under otherwise ideal conditions. the enterotoxin is not heat resistant . The D-value for C. lower concentrations may be effective in combination with other factors. At cold temperatures. etc. Salt concentrations of 6 . however.it is destroyed by heating at 60 ºC for 10 minutes (14. perfringens strains. Some studies indicate that the presence of 3% NaCl delays growth of C. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. Recorded heat 228 .45 ºC. although C. perfringens can vary quite considerably in their heat resistance. It grows best at pH values between 6 and 7 (the same pH as most meats). Its optimum temperature for growth is 43 .97 (glycerol and sucrose respectively) depending on the solute used to control aw (13.5 pH C. type of solute. 13). perfringens. the limits for aw are affected by temperature. vegetative cells die rapidly (12). 10.7 Characteristics of C. 0 . 18). 10.6 Aw/Sodium chloride C. In addition.8% inhibit growth of most C.FISH AND SEAFOOD 10.3. 10.5 minutes (17). will survive greater extremes of pH (and aw) (12.3.

3 is the lowest value where growth has been recorded. so can be destroyed by pasteurisation. the lowest limit for growth is likely to be considerably higher .4 ºC is probably a more likely minimum for growth in foods (19).23. being potentially capable of growing . monocytogenes is reported to be 45 ºC (20). monocytogenes in crawfish tail meat at 55. 10. 10. and temperature. D-values for L. In foods.albeit slowly . Morphology Gram-positive short rods. 10. 0.especially at refrigeration temperatures. Its optimum growth temperature. however. 1.2 has been suggested as the lowest working limit (22).4.4. 60 and 65 ºC were reported as 10.2.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic.64.5 pH The ability of Listeria to grow at different pH values (as with other bacteria) is markedly affected by the type of acid used. or even below 0 ºC. requiring days or weeks to reach maximum numbers. However.2 Oxygen requirements Aerobe or microaerophilic. the organism is able to grow at pH values well below pH 5 (pH 4.0 µm. -0. is between 30 and 37 ºC. respectively (21).0.4.1 Listeria spp.98 and 0.3 . It has been reported to have slightly greater heat resistance than certain other foodborne pathogens. however. using hydrochloric acid as an acidulant).at refrigeration temperatures down to.4 10. pH < 5. The upper temperature limit for the growth of L. 229 . monocytogenes is not a particularly heat resistant organism. growth at low temperatures can be very slow. and where acetic acid is used as an acidulant.8 minutes for heat sensitive spores (12).0 minutes for heat resistant spores to 1. it is not a sporeformer. 10.4 .19 minutes.4.4.4 Heat resistance L.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. 10.5 . Under ideal conditions.6 .2.5 x 0.

7 x 1. 10.8 .12% NaCl or more). 10. It grows best at aw of ≥ 0. 10. 25).1 Plesiomonas Morphology Gram-negative.45 ºC. 0.0 .0. 10.3 µm.0 µm.5. 230 . 0.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24). 10. Morphology Gram-negative short rods. 10.1. It is likely to survive.90.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24. but has been shown to be able to grow at an aw level of 0.5 .5.FISH AND SEAFOOD 10. or even grow.5. 10.5 10.0 x 1 .5 (24. peritrichous flagella.5.4.5 pH Plesiomonas has a pH range for growth of pH 4.5 .3 Temperature Plesiomonas can grow in the temperature range of 8 .6 Aw/Sodium chloride L.6 10. at salt levels found in foods (10 .1 Salmonella spp. monocytogenes is quite tolerant of high NaCl/low water activities.2 Oxygen requirements Facultative anaerobe.83 (20).6. The bacterium may survive for long periods at aw values as low as 0.8. 25). the optimum being 30 ºC (23).

therefore. 231 .6. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. Furthermore. Salmonellae are quite resistant to freezing. pasteurisation and equivalent treatments will destroy the organism under normal circumstances. or possibly 0. and appropriate heat treatments must be determined experimentally for low-aw foods. However. Growth is slow at temperatures below about 10 ºC. some strains are able to grow at temperatures as low as 4 °C (26).8 . Salmonella senftenberg 775W is about 10 .2 Oxygen requirements Facultative anaerobe.5 . The optimal pH for Salmonella growth is between 6.5 ºC. Salmonella has the potential to grow at aw levels as low as 0.48 ºC.3 Temperature Salmonellae can grow in the temperature range of 7 .20 times more heat resistant than the average strain of Salmonella at high aw (27). depending on the type of acid and temperature.93 (as reported in dried meat and dehydrated soup).0. substrate. 10. although salt tolerance increases with increasing temperature (28).945.7. Salmonellae are quite resistant to drying.9. 10.6.4% NaCl.5. 10.4 Heat resistance Salmonella is not a spore-forming organism. However. It is not. under otherwise ideal conditions. D60 ºC values normally range from about 1 to 10 minutes.5 pH Salmonella has a pH range for growth of pH 3. temperature and pH. The growth of Salmonella is generally inhibited by the presence of 3 . 10.6.6. with a z-value of 4 .6. the optimum being 35 . depending on serotype. Some death will occur at pH values of less than about 4. a heat resistant organism.PATHOGEN PROFILES 10.6 Aw/Sodium chloride Where all other conditions are favourable. strains vary in their ability to withstand heating. and with an appropriate acid.37 ºC.5.

5). 10.37 ºC (29). 232 . NB: The growth of Staph. aureus viability. 10. aureus than citric acid). but may cause some cell damage (30). 0. Staph.7. depending on aw. aw and temperature (sensitivity to acid increases with temperature).2 to 9. It can survive well at low temperatures. the organism is heat sensitive and will be destroyed by pasteurisation. the D60 ºC value is 2 .9 µm diameter.0.0 .7.3 Temperature Under otherwise ideal conditions.FISH AND SEAFOOD 10. 10. 10.7. aw. under otherwise ideal conditions (29. however.7 . etc. Under most circumstances. The following relate to limits for growth only.3 (optimum 7.20 minutes. aureus can grow within the temperature range 7 – 48.1 Staphylococcus aureus Morphology Gram-positive cocci.7. Most strains of staphylococci can grow within the pH range 4.). As is the case with other bacteria. aureus is more limited under anaerobic than aerobic conditions.7.5 pH The pH at which a staphylococcal strain will grow is dependent upon the type of acid (acetic acid is more effective at destroying Staph. stressed cells can also be less tolerant of heating.2 Oxygen requirements Facultative anaerobe.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH.5 ºC. Freezing and thawing have little effect on Staph. with an optimum of 35 . The limits for toxin production are also narrower than for growth. 31).7. fat content. In meat macerate.7 10.

especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.8. In general.8 10.4 .0 .86 is the generally recognised minimum in foods (32). pH.7. aureus can grow in 7 . Staphylococci are more resistant to salt present in foods than other organisms. but certain strains can grow in 20% NaCl. 34).>0.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5.10% NaCl. Staph. 10.2 . atmosphere.8.99 aerobically under otherwise optimal conditions. 0. strain.6 Aw/Sodium chloride Staph.7.1 Vibrio cholerae Morphology Gram-negative curved rods. 31) between 0.7. 10.8 x 1. * dependent on temperature.9.87 and > 0.83 . An effect of increasing salt concentration is to raise the minimum pH of growth. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.2. 233 .PATHOGEN PROFILES 10.0. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31.2 Oxygen requirements Facultative anaerobe.3 Temperature The optimum temperature for growth has been reported as 30 ºC. with a temperature range for growth of 10 . an aw of 0.99 little or no toxin production in anaerobically packed foods.7 Limits permitting toxin production 10 .5 .6 µm 10. It can grow over the aw range 0. In general. and solute.5) (29.43 ºC (35).0 (optimum 7. However. 10.8.

10. V. cholerae in crab meat homogenate at 60 ºC is 2. cholerae is sensitive to heat at temperatures of >45 ºC. 37. cholerae may persist on frozen fish for >180 days (36).2 Oxygen requirements Facultative anaerobe.4 Heat resistance V. cholerae can grow over the aw range 0. 38). with optimum at 7. there is evidence that V.8. 234 .1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. and it is sensitive to refrigeration temperatures (0 . 10. declining in number during storage.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media. under otherwise favourable conditions.0. 10.97 . with an optimum of 0.8. are 5 and 43 ºC.5 pH This organism is tolerant of high pH values.998.984 (35). it has a pH range for growth of pH 5 . The D-value for V.9.8.6 Aw/Sodium chloride V.FISH AND SEAFOOD Also. parahaemolyticus is easily destroyed by drying. It can grow in the presence of 0.9 10.0% NaCl (35). but not acid conditions. minutes (35).5 ºC). with an optimum of 37 ºC (39). respectively.6. 10. 10. but only moderately sensitive to freezing (40).1 . 10.6 (35.

9.10. parahaemolyticus will grow at aw levels as low as 0. with an optimum pH of 7.2 Oxygen requirements Facultative anaerobe.3 Temperature The optimum temperature for growth has been reported at 37 ºC. depending on solute.PATHOGEN PROFILES 10.922.10.4 Heat resistance V. 10.6 Aw/Sodium chloride V. 10. 235 . The D-values at 55 ºC for clam and crab homogenate are 0. parahaemolyticus is unable to grow in the absence of salt. with a temperature range for growth of 8 .5 minutes. 10. but the optimum concentration is 2 .98 (35.8. vulnificus is heat sensitive. 10.11.8 .6 (41).4 Heat resistance V.10. and 2.1 Morphology Gram-negative curved rods.43 ºC (42).0.6 . respectively (39).9. V. 10.5 pH The pH range for growth is 4.9.02 . it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43). and are readily destroyed by cooking. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.10 Vibrio vulnificus 10. The organism can survive for long periods at freezer temperatures (42). 41).10. 10.4% (39. The optimum concentration of NaCl equates to aw of approximately 0. parahaemolyticus are not heat resistant. 41).5% NaCl.29 minutes. respectively.

W.. Dodds K. 279-303.H.M. Principles of control.N. Eds.L. ASM Press. 9. 2001. Bier J. Dodds K. Gorham J.O.P. Marcel Dekker. Murrell K.. Volume 1.H. Gaze J. Smith J. Great Britain. vulnificus grows over the range 0.10. 6 (1). Stelma G. New York. Volume 1: Diseases Caused by Bacteria. 5-19. 97-131. 1989. International Commission on Microbiological Specifications for Foods. Eds.. Ed.. in Foodborne Disease Handbook. Doyle M.J. Miliotis M. 1-19.6 Aw/Sodium chloride V. Hui Y.. Kovacek K.L. Caister Academic Press.N. 7. 357-67.H. Clostridium botulinum and Clostridium perfringens. Ed. with an optimum pH of 7.11 1. in Foodborne Pathogens.K.A. 3.L. Marcel Dekker. Murrell K. in Microorganisms in Foods. Juneja V. 8. Eds.. Food Control. Stelma G.P.M. Hauschild A. Marcel Dekker. Gorman J. 2.5. in International Handbook of Foodborne Pathogens. 10. Hauschild A.. in Clostridium botulinum: Ecology and Control in Foods. References International Commission on Microbiological Specifications for Foods. in Foodborne Bacterial Pathogens. 2003. Cliver D..W.H. 1994. Beuchat L.5 pH The pH range for growth is 5 . Kim J. Eds. Novak J. 10..E. Foegeding P. with an optimum of 2.FISH AND SEAFOOD 10.. 383-408. 1993. Aeromonas.P.W. 1996. 10.C. Bhunia A. Doyle M. 57-63. Marcel Dekker Inc. Abeyta C. Notermans S. London.D. New York. Dodds K. 111-89. 1-27. in Clostridium botulinum: Ecology and Control in Foods.8 (42). Austin J.R..10.. Aeromonas species. Lund B. in Foodborne Disease Handbook. Volume 5: Microbiological Specifications of Food Pathogens. Fratamico P. 1989. Hauschild A.H. Microbiology and Molecular Biology. 1994. Aeromonas hydrophila. Eds.. Faris A.W. Marcel Dekker.H. Eds. 2005... 236 . Peck M... Clostridium botulinum. New York. Eds. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.L. Washington D.O. Cliver D. New York. Hui Y. New York. 1995.5% (42).D. in Food Microbiology: Fundamentals and Frontiers. Doyle M.5 . Johnson E. Clostridium botulinum. Ed.R. 11.M. Clostridium botulinum.R.. Palumbo S.... Blackie. 121-76.D. Potential hazards associated with REPFEDS. New York.10. Marcel Dekker. 4. 6. 1993.0% NaCl. Betts G.W. Diseases Caused by Bacteria.D. Marcel Dekker. in Foodborne Bacterial Pathogens.. Montville T. Aeromonas hydrophila group. 5.. 329-49. New York.

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

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

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


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

com Web site: www.org/fishery/ European Association of Fish Producers’ Organisations H.fiskeriforskning.uk Web site: www.is Web site: www.com/ Danish Institute for Fisheries Technology and Aquaculture (DIFTA) illemoesvej The North Sea Centre DK-9850 Hirtshals Denmark Tel: + 45 98 944300 Fax: + 45 98 942226 Email: difta@difta.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.suite.fao.dk Danmarks Fiskehandlere H.is/ 242 .gov. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo.bim.gov.asp Food and Agriculture Organization of the United Nations Fisheries and Aquaculture Department Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57053152 Email: FAO-HQ@fao.eapo.ie/templates/homepage.uk/index/fisheriesfarming-and-food.matis. Andersens Boulevard 37.dk Web site: www.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: post@fiskeriforskning.FISH AND SEAFOOD Department of Agriculture and Rural Development.no Web site: www.htm Irish Fish Processors and Exporters Association 25 Kincora Avenue Clontarf Dublin 3 Eire Ireland Tel: + 353 1 833 7882/ 02770249 Fax: + 353 1 833 7882 Email: tormg. Northern Ireland Fisheries and Rural Policy Division 4th Floor North Dundonald House Stormont Estate Belfast BT4 3SB United Kingdom Tel: + 44 (0) 28 90524999/ 4991 Fax: + 44 (0) 28 90378323 Email: dardhelpline@dardni.C. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: matis@matis.dk Web site: www.ifpea@eircom.dardni.fiskehandlerne.org Web site: www.difta.dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .

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.ifr.nz/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.gov.cmar.ifst.org.au Web site: www.com Web site: www.csiro.uk/en/index.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.ac.dh.seafood.org.uk Web site: www.uk Institute of Food Science and Technology (IFST) 5 Cambridge Court 210 Shepherds Bush Road London W6 7NL United Kingdom TeI: + 44 (0) 207 6036316 Fax: + 44 (0) 207 6029936 Email: info@ifst.co.gsi.fdf.bsi-global.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.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.nz Web site: www.org.gov.hpa.org 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.com/ 243 .uk Web site: www.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.co.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.

asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.int/en/ WHO Regional Office for Europe Scherfigsvej 8 DK-2100 Copenhagen Ø Denmark Tel: + 45 39 171717 Fax: + 45 39 171818 Email: postmaster@euro.int.euro.co.se/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: fao-who-cc@bgvv.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: hpsenquiries@hps.who.de 244 .who.int/ SIK .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.who.hps.uk Web site: www.net/web/indexen.codexalimentarius.int Web site: http://www.sfam.int/foodsafety World Health Organization (WHO) Headquarters CH .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.who.scot.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.co. Web site: www.who.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. jsp Food Safety 10 .org Web site: www.nhs.int Web site: www.uk/index.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: foodsafety@euro.nhs.org.uk/scieh.sik.se Web site: www.sofht.euro.uk Web site: www.

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


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. growth and survival in crustacean shellfish 86 Aeromonas sobria. pathogenic microorganisms in molluscan shellfish 59-60 as contaminant in enrobing process 37 as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45 growth in vegetable and rice 247 . as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. 69-70 Aeromonas hydrophila. in crustacean shellfish 86-7 ATP. source of PSP toxins 142 Alexandrium ostenfeldii. 102 Bacillus spp. as autochthonous bacterial flora of fish 3 Aeromonas spp. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. planktonic source of spirolides in shellfish 163 Allergens. preserved by dry/wet salting 99 Anisakis spp. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes.INDEX Achromobacter spp. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. causing spoilage of dried fish 110. 111 Astroviruses. in crustacean shellfish 87 Alteromonas spp. to test microbiological criteria 19 Aeromonadaceae. 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 in fish sauce 127 Bacillus cereus.

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

of fish – different processes 94 Dun. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. 152 Diphyllobothrium. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. 37 as spoilage organisms in chilled products 43 as spoilage organisms in tropical crustacean shellfish 81 as spoilage organisms in vacuum-packed dried fish 111 growth and survival in cured. 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. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. 106 Contacts 241-5 Conway method. source of azaspiracids 159 source of DSP toxins 148-9. as contaminants in scampi 36. smoked and dried fish 114 Enzymic spoilage. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. growth and survival in crabmeat 84 growth and survival in cured. to determine fresh fish spoilage 11 Corynebacteria.INDEX pathogen profile 226-7 Clostridium perfringens. to determine fresh fish spoilage 11 Eels. adverse condition on poorly preserved fish 110 EC grading scheme. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. 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. hot-smoking 107 ELISA assay. as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45-6 pathogen profile 227-9 Coated fish products. to detect NSP toxins 155 ELISA kit (Biosense). processing 334 Cockles. for detection of DA 148 ELISA kit (DSP-Check). smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . 149-50. as autochthonous bacterial flora of fish 3 Crabs.

source of PSP toxins 142 Gyoshoyu. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. as source of ciguatoxins 156 Garlic. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. causing pink condition on fish surface 110 in fish sauce 127 Handling. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. 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. 130-5 Fish-spoilage bacteria 12 Fishtester. definition of terms 176-7 implementation and review of plan 184-6 in fish and seafood product manufacture 175-88 seven stages of system 17784 Halobacterium spp. fish sauce 123 HACCP. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. concerning crustacean shellfish 87-8 European Grading Scheme. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. as source of inulin in lactic acid fermentation in fish products 134 Garos. of prepared fish products 44 Gambiertoxins. of fresh fish spoilage 911 Evisceration. for evaluation of fish quality 9 Eurotium. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. preserved by marinating 101 Gravlaks. of fish – effects on microflora 5-6 Hepatitis (non-A. processing 35 Fish component. fermented fish product 126 Green cured split cod 97. 102 Gymnodimines. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. member of cyclic imine group 161 Gymnodinium catenatum. to measure fish skin resistance 10 Flavobacterium. specific EU hygiene rules 196-224 Fresh fish. of fish at sea 5 Fatty fish. mould causing spoilage of dried fish 110 Evaluation. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes.FISH AND SEAFOOD EU legislation. non-B) viruses.

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

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

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

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

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

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

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