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Leatherhead Food International Leatherhead Food International
MICROBIOLOGY HANDBOOK FISH AND SEAFOOD
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
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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
I also acknowledge and thank Jackie Apps. which has been the chief factor in ensuring the completion of this book.FOREWORD The Microbiology Handbook series includes Dairy Products. and a detailed list of contacts for various food authorities. Rhea Fernandes Leatherhead Food International v . guides to HACCP. The books are intended as a source of information for microbiologists and food scientists working in the food industry and responsible for food safety. My gratitude is extended to Dr Peter Wareing. and Meat Products. both in the UK and elsewhere. All handbooks also contain a review of the related legislation in Europe and UK. Fish and Seafood. Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product. thank you for all the support. for his scientific input and Victoria Emerton. The books in the series are designed as easy-to-use guides to the microorganisms found in foods. Training Manager. and growth characteristics for key pathogens associated with the product. Technical Team Leader for her valuable editorial contribution. published by Leatherhead Food International and RSC Publishing. Alison Turner. I dedicate this book to my husband . and Ann Pernet for the work put into typesetting and indexing. Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged. potential hazards associated with the consumption of a range of products. I thank the contributing authors for their collaboration.Goldwyn.
2 Initial Microflora 1.7 References 3.3 Processing and its Effects on the Microflora 3.5 Pathogens: Growth and Survival 4.7 Published Microbiological Criteria 2.4 Processing and its Effects on the Microflora 2.7 Published Microbiological Criteria 1. vii .7 References 4. CHILLED AND FROZEN RAW FISH 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 2.8 Further Reading CRUSTACEAN SHELLFISH 4.4 Spoilage 4.1 Definitions 1.1 Introduction 2.8 References MOLLUSCAN SHELLFISH 3.2 Initial Microflora 4. 4.6 Pathogens: Growth and Survival 1.5 Pathogens: Growth and Survival 3.2 Definitions 2.5 Factors Affecting Fresh Fish Spoilage 1.1 Definitions 3.3 Processing and its Effects on the Microflora 1.1 Definitions 4. 3.6 Pathogens: Growth and Survival 2.4 Spoilage 1.3 Initial Microflora 2.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.2 Initial Microflora 3.3 Processing and its Effects on the Microflora 4.6 Published Microbiological Criteria 3.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.4 Spoilage 3.8 References 1.
8.7 Further Reading FISH AND SHELLFISH TOXINS 7.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.5 Pathogens: Growth and Survival 5.7 Food Hygiene (England) Regulations 2006.9 Azaspiracids 7.2 Initial Microflora 5.1 Introduction 7.5 References 93 93 95 96 108 113 116 117 120 123 123 126 127 135 136 137 140 141 141 142 144 146 148 151 153 155 158 160 165 166 175 175 176 177 184 186 6.6 Published Microbiological Criteria 5.1 Definitions 6.5 Regulation (EC) No.1 Definitions 5.10 Cyclic Imines 7.3 Tetrodotoxin 7.4 Spoilage 5. CURED. 9. 2006 No.11 Conclusion 7.5 Pathogens: Growth and Survival 6.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.3 Processing and its Effects on the Microflora 6.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.1 Introduction 8. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.6 Regulation (EC) No. SMOKED AND DRIED FISH 5.2 Legislative Structure 190 9.6 References 6.3 Processing and its Effects on the Microflora 5. EU FOOD HYGIENE LEGISLATION 189 9. 852/2004 on the General Hygiene of Foodstuffs 191 9.10 Further Reading 223 viii .9 Other Relevant Legislation 223 9.4 Regulation (EC) No.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.8 Guidance 222 9.I.1 Introduction 189 9.4 Spoilage 6.6 Lipophilic Shellfish Toxins (LST) 7.7 References 5.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. 14 (Hygiene requirements specific to the UK) 221 9. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption 212 9.2 Definitions 8. 7.2 Initial Microflora 6.5. S. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.3 Stages of a HACCP Study 8.3 Regulation (EC) No.8 Further Reading FERMENTED FISH 6.8 Ciguatoxins 7.4 Implementation and Review of the HACCP Plan 8.
11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix . 10.9 Vibrio parahaemolyticus 10.7 Staphylococcus aureus 10.10 Vibrio vulnificus 10.3 Clostridium perfringens 10.6 Salmonella spp.1 Aeromonas spp.4 Listeria spp.2 Clostridium botulinum 10.10 PATHOGEN PROFILES 10.5 Plesiomonas 10. 10. 10.8 Vibrio cholerae 10.
Poultry. smoked and dried fish. Cross N. in Handbook of Food Products Manufacturing: Health. fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs. 94767. Luten J. the consumption of fish and seafood has increased. Eds. Milk.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish. Oehlenschlager J. and fish and shellfish toxins. Chandan R. Clark S.. Because of their health advantages over red meats. Seafood and Vegetables. Safety and Processing of Wild and Farmed Fish. Nollet L. Hui Y.H. Saebo A. 2007.. The second edition of this handbook is a review of the entire book for currency of information. Seafood Research from Fish to Dish: Quality. Catches can be gathered from seas.A. thus. chilled and frozen prepared fish. Jacobsen C. Often contamination is from human and animal sources. Hoboken..M.. Meat..INTRODUCTION Fish and seafood are a main source of animal protein in the diet. 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.. xi . Further Reading General Fish/Seafood Microbiology Cross N. Seafood processing: basic sanitation practices. season.B. Bekaert K. rivers and lakes whose water can range from pristine to contaminated. The Microbiology Handbook. cured. Wageningen. Wageningen Academic Publishers. 2006. for fish... Geographical region. and. molluscan shellfish. crustacean shellfish. fermented fish. Wiley.
Oxford.K.... Noordwijkerhout... Gorham J. 2002. Dalgaard P.. Gram L. Huss H. Control of indigenous pathogenic bacteria in seafood.M.H. Vo1ume 39. 245-66 Bremner H.S.M.H.H. 459-72.International Commission on Microbiological Specifications for Foods. Academic Press. 200-23.A.Y. Dalgaard P.. Blackwell Publishing.. Blech Z. Salmonella spp. November 1995. 2004. Ashie I.R. 33 (1). Kluwer Academic / Plenum Publishers. Cambridge. Spoilage and shelf-life extension of fresh fish and shellfish. Blackie. CRC Critical Reviews in Food Science and Nutrition. Ed. 1997. in Food Plant Sanitation. 2002. Predictive microbiological modelling and seafood quality. Eds.L. Smith J.D. Woodhead Publishing Ltd. Bengtsson N. Fish and fish products (Microorganisms in fish products). Huss H. 329-65. 1997.A.. Oxford. Hafsteinsson H. 2004. Smith J. Ed. 36 (1-2). 2005. Gudmundsson M. Hui Y. Blackwell Publishing. 87121. Ed. 413-30. 1997. in Minimal Processing Technologies in the Food Industry.H. 167-77. xii . Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. Elsevier.. Gram L. in Advances in Food and Nutrition Research. London. in Kosher Food Production.. 61 (3). Eds. 372-6. Luten J. Rawles D. and Clostridium botulinum in smoked fish and shellfish. in Fish Processing Technology.B. Safety and Quality Issues in Fish Processing. Ed. New York. 1997. Microbiological spoilage of fish and fish products. Bruinsma B. Stanfield P. McCarthy S.R. Martin R. November 1995. Food Control.Y. 91-8.S. Huss H. Seafood processing: basic sanitation practices. New York.L. Seafood: processing. 1996. in Food Processing: Principles and Applications. 174-249. Flick G. Hall G. Proceedings of the International Seafood Conference. basic sanitation practices. Ed.. 318-23. Proceedings of the International Seafood Conference. The incidence of Listeria spp. 431-43. Modified-atmosphere packaging of fish and fish products. Journal of Food Protection.N. 8 (2). Ed. Biogenic amines in fish and shellfish.. in Seafood from Producer to Consumer: Integrated Approach to Quality.. Nip W-K.A. 1998.B. Amsterdam. Stanfield P. London. 121-37. Taylor S. Davies A.P. Amsterdam. 1997.. 1996. in Seafood from Producer to Consumer: Integrated Approach to Quality. Eds.E. International Commission on Microbiological Specifications for Foods.L. in Microorganisms in Foods 6: Microbial Ecology of Food Commodities. Noordwijkerhout. The fish industry.J. Journal of Food Protection.. 543-61. Tong P. Heinitz M. Microbiology of fish and fish products. International Journal of Food Microbiology. 2003.. Ventresca P. 60 (4).. Cambridge. Johnson J. Hui Y. Simpson B. Marcel Dekker. Woodhead Publishing Ltd. Ohlsson T. 1996. Luten J. Minimal processing in practice: seafood.H. Elsevier. Blech Z.
359-403.R. Chapman and Hall. xiii . guidelines. 1995.D. 118-20.. Seafood toxins of microbiological origin. 1994. 1-17. Evaluation of the bacteriological quality of seafood. Dalgaard P. 26 (3). International Journal of Food Microbiology. 5662. International Journal of Food Microbiology. PHLS Microbiology Digest. Ward D. Institut fur Veterinarmedizin. 93-105. 54-61. 121-8. 1996. Bonnell A. Volume 5: Microbiological Specifications of Food Pathogens. London. 16 (1). Blackie. Eds. Chapman & Hall. Gram L. 1994. 166-73. 26 (3). Blackie. Olley J. 1989. 2257. 1992. 265-79. 1994. Bacterial pathogens transmitted by seafood. 1992. Twigg B. 1990. Elsevier. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. A new look at microbiological standards. The microbiology of seafood products. Jakobsen M. Huss H. Sterling Publications Ltd. in Quality Assurance in Seafood Processing: A Practical Guide. Application of predictive microbiology to assure the quality and safety of fish and fish products. 1994. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Glasgow. Modelling of microbial activity and prediction of shelf-life for packed fish. Ross T. 44 (12). August 1991. Berlin.. Garrett E. Bacterial spoilage of seafood.H. in Fisheries Processing: Biotechnological Applications. Food Technology. August 1991. Microbiological quality of fishery products. 1991. McMeekin T. Huss H. Ed. Fish spoilage and safety. International Commission on Microbiological Specifications for Foods. Hobbs G. International Journal of Food Microbiology. in Public Health Aspects of Seafood-borne Zoonotic Diseases. London. Copenhagen. London. Liston J.H.M..N. Amsterdam. Copenhagen. Evaluation of the bacteriological quality of seafood. 90-106. Poultry and Fish Products. Dalgaard P.. Ed. 1990.. Amsterdam. Food Science and Technology Today.. 25-39. 305-17. Gibson D. Amsterdam. Martin A.M.International Commission on Microbiological Specifications for Foods. Huss H. Eds. Bonnell A. 1992. Gilbert R.D. in Quality Attributes and Their Measurement in Meat. Activities Report of the R&D Associates. Microbial growth and its control in meat. Gram L. Microbial hazards of seafood consumption. Fish: microbiological spoilage and safety. Qualitative and quantitative characterization of spoilage bacteria from packed fish. Sofos J.H.. and specifications for seafood products.J..A. in Microorganisms in Foods.. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Liston J. Ward D. Microbiological indicators and seafood processing. 46 (1). Elsevier. Eds. August 1991. Liston J. 1994. Ed.R. Jakobsen M. 459-78. 5 (3). poultry and fish. Elsevier.A. 1992. Eds. 319-33. Ed.M. Liston J. Bernoth E-M. 11 (2).R. in Food and Beverage Technology International: USA. 269-82. 1995. Pearson A. London. Copenhagen. Liston J.S. Dutson T. Jakobsen M. Ed.
The bacteriology of fish handling and processing. Food Technology. 104-9. Chichester. Journal of Food Quality.spoilage of fish. Regenstein J.Pedrosa-Menabrito A.C. 42 (3). Part I . CSIRO Food Research Quarterly. 71-117. in Developments in Food Microbiology. and specifications and inspection of seafood products. Hodgkiss W. 117-27. Elsevier Applied Science Publishers. Dicharry A. Seafood-borne bacterial pathogens of marine origin.R.R. Eyles M. Hackney C. 1988. Microbiology of fish. in Microbiology of Frozen Foods. Davies R. 1985. Hobbs G. xiv . 199-226.K. 169-208. Microbiological standards. Volume 1. Lamprecht E. 11 (2). Applied Science Publishers. 90-3. 103. 1982. 46 (1). Ed. London.J.S. 8-16. Norris J.. Food Technology. Garrett E.M.a review. Microbiology of frozen fish and related products. Wiley. Robinson R. Microbiological hazards associated with fishery products. 1988. 1986. Ed. Hobbs G. Shelf-life extension of fresh fish . London. 1987. Ed..K. Simmonds C. 1988.. guidelines. 42 (3). in Essays in Agricultural and Food Microbiology..
1.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. and is mostly interspersed between the muscle fibres. Fish fillets are a poor source of carbohydrates. Fish are an important part of a healthy diet since they contain high quality protein. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. Protein content in fish fillet varies typically from 16 .2 . migratory patterns. The lipid fraction is the component showing the greatest variation. CHILLED AND FROZEN RAW FISH Associate Prof.20%. 1 . Fish fillet composition can vary significantly within the same species due to feed intake. ‘fish’ also refers to the flesh of such animals used as food. typically 3 and 15% of total weight. The connective tissue content of fish is also lower than that found in meat. In addition. Fish can be divided into fatty and lean fish. In this chapter. The lipid content. This super class of vertebrates includes all the bony and cartilaginous finfish. lean fish are those fish that store most of their fat in the liver. Fish flesh is dominated by the abundance of white muscle in relatively short segments. giving it its characteristically flaky structure. In addition. typically fluctuates between 0. and spawning season.21%. “fish” will be used for fresh and seawater finfish. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features.5% (1). However. and other forms of aquatic animal as part of their ‘fish’ definition. offering less than 0. and excludes molluscs and Crustacea. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. crustaceans. it shows a typical season pattern especially in migratory species such as herring or mackerel. while fatty fish have fat cells distributed along their bodies. respectively (1). Muscle composition and structure of fish also differ from those found in other meat. but typically present a low fat percent when compared to other meats. which can be up to 67%. most fish contain omega 3-fatty acids and other essential nutrients. fins and a streamline body.
Spoilage describes changes that render fish objectionable to consumers. usually carbon dioxide (CO2). Fresh fish is raw fish that has not been processed. hence. colour.FISH AND SEAFOOD Chilled fish is fish that has been cooled to. Evisceration is the removal of the viscera from a fish. 2 . odour and texture. Controlled-atmosphere packaging (CAP) refers to packaging in an atmosphere where the composition of the gases is continuously controlled during storage. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. DMA is dimethyl amine. firmness. but no further control is exercised during storage. through their development on fish. nitrogen (N2) and oxygen (O2). at -2 to -4 °C. Organoleptic refers to qualities such as appearance. juiciness. and maintained at temperatures just below the freezing point. Quality refers to palatability and organoleptic characteristics such as tenderness. and maintained at or below -2 °C (normally below -12 °C) during storage. spoilage microflora describes an association of microorganisms that. and flavour based on the maturity. transportation and sale. Frozen fish is fish that has been cooled to. during storage. marbling. and texture of the fish. The proportion of each component is fixed when the mixture is introduced. colour. Superchilled fish is fish that has been cooled to. This technique is primarily used for bulk storage. but not below 3 °C during storage. and maintained at or below 7 °C. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. transportation and sale. renders that fish objectionable to consumers. transportation and sale. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases. frozen or preserved.
However. Shewanella and Pseudomonas. some species (most Vibrios) require sodium chloride for growth. For example. are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest. potential human pathogens may be present in both scenarios. gills and the gastrointestinal tract of fish. These pathogens can be divided into two groups: organisms naturally present on fish (Table 1.I). 1. Micrococcus. Gastrointestinal tracts and gills typically yield high bacteria numbers. 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.2 Initial Microflora The subsurface flesh of live. Gram-positive organisms such as Bacillus. In fact. posing a concern for seafoodborne illnesses (2). microorganisms colonise the skin. Clostridium. healthy fish is considered sterile and should not present any bacteria or other microorganisms. whenever possible. 3 . It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish. Flavobacterium. and those that although not autochthonous to the aquatic environment. since they heavily depend on the type of sample analysed and the protocol used for isolation. In general. as with other vertebrates. the natural fish microflora tends to reflect the microbial communities of the surrounding waters. It is difficult to estimate how many microorganisms are typically associated with fish. processing or storage (Table 1. Human pathogenic bacteria can be part of the initial microflora of fish. Moraxella.) are also common aquatic bacteria. It must be noted that mesophilic bacteria can rapidly overgrow psychrotropic organisms. The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. On the contrary. The number and diversity of microbes associated with fish depend on the geographical location. molecular-based methods have not yet been used to address this issue. Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp.CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed.II) (3). several culture media containing sodium chloride and more than one incubation temperature should be used. the season and the method of harvest. More accurate. Lactobacillus and coryneforms can also be found in varying proportions (1). and typical of the fish flora. Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters.
numbers of pathogenic bacteria in raw fish tend to be low.>106 Salmonella spp. 105 . 4).37 °C High for most species.FISH AND SEAFOOD TABLE 1.37 °C Unknown Unknown TABLE 1.106 cells/g (exception: V.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. and risk associated with the consumption of seafood is low (2.II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.103 High (107 .14 .102 101 .1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.19 µg/kg bodyweight from < 102 . In addition. Toxin levels 0. Plesiomonas shigelloides 3 . However. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp.35 °C 8 . estuarine environments. vulnificus) 5 . 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. birds.106 cells/g. sewage. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.0.26 °C 10 . Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. stream water.109 cells/g) * Some sources considered L. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . and mud Ubiquitous. 4 . Escherichia coli Yersinia enterocolitica 101 .1 .
chilled brine or refrigerated seawater at -2 °C. in which the health of each animal can be assessed prior to slaughter. in recent decades. with the production of off-odours. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating. the cuttings provide areas of exposed flesh that are open to microbial attack. human hands and clothing. and their re-use will increase the crosscontamination of other fish with such microorganisms. If evisceration is carried out at sea. The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea. birds and soil.3 1.3. potential contaminants added during handling and processing. boxes and/or baskets. and during harvest. In addition. 1. Fish may be eviscerated prior to storage at sea . These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans. During capture and storage. cortisone levels increase during prolonged stress and can alter the fillet quality. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. The length of time that set nets have been in the water or the time trawlers’ nets are towed.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. and especially when fish is stored on board for longer periods. handling and processing Wild finfish are usually caught by net. icing or freezing. finfish will almost invariably come into contact with nets. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria. mackerel or herring. Increasingly. has an effect on the amount of stress and physical damage that the fish will suffer during capture. The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. hook and line. Physical damage such as loss of scales. offering better control of fish health prior to. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating.1 Processing and its Effects on the Microflora Capture. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms. However. After capture. with very little control over the condition of the fish at the time of death or the duration of the killing process. digestion and off-flavours in uneviscerated fish. decks. and the killing process is designed to minimise stress. and spoilage. aquaculture practices have been expanding worldwide. ropes. in which the bacterial load 5 . and conditions of storage.a practice that may have both advantages and disadvantages. with fish such as tuna and cod being more commonly eviscerated than sardines. or traps. In eviscerated fish. This contrasts greatly with the meat industry. however.
FISH AND SEAFOOD can be substantial. ferric ion (Fe3+)). Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. in time. By vacuum packing foods. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. and of the quality of the water used for ice and to wash fish. These containers are also used for displaying the fish during auction at the quayside. Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. 7). What is more. often in the absence of adequate refrigeration. from water activity (aw) through to physical damage.). cleaning of the work environment. partial fermentation of sugars occur leading to lower pH. not part of the normal flora of the meat animals (8). use of effective detergents and disinfectants. The atmospheric conditions surrounding a product may be passively or actively altered. careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. As with all foods. but isolate the food in an environment different from the bulk atmosphere. for example. In MAP.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. In the absence of oxygen. but if applied properly should extend the life of the product. and to limit the loss of quality (5). Physical barriers not only protect from physical damage. fish have a higher post mortem pH. MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. a reduction in the oxygen tension is achieved which. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO). and minimising handling will all reduce microbial cross-contamination. they are usually process contaminants. 1. however. These changes can be deleterious. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. nitrogen. 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. Packaging changes the intrinsic and extrinsic parameters affecting a product. Anaerobic atmospheres have less effect on fresh fish shelf life. allowing more growth of spoilage organisms. if there is some oxygen demand from the product. by actively altering the composition of the 6 . oxygen etc. will result in fully anoxic conditions. However.3.
sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). slower-growing flora (11). In the United States. VP is one of the oldest forms of altering the interior gaseous environment of a pack. Micrococcus and Moraxella). gutted or filleted fish.CHILLED AND FROZEN RAW FISH surrounding gas. The microflora of meat is not the same as that of whole. Gases used in MAP of fish most commonly include carbon dioxide and nitrogen. without overt signs of spoilage. although less information on their effectiveness is available. the higher pH and reduction potential (Eh) of the fish muscle. including salt. discolour pigmented fish (11). but only in recent years has it seen a marked expansion in use and market share. the inhibition of common 'spoilage' bacteria (Pseudomonas. Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). as they are able to grow at temperatures as low as 3. the pathogens that may be able to grow before spoilage occurs. after additive application. the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . botulinum. Many additives have been tried in conjunction with changed atmosphere. The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. and the promotion of a predominantly Gram-positive. The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). Flavobacterium. High concentrations of carbon dioxide have the most pronounced microbial effect. a modified-atmosphere may contain any gas necessary for the desired effect. and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. may not be considered ‘fresh’ fish. The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains. which are able to grow rapidly at low temperature.3 °C and produce toxins. This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). phosphates. nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). Modified-atmosphere preservation of fish was first reported in the 1930s. Sulphur dioxide. and increase in-pack drip (12). The products. does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. Replacement of oxygen with nitrogen. and the problems of muscle structure damage by the modified-atmosphere (11). an inert and odourless gas. which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). but can dissolve into fish liquids and deform packages.
TMAO) that can be reduced to produce 8 . as already stated. botulinum.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). use of high-quality raw fish. In the United Kingdom. this problem occurs to a lesser extent with meat. grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. For fatty fish. however. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1. Only the highest-quality fish. adequate product labelling in respect of storage temperature. Subsequent storage must be at or below 3 °C. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred.g. adequate shelf life and cooking requirements. 3. from both a microbiological and a chemical standpoint. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. The addition of high levels of carbon dioxide to these fish. at all times. trimethylamine and sulfides). 5. should be used. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). 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. and the use of a HACCP plan. greater advantages are seen.g. 4. Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. These conditions include storage of the product at ≤ 3. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. For other less fatty marine fish.3 °C at all points from packaging onwards. owing to the dissolution of cellular structures. The problem of water loss in MAP fish is common. however. but is recognised as a drawback of carbon dioxide application. The growth of these bacteria is reduced by high carbon dioxide concentrations but. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13). the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). 2. The replacement gas should contain at least 40% carbon dioxide. Although all fish species need to be treated separately. may result in unacceptable colour and textural change and hence little shelf life extension (11). The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. 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).
the gills from pink and shiny. these are summarised in Table 1. as traditional spoilage patterns of the external organs are typically very distinct. to brown and slimy with an intense off-odour. and the flesh turns from bright and elastic to dull and soft (1). Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . and this pattern is the basis of freshness grading schemes. Various methods for whole fish freshness evaluation by a trained panel have been proposed. Over time. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. When working with fillets. 18) and are probably the most widely used. 17.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. B. 1. the skin turns from iridescent to dull and bleached with bacterial slime. or for that matter evaluation of the taste of the fish. so lower carbon dioxide concentrations may have better preservative effects. A. texture. C (C=reject) Significantly. smell and 9 . with no smell. Loss of freshness and spoilage cannot be separated as processes.III. TABLE 1.CHILLED AND FROZEN RAW FISH off-odours.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).4 1. reactions caused by activities of the fish’s own enzymes. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids).4. The eyes turn from convex and clear to concave and opaque. Degradation of whole fresh fish stored in ice generally follows a set pattern.III Fish-quality scales Name of scale Torry Scale Quality Index Method European Grading Scheme Scoring system 10 to 0 (≤ 4 reject) 0 to 24 (arbitrary reject) Extra. and metabolic activities of microorganisms present in the fish. Taste panel evaluation of cooked fish may be used to score or grade colour.
Generally. the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. thus. To a varying extent. From being flaccid. After some time. this will generally correlate with the length and severity of rigor. chemical. a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. rigor resolves. 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. possibly slightly metallic. fruity and/or bitter off-flavours. ii) Neutral taste. During the initial storage. rigor mortis is the first noticeable change in the fish. 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. post-rigor values are typically in the range of pH 5. little flavour. As the fish degrades. AMP. RT Freshmeter or Fishtester).8 to 6. sulphurous and/or rancid smells. texture becomes soft and watery or hard and dry. 23).g. the pH drop of the muscle is correspondingly less. 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. By measuring the concentrations of the six components. the muscles relax again and the fish returns to a flaccid state. development of sickly sweet. After death. ADP. an anti-freeze. biochemical and microbiological actions. Because fish tend to have relatively little residual glycogen compared with mammals. TMAO permits a high Eh to remain in the muscle tissue. as little endogenous reduction of TMAO occurs (18). 19). However. reduction in ATP and increase 10 . HxR and Hx) gives a quantity (K-value. The process of ATP breakdown is used as an indicator of fish freshness (24). These processes are strongly influenced by the physical conditions of storage. the conductance generally increases.FISH AND SEAFOOD taste of cooked fish.5 (18. an osmoregulator. IMP. spoiled and putrid. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical. which increases from 0% towards 100% with time. the Torrymeter. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. iii) Traces of sour. cabbage-like. After death. sea-weedy taste. bacterial reduction of TMAO to TMA. ammoniacal. or its remaining shelf life (1). or simply a waste product present due to bioaccumulation (22). %). iv) Enhancement of the spoilage characteristic of phase iii. an intense odour compound. the Eh of fish muscle remains relatively high (20). the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. all marine fish use/have TMAO (21).
which may also be formed by bacteria (25). however. the importance of microbial action increases with time (5. 11 . and a segregated weak acid to absorb them. does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection. Neither does the change in pH during storage correlate well with the production of TVB-N. colour changes. Quality indices based upon the products of microbial metabolism do not explain changes in quality until microbial growth produces measurable changes in the fish. however. and are especially important in the spoilage of frozen fish. 18). Chemical degradation continues after the initial post mortem phase. 31). the residual acid is then titrated. hypoxanthine. For some fish species. However. The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. also termed TVN). as microbial spoilage is limited by the low storage temperature (1. The Conway method (and variations of it) uses a strong inorganic base to volatilise the bases in the fish sample. 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. so the measurement is effective only during the initial loss of freshness. 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). using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. but rejection of whole fresh fish by sensory methods such as the EC grading scheme (32) is based upon non-specific odour detection and physical appearance. Lipid oxidation and other oxidative changes lead to oxidative rancidity. During spoilage. 26). as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. therefore. these measurements are usually used to quantify the amount of spoilage. or a specific fraction of the volatile bases. for example the TMA fraction. Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. Comparing results for different fish species. Evaluation of Total Volatile Base-Nitrogen (TVB-N. is later significantly reduced by bacterial action. 7).CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). Oxidative rancidity is known to reduce the quality of fatty fish in particular (19. a correlation can be made between the spoilage of the fresh fish and the production of TVB-N. 27) produced by aerobic respiration and other forms of metal ion reduction (28). not to describe freshness.
For fresh fish. 5).2 Fish-spoilage bacteria The parameters affecting the multiplication of microorganisms in foods have been categorised into two general groups: intrinsic (inherent qualities of the food) and extrinsic (qualities of the food environment) (35). the lower the temperature of storage.and temperate-water fish spoilage rates may be many. For example.1 Factors Affecting Fresh Fish Spoilage Temperature By far the most effective method of reducing the rate of whole fresh fish spoilage is temperature control (1. the results of altering one parameter may have far-reaching consequences on others. yoghurt spoilage by yeasts and clostridial spoilage of cheese (5) are examples where it has been recognised for many years that a particular minority of the microbial flora present in the product is responsible for its spoilage. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. Fish spoil as a result of the chemical.5. by altering the gas atmosphere (extrinsic). 34). For fresh fish iced immediately after capture and continually stored in ice. careful temperature control during storage is not only important in terms of quality loss. The sum of these changes may result in a different spoilage profile. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity.5 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. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. the slower the spoilage processes proceed (within limits). but also crucial for the assurance of consumer safety. 1.FISH AND SEAFOOD 1. The reasons for the differences between tropical. Also.5. biochemical and microbiological reactions taking place within and on the fish. The factors are interactive and cannot be completely isolated. 1.3 Recognised specific spoilage organisms (SSOs) The degree of spoilage leading to sensory rejection of fish is partly dependent on the perception of the consumer.5. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 . a minority are often associated with the majority of the spoilage. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33.
and proteolytic and lipolytic enzymes that may affect the quality of fresh and. although only since the late 1960s has it been realised that specific metabolites of Sh. Sh. These organisms are described as potential spoilers.CHILLED AND FROZEN RAW FISH started in the 1940s. obligate respiratory bacteria. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. motile with polar flagella. plus other characteristic compounds of the species responsible for spoilage. produce hydrogen sulphide (H2S) from cysteine. (9) and they are considered SSO of iced freshwater fish (35). the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. They are generally characterised as Gram-negative rods. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. aldehydes. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. 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. or faster than the rest of the flora of ice-stored fresh marine fish (9). more especially. the spoilage of a product may be strongly influenced by the conditions under which the product is held. 13 .10% of the total flora of fresh fish from temperate marine waters.1 Pseudomonas spp. 35). putrefaciens spoilage of fish is due to its biochemical action on muscle. As previously noted.3. mediated spoilage is characterised by ‘fruity’. Pseudomonas spp. which produced the characteristic spoilage compounds of the fish (30.5. catalase-positive. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp.3. and may play some role in the spoilage of freshwater fish. 35. processed (e. therefore. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine. ‘oniony’ and ‘faecal’ odours from the production of ketones. During the 1970s.g. frozen) fish products.5. putrefaciens growth may be used as indicators of spoilage (30). It is able to grow as fast as. it is often isolated as about 1 . It is also present in fresh water. 36). The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. 1.e. on fish are diverse and in many cases species-specific. oxidase-positive. It is thus an important spoilage organism of gadoid fish such as cod. its ability to reduce TMAO to TMA. Members of the genus are able to produce pigments. The importance of Sh. i. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms.2 Shewanella putrefaciens Sh. 37).
and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. Sh. This type of intoxication occurs when bacteria convert the histidine 14 . It is also able to produce hydrogen sulphide and a range of other off-odour compounds. Research has shown that approximately 107 cfu/g of Ph. like all members of the Pseudomonadaceae. with several new species being described. phosphoreum sufficient to cause spoilage was found. as the ferric iron is insoluble. putrefaciens strains are characterised as microaerophiles or anaerobes. Shewanella affinis. Sh. Shewanella waksmanii. owing to its cell size and activity. Shewanella oneidensis. 41). The prevalence of the bacterium in so many environments. Ph. and Shewanella profunda were described. whereas >108 cfu/g Sh. phosphoreum were required for 50% of taste panellists to reject a sample. at the time of rejection. putrefaciens were identified as Shewanella algae. Shewanella frigidimarina. Shewanella livingstonensis. 38). In terms of its taxonomy. In 50% N2/50% CO2 MAP cod. baltica has been identified as the main H2S producer in cod during cold storing (37). 1. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39).how to reduce a molecule that they are not able to take into their cells efficiently. Another distinct feature of the physiology of Sh. does not show carbon-source chemotaxis. mesophilic strains of Sh. Sh. Shewanella gelidimarina. 41). and recent taxonomic description using modern molecular methods bifurcated this species. Later. the bacteria have a significant problem to overcome . The ferric reductase activity of Sh. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12).5.3. it was proposed that this organism. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). although. putrefaciens has been studied. putrefaciens were required. but Sh. putrefaciens growth has been collected (12.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. Shewanella olleyana. This wide-ranging respiratory capability is thought to be unique (39). they are heterogeneous. unlike many other motile bacteria. putrefaciens lies in its chemotaxis: the bacterium. phosphoreum has also been identified as responsible for histamine fish poisoning. additional psychrotrophic species such as Shewanalla baltica. it is strictly respiratory.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). and Shewanella aquimarina. a population of Ph. Initially. Shewanella denitrificans. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). putrefaciens was not present in such numbers (23. In fact. 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. Ph. Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection.
Huss (1.6 Pathogens: Growth and Survival Endogenous chemicals. 5) reports that fresh fish consumption has never been shown to cause human botulism. previous exposure and access to supportive treatment.CHILLED AND FROZEN RAW FISH present in fish into histamine. However. Fewer still are capable of growing on fresh fish.6. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. C1. B. B. algal toxins. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42).1 Clostridium botulinum C. 5). Adult disease is caused by the production of the botulinum toxin. 1. 1. C2. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions.4 Brochothrix thermosphacta and lactic acid bacteria B. Acetate production has been reported as a good indicator of spoilage by this organism. botulinum recorded up to 1963. lightly preserved fish products are associated with botulism. (and very rarely F and G) have been reported to cause human disease. 75% (with a 35% mortality rate) occurred in Japan. it is bacterial risks that increase after capture of the fish. they do not produce significant product spoilage). The heterogeneity of the types (A. When inoculated into VP corned beef and sliced ham. Of 404 intoxication outbreaks of type E C. presents a potential risk. types B. botulinum. thermosphacta in the spoilage of some MAP fish. 6).5. bacteria and higher parasites all present some risk associated with fish consumption (1. human viruses. Some Ph. Review of outbreak data suggests fresh fish to be safe. F and G) is well documented (44). thermosphacta did not produce off-flavours until 2 . D. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. Growing evidence suggests a role for B. including antisera. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. 1. this is probably due to spoilage occurring before toxin elaboration. these risks are substantially reduced (1. Recent studies have investigated the dominance of B. where a particular 15 . but with proper cooking. a convenient but diverse species of bacterium with a number of different types. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). 5.e. age. E. E and F include bacteria with minimum growth temperatures of approximately 3. E.3 days after having reached 108 cfu/g (42).3. Types A.3 °C (Group II) and these are non-proteolytic (i. B. Of these. especially type E botulinum intoxication. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment.
vulnificus is associated with warm marine and estuarine waters. whether stored aerobically or in modified-atmospheres. 1. were associated with finfish between 1978 and 1998 (46). Other lightly or semi-preserved (e. including epidemics in India. storage and preparation (11). Russia. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. and when in contact with contaminated water (no CDC reported cases from fish.g. Proper chilling. sushi) (5). again. Southeast Asia. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. Despite the ubiquitous nature of type E C.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . A unique clone of V. 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. 5). In Japan. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. and proper cooking (boiling for 1 minute. The bacterium causes primary septicaemia. there is no specific guideline that describes a minimum level of V. a Japanese boiled and semi-dried sardine dish.3 °C) will eliminate all risk. is small. I-sushi. Since 1996. parahaemolyticus O3:K6 is responsible for many of the recent V. V. Only about 1% of marine isolates produce a thermostable haemolysin. the majority are associated with products not cooked immediately before consumption. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47).g. The first report of the organism being a foodborne agent was with shirasu. Generally. which is believed to be required for virulence. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45).6. accounted for the majority of cases (1. which can cause human gastroenteritis. and North America. the majority of outbreaks are caused by consumption of raw fish products (e. 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. This strain has been responsible for 50 to 80% of all V. 11). USA. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. Japan. Currently. 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%) (2.2 Vibrio parahaemolyticus and other vibrios V. the risk posed to the consumer of fresh fish. parahaemolyticus is a marine organism. 1978 . Proper temperature control (<3. due to the cultural preference for raw fish dishes. France. salted or pickled) fish have also been associated with botulism.2005). parahaemolyticus cases have increased across the world. V. parahaemolyticus outbreaks. smoked.
1.6. in assessing the risk of fishborne infection by P.. infection. in its habitat. but the bacterium is known to survive and grow in the shallow marine. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. Despite this. is historically associated with faecally contaminated water. their potential to produce disease-causing exotoxin in food has not been fully examined (48).000 17 .20 °C. Chill storage may not eliminate growth. cholerae O1. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. cytotoxic.55 cases per 100. P. L. Again. monocytogenes has been well documented as a foodborne human pathogen. are present on fish and can grow to significant numbers during storage (1. 17). are associated with human diarrhoeal illnesses. sodium channel blocking and haemolytic activity (48).4 Listeria monocytogenes L. There is no risk associated with the consumption of properly chilled and cooked fresh fish. Similar problems as those encountered for Aeromonas spp.26 to 0. Some isolates are true psychrophiles. coli on isolation media (50). possibly due to under-reporting because of their likeness to E. monocytogenes incidence has been between 0. 1. and especially Aeromonas hydrophila. and fish and shellfish are probably the major reservoirs for the organism (50). Although the psychrophiles are unable to grow at body temperature. 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. They are aquatic organisms with an epidemiology that is yet to be fully understood. In the US. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. the causative agent of cholera. others are psychrotrophic mesophiles (49).6. but proper cooking should substantially reduce risk of infection. there is no risk from the consumption of properly handled and cooked fresh fish. proper chill storage will eliminate any risk associated with this organism.3 Aeromonas Aeromonas spp. and especially estuarine environment.CHILLED AND FROZEN RAW FISH disorders). shigelloides is similar to Aeromonas spp. shigelloides are encountered. Most cases have been sporadic rather than associated to large outbreaks. with minimum growth temperatures around 0 °C and optima of 15 . V. and proper cooking will substantially reduce risk. 5. The major virulence factor appears to be the production of toxins with enterotoxic. which is often fatal (>50%). It is known that Aeromonas spp. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. however. listeriosis cases have been reportable to the CDC. Since 2000.
Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 .6. Photobacterium spp. 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. If fish are not properly refrigerated. Infections are mostly associated with the consumption of raw or 18 . at risk groups .6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. Klebsiella pneumoniae and Hafnia alvei. Clostridium spp. 1.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity.. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). 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. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. especially pickled fish. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. leads to the production of histamine.. there is a possibility for growth of the organism on chill-stored fish. it is not apparent to the consumer and it cannot be destroyed by cooking. The organism poses a serious risk in chilled products not cooked before consumption.. 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. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. 1. D60 of 1. and Lactobacillus spp. Like other toxins. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. Lightly preserved products. alcoholics. some Vibrio spp. but proper cooking will reduce any risk. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54).2002 (55)) and is also common worldwide.48 minutes in salmon have been reported (53).98 minutes in cod and D60 of 4.FISH AND SEAFOOD persons. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. neonates. but cooking will significantly reduce the risk of infection. but more especially by Morganella morganii. which has a maximum permissible level (for fish products from fish species associated with high amount of histidine) of 200 mg/kg fish in the EC. full risk analysis is not possible (52). foetuses. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. Fortunately. Being a psychrotroph.pregnant women. It is regularly isolated from seafoods (51).6.
H. which in many cases will eliminate the risk. especially in warm-water areas and where fish are to be consumed raw. FAO. EC. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. parahaemolyticus (see Table 1. coli counts where this method is preferred.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. 2003. including parasites and/or quantity of their toxins / metabolites per unit of mass.IV) (57). Gram L. 1. it may be desirable to test for Salmonella and V. Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. 444. Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF). Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it.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. TABLE1. In summary. Ababouch L. area or lot (CAC. parahaemolyticus Staph. For fish from inshore or inland waters of doubtful microbiological quality. volume. Faecal coliform counts may be used instead of E.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . No. 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. long storage under refrigeration or one of the former prior to freezing. 1997) Huss H. Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease. Rome. The most widely accepted microbiological criteria for chilled and frozen raw fish are those set for aerobic plate counts (APC) at 25 °C and E.1997... A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56).
9.T. Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms. Codex Alimentarius Commission. 1994. 1993. 5 (1). References Huss H. The role of seafood in bacterial foodborne diseases. Rome. Microbiological spoilage of fish and fish products.. 13. 2.. Food Control. Kirby R.FISH AND SEAFOOD 1. Rhodehamel E. International Journal of Food Sciences and Techonology. Ashie I. Gram L.. International Journal of Food Microbiology. 12. Ahmed F.8 1.K. 2... Smith J. 67-71. 283-94.K.. Microbes and Infection. 91-8..J. 12. 37. 291-304. Food and Agriculture Organisation. 14. 87121. Dalgaard P. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth. 107-27. Nychas G. Huss H. 24. Capell C. International Journal of Food Science and Technology.R. 8 (2). in FAO Fisheries Technical Paper No. International Journal of Food Microbiology. Incidence and growth of psychrotrophic Clostridium botulinum in foods.E. Food Control. Ed. Davey K. 19. 87-118. Gibbs P... Jeksrud W. Reddy N.R. 6.. Huss H. Food and Agriculture Organisation of the United Nations. 36. Italy. 1995. Huss H. 4. Quality and quality changes in fresh fish. 1651-60. 334.S. in FAO Fisheries Technical Paper No.H.J. Codex Alimentarius Commission. 8. Rome.. 11. Jehanno D. and natural toxins. 348. 3. M.E.. Incidence of foodborne pathogens in European fish. parasites. 27. 2002. Huss H. Assurance of seafood quality. 1992. Simpson B. Food Control. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). Critical Reviews in Food Science and Nutrition. 7.A. 2003. Food and Agriculture Organisation. Feldhusen F. Ed. 2000. Spoilage and shelf life extension of fresh fish and shell-fish.. 5-7. Food and Agriculture Organisation of the United Nations. Fletcher R. 1996. 12...H. 10. Sivertsvik M. 1994.H.R. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres. Control of indigenous pathogenic bacterial in seafood. 1992. International Journal of Food Microbiology. 2001.P. Kautter D. Davies A.H. 33. 5. Rosnes J.R. Gram L. Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review.N. Armstrong D. 20 .A. 1997.J.A.H. Journal of Food Safety. 1996. Davies A. Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef. 121-37. 243-60. 1989. activities and safety..
. Khayat A... Ed. Microbial and biochemical spoilage of foods: an overview. Slade A. Rome.H. 1992. http://seafood. Byrne A. Microbiological spoilage of fish and fish products. Edinburgh. Rome.CHILLED AND FROZEN RAW FISH 15.R. 1971. 22-36. Lancaster. AVI Publishing. National Advisory Committee on Microbiological Criteria for Foods. 26.. Saito T.H. 1995.ucdavis. Letters in Applied Microbiology.. The micro-determination of ammonia. 1995. Eds. Westport. 1982. Journal of Applied Bacteriology. Ackman R. Technomic. Autolytic changes in fish post mortem. 28... 21 16. Huss H. Part I . 1988.. Ed.M. 41-50. 1-18. 19-22.J. Hebard C. Non-enzymatic oxidation of seafood lipids. Surette M. 354-8.E. 26.M. Arai K.. Adopted. Biochemical Journal. 25. Gill T. 1959. 30. Leblanc P.edu/haccp/compendium/chapt08. in Fresh Fish.J. Bacteria active in the spoilage of certain seafoods.A. 24. 20.. Gram L. Ratnayake W. Flick G. Gill T.. Sea Fish Industry Authority. 27. 18. 419-29. 19. 184. Huss H. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage. Yajima T. 1996. Schwall D. Shewan J. Børresen T. Gibson D. Huis in't Veld J.. March 20. Flick G. Food and Agricultural Organisation of the United Nations. 1985.J. International Journal of Food Microbiology. International Journal of Food Microbiology.H..E. Reginstein J. Nature. Washington D..A. 21. 117-27.E.spoilage of fish. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products. Davies A. Flick G. 1995.E. Qualitative and quantitative characterisation of spoilage bacteria from packed fish. 29.E. An absorption apparatus for the micro-determination of certain volatile substances. Dalgaard P. Martin R.. 1. 42-54. 305-17. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere. 36.. 27.S. 1415. International Journal of Food Microbiology. 17.htm Anon. 121-37. Eds. Chemical composition. Martin R. 1996. 1983. 22. 23.N.M. Herbert R.M. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout. in Chemistry and Biology of Marine Food Products.E.J.J. Quality and Quality Changes. 1933. 11. 2007. 33.J. in Advances in Seafood Biochemistry.. 245-67. National Advisory Committee on Microbiological Criteria for Foods. 1995. 21. 1992. Hebard C.J. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish. 33.a review. 1987. 37. Quality and Quality Changes. Huss H. 34. Hendrie M. Lipid oxidation in seafood. Journal of Agricultural and Food Chemistry. in Fresh Fish. Pedrosa-Menabrito A. Shelf life extension of fresh fish . Conway E.J.G. 130-2.H. Food and Agricultural Organisation of the United Nations. Martin R... Food Technology.C. Changes in purine nucleotide of red lateral muscle of rainbow trout. Journal of Food Quality.
Role of proteolytic bacteria in fish spoilage. 39. Rome. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere.. 4. 9.. Dalgaard P. and an Achromobacter species. Gram L.F. 174.M. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish. 40.. Nychas G. Journal of Applied Bacteriology. Dalgaard P. 1989.M. 38. in Fresh Fish. 32. 33. Fish spoilage bacteria – problems and solutions.. 1984.R. 42.. International Journal of Food Microbiology. Brochothrix thermosphacta. Saffarini D.. 67. Kell D.europa. 51-62. 41. 48. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens. Environmental Biotechnology. Food and Agricultural Organisation of the United Nations. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches. 44. Dixon N. 37. 1984. 1995.. 334. Occurrence and importance of proteolytic bacteria as spoilage indicators. Sumner J. 1985. 1973.H..H. and regulation. Karnop G. Annual Review of Microbiology. 262-6. 1982. 61-6.H. Italian Journal of Food Science. 13.J. 26. physiology. Gram L. Magno-Orejana F. 36. 323-9. Vogel B.J. Cohen D. 71. 34.FISH AND SEAFOOD 31. Scanlan R. 2005. 36. Pseudomonas fluorescens. Lee J. Myers J. 317. 72-80....E. Oxford.L. 33. FAO Fisheries Report. Jørgensen B. Gorczyca E. Brady P. II. 328-9. 18-21. 1996.R. Blackwell Scientific Publications. Venkateswaran K. 1992. Drosinos E. Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens.S. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. Nealson K.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J. 6689-97. 3429-38. Huss H. Satomi M.. Growth and activity of Shewanella putrefaciens isolated from spoiling fish. Myers C.M.. Taussig M. Archiv für Lebensmittelhygiene. Libbey L.. 35. The effect of storage temperature on shelf life.B... 109-36.A. Huss H. Quality and Quality Changes.eu/LexUriServ/LexUriServ. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia. 62-70. Rome. Applied and Environmental Microbiology. 1994. 1989. Processes in Pathology and Microbiology. The inhibition by CO2 of the growth and metabolism of microorganisms. Iron and manganese in anaerobic respiration: environmental significance. 22 .H. 311-43. Applied Microbiology. 43. 2002. Miller A. Journal of Bacteriology. Ed.
Bean N. Lehane L. International Journal of Food Microbiology.. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets. Morbidity and Mortality Weekly Report. 100-3.K. USA.. 20. E and F in salmon fillets stored under modified atmospheres at low and abused temperatures.C. 1991. The International Commission on Microbiological Specifications for Foods.C. 1-37. in Microorganisms in Foods 6. 49. Slutsker L.. 47. 1-34. Mac Rae I. 1993.P..L. 53. 2005. 23. 17-34. 12.. Wadström T. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures. Majeed K.J. Journal of Infectious Diseases. Miller A. New York. 51.A. Microorganisms in Foods 2. 54. Thompson S. 2000.H. Blackwell Scientific Publications... Altekruse S. 181-8. The International Commission on Microbiological Specifications for Foods.M. Applied and Environmental Microbiology.... Ljungh A.. Genigeorgis C. 46. MacKinnon L..N. Embarek B. Embarek B. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico. Ray B.. International Journal of Food Microbiology. Olley J. 23 . Bishop R. Modern Food Microbiology. 1661-6. Lindroth S. Golden D. Surveillance for Foodborne-Disease Outbreaks . Fish and Fish Products (Microorganisms in Fish Products). Braden C. Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B. Wilson S. 2194200. 56. Daniels N. 51 (4). Huss H. 1998-2002. 52. 172-249. 57. 303-12. Garcia G. Painter J. Panicker G. International Journal of Food Microbiology.M. 181-96. Whiting R.CHILLED AND FROZEN RAW FISH 45.. Aeromonas and Plesiomonas as food. 1986. Myers M. Smith J. 2000. 1973-1998. 181. Jay J.M. 2003.. 12. Woodruff R. detection and growth of Listeria monocytogenes in seafoods: a review. 2005.P. Griffin P. Kluwer Academic/Plenum Publishers. Food Technology. 1987. 55 (SS10).. 1994.and waterborne pathogens.. 58. Loessner M. 50. Oxford. 48. 1997. 55. The International Commission on Microbiological Specifications for Foods.United States. Lynch M. Presence. Ed.. Use of risk assessment to reduce Listeriosis incidence.. 1991.K.H... Bej A. International Journal of Food Microbiology..L. Histamine fish poisoning revisited. Journal of Food Protection. 2006. Microbial Ecology of Food Commodities. 330-6.W.. Sampling for Microbiological Analysis: Principles and Specific Application. Hammond R. 85-95.J. Vibrio parahaemolyticus Infections in the United States. 69.. Springer. International Journal of Food Microbiology.A.. 50.K.
Eds. Fisheries and Food. Journal of Food Protection.P. 15. Food and Agriculture Organisation of the United Nations. Shinano H. Trolle G. Technomic Publishers. Torry Research Station of the Ministry of Agriculture.G. Ed. Bulletin of the Faculty of Fisheries (Hokkaido University).D. 1985. Huss H. 1984. International Journal of Food Microbiology. Cann D. Aberdeen. 741-50.9 1. Huss H.9.. Jones Y.. in Seafood Science and Technology. Simard R..9. Academic Press. Handling and processing of scad. Picard G.A..E. 73-89. 1997. Gram L. Labuza T. Ross T. 1961.. Villemure G.H. Ravn Jørgensen B.M. Food and Agriculture Organisation. Journal of Food Science.). Sumner J. 65-72. 62-70. 55.C. The Microbiology of Seawater Fish. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. 6. 1992.. 35. Kitts D. Magno-Orejana F.. Application of predictive microbiology to assure the quality and safety of fish and fish products. Aberdeen. Houston N.. Fisheries and Food. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches... Borgstrom G. Olley J. Dalgaard P... 1. 1988.. 1984.. 1992.D. Cho Y. 1986. 51.M. Huss H. Ogden I. Akiba M. Gibson D. in FAO Fisheries Report No. Smith G. International Journal of Food Microbiology.L. Gibson D. Torry Research Station. in Torry Advisory Note No 93. Changes in microflora and chemical compounds in mackerel stored by partial freezing.1 Further Reading Processing Anon.2 Spoilage Shewan J. 295-307. Estimation of the bacteriological quality of fish by automated conductance measurements. 1989. in Fish as Food. Rome. Microbiological quality and shelf life prediction of chilled fish.. 1.FISH AND SEAFOOD 1. 4. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. International Journal of Food Microbiology.H.M. McMeekin T. 13-32. Lancaster. Hobbs G...S. 317 (Suppl.1983. 1. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging.H. Florida. 24 . Ed.. 271-85. Torry Research Station of the Ministry of Agriculture. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere.. 1989. International Journal of Food Microbiology. 317-21. 127-34.. Shahidi F. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. Taoukis P.
Effect of culture age. 2197-203. Jacobsen M..europa. 1987.J. 25 .H.. 1997. International Journal of Food Microbiology. 479-88. 1987.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. Garcia M.. A rapid method for detection of histamine producing bacteria.. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. 1992. 72.R. Gram L.eu/LexUriServ/LexUriServ. 1993. 59.M. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. Amsterdam. Hall G.. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C.K. Otero A. Blackie Academic & Professional. 322-6. Huss H.. Eds.. Journal of Applied Bacteriology. Modified atmosphere packaging of fish and fish products. 1. International Journal of Food Microbiology.. Huss H. 5. Huss H. 137-46. Applied and Environmental Microbiology.9.H. in Fish processing technology.CHILLED AND FROZEN RAW FISH Einarsson H. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Trolle G. London. Elsevier Science. 1992. Gram L. Klausen N. Sala F.9. 4. in Quality Assurance in the Fish Industry. Liston J. 1.L.3 Pathogens Condon S. 20023.H. http://eur-lex. 65-72. Ed.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A.
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. 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. Sushimi tuna).g.). smoked (hot) mackerel. 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. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr. Even though heating prior to consumption will eliminate pathogenic microbes. clarity of eyes.g. emphasising the high quality of the seafood and the gourmet nature of the prepared products. Few seafood products are sold ‘ready-to-eat’. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2. This is no longer the case with many processed products now available in stores. The same used to be said for the difference between fresh and frozen products. smell etc. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e. products of lesser quality. it does not necessarily apply to the toxins that may have been produced. but still acceptable and meeting all safety criteria. or could be disguised by sauces.2. the exceptions being Sushimi (high quality raw fish). seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. colourings etc. The majority of seafood products produced require a cooking stage before consumption. due to the early cessation of spoilage. smoked (cold) salmon. and crustacea such as shrimp and crabs.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. colour of gills. although recent developments in improved supply chains. 27 .
They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. Nevertheless. Shelf life on ice. and these are increasingly being presented with an ever-expanding range of accompaniments. Frozen fillets may have a surface layer of 28 . they are usually of a better standard than species such as tilapia. Doubts have been raised over the microbiological status and handling practices of reef-caught fish.2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. and pathogenic Vibrio spp. Prepared natural fillets are fillets of fish without any additional coating. but these are now finding favour as products in their own right. where the preparation has already been carried out. molluscs. rather than frozen. in particular. often using the simplicity of a microwave oven. 2. from both wild capture and aquaculture sources. 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. they are usually supplied filleted with head on. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. At the same time. etc. which can be farmed. or in MAP depends on catch conditions and treatment. and the consumer only has to heat (cook) the product.FISH AND SEAFOOD Recent trends in the market for food and seafood products. prawns. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. Seafood is the term used descriptively to include all categories of product. Listeria spp. This includes all categories of seafood including trout and other freshwater species. Microbiological hazards include Salmonella. and this has produced a greater availability and diversity of products in the marketplace. but should be equivalent to cold-water products. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish. The expected shelf life attainable for this format is usually up to 8 days. They are typically flown into export markets. and are well known for their diet consisting of waste materials. shellfish and molluscs. There is also a limited market for filleted material. as they tend to lose colour and sheen after defrosting. have shown the increasing importance of convenience foods. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein. the processing industry has developed a wide range of value added products.. including freshwater fish.. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP).
salmon. a protective glaze may be added to the frozen steak. 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. 29 . and salmon. The result is fish with a typical residual salt content of 1. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings.5%. but may be increased (up to 1 month) by packaging methods like MAP or VP.2.g.g. be flavoured with herb dips or peppercorns. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product. sawdust with other woods adjusted for flavour profile and burn duration).g.2 days) longer than that of non-prepacked fish. Pâtés are prepared with raw fish. resulting in a product that is ready-to-eat without any further cooking. Again. The shelf life for MAP fillets is typically up to 5 days.. Cold smoking is essentially a flavouring process. or pre-packed frozen with added glaze. or pre-packed in MAP or vacuum packaging (VP). Fresh fillets are sold from wet fish counters. The expected shelf life of these products is up to 8 days (or longer for some chilled. Cold-smoked natural fillets are prepared from a range of cold water fish including herring.g. in VP or through mail order. mackerel. which is approximately 30% (or 1 . A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. cod). etc. and the product then shaped and heat-processed to cook the product before being chilled. natural fillets of fish such as herring. They are sold from wet fish counters. the ingredients premixed. swordfish steaks).CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. Hot-smoked fish are prepared from brined. Added colours can be included with the brine solution to meet specific requirements for products e. with a minimum residual salt content of 3. Some products may be salted for flavour only (e. annatto with smoked haddock. tuna steaks) or band-sawn from a frozen block (e. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes. which may. in addition. mackerel. Fish steaks can be cut from a natural fillet or loin (e. 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.5 .5% in the aqueous phase. 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. VP products) because of the high salt content acting as a barrier to spoilage. This process in effect cooks the product and reduces or eliminates microbial contaminants. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. haddock.
jellied eels. etc. long-life seafood mixes using sorbate and benzoate as preservatives.or a five-stage process. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process. Prawns. Other. salmon) coated and sold as chilled products. and seafood mixes. and formed into round cake shapes.) Raw material is typically fish fillet.. rice. and heat-fixed. tuna. e. There is an increasing market for high value fresh or thawed fish (e.g. Band-saw dust and fish trimmings from fishblock preparation materials may be used. fish nuggets/bites with dips and seafood-based snack products.g. The protein source can be fish fillet or blocks. The cakes are usually coated using a three. 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. binders. rice or pasta. haddock. addition of ingredients such as colours. Surimi is the insoluble protein fraction of fish after washing and processing. or cut from a preformed frozen fish blocks into shapes. Natural prawns from cold or warm waters can include those peeled. which may include molluscs. Other ingredients may be natural or reconstituted.g. 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. mixed whitefish. pasta or potato. cephalopods. They are pasteurised products and treated as high risk raw materials. The majority are products made from various types of whitefish (cod. flavours. ‘fingers’ before being coated in batter and/ or breadcrumbs. e. it can be used as a seafood analogue. Scampi are tails of Dublin Bay prawn or langoustine. Breaded fillets/steaks and battered fillets are either natural or shaped fillets. Fish cakes are products produced by mixing fish with potato and herbs. and may also include additional ingredients such as fresh herbs and spices. coated in batter or breadcrumbs. It is often used as a protein supplement. taramasalata. with an expected shelf life of around 7 days.FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate. 30 . With further processing. and in the shell.
3. searing or bar marking). skin. After capture and during any subsequent processing. The edible tissue of some species of fish are more susceptible to gaping than others.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. and the potentially toxic by-products of histamine-producing bacteria (5. 6). see Table 2.I. and ingredients added to the final product. Contamination will. The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. will the saprophytic flora start to invade the surface flesh (1. fat content. is not used for sushi or other high risk (and high value) products.3 2. which may become a focus of contamination deep within the muscle. Dalgaard et al. storage. or visible damage from gaff marks (4). Tuna that does not meet the highest of grades based on colour. i. 2). sensory assessment of freshness etc. In summary. Only after death. and pathogenic flora associated with seafood. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). For a summary of background information on the hazards associated with fisheries products and ingredients. on the whole. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. species (Bluefin>Bigeye>Yellowfin>Albacore). remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. product handlers.e. 31 . Grading is based on size of fish (>25 kg).1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. as a result there is an increase in spoilage rate thereby reducing the product shelf life. Recent public opinion has favoured the use of dolphin. handling or preparation stage. Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). and intestines of the fish. including seasonal variation. and have emphasised the origin and sustainability of the fish as key marketing points.and turtlefriendly long lining to the less discriminatory netting. the microbial flora of seafood is associated with the gills. A primary role in fish spoilage has been assigned to this organism. it may be used for other products or markets depending on the product specification. whilst gaping is also dependent on a range of other factors that effect the muscle composition. and processing conditions such as dehydration or poor filleting practices.
staphylococcal toxin. however. for chilled products. Moulds Enterobacteriaceae Salmonella. 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. Aspergillus. toxin. anaerobic spore-formers. product assembly. moulds Bacillus. intermediate processing. yeasts Pathogenic microflora/toxins Salmonella.4 2. moulds.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse.I provides an overview of common raw materials. and associated spoilage and pathogenic microorganisms. be considered at an early stage during new product development. it is essential that during new product development a full hazard identification and risk analysis be conducted to ensure that the microbial risk for specific ingredients are identified.3. cereus Staphylococcus aureus. 32 . streptococci. Shigella. Enterobacteriaceae.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. shelf life determination. and. Bacillus cereus. Staph. raw materials. Clostridium botulinum. and so are the potential microbial contaminants associated with them. moulds Bacillus Bacillus. nuts and fruit Associated microflora Lactobacilli. Pseudomonas. moulds Bacillus. Listeria. spore-formers. Table 2. Penicillium. enteric viruses 2. protozoa and viruses B. pseudomonads.4. coliforms. TABLE 2. Clostridium Lactobacilli. cereus Bacillus licheniformis.FISH AND SEAFOOD 2. B. Fusarium. coryneforms. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter. cereus B. Aeromonas. Bacillus subtilis B. histamine Cryptosporidium. mushrooms.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. In addition. Staphylococcus. by the HACCP team or by a hazard and risk assessment programme. Clostridium perfringens. Vibrio cholerae. Escherichia coli. yeasts Lactobacilli. cream. and milk Cheese Water/Ice Bacillus.
as the full complexity of the processes used for specific products cannot be covered in a general text. defined holding time at a chill temperature (0 – 3 °C). Examples of tempering techniques include the use of heated air. and contamination from equipment to clean fish muscle. there are a number of distinct methods used. as a heat process is often required for functional or organoleptic reasons. depending on the size of the fillet 33 . and potential opportunities for growth of foodpoisoning bacteria.4.2. this may be an incidental effect. the core temperature. up to -5 °C so as to improve the cutting and handling properties during processing. Rapid controlled cooling of heat-processed components will help to minimise spoilage. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. It is critical that the time and product temperature during tempering be controlled to avoid elevated temperatures (>5 °C) on the product surface allowing microbial growth or the onset of spoilage in raw material. 2. It is recommended that the principles of HACCP are applied to novel processes. 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. Where the raw materials are frozen.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. but it is assumed that any band sawing or dice process will introduce heat from blade friction. 2.1 .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. but it also poses a risk of cross-contaminating other products undergoing tempering.4. or the use of microwave technology. For some components.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing.3. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components.4. to evenly raise the temperature of the entire frozen product from -20 °C or lower. All have the same intention. but will also need to ensure that the optimum shelf life can be safely obtained. Generic examples have been used to illustrate only the major points. Effects on the microflora are minimal during tempering (7). line extensions and process reviews.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes.3 Preparation of fish component of chilled and frozen recipe dishes 2. Figures 2.
1B .1.1A .FISH AND SEAFOOD processed.1C . 2.7 days. Raw material preparation stages for chilled or frozen coated fish products 34 .3.Shaped Fillets. after landing.Scampi. 2. drained and then refrozen for storage until required for further processing. 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.4.4. Fig.2. frozen in a nitrogen blast freezer and weighed into sacks for later processing. The “blown” meat is graded. Frozen cores are usually batter-enrobed. It is then size-graded. chilled or sold chilled on defrost with an expected shelf life of 6 .2).1. may not rise above 0 ºC during processing. On arrival at the processing plant. with the coating heat-fixed.3. the scampi is washed in potable water to remove foreign bodies.1.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. by mincing and mixing with a binding agent (as described in section 2.Fish Cakes). Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. quickfrozen. 2. The processing of three examples types are described in Figure 2. are placed into plastic boxes and covered in ice to preserve their condition.1 (2. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.
Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. and water is added to rehydrate the components. Therefore. or those where the trimmings. additional spoilage organisms may be introduced. such as aerobic spore formers and Staphylococcus spp. The mix is shaped and formed ready for packing or for enrobing. or promptly processed to avoid further quality loss. spoil more rapidly than the dry ingredients. material will need to be transferred swiftly to a chiller.. as the physical structure of the material has been destroyed.1. Waste material produced from this type of processing is often reused in the third category (fish cakes). pathogens such as Salmonella and Listeria. The remaining dried ingredients are mixed with the fish.3.3. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. ii) Fish-filleting waste . but.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. those that use frozen blocks of whole fillets. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. fish fingers).off-cuts or belly flaps.2 Shaped products This type of product falls into two categories. with the coating heat-fixed. 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. iii) Fresh/fillet/minced fish block. Fish cake cores may be batter-enrobed.7 days. The fish component of the cake will.g. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. 2.block-sawn dust. quickfrozen.4. chilled. or sold as a chilled product after thawing. from potato flake.1. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. 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. that are to be reused for other foods. are handled and stored as any other food product. more seriously. once the dry components have been rehydrated. with an expected shelf life of 6 . because of its nature. It is for this reason that all by-products from primary processing.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.4. unless it is poorly handled. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. 35 .
vibrios and other fishassociated spoilage organisms.4 Battering/enrobing process Figure 2. The first stage. any excess is removed with an air knife. Over time. because of the intensive handling required during processing. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. from flour. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. and to carry flavourings where required. lactobacilli from the milk powder and flour. Natural fillets d. which is product-dependent and not always required. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. e. e. starch. 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. chilled. Breaded or battered fish fillets are fully heat-processed before consumption. and may include milk powder. some of this initial flora 36 .g. spices. raising agents. and if required. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process.or fivestage) frozen or chilled. as well as ensuring that there is minimal contamination from human processors.FISH AND SEAFOOD In all of the above examples. Tempered band-sawn materials b. Fish products. i. Batter contains water. absorb any excess moisture on the surface of the fish portion. with the coating heat-fixed. a characteristic flora will develop from the batter components. Initially. perfringens could. such as Listeria and Vibrio spp.2 illustrates the typical stages of enrobing or breading. a.. Products are preformed and presented to the process (either three.4. with an expected shelf life of 6 . 2. is the addition of a pre-dust. rapidly multiply if strict temperature control was not maintained. which can occasionally be isolated from protein-handling processes. Bacillus spp. rusk or crumb. If a crumb coating is required. fish cakes Portions or fillets may be batter-enrobed. which typically consists of flour. IQF/pre-formed fillets c. flour. quickfrozen. milk powder and spices. and possibly Staphylococcus spp. Spore-forming anaerobes such as C. The function of this is to improve the adhesion of subsequent coatings. or sold as a chilled product after thawing. potentially. A large variety of product formats and types can be batter-enrobed.g.7 days. and Enterobacteriaceae from flour.e. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. egg.
dry batter mixes are all potential sources.2. aureus and B.2. and both of these can produce heat-stable toxins that can survive a batter heat-fix process. With inadequate temperature control of batter systems.CHILLED AND FROZEN PREPARED FISH PRODUCTS associated with the ingredients will be diluted out. time-limitation or production run temperature control. in the case of B. The FDA has issued clear guidelines on acceptable 37 . this is not of great significance to the microbiological standard of the finished product. replaced in part by process contaminants and bacterial flora from seafood. but because batter is heat-set. Staph. 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. but it is believed that poor hygiene regarding product and food handlers. and.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. This can be achieved by frequent clean-downs and adequate hygiene. cereus. cereus can proliferate under certain conditions. Fig. contaminants such as Enterobacteriacae can proliferate and develop during the process. Flow diagram outline of a typical three.
2. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. as this could introduce off-flavours and reduce shelf life. Flow diagram outline for recipe dish ingredients preparation 2. For example. A temperature no greater than 10 ºC for 12 hours cumulatively or 21.g. The vegetables must withstand processing.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe.1 ºC for no more than 3 hours is recommended (1). but they are supplied without a source of carbohydrate e. 38 .5. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7).1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal.2. Fig. and must not show evidence of disease.3.4. rice. canned fish may be used in their preparation owing to it having wider availability and low wastage.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes.4. pasta or potato. to produce a specific product. Figure 2. The protein source can be fish fillet or block. There are two main categories.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products.
39 . the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. rice or pasta. haddock. However. for example. not just as accompaniments. 8) will. all-year-round fresh vegetables are preferred.4. Salmonella typhi. Raw material is typically fish fillet. (7. encourage more rapid decay and loss of structural integrity.5. The opportunity for growth of L. For example. treated as a highrisk ingredient. chopping and mixing of material. 2. and mixed whitefish). i. even those with pH-modified dressings.e.3 log reduction in bacterial numbers on the surface of vegetables. spore-formers. The majority are products made from various types of whitefish (cod. For example. In an attempt to improve washing and reduce chlorine taint. will disperse the saprophytic flora of vegetables . However. coryneforms. pastry. A number of examples of direct faecal contamination of vegetables exist in the literature (8). and protected from any post-processing contamination. coli O157. Any assumptions regarding the application of good agricultural practice to produce should be made with caution.lactobacilli. and reduce spoilage organisms and potential pathogens. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. Products must be washed thoroughly to remove pests and soil.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. and may include sauce accompaniments. any processing stage. Washing with a biocide will at best achieve a 2 . This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. Although the processes are claimed to be superior to using unmodified hypochlorite for washing. if the process is not appropriately controlled. but also as components in recipe meals. they should be stored under temperature-controlled conditions. with cases of foodborne disease traced to Listeria monocytogenes. coliforms and pseudomonads throughout the mix. frozen block or canned fish. loss of quality. The presentation of vegetables has a significant effect on the product quality. and may include molluscs or prawns that may or may not have undergone a minimal heat process. Blanching of vegetables can be an effective means of reducing or removing surface contamination. or breadcrumbs. and E. colour and water will occur. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components. Once vegetables have been blanched. control is more onerous. with the accompanying release of additional nutrients by processing. For premium products.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings.
can germinate and rapidly multiply. and an increased risk of contamination from L. spore-formers can proliferate if adequate control of temperature is not maintained.) to the product (12. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. Products or dried ingredients contaminated with either spores of aerobic bacteria (e.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. Sauces can be multi-component preparations or more simple preparations. Potato toppings made from flake or powder will normally have little associated spoilage flora. 13). but temperature-abused pasta can also spoil rapidly by mould growth. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. rely on the use of microbiologically sound ingredients. Appropriate handling facilities or equipment will be required to protect the product.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. and then cooled slowly allowing germination and re-growth. B. especially in those products that have a vegetable component or rice accompaniments.6. cereus are well known and are still a common issue in the UK (9).g. such as butter garnish with black pepper. If the sauce contains modified starches for freeze/chill stability. 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. For this reason all products.4. Placing such products in a standard chill store to cool down could result in condensation. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. aureus toxin is well documented (10. On hydration. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. a process equivalent to at least a 5D Listeria kill will be necessary. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. Sauces or marinades are added to fish to broaden appeal or to accompany components. An incident involving Staph. cereus). Alternatively. Typically. where composite meals are heat-treated.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide.4. 40 . Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. 11). cereus. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. with the range always expanding as new tastes and cuisines are developed. This is a particular issue especially where products are heat treated to eliminate vegetative cells. 2.
etc. respectively.herbs. the sauce component will spoil rapidly and the possibility of vegetative pathogens from post-process contamination or raw components growing to significant numbers cannot be ruled out. flour products. Marinade-type products are becoming more popular. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. for instance. and handling of processed sauce can be less rigorous than. 13).4. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. 41 .4 and 2.5 illustrate the recipe dish ingredients preparation and assembly and packaging. 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. The format uses flavour compounds . spices. suspended in an acidified sauce with an oil component. that of a dairy-based sauce. 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 (12.2. Figures 2. Without barriers.
7. 42 . Flow diagram outline assembly and packaging of recipe dish 18.104.22.168.4. and tuna loins are packed by placing the product in a plastic pouch and removing all the air. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. which may restrict either microbial growth or enzyme activity. These include: 2.4.2 Modified-atmosphere packaging Originally developed for the red meat industry. as it dissolves in the surface moisture to form a weak acid.2. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. 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.FISH AND SEAFOOD Fig. the method replaces air in the sealed packaging with a defined mixture of gases. botulinum to grow.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions). generally carbon dioxide and nitrogen. The oxygen-free conditions will however allow other anaerobic bacteria such as C. unless the temperature is strictly controlled to below 5 °C throughout the supply chain. 2.
the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. 43 . temperature control is important in preserving chilled integrity of ingredients. phosphoreum. The fish component of recipe dishes. but is not limited to. followed by moisture loss. Not only will these organisms have a numerical advantage over other proposed spoilage organisms. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. Aeromonas spp. initially. Beyond this. Spoilage will be most readily detected in proteinaceous components of ready meals. Nonetheless. This is also true of chill storage and the distribution chain. is typically uncooked or at best lightly processed to retain eating characteristics. The case for growth and contamination of products with Aeromonas is stronger. (20). It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage.10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18).. a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. at slightly higher temperatures (10 ºC). by a loss of texture. Vibrionaceae will succeed (17 ). where the protein component is nearly always heat-processed. and finally flavour loss.5 Spoilage During processing. and in influencing the final eating quality of the product. This is considered to be the one major difference between fish and meat products. At refrigeration temperatures (5 . Spoilage of this material is not expected to differ greatly from the processes described elsewhere. and Yersinia spp. but little on in-pack spoilage. However.1 Chilled storage For chilled products or chilled components of frozen products. The literature has good information on spoilage of individual components of recipe meals. in numerical terms. both organisms are capable of growth at chill temperatures. Shewanella putrefaciens and Ph. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15).5. except for some crustacea. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). as this organism is a common contaminant of freshwater fish species (19). 2. but they will also be readily adapted to the nutrient source and conditions.
within dairy products are well known. 2.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10).. Bacillus spp. By retaining the intrinsic spoilage flora of seafood. As a note of interest.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. where the variety of components and ingredients used in manufacture is varied and complex. 44 . enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage.5. auto-oxidation of fats/oils).5.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). Serratia spp. These have been proved to impart off-flavours. The conditions and freezing process most favourable for good-quality products will. and Aerobacter spp. the shelf life of products is limited by the natural spoilage process described earlier. however. Pseudomonas spp. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24). Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp.FISH AND SEAFOOD 2.. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes.. For example.g. eliminating nematodes in products that do not have a heat treatment step. this should also take into account shelf life of product components used in the assembly of the final product. freezing also has a protective role in improving the safety of raw materials by. also be those that have the least effect on the viability of microorganisms (21). The type and nature of the enzymes and their activity will depend on the bacterial species. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. Products that are developed with an extended shelf life beyond the 6 . This reinforces the need for prompt processing and temperature control of ingredients and finished products. 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. producing characteristic off-flavours. botulinum. which would warn the consumer of the potentially unsafe nature of the product. 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. for example. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. However. Still.
perfringens contaminating seafood will depend on the source of raw material.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.3 Clostridium perfringens A study examining C.1 Bacillus cereus This is a common contaminant of dust. etc.104/ g should be treated with some concern (26). botulinum when held in favourable conditions for growth (28) i. water. air. along with previous discussion. or a heat shock followed by holding at a temperature suitable for growth. low oxygen or anaerobic conditions. Incidents in such diverse materials as cream.6. Vibrio spp. 27). have been linked with food poisoning and should also be considered.6. and Campylobacter are recorded in filter feeding molluscs such as mussels. Counts of between 103 .6..e. Additional hazards from marine bio-toxins. and many raw or processed foods (26). This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. Hazards attributable to the non-seafood components are listed below and this information. agricultural run-off or sewage can contaminate in-shore species. and some effects related to product type and time to toxin production have been noted.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. especially molluscan filter feeders. Inappropriate holding times and temperatures will.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. Therefore. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). 2. typhi. spices and herbs and vegetable shoots have been reported (12. farmed or free-living crustacea (30). permit rapid growth of B. 2. Cases quoting contamination of seafood with organisms such as S. in some cases. 2. cereus. with pathogenic microorganisms or viruses. potato. may be of benefit within any risk assessment for chilled and frozen prepared fish products. A number of other Bacillus spp. any risk of C. are dealt with elsewhere in this book. histamine intoxication. Germination of spores requires favourable conditions. These are C. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. 45 . 13. and this issue may be of greater consequence for warmwater. In addition. botulinum type E and Vibrio parahaemolyticus (25). soil.
6. which is not affected by any subsequent cooking process. and this is believed to be due to a tolerance to brine used in the smoking process (32). indigenous or other. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. 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. but under the appropriate conditions. These include inadequate refrigeration and preparing foods far in advance of planned service.6. However. L. and inadequate cooking or holding within the temperature growth range (27). 46 . if any other potentially pathogenic organisms. aureus enterotoxins in foods. Shigella spp.4 Listeria monocytogenes Interest in this organism has been characterised by a number of very serious incidents associated with a variety of product types (27). the restrictions on growth rate seen with seafood will not apply. It is unclear. These conditions will allow the production of the toxin. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). infected food handlers. will colonise factory environments in a similar fashion except for possibly Vibrio spp.5 Shigella spp. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). Chilling may help Shigella to survive. 2. growth may occur in foods (26).6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. An acceptance has now developed in the UK of the presence at low levels of L. when introduced or inoculated into sterile components of chilled prepared recipe dishes. and will therefore cause “food poisoning” even in a product that is cooked before consumption. 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). monocytogenes as a contaminant in foods (31). Sources of contamination are many because of the widespread distribution of this organism in the environment.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. 2. at the present time. 2. aureus (35). monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. L.6. but.
7 Published Microbiological Criteria Due to the complexity of many processed food products. Microbiological specifications do not necessarily relate to product safety. It should be remembered that 47 . unless the criteria is for a pathogenic organisms where only non detection is acceptable. aureus enterotoxins and B. it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur. For instance. The following tables (Table 2. 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. with respect to the number and types of ingredients used. 2. etc. the fresh product is often consumed before the results are obtained. pasta. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40).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. Microbial guidelines. are not mandatory. as published by for example. and not to final products. but these may be eliminated by a subsequent heat-treatment stage. although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”. 2. and their association with similar materials. cereus emetic toxin are often confused owing to similar symptoms.II and 2. (36). pastry. and as such the monitoring is for verification purposes rather than control. 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. such as batter mixes.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts. 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. .III based on IFST Guidelines (39)) provide example guidelines for seafood products.CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph. Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems.6. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). and are a recognised means of providing data on manufacturing standards. but provide the industry with an indication of the microbial level at which action is required to ensure product safety. although due to long analysis times required.
batter etc) should also be consulted when establishing the specifications for raw materials and final product. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp. coli GMP* <102 Maximum 104 TABLE 2. monocytogenes B. Maximum= Levels at end of stated shelf life 48 . pasta. coated fish.g. fish cakes Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Pathogens GMP* Maximum Criteria for absence not generally applicable Indicators and spoilage organisms Organism E. aureus V. fish fingers.fish products.FISH AND SEAFOOD the microbial criteria for other products (rice.III IFST Guidelines for processed foods . parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E. cereus Staph. L. 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.II IFST Guidelines for part-cooked foods – e. TABLE 2.
. 8. 1. Ltd. Ward D. Fish: microbiological spoilage and safety.. 7. 1993.. Christiansen T.L. Communicable Disease Report. Kumeta H. 13. Huss H. 166-73. Proper processing and handling of raw tuna. 1962. Powers E. 233-7. 2005. 67-73.G. 1976.. Microbiology of processed spices.M. Murrel H. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. Hackney C. Bartlett C. 70-2. in Fish Inspection Quality Control and HACCP: A Global Focus. 15.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). 1311-7. Tu A. 49 4. 5 (3). Pathogenic microorganisms associated with fresh produce. Kluwer Academic. Oxford. Bartram P. Journal of Milk Food Technology. Powers E. 1997. Matsuyama H. Pvt. 25 (1). 14.M. 663-87. The bacteriology of fresh and spoiling fish and some related chemical changes.. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. Cambridge. New York. Incidence and levels of Bacillus cereus in processed spices. Food Science and Technology Today. Collette R. 1976.8 1. Woolaway M. 10. Animals and Spoilage. Eds.. Aureli P. 2.E. 3.L. Journal of Food Protection. Outbreaks of foodborne illness in humans. 1997. Sakamoto T. Ishizaki K.. Behling A. 5.C. 7 (24). 668 -70. 1982. Beuchat L. 24 (5). Journal of Milk Food Technology.. Bacterial histamine production as a function of temperature and time of incubation. Recent Advances in Food Science. 1997. 11.H.. 6. . 47. References US Food and Drug Administration. Masuoka Y. England and Wales Quarterly Report. Washington D.L.C. Taylor and Francis. London. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. 12. Yumoto I. Hoshino T. 2. IBH Publishing Co.J.. 96. 59 (2).A.T. Mejholm O.. 1997.. 2001. 1991. Dalgaard P.R. Communicable Disease Report.R. Letters in Applied Microbiology.. London. Wienke A.. 39 (10). Van Nostrand Reinhold. Martin R.. 38 (11). Ohgiya S. Letters in Applied Microbiology. Taylor S. Shewan J...M. Journal of Hygiene.W.. 9.R. Hobbs G.. Journal of Food Science.. 204-16. Gilbert R. Fish and Fisheries Products: Hazards and Controls Guidance. 1986. 215. 1991. 373-8. Lawyer R.C.K.J. Latt T. Importance of Photobacterium phosphoreum in relation to spoilage.R.R. Food and Drug Administration... 167-93. Microbiology of Marine Food Products. International Commission on Microbiological Specifications for Foods. Brown T. Toxin Related Diseases: Poisons Originating from Plants. Slavin J.
Leistner L. Jemmi T. 31. 24.. Lund B. Is Aeromonas spp..C..M. Halligan A. a foodborne pathogen? Review of the clinical data..R. Leatherhead Food Publications. 19. 1988. Walker S. Fresh and processed fish and shellfish . Modern Food Microbiology.. Eds. 26.R. Ed. 50 17.W. What problems does the food industry have with spore forming pathogens B. 1977.J. Food Taints and Off-flavours. 4 (1).Yersinia. 655-60. Loessner M. Matches J. 18. 309-16. 25 (2). Liston J. Blackie Academic and Professional. 1994. 29. Morgan D. Journal of Food Safety.H. U. Granum P. 11. 28.E. Huss H.. Ronner U. 1996. International Commission on Microbiological Specifications for Foods. Food Research International. 32. Aeromonas and Verotoxigenic E.J... Connel J. 333-6. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service. Blackie Academic Press.J. 1996.FISH AND SEAFOOD 16. Jay M.V. perfringens? International Journal of Food Microbiology. in Advances in Fish Science and Technology. 23. Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants. Ray B. 1979. Curran D. Effects of freezing and storage on microorganisms in frozen foods: a review. 4. Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens.. . 2000. 155-9. in The Microbiological Safety and Quality of Food. 27. Fishing News Books Ltd. Survey of the incidence of Aeromonas and Yersinia species in retail foods. PHLS Microbiology Digest. Barnes E.K. Aspen Publishers. 59-72. Advisory Committee on the Microbiological Safety of Foods. Andersson A. Gaithersburg. Food Control. Lund B. Golden D. Food preservation by combined methods. Oxford. London. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures. Wood L. in Anaerobic Bacteria in Habitats Other than Man. 351-72.. 2005.C. 51-158 Saxby M.C. cereus and C. Anon. 472-506. 28 (4). Liston J. Leatherhead.H. London. 34-40. The Emerging pathogens .J. 30.. Springer. Mead G.. 1996.L.A Literature Survey. Speck M. 1986.. 1987. coli (VTEC) . Clostridium perfringens in the environment. 21. 1990.. 22. Gould G. 13 (1). Journal of Food Protection. 9. 65. Food Microbiology. International Journal of Food Microbiology. Baird-Parker T. Anaerobes in relation to foods of plant origin. Trolle G. 1992. 25. Brooks J. 1992.M..A. SAB Symposium Series No. 20. Her Majesty’s Stationery Office. New York. 274-86. Huss H. Gram L. Report on Vacuum Packing and Associated Processes. Blackwell Scientific Publications. Gram L..J. 28. Glasgow. 41-3. 1974. Applied Microbiology. 13. Microbiology in fishery science. Eds. 40 (5). 1993. 1995. Keusch A.
1991. British Retail Consortium. 35. 1987. Cambridge. 34. London. Foodborne Pathogens an Illustrated Text.H..pdf Shapton D.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33.A.G. 1997. 36. 60 (4).co.L. http://www. London. Evans M. Shigella as a foodborne pathogen. Woodhead Publishing Ltd.F. 39.A. 38.eu/LexUriServ/ LexUriServ. Principles and Practices for the Safe Processing of Foods.chilledfood. 1992. 372-6.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. Gloucestershire. Wolfe Publishing. CRC Press. Shapton N. Institute of Food Science and Technology.2. 2006. http://www. U. Phil Voysey. Chilled Food Association.K. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs.europa. Smith J. 1998. Varnam A. 40.A. Journal of Food Protection. McCarthy S. Development and Use of Microbiological Criteria for Foods. Kraft A.. Journal of Food Protection. 37.campden. 41. http://eur-lex. U. Campden & Chorleywood Food Research Association. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. 788-801. 51 . Microbiological Criteria. 1999.K.uk/cday/posters2008/npd6. 50 (9). Chilled Food Association.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology. Psychrotrophic Bacteria in Foods: Disease and Spoilage.
and scallops. Pelecypoda. chitons. and 2. The phylum consists of five classes: Amphineura. 4). oysters and clams can process about 3. can hold the shells tightly shut against enemies (1). O. they also trap bacteria. the adductors.1 Definitions General features of the mollusc The phylum Mollusca. abalones. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). P. Special tracts of cilia move this mucus toward the mouth.1 3. In these forms. Pelecypods have little need for locomotion. where it is eventually swallowed and digested. Food and Drug Administration 22201 23rd Dr. Scaphopoda. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. snails. Many microorganisms ingested by shellfish survive the digestive process (3. In most species.S. lying well within the mantle cavity. Strong muscles. oysters. slugs. For example. chemical contaminants and other impurities on the mucus of the gills. mussels. while 53 . The class Pelecypoda includes all of the bivalves such as clams. respectively. the foot is compressed to form a muscular spade for digging and the head is greatly reduced.6 litres. scallops. clams. Box 3012 Bothell WA 98041-4421 United States of America 3.3. MOLLUSCAN SHELLFISH Carlos Abeyta. octopi. mussels. and shrimp. whelks. the shell. composed of two valves hinged together dorsally. Jr. Pacific Regional Laboratory Northwest U. The class having species of economic and public health importance is the Pelecypoda. cockles.E. cockles. completely encloses the body. Other species of shellfish such as crabs. which includes oysters. The term shellfish in this context is limited to oysters.1. clams. lobster. mussels. Gastropoda.7 litres of water per hour. squid. and Cephalopoda. cockles and scallops. viruses. S. Plankton and algae form the bulk of their diet from seawater. In the process of filtering water. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells.
3. but maintain contact with the water by means of a siphon tube (5). and usually at higher concentrations than found in their habitat.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 . dwelling on the bottom or attached to structures in the water column. are not as vulnerable to contamination through pollution of their beds as the bivalves. Clams are benthic and live burrowed in the mud.I).North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp.South America Pacific – Japan Quahaug (also known as hard clam and hard shell clam) Coquina clam Hen clam Butter clam (also called the Washington clam) Mogal clam Little neck clam (also called rock Cockle Pacific littleneck) Atlantic – North American/Europe Atlantic – Europe/North America Japan Atlantic/PacificNorth America Donax spp. many species of bivalves are edible (Table 3.2 International commercial products Globally. Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 . The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3.1.FISH AND SEAFOOD commercially valuable. Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific . Whatever contaminants are in the water will eventually get into the shellfish. The habitat of oysters and mussels are sessile.
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 .
1 Clam Clam is the common name of a large group of often edible. hard-shell or soft-shell clam. 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 3.2. The muscular foot is used to burrow in mud or sand. and other names. A mantle tissue encloses the body of the clam. laterally compressed.1. Different species are variously called quahog. or siphons. Most clams are only a few centimetres in their maximum dimensions. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint. but 56 . littleneck. geoduck. cherrystone.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. mostly marine bivalve molluscs. Buried clams leave tubes. extended above the surface to maintain water currents needed for respiration and feeding.
Semi-preserved: meats cooked and packed in spiced vinegar. sun-dried. clam madrilène and clam liquor. trimmed and packed. the meats are smoked and packed in oil in cans or jars. oyster soup. Dried: shelled meats. either whole or minced. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. Live: in shells. Smoked: meats. tidal waters or in brackish water in the mouths of rivers. or shelled meats. in cans with either brine or clam liquor. Frozen: shelled meats.1. all in cans. hermetically sealed. 57 . skewered with bamboo sticks.2. Canned soups include clam chowder.MOLLUSCAN SHELLFISH the giant clam of the Indian and Pacific oceans may be nearly 130 cm long and weigh 227 kg. uncooked. Canned: meats removed from the shell by steaming. after precooking and brining. smoked meats are also canned.2 Oyster Oysters are marine bivalve molluscs. or shelled meats. uncooked meats packed unprocessed. Frozen: in shell. These molluscs feed on minute plants and animals carried to them by the current. and no siphon. Soups: oyster stew. 3. oyster bisque. Fresh: in shell. uncooked (shucked). washed. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. packed in weak brine. Oysters lack a foot or have a reduced foot. having rough and irregularly shaped shells that are closed by a single adductor muscle. Canned: clams are steamed open and the meats removed from the shells. Dried: Hoshigai (Japan) shelled meat. Live: in shell Fresh: in shell or shelled meats. then heat-processed.
The familiar marine mussel is asymmetrical. smouldered and afterwards dried (Japan). The shells are fluted and round with a broad. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. flattened ear or wing at the hinge area. and held apart. Fresh: shelled meats (often only the adductor muscle and the roe are eaten. or packed with jelly. The adductor muscle is the only edible part of the animal. with the posterior ends of the shells (valves) broad and rounded. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves).1. and the anterior end smaller and pointed. Live: whole with shells. vinegar solution. Smoked: meats may be canned in edible oil. Fresh: cooked meats. are united. or filaments. Bait: used live extensively for baiting fish hooks for line fishing. mussel paste. in mussel liquor.) Dried: peeled. scallops possess a single large adductor muscle.2. They have filibranch gills. by interlocking tufts of hair-like cilia.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. scallops can swim or leap by a series of rapid closings and openings of the valves. The posterior halves of the valves contain a large adductor muscle that holds the valves closed. gutted. boiled. mussel in butter sauce. The common scallop is about 5 cm long. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves. also with other ingredients.FISH AND SEAFOOD 3. brine. 58 . sauces. in which the individual branches. Semi-preserves: cooked meats with vinegar-acidified brine and spices. Internally. Unlike other sessile bivalve molluscs.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms.2. Salted: cooked meats bottled in brine. cooked meats packed in dry salt for transport. In some species. Canned: cooked meats.1. 3.
also bottled in brine. Fresh: meats removed from the shell by boiling. Vinegar cured: bottled in malt vinegar after brining. Cockles range from about 1 to 15 cm in length and have two valves of equal size. Live: in shell.MOLLUSCAN SHELLFISH Frozen: shelled meats. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms. in sauce.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. which affects the temperature. clams and mussels. Whatever contaminants are in the water will eventually get into the shellfish (7). nutrient concentration and pollutants of the water column. Canned: in brine. The microflora are dependent upon a number of factors such as season. The following pathogenic microorganisms causing illness in humans are characteristically associated with molluscan shellfish: Vibrio parahaemolyticus Vibrio cholerae O1 Vibrio fluvialis Vibrio hollisae Vibrio vulnificus Vibrio cholerae non-O1 Vibrio mimicus Vibrio furnissii 59 . mostly in oysters. 3.1. are vectors of disease transmission (8). Canned: in own juice. pH. and butter. 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. 3. they accumulate pathogenic microorganisms from polluted waters. Salted: meats either lightly or heavily dry salted. depending on length of journey.2. Raw and partially cooked shellfish. and the environment. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding. Because molluscan shellfish are filter feeders. salinity.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves.
13).3. observed microscopically. (9.000 /g. Temperature. and shucked shellfish that is shellfish.3. mussels. Enterococci spp. 3. whole or in part.. There are microorganisms.1.2. but have never been cultured (14. shellstock (live) which is shellfish in the shell.. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. salinity. Micrococcus spp.3. Acinetobacter spp. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish.. In summary. spirochetes of the genus Cristispira and Saprospira. Pseudomonas spp. For example. Bacillus spp. 12.. and cockles The processing of bivalves takes two forms. Water temperatures affect the concentrations of microflora in the water. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. Other microorganisms found in lesser numbers are the Achromobacter spp.. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. rakes.1 and 3. and Aeromonads. Moraxella spp. the initial flora composition and concentration of bivalves are highly variable. Alcaligenes spp..1 Shellstock 3.1. and yeast cells of Rhodotorula rubra and Trichosporon spp. 10. Vibrios.1 Processing and its Effects on the Microflora Oysters. that have never been cultured on laboratory media. or by 60 . non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. 3.. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. dredges.1.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs.. The largest groups found are the Pseudomonads. Flavobacterium spp. mechanical harvesters.. clams. 15). Campylobacter coli Bacillus cereus Hepatitis non-A. At harvest. responding to the environment in which they are harvested. 11.3 3. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. nutrients. They are termed viable non-culturable microorganisms. from which one or both shells have been removed.
Secondly. Shellstock are placed in drycooler storage containers in the facility. after harvesting.1). wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. usually in open-mesh containers and/or burlap sacks.1. The shellstock is transported dry. Water used for shellstock washing is from approved growing areas. by truck or boat. There are three primary reasons for wet storage: first. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. wet storage also may be used to de-sand those shellfish species that tend to accumulate sand in their mantles and gills thus making them more palatable. Thirdly. or immediately placed in refrigerated trucks. Generic flow diagram of live shellfish from harvester to point of sale 61 . shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. or from other sources approved by the State Shellfish Control Agency (SSCA). or in bay-approved shellfish-growing waters prior to sale or processing.3. in containers from which they can be retrieved easily. for transportation to the point of sale.MOLLUSCAN SHELLFISH hand (Figure 3. If by boat. Fig. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock. Shellstock may be wet-stored at the processing facility in tanks of clean seawater.
vulnificus. shellstock are placed in refrigerated trucks for transport or placed in wet storage. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). to avoid possible contamination of the shellstock. for a considerable length of time. synthetic seawater for the purpose of de-sanding and salt uptake. An outbreak of infectious hepatitis in Sweden in 1956. Harvest conditions are another factor that influences the growth of bacteria in shellstock.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. 25). Over a period. Klebsiella. there is evidence that in the absence of adequate refrigeration. with prolonged storage the microbiological quality of the bivalve decreases. and Citrobacter species) and Vibrios. 22). There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. and temperature storage of shellstock to point of sale. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7.3.1. including V. 16. when it reaches the processing facility. The washing of sediment and detritus soon after harvesting will prevent further contamination. 17). shellstock are not refrigerated on the harvest boat. some pathogenic organisms will survive and multiply in shellfish (20. coli. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. with the exception of A. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. hydrophila (18). 26). If long delays occur before reaching the processing facility and/or refrigeration. 23. Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. Generally. are prevented. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. bivalves tightly close their shells thus trapping water and microflora. and the sanitary practices applied during harvesting and shipping. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. The most important factor in controlling bacterial growth in shellstock is temperature. Enterobacter. At this temperature the multiplication of faecal coliforms (E.1. without adequate refrigeration. 21. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . Generally. At harvest. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. handling. the bacterial counts in the meats will increase.FISH AND SEAFOOD 3. Water used for shellstock washing should be of good sanitary quality. and thus contaminating the shucked meat at the point of sale. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). Furthermore. 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. 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). 24.
In the blower-washer.1.1. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. or subjected to splash from foot traffic. the finished product may become contaminated. shellstock is subjected to contamination when stored on the floor in standing water. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank.3. 3. If shellfish are not reasonably clean at the time of processing. Due to the nature of the shucking operation. lobsters.2.2 Effects of microflora during processing The same considerations as previously discussed. thus contributing to high bacterial counts in the final product (28). washing and culling shellfish prior to storage is essential.2. Shuckers routinely use either cotton or rubber gloves during the shucking operation. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported. clothing becomes very soiled. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). 3. 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). for the effects of microflora during the processing of shellstock. To remove the shellfish attached to the remaining valve. Containers are placed in packing boxes for shipping at refrigerated temperatures. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station. In addition.2). The process of shucking involves forcing a knife between the valves and cutting the adductor muscle of one valve freeing the shellfish from one valve.1. through which the washed meats are discharged for rinsing and then packed in containers.3. the tank is opened through a drain gate and chute. washing.3. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. After washing of meats.2 Shucked shellfish 3. Studies have shown the use of rubber gloves reduces the contamination level. and fish. are applied to the shucking operation of shellfish. and packing shellfish into containers (Figure 3. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. Under these conditions if the shuckers enter the packing room.1 Processing description The processing of shucked shellfish is limited to washing of shellstock. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. removal of shellfish (shucking). whereas cotton become wet and 63 . a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. Thus.
2 ºC or less. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. Shucking containers are rinsed clean with running water. 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. Fig 3. within one hour. which may lead to contamination of product. Shucked meats are thoroughly drained.2. and sanitised before each filling. 2. 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. 3.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). There is a tendency to store such articles on the shucking benches or in packing rooms. and gloves help prevent the contamination of shellfish. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. 4. aprons.
47 to 6.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. As described previously. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. These organisms may play a significant role in the spoilage processes. these include lactobacilli. anywhere during the process of shellfish from harvesting to the point of sale.2 ºC or less. rough.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. Temperature abuse will result in high bacterial counts. where there is a breakdown of temperature control or prolonged storage of shellfish. and packing the shellfish in a timely manner. at the turn of the century. Shucked shellfish are held and transported at temperatures of 7. particularly oysters stored at 7 ºC. are an excellent medium for growth of bacteria. Shucked shellfish are an excellent medium for the growth of bacteria. 36). Refrigeration and shipping of shucked shellfish in wet ice is highly recommended.8% (37). Shellfish. spoilage will occur. and yeasts. Glycogen. 34. in oysters. 65 . particularly shucked meats. The spoilage process in shellfish may be divided into three stages: increase of acidity. a carbohydrate. and proteolysis (10). 31. abundant gas production. 33. 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. Much of the work has been done with oysters and clams. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. Alcaligenes. Earlier studies. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. Proteus. showed that organisms such as Achromobacter. In summary. Pseudomonas. 3. Micrococcus. The environment has an effect on the bacterial population found in shellfish. Flavobacterium. Lactobacilli seem to be the predominant microflora in shellfish. Glycogen is hydrolysed by the initial flora found in shellfish. shucking pails. All equipment coming in contact with the shellfish such as colanders. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. For example. whereas bacterium such as Pseudomonas. Achromobacter and Flavobacterium levels decrease during storage. blowers and other utensils that have cracked. 35. 32).2 ºC. glycogen levels range from 0. The initial flora found in shellfish is the major factor of spoilage in bivalves. This process changes the bacterial profile to microorganisms favouring low pH. Controls that should be applied include using clean packing equipment. thus producing acid and gas (38). streptococci. is stored in the bivalve as a source of energy. skimmers. clean gloves.
parahaemolyticus. primarily in brackish or saltwater. It is a common marine bacterium.5. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8.FISH AND SEAFOOD 3. Diseases caused by Vibrios are enteric in nature. coastal areas). however. there is a recent report of low levels of V. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. parahaemolyticus possibly producing illnesses (40). lot and retail outlet. This bacterium is present in all the major processing steps of a shellfish plant.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. Between May and September 1997. diarrhoea. They can be introduced to the shellfish from the environment or added during processing. One of 250 cases reported involved shucked products with low levels.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. parahaemolyticus multiplies rapidly at temperatures above 20 ºC.1 Vibrio parahaemolyticus V. V. 3. ranging from epidemic cholera to sporadic cases of diarrhoea. 3. and to the point of sale.1. during processing.5 cases/100. parahaemolyticus causes gastroenteritis lasting 24 . and estimates based from a hospital study show less than 0. The incidence of V. Products involved were from the same processor. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. and very common in Japan. widespread in both polluted and unpolluted waters.5. shellstock and packing of shellfish in jars) (29). Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. Fortunately.000 population/year (46). only a small percentage of strains in the marine environment are potentially pathogenic (45). The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. V.e. nausea. 66 . Major outbreaks are associated with the warmer months. Sporadic cases are frequent along the coast of the United States. where outbreaks occur regularly. 41). Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). headache and fever. These organisms can be present in the shellfish from the point of harvesting. and harvesting of shellfish in the cooler months. 44). prevention of cross-contamination. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42).48 hours with abdominal pain. approximately 23 cfu/g. parahaemolyticus from shellfish is low. from harvesting to finished product (i. although non-hemolytic strains have been associated with illness (43. of V.
The organism attaches to the intestine and release of a toxin is suspected. cirrhosis.5. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. and vomiting. chills. or immunosuppression. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. cholerae non-01 (51).MOLLUSCAN SHELLFISH Control of V. Healthy individuals are susceptible to gastroenteritis.7 days. cholerae non-01 has been associated with septicaemia. and rapidly become inactive at low storage temperatures. 48). with a fatality rate of 50% for individuals with septicaemia. leukaemia. parahaemolyticus growth in shellfish meats is temperature dependent.4 . nausea. Primarily. or primary septicaemia. fever.0. 3. and preventing cross contamination once the product is cooked. are particularly susceptible to primary septicaemia. and nausea within 24 . the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). Control of V. parahaemolyticus the estimated annual incidence rate is very low. 57. All isolates are considered pathogenic. cholerae non-01 is ubiquitous in the estuarine environment. High-risk individuals. vulnificus is by rapid refrigeration during warm weather months. 56). diabetes. They are sensitive to cold. 3. 58). Illnesses are usually associated with the consumption of raw oysters as described in Table 3.3 Vibrio cholerae non-01 Illness caused by V. V. in the range of 0. vulnificus are fever. It causes diarrhoea. 49).5.II (8. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). gastroenteritis. these individuals are advised not to eat raw shellfish. Oysters are the most hazardous food associated with V. Onset of symptoms occurs in 48 hours and can be severe. The bacterium is an estuarine species that causes wound infections. Control of oyster-associated 67 .8 case/100. cholerae. 54. abdominal cramps. lasting 6 . cholerae non-01 is generally a less severe gastroenteritis than that caused by V. 23 of 24 major USA-West Coast estuaries were positive for V.000 population/year (8.48 hours of onset. however. Hazards from this bacterium can be controlled by thorough cooking of shellfish. like V. In one study. The occurrence of V.2 Vibrio vulnificus The symptoms of V. Inadequate refrigeration will accelerate the growth of V. Again. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. which have been implicated in shellfish-related disease outbreaks (31). 46. those who have liver disease.1.1. 53. V. 46. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. V. 55. parahaemolyticus. vulnificus. the organism has been found in the Gulf of Mexico. It is one of the most severe foodborne infectious diseases. Death has been reported to occur within 36 hours from onset.
mimicus.5.5 Other Vibrios spp. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. Onset of disease is 6 hours to 5 days. or cross-contaminated shellfish (57. preventing crosscontamination of cooked seafood. Illness can include abdominal cramps. parahaemolyticus and V. There is evidence of strains of V. these organisms cause gastroenteritis lasting 24 . particularly in oysters. hollisae. 3. V. 63). nausea.5%) and 2 . drying. imported product from a country experiencing an epidemic. associated with shellfish-borne illness outbreaks. fluvialis.1. undercooked. headache and fever. cholerae 01 is transmitted by faecal contamination of food or water. Poor sanitation and contaminated water supplies will spread the disease. In addition. is the same as previously discussed for the non-01 strains. salinity. They are common marine bacteria. fever. and V. cholerae 01 establishing in the U. dehydration. The presence of these strains appears to be in low numbers and non-virulent (59).5. however. (8.FISH AND SEAFOOD V. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). cholerae 01 in molluscan shellfish. V. including V. nausea. jejuni requires reduced levels of oxygen (3 .10 days. and illness lasts 7 . processors should know the product source. 3. Death may occur after severe fluid and electrolyte loss.10% carbon dioxide for optimal growth conditions. vulnificus.5. 62. The frequency of infection of these organisms is no different from other Vibrios. 60). widespread in both polluted and unpolluted waters. and the transmission of the disease by consumption of raw. The control of V. 3. Onset of illness is from 3 to 5 days after ingestion of contaminated food. In developing countries. V. cholerae 01. and nausea.2 Campylobacter jejuni C. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. cholerae non-01 illnesses is by cooking seafood thoroughly. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. Gulf coast environment.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. In general.S. Control of infection from these organisms is no different from the other Vibrios. and shock. abdominal pain. and southern Asia including India and Bangladesh.g. There are several other Vibrio spp. low pH. the pathogenic severity is lower than V. respectively.1. It causes diarrhoea. vomiting. diarrhoea.48 hours with abdominal pain. acidity and heat. e. 68 . Illnesses are associated with the warmer months. 61.
V. Human feeding studies suggest that about 400 . cholerae 01 and non-01. parahaemolyticus. 85). hydrophila group (75.MOLLUSCAN SHELLFISH TABLE 3. Campylobacter spp. 76. In one case. C. control of C. 69. 71). however.3. 68.500 cells/gram of Campylobacter may cause illness (64. from Pacific shellstock oysters purchased from a retail grocery store (74). infection occurred among participants of a firemen's banquet. 81. To reduce the risk of shellfish-borne campylobacteriosis. 70. shigelloides (79. 83.4 to 114 cells. 73). They have been implicated in limited shellfishassociated gastroenteritis cases (8. have been isolated from shellfish beds throughout the world (67. jejuni begins in the harvest of shellstock from approved waters. of Incidents 4 279 0 1 0 No. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). although their role as a pathogen is highly controversial and circumstantial. 78) and P. ranging from 0. 65. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. of Cases 4 362 0 1 0 a Includes V. Recently. jejuni were recovered. 3. 84. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68.5. V. 82. hollisae Infective dose varies with host susceptibility and virulence of the strain. This finding is significant in that C. and their immediate proper refrigeration. V. mimicus. they survive well at refrigerated temperatures. 77. 80) are ubiquitous in the marine environment. fluviallis.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish.5. Although. The seasonality of 69 . C. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. 66).3 Other organisms of concern 3.II Molluscan shellfish associated outbreaks caused by Vibrio spp. and V. jejuni does not multiply in shellstock at refrigerated temperatures. V. Campylobacter appear to survive well in oysters during storage. however. 72. vulnificus. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. whereas at 10 ºC Campylobacter survive only for 5 days (71).
cholerae V. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. Caution should be exercised by individuals with haematological malignancies (88). Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. FDA/Administrative Guidelines 3 Source (91).600. prepared sandwiches. vulnificus in molluscan shellfish. 87) of the outgrowth of A.000 to 3. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. negative for heat-labile toxin (LT) or heat-stable enterotoxin (ST) 1 Negative for the presence1 Negative for the presence of toxin producing 01 or non-01 organisms 1 <10.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. 3. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. hydrophila group stored at refrigeration temperatures. 70 . coli Salmonella V. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. 2 Source (90). FDA/ Administrative Guidelines. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures.000/gram (29). They are easily destroyed with mild cooking temperatures. jejuni V. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. Test Standard Plate Counts E.0001 negative for the presence3 Under review4 Source (89). coli Enterotoxigenic E.000 cfu/g1 <230/100 g2 <1000 per gram. Note: Although molluscan shellfish are not included in the selected high risk food. In one study. parahaemolyticus Staph.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. Control of these organisms in shellfish products is limited. dried milk/whey drink powders. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. Similar results were seen by other investigators (86. and where indicated are intended to be used for official control purposes. non-dairy frozen desserts. being isolated more often during the warmer month. soy products. aureus C. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation.
MOLLUSCAN SHELLFISH 3. 231-51. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). Infectious diseases associated with molluscan shellfish consumption. Eliot C. Old H.S.. Van Nostrand Reinhold. 4. 1940. 29. Washington D.. 7. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. 1964. Applied Microbiology. Philadelphia. Colwell R. 8. 1973. 64.W.N. Journal of Milk and Food Technology.D.. 1936. 72-8. 1970. Gill S. Rowse A. 544-8. Microbiological characteristics of Dungeness crab (Cancer magister).C. Saunders Company. 105-8. Lee J.. Jr. in General Zoology. Ed. Applied and Environmental Microbiology.L. American Journal of Hygiene. 447-52. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. Ward D.. Applied Microbiology. USA. 1981. Ray S. American Journal of Public Health.F. Fleet G. Liston J. Microbiology of bivalve molluscan shellfish.R. Fate of enteric bacteria in estuarine sediments and oyster faeces. 71-6. 12.. 1968. 33643. water and sediment from Galveston Bay. 7.. Effects of harvesting waters and storage conditions on yeast population in shellfish. 6. 17. 10. Hackney C. Hood M. Fish and Wildlife Service.. 1975. 36. 5. Tubiash H.. Lovelace T. 1960. 1983. 8. 1995.7 1. Jr.E. 30. 13.A. 46.. Walker W. 633-40. Eds.R. 11.K.. 2. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. American Journal of Public Health. New York. in Microbiology of Marine Food Products. Eds. 755-76.S.. Applied and Environmental Microbiology. 44. 1963. Nauman R. 19-34.B. Bacterial flora of the market oyster. 1982. 104-9. Villee C.S. U.M.R. Galtsoff P. in Fishery Bulletin 1964 No. 419-25.H. The American Oyster. 1984. 1991. Smith F. 16.W.R. Bernard F. Rippey S. Tall B. 58. 6. A typhoid fever epidemic caused by carrier bootlegging oysters. References The Phylum Mollusca. 33-6. 1926. W. Biology of the oyster in relation to sanitation.S. 14. 42. 82-7.R. Thompson C. Clinical Microbiology Review. Schmidt C. Vanderzant C. Viability and release Salmonella charity and Escherichia coli from oyster faeces. Colwell R. Galtsoff P. 245-7.C. 30. in Multilingual Dictionary of Fish and Fish Products. 9. Cook D. Cook D. 15. 3. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas).E. Journal of the Mississippi Academy of Sciences.R.A. 13. Pfeifer D.A. 1994. Journal of Food Protection. Proceedings of the National Shellfisheries Association. Government Printing Office. Wiley-Blackwell. Organisation for Economic Co-operation and Development. 26. Veliger.K.. 71 .J.
26. American Society for Microbiology. Washington D. Vasconcelos G. 1961.. Journal of Food Protection. U. Journal of Food Protection.S. 22.M. Olympia oysters (Ostrea lurida). 1964. Food and Drug Administration. 27.FISH AND SEAFOOD 18. and storage.S.A. Timetemperature effects on the bacteriological quality of stored shellfish.. 4. 324.. Presnell M.. 1490-5. Applied and Environmental Microbiology. 53. F61-9..B.W. Part II Harvesting. Abeyta C.D. Colburn K. Beck W. 31.A.E. LaRock P.H. 72 . Jones W. Sakazaki R. Journal of Food Science.W.. 1995. Kaysner C..A.C. 24. Ltd. re-laying. Roos B. 1205-10. Establishment of critical control points in west coast oyster processing plants. Saikon Publishing Co. Son N.S. and Manillal clams (Venerupis japonica).B.. Arcisz W. Ed. 23. Food and Drug Administration. Koburger J. 343-5. 50. 123-8. Bacteriological changes in live shellfish: Pacific oysters (Crassostrea gigas).. Maryland. 19. 21. Public Health Report. 45. Public Health Service Technical Report. Chesapeake Science. Special Report U. 32. 20.T. 5. 1971. Applied and Environmental Microbiology.B. 343-9. Kelly C. Incidence of Samonellae in clams. Presnell M. in International Symposium Vibrio parahaemolyticus. 25. 29. seafood-handling procedures on Vibrio parahaemolyticus. Rodrick G. 1221-8.J. Bacteriological survey of an oyster bed in Tangier Sound. Svenska Lakartodningen. during depuration. Public Health Service. Influence of U. Williams L.. in National Shellfish Sanitation Program Manual of Operations. Effects of storage on microbial loads of two commercially important shellfish species. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. 1214.G. 1956. Temporal occurrence of Vibrio species and Aeromonas hydrophila in estuarine sediments. Hepatitis epidemic conveyed by oysters. 994-1002. Trost P.W. Ruple A. 1973. Bacteriological studies of commercial shellfish operations on the Gulf Coast. 1989. 1967. Crassostrea virginica and Mercenaria campechiensis. Cook D. Crassostrea commercialis.A. 47 (5). 989-1003. 1984. I.. Wekell M. Hoff J. U.. Vaughn M. Sadaguchi G. Eds. Tokyo. 1985. Survival of enteric organisms in shellfish. Stott R.. 167-71..C.. Applied and Environmental Microbiology. Kelly C.W. The influence of time and temperature on the bacteriological quality of shell oysters during processing and shipping. 1954. Behaviour of pathogenic bacteria in the oyster. 28. crabs and mullet.H. Food and Drug Administration.W. FDA Gulf Coast Technical Services Unit. 40. 30. oysters.F.J. 69. Fleet G. Takeda Y.. 1980.. U. Jr. Hood M.S. Fujino T. 52. P-1.E. Ericksen T. 1983..A. Blake N.A. Handling and Shipping Shellfish... Ness G.. Native Littleneck clams (Protothaca staminea).J. Fraiser M. 1992. Liston J.S..
204-5. 60. 73 . Fieger E. 1976. Jr. 1349-51.N..A. Vanderzant C. 1989. The bacterial flora of market oysters. Hoge C. U. Ward W. Jacobson M. 33. 133-47..A. Berry F. 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. Abeyta C. Bacteriological control of oysters during processing and marketing. 47. Academic Press. Powers C. DePaola A. Miyamoto Y. 38. West Coast States and British Columbia. Jr..M... 1961.T.F..G. 27. 97. Takikawa I. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States west coast. 2891-3. in Fish as Food..M. 42. 69. 1997..S. Journal of Bacteriology..W.. Takeda Y. Vibrio infections associated with raw oyster consumption in Florida. 39.M. Arcisz W. Yamai S. Abbott S. New York... 353-7. Thompson C. Joseph S.. Serological and hemolytic characteristics of Vibrio parahaemolyticus from marine sources. Bacteriological examination of oysters stored at low temperatures.A. Takizawa K. In vitro haemolytic characteristics of Vibrio parahaemolyticus: Its close correlation with human pathogenicity. 1997. Public Health Reports. Ishibashi M. Obara Y. Stroh E. Food and Drug Administration. Journal of Infectious Disease. Peeler R.M. Sidwell V. 41.D. U. Journal of Food Protection.. 1147-9.G.W. 1937. Shellfish Specialist. Microbiology of shellfish decomposition.. 38. Bothell..M. 36. Borgstrom G. 1.. 160. 100. 37. 44. 2. Ed. 45. 1958.. Grodner R. 273-80. Akiyama S. Epidemiology and spectrum of Vibrio infections in a Chesapeake Bay community.MOLLUSCAN SHELLFISH 32. Journal of Infectious Disease.. Infectionskrankheiten und Hygien. Bulletin of the Johns Hopkins Hospital. Hlady W. 313-22..L. 41. Leitch J. Morris J. Kelly M. Parasitenkunde.B. 1989. 35. Libonati J. 128-31. Applied Environmental Microbiology. Studies on pathogenic halophilic bacteria. Bacteriological findings in Baltimore oysters. 43. 1926. Wekell M. 25. 1979. Yokohama Medical Bulletin. 107-8. Tanikawa E. 27. Joseph M.. Watsky D. Kaysner C. The bacterial content of market oysters... 13-7. Sample T. Journal of Clinical Microbiology... 1914. Washington Report October 14. kanagawa-negative Vibrio parahaemolyticus from patients and the environment in the Pacific Northwest. 1916. 985-93. (2). Geiger J. 40. Marine Fisheries Review...E. Kelly C. Journal of Bacteriology. Loomis A. 46. Janda J. Journal of Food Science.P. 2820-2.. Crassostrea virginica.L.D.. 1987. 1954. Urease-positive. Volume 1. 41. Stott R. Zentralblatt für Bakteriologie. Kaysner C.A. Israel E. Vibrio parahaemolyticus illnesses.S. Kato T. Novak A. Journal of Clinical Microbiology. 1969. 53.. 34. Geographic and monthly variation in composition of oysters. 1989.C.
Hug A. Jr. Zywno S. cholerae in Louisiana. Blake P. epidemiologic. Gulf Coast. Duran W. Becker S.. Weaver R. Annals of Internal Medicine.. Wilson T.. 10. 1988.L.T.... Jr. 51..B.. Snyder J.W. Hickman F.E. Applied and Environmental Microbiology. Becker R.. Non-01 Vibrio cholerae bacteraemia: Case report and review. 1981. New York. 1985.M. 305-9.T.M. 49.. Puhr N.M.. Allegra D. Leitch J.O. Colwell R..A. 1979. New England Journal of Medicine..A. serology. Blake P... McCay C. McFarland L. Kaper J.R. Feldman R.G.G.M..B. Ecology. Conte J. Tacket C.C.J.A. 1984. 1982.A possible endemic focus in the United States. 54... Vibrio vulnificus: Man and the sea.. McPhearson R. 59. Riddle C. 60. 802-6. Wachsmuth I. Applied and Environmental Microbiology. Kaysner C. Review of Infectious Disease. 343-50... Applied and Environmental Microbiology..F.A. 91-103. Maden J.J. Johnston J.. Kaper J. 52. 94.. Barrett T. Colwell R. Twedt R.G. 8.. Lancet. An epidemiological study of V...G..D... Twedt R.R. Plaard R. 61. Pons V.M. Ecological relationship between Vibrio cholerae and planktonic crustacean copepods. 58..T. 1475-8.. Rahman R. 1. Craig J. 55. Non-01 Vibrio cholerae in shellfish.. Wekell M.P. 1983.. 46. 1980. Lockman H. and laboratory characteristics of sporadic cases. Jr.. 305. 312. Bradford H.. New England Journal of Medicine. 1981. Hug M. 1012-7. 5972. Adams M.. Jr.J. 56..A. 1984..G.F. Applied and Environmental Microbiology. 45. Morris J. Hunt J. Roberts N. McFarland L... and enterotoxin production of Vibrio cholerae in Chesapeake Bay. 41. Hollis D. Hebert S. sediment and waters of the U. 1344-8. Cholera .M. Colwell R.M.B. 149. Lee J.. 2050-3.W.. Spite G. Jr. 253.. John Wiley & Sons Inc. Clinical features and an epidemiological study of Vibrio vulnificus infections.G.G.R. 558-61. 299-312..A. 1987.G. Blake P. Peeler J.. Morris J.E. Wilson R.V. 53. Jacobs R.K.. Illness caused by Vibrio damsela and Vibrio hollisae. Journal of Food Protection. Characterisation of Vibrios cholerae isolated from oysters.. 275-83. Davis B.R. Blake P. Siebeling.O. DePaola A... Abeyta C. Brenner F. Morris J. Morris J. 1983.. Presnell M. Miller H.. Jr.T... Caraway C.M. Roderick G. 1985.T. 53.C. West P.I. DePaola A. 57. Motes M.. Non-O group 1 Vibrio cholerae gastroenteritis in the United States: Clinical. Small P. Jr. Ed. 37. Safrin S.A. Journal of Infectious Disease..A. 50... 1294-7. 1982. Tacket C.M. Microbial Ecology. Journal of the American Medical Association. Wathen H.F.S.N.D.. 656-8. Bradford H.W...W..A..E. R. Incidence of Vibrio cholerae from estuaries of the United States West coast. Black R. Vibrios in the Louisiana Gulf Coast environment. Feeley J.FISH AND SEAFOOD 48. Stott R. Wazenski T.. 74 . in Vibrios in the Environment. Joseph S.. Cholera and other vibrioses in the United States.. Francis D.O.E.
J.G... 101-4.. in Campylobacters.A. Microbial Ecology in Health and Disease.A. from patients with diarrhoea.. Infective dose of Campylobacter jejuni in milk. 1980. Feldman R.. Eds. 1996. Davis B.E.M.A. Blaser M. 1991.. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster.R.M.S. Huges T. 157... International Journal of Food Microbiology. 13. Blake P.S. 6-9 September 1983. Jr. Griffin M. Rowe B. in Campylobacters. Black R.. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N.. Abeyta C. Jr. Isolation of vibrio-like group. 1584. Eds. Austin S.. S41. 11.J.. 66. 4..H... Black R.. 4.F. London. Wilson T. 4...L.. 256. 80.I. Blaser M. Disease from infection with Vibrio mimicus. Shandera W. Moore J.R. Robinson D. Journal of Clinical Microbiology. Abeyta C. 99.. Moore J.M. Ketley J. 1993.. Journal of Infectious Disease.E.J. 282.B. Plenum Press. 169-71. Ireland.R.J..A.P. 1983. in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections. Wilson I.. Wareing D.M.. infections in man.M.D.A..M. Brenner D. A method for isolation of Campylobacter spp. Helicobacters and Related Organisms.R.L.A. 323-5. C. Morris G..X. Kaysner C.G.G..M. 1980. Campylobacter gastroenteritis associated with raw clams. 1981. British Medical Journal. 64. 69. Journal of Food Protection.A.R.D. 73.E. Eds. 68. Levine M. Jones D. Brussels. Stott R. Abeyta C. Jr. Helicobacters and Related Organisms. Newell D. 56.. Deeter F... Proudford R. 51-64. Wilson T. Huq M. Clements M. Alam A. Skirrows M. (jejuni. 1995. Wilson I.A.. Newell D..K. New York. Fitzpatrick M. 257. Murphy P. New York.. Kaysner C. 75 . Feldman R. coli and lari) from shellfish and the marine environment. Kaysner.K. Plenum Press. 621-4. Wekell M. 67.M.E. 607-9. Dalley E.G.A.M. 71.. Wekell M. Studies of Campylobacter jejuni infection in volunteers. 63.P.K. a newly recognised Vibrio species. Levine M. Journal of the Medical Society of New Jersey. 1983...W.. Matsuda M.G. Journal of Rapid Methods and Automation in Microbiology. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness. 472-9.. Ketley J. Davis J. 1996. Hughes T. Hunt J..MOLLUSCAN SHELLFISH 62. 1987. Pearson A.G. Wareing D.G. Johnston J. Public Health Laboratory Service. 65. Arumugaswamy R. 72.. 1988. EF-6. Annals of Internal Medicine. Experimental Campylobacter jejuni infection in humans.. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp. Clements M.. 70. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation. Murphy P.
1988. Journal of Food Protection.M. 1986. 343-9.L. Tenge B. Journal of Food Protection. 1985.. GA..M.. Wekell M.W. 75. Tzipori S.H. Wekell M. Wachsmuth I. 1987.. Van Damme L. III. Robinson J. Applied and Environmental Microbiology.. Journal of Food Protection.A. Trost P. Atlanta..D. 731-8. Tall B. 82. 1981. Lockman H. 39. Hazan T.R.N. 48. 7-12. Robins-Browne R. 27.. Petersburg. Partridge K. 5. Kaysner C. 76 ..L.. Fla.T. Holmerg S. The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish.B..M..P. Applied and Environmental Microbiology. 52... 223-47. and treatment of Plesiomonas shigelloides diarrhoea. Fliermans C. Bark D.. Sullivan J. Stelma G.. 76. Oyster borne disease outbreaks in St. Department of Health and Human Services/Public Health Service/Food and Drug Administration. Farmer J.W.J. Blake P. 77. 53. 690-4... 84. 849-55. Vandepitte J. Jr. 79. 85.M.D. 36. Baker R.F.A.. vibrios and Aeromonas hydrophila. Hood M. 55. Hickman-Brenner F. 359-77. Plesiomonas enteric infections in the United States. Abeyta C.J. 47. Stott R.C... Gracey M..A.A. 81. 598-601. 1989. Journal of Clinical Microbiology. 1997. 449-57.. Annals of Internal Medicine. Hirsch R. Singelton F.R. 1146-9. Ruple A.. Jr. Herrington T. Burke V. Aeromonas hydrophila: ecology and toxigenicity of isolates from an estuary. Prevalence and distribution of Aeromonas hydrophila in the United States. Journal of Food Protection. Abeyta C.J.K. Journal of Food Protection. Infection and Immunity. Kaper J. 83. 86. Clinical features. Koburger J.A.L..M. Kaysner C.J.FISH AND SEAFOOD 74. Frequent isolation of Edwardsiella tarda and Plesiomonas shigelloides from healthy Zairese freshwater fish: A possible source of sporadic diarrhoea in the tropics.. Cooper M.A. epidemiology.. 80. 475-9... Seidler R. In vitro and in vivo pathogenicity of Plesiomonas shigelloides. Peterson D.. 47...C. Journal of Rapid Methods and Automation in Microbiology. 1990.. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness.A. 105. Herrington D. Beaman J.M. 1984. Cook D.M. Jr.. Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species.. Miller M. 979-85. 78.W... 1984. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. Levine M.. Colwell R. 1978.A. 1980. Plesiomonas shigelloides: An opportunistic food and waterborne pathogen.. 50. Incidence of motile aeromonads from United States west coast shellfish growing estuaries. Kain K. 998-1001.D. Applied and Environmental Microbiology. Kelly M. Journal of Applied Bacteriology. Abeyta C.. 87. Effects of processing and storing oyster meats on concentrations of indicator bacteria. Hunt J. 1989. Wekell M.. Esch G. Report (1/13/84). 52. Kaysner C.
S. U. fresh or frozenadulteration by bacteriological contamination Sec. Government Printing Office . in U. 89. Program 7303. Sherlock C. 77 .8 Further Reading Ahmed F.. Seafood Safety. Ed. National Academy Press. Burdge D.. 1987.S.R..030.W. Clams.S. 1995.. oysters. 1995.S. Smith J. Chapter 5: Foods.H. U. Technomic Publishing Co. Quality Control. Food and Drug Administration.L. 3.A. Washington D. Aspen Publishers Inc.C. Martin R.S.. Washington D. Baird-Parker T. 2000.. 63-8. Lancaster. Inc.C. Lund B.. Part I Sanitation of Shellfish Growing Areas.E. Ed. mussels. Fish Inspection. Food and Drug Administration Compliance Program Guidance Manual. 1997. Food and Drug Administration. U.S. Food and Drug Administration. and HACCP: A Global Focus. Food and Drug Administration. Food and Drug Administration. The Microbiological Safety and Quality of Food.1987. 1991. U. 90.S. Gaithersburg.E.S.MOLLUSCAN SHELLFISH 88.Pathogen monitoring of selected high risk foods (FY 88/89). Slavin J. Food and Drug Administration. Food and Drug Administration. in U.C. Food and Drug Administration Compliance Policy Guides. Washington D..C.W. U. Collette R. in National Shellfish Sanitation Program Manual of Operations. Food and Drug Administration.M.. 91. U. 560. Gould G. Ed.12).600 (CPG 7119. Pittsurgh. 7. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease.
4.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. e. Peeled and cooked shrimp are ready-to-eat after controlled thawing.5 kilo pack (boxes or bags). Peeled ready-to-cook shrimp are raw.5 and 1. packed and frozen in 0. the front pair ends in claws. peeled product. They possess a segmented body. jointed limbs and a chitinous exoskeleton. Species eaten from tropical waters include Penaeus spp. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. Shrimps. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi).. crabs. Crayfish. 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.0 kg polythene bags. and available in 0.g.. and Trachypenaeus spp. Pandalus borealis (deep-water shrimp). CRUSTACEAN SHELLFISH Linda Nicolaides. (deep-water shrimp). Such products are usually only partially cooked before freezing. and are usually aquatic. crayfish and shrimp (1). Pandalus montagui and Crangon crangon (inshore shrimp). lobsters and crabs have ten legs. and frozen in 1 and 2. Parapeneopsis spp. with additional water to form a glaze.0 kg polythene bags.Ph. M.5 and 1.. lobsters. 79 .
traditionally. However. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed. tropical waters. only live carcasses are processed. Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. during the peeling of cooked shrimp. Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors. Shewanella putrefaciens and 80 .2 Initial Microflora As with finfish. so that spoilage may occur during landing and transportation to the processing plant. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp.g.g. which means that the haemolymph can be a reservoir of bacteria. Following capture or harvest. In comparison. spoilage bacteria are of little importance as. the season. packed into polythene bags. lobster tails and cooked crabmeat presented decoratively in their shells. particularly for members of the genus Vibrio (2). for crabs and lobsters. with the increasing demand for ready-to-eat/cook crustacean products. hence.FISH AND SEAFOOD Crabs. boiled to kill the crustaceans and cook their meat prior to consumption. it is important that shrimp are handled and iced correctly in order to keep them in as fresh and uncontaminated a state as possible. For example. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria. If these products are subjected to temperature abuse. Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP). storage above the recommended storage temperature. shrimp die immediately after capture.. Cooked and chilled. the potential exists for crosscontamination from both the processor and from the environment. or frozen crab. their feeding and living habits. lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption. Pasteurised crabmeat is edible meat removed from the exoskeleton by hand. particularly after it has been cooked. stored and transported as frozen product. e. As with finfish. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. The structure of the circulatory system of crabs is not closed. which include the environment from which they have been captured or harvested. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance. 4. the geography of the area they are captured from or are grown in. e. and the temperature and quality of the waters in which they exist.
1. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. A large proportion of shrimps are produced by aquaculture. chilled sea water. Shrimp might be further processed with the shell on. The claws are usually bound together to prevent the animals fighting and causing damage to each other. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. up to 16 days for tropical species compared with 8 .1 Prawns and shrimps An example of the process used for preparing frozen. or are found in brackish water around the shores. Shrimps can be processed whole. Hence. gills and thorax. After capture and transport to the processing plant. They are then taken to market alive in either holding tanks. Vibrio spp.. from the time of capture and processing. graded according to size. In contrast. uncooked prawns and shrimp is presented in Figure 4. or with the shell and central vein removed. glazed and frozen (see Figure 4. or cooked. Shrimp will then be glazed and frozen in a raw state.1).. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected. lobsters and crayfish are harvested from cages or pots containing bait. the rate of deterioration of the crustacean shellfish by these organisms is reduced. whether GHP are in place. 4. thus. they are washed. When the shrimp arrive at the processing plant they are washed and then sorted into size. 4. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . These animals can survive out of water if the surrounding environment is kept cool and damp. As with finfish.for example. In comparison shrimps die quickly after capture.10 days for temperate water species (3). Micrococcus spp. Pseudomonas spp. 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. and Achromobacter spp.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. however. The rate of spoilage will be a function of time and temperature. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots. Alcaligenes.3. bacterial contamination and spoilage are inevitable. especially if they are stored in ice or iced water. the genera associated with tropical species include Enterobacteriaceae.. with “head on” or “head off” whereby the edible tail is removed from the head. corynebacteria. chilled 81 .3 Processing and its Effects on the Microflora Crabs. so they are either sold on the local market or are processed in local processing plants. or transported directly to processing plants. peeled and cooked. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed.
frozen uncooked prawns 82 .FISH AND SEAFOOD frozen. Fig.1. and packed to meet the customer’s specification. spices and cooking batter. After cooking. 4. Flow diagram . particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour. Good Manufacturing Practice (GMP). 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. Breaded products introduce an added risk of spore forming bacteria. including GHP. should be followed to ensure that the product is not re-contaminated with microorganisms present in the processing environment or from the workers.
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. 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.CRUSTACEAN SHELLFISH 4. whilst boiled crustaceans have a pleasant slightly sweet smell. wet and ‘soapy’ or ‘jammy’ to the touch. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. slightly iodine smell. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. With spoiled crabs and lobsters. 83 . with the growing demand for ready-to-eat products from crab and lobster meat. However. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. The shell will be soft. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. 4.3. and have a sweetish.4. for crabs. If hygienic procedures are not followed.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. the joint between the tail and carapace is open and the limbs hang loosely. viruses or parasites.4. The shells have a crisp. the meat is either repacked and sold in the shells. they include Sh. spoiling prawns and shrimps have shells that are soft. or in hermatically sealed cans that have undergone an appropriate heat treatment. there is no indication of life. Once removed and cleaned. since crabs and lobsters are traditionally consumed immediately after cooking. immediately prior to boiling. putrefaciens. 4. bright and dry appearance. An offensive odour is also produced. putrefaciens and Pseudomonas spp. If boiled. Heat is produced if the shrimps decompose in a confined space. or packaged and processed. there is no associated spoilage problem. contaminating microflora may include enteric bacteria.2 Cooked crabs and lobsters As indicated earlier. If the tail drops. 4. In comparison. dull and sticky.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. and a strong ammoniacal odour is produced.
shrimp and shellfish (11). monocytogenes in cooked ready-to-eat foods. in domestic/imported shrimps (15). later surveys in 1990 recovered the pathogen from retail raw. Although foodborne illness had not been associated with the product. 5. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. 7). Further surveys identified Listeria spp. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. 6. animal manure. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). although efficient cooking will quickly eliminate it (6). The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10).5. Hence.5. The presence of L. nearly 30 years elapsed before the potential hazard of L. 4. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. An import alert was put in place in June 1987. shrimps and cockles sold (17).1 Pathogens: Growth and Survival Salmonella spp.5% of frozen raw shrimps from different countries (13). in 24. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). as well as for Salmonella. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. Salmonella cannot be completely removed from raw shrimp during processing. Moreover.cooked prior to consumption. cooked and smoked fish. 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. even if GMP is followed. a common pond fertiliser used in aquaculture.5 4. Following this. and usually for both. monocytogenes was absent from 40 samples of cooked prawns. 16).FISH AND SEAFOOD 4. fish fingers. The above results prompted the FDA to develop a compliance programme for the pathogen. However. Aquatic birds and the hands of workers have also been identified as sources of the pathogen.2 Listeria spp.
70% of healthy individuals carry toxin-producing strains of Staph. Control should be achieved by promoting GHP and staff training programmes. V. 85 . which can become colonised by V.5. The former.5. However. V. about 50 . Researchers have demonstrated that inoculated shrimp supported toxin production by C. Once the competing microflora has been removed by a heat treatment.g. 4.5 Vibrio spp. cholerae O1 and V. 22) as well as from undercooked crustacea when harvested from contaminated waters (23. 4.3 Staphylococcus aureus Staph. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. 19). therefore. no incidence of foodborne illness caused by L. as part of the natural bacterial flora. pregnant women and the immuno-compromised. cholerae is divided into two groups. is widely distributed in aquatic environments. especially in products that have been vacuum packaged. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. Crustaceans have been linked with food intoxications attributed to Staph. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. 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. aureus because of the ubiquity of this organism in the processing environment. The main source of contamination is the hands of processing personnel.5. 24.CRUSTACEAN SHELLFISH old. An epidemic that began in South America in 1991. The pathogen has resistant spores. cholerae. botulinum type E at 10 °C but not at 4 °C (20). the causative agent of cholera. e. 25). the pathogen can grow at temperatures as low as 3 °C. Vibrio cholerae. particularly crabmeat (18.4 Clostridium botulinum C. monocytogenes has been attributed to crustaceans. Crabs are covered in a chitin exoskeleton. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. although contamination by types A and B may occur from the environment. cholerae non-O1. pasteurisation and cooking. combined with the degree of handling of these types of product. aureus on their hands. To date. botulinum type E is commonly associated with fish and fishery products. so is able to survive heat treatments that eliminate vegetative cells. aureus is not a member of the intrinsic flora associated with crustacean shellfish. whilst boils and cuts are also common sources of the pathogen. was associated with seafood consumption. 4.
jejuni has not. Quantitative levels were below limits of detection in all cases (<0. 4. however. been implicated in a case of foodborne infection associated with crustaceans.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. Parasitic infections are controlled by either cooking infected material prior to consumption. 32). to date. V. hydrophila is able to grow well at refrigeration temperatures (0 . e. Japan. 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. Norovirus. 4. Caliciviruses. V. However.5. 4. Hepatitis A virus presents the most serious hazard. whilst 5. particularly trematodes (6). Astroviruses and non-A.8% of samples contained C. although A. but has the potential to cause fatal foodborne illness (26). parahaemolyticus. hydrophila has been isolated from prawns (29). Of these. in a survey carried out in the United States between 1977 and 86 . 4. the potential hazard exists as C.30 MPN/g) (27). However. It is more commonly implicated in countries where crustacean shellfish are eaten raw. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28).8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. occurs in crustaceans from warm environments. a marine halophile. A. vulnificus is an invasive and lethal pathogen associated with wound infections. 31.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States.g.7 Aeromonas hydrophila and Aeromonas sobria A. non-B Hepatitis viruses (31).5. hydrophila and A. South America. or freezing at a temperature of -23 °C for 7 days (30.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A.6 Campylobacter jejuni and Campylobacter coli C. coli. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat.5.5. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture.
produced or imported into the European Union (EU). The toxin causes amnesic shellfish poisoning (ASP) when consumed.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. Selected parts from this regulation is presented below.5.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). although it is one of the European Union (EU) legislated allergens. are included in the Commission Regulation (EC) No. They can survive low pH levels.6. which results in a persistent and apparently permanent loss of short-term memory (36). or chlorine concentrations similar to those found in drinking water. from farm to fork (37. 2073/2005.CRUSTACEAN SHELLFISH 1981.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). which could be transferred to the meat during unskilled removal from the shell (35). 87 . 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.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. Norwalk viruses can also withstand heating to 60 °C.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. 4. consistent with that in the stomach. 38.5. 4. 4. The risk of such hazards is far higher with fin fish and molluscan shellfish. A summary of microbiological criteria for crustacean shellfish is listed below. 39). 4. only 0. No foodborne incidents have been attributed to this source to date. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. Provisions laid down concerning the use of additives would also apply to these types of products.
FISH AND SEAFOOD
22.214.171.124 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 126.96.36.199.1 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. 188.8.131.52.2 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. 184.108.40.206 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
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.
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.
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.
and a preliminary operation to some smoking. dependent on the country or the type of product. 93 . and before significant spoilage takes place. Usually. It includes the salting. SMOKED AND DRIED FISH Dr. and the extracted moisture is allowed to drain away. and salting is done by stacking the fish in layers. Wet salting or brining. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. fish is immersed for a few hours in a relatively low concentration brine. drying and smoking processes. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. CURED. The fish is headed. drying.5. and marinating processes. rather than for preservation itself. Fish curing is practised worldwide. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. both to preserve and to create a variety of new products.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 can be salted in three different ways or their combination: Dry salting. The processes take anywhere from a few hours to a few days or even months. alternating fish layers with a layer of salt. where the salt replaces moisture in the fish flesh. their combinations and/or variations. This can be called ‘kench’ curing and is mainly used for non-fatty white fish. and is sometimes known as ‘pickling’. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. Salting is both a method of preserving fish. split and opened out flat.
under normal pressure. The fish is mixed with dry salt but in a watertight container. After the freezedrying process. the ice sublimes and the vapour is removed from the fish by vacuum pumps. It has found limited application with fish. are added to the marinade. as concentrated brines are formed. where an organic acid. or boiling it with dry salt and water. this was done naturally by exposure of the fish to the air. Thus. prior to or during the boiling process. It is not commonly used for fish products. Freeze drying. Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. The products are usually consumed raw. it can also be named ‘pickling’ or ‘blood pickling’. thus delaying spoilage for a few days. Food products can be dried in three different ways: Air or contact drying. this ‘brine’ can be collected and placed again over the fish several times. As the water is drawn out of the flesh by the salt a brine is formed. Smoking is an ancient process for preserving and flavouring food. evaporated water is removed by vacuum pump. When the process is running in a closed system. the preservative effect is due to the low pH used in the process. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms. Marinades are acidified brines. depending on the type of product. is used as a preservative instead of salt. This is a mixture of a dry and a wet salting process. the product rehydrates quickly but does not retain reabsorbed moisture on cooking. and the fish can be immersed in such brines for long periods. Salted-boiled fish are the products obtained on either boiling fish in a brine. Traditionally.FISH AND SEAFOOD Dry / wet salting. Included in this group are some lightly preserved products where no acid is added to the marinade. as well as other flavourings (sugar. without other preservation methods applied after boiling. This is the common method of drying fish. Longer shelf life is achieved in the products when salt is added. Heat is transferred to fish from heated air or a heated surface. thus reducing the water activity of the product. usually vinegar. thus making the product unsuitable for microbial growth. The product is dried by contact with heated surfaces or by radiation. Marinating or pickling. such as ‘gravad’ fish. Salt-boiling. and the air movement removes the vapour. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. Vacuum drying. spices). Salt. Fish in contact with refrigerated plates freeze. 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 . Usually.
process temperatures are over 60 °C. leaving only elevated levels of carbon monoxide. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. or by means of smoke-flavouring processes with liquid smoke. Hot-smoking. or liquid smoke. Smoked fish is prepared by subjecting the fish to the direct action of smoke from wood or wood sawdust burning. Clear smoke. or by means of smoke flavouring processes. The different spices used in the immersion brines are also a potential source of microorganisms. In this process. such as injection or dipping in a solution of liquid smoke. With the exception of some Asian countries. Cold-smoking. However. which impart the flavour of smoke to fish. temperatures below 30 °C are used. Smoked-flavoured fish. It can be injected into the product. by exposure to the wood or sawdust smoke. fish can be smoked by means other than the direct action of smoke. the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. It is commonly used in place of traditional smoke. by exposure to the wood or sawdust smoke. or added to the brine with other flavourings. sporeforming bacteria or osmophilic moulds. It can be smoked either traditionally. and the preservative effect is only slight. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour.CURED. including pathogens. Usually. Smoking can be either traditional. the salt used in the processes may contain halophilic bacteria. the fish is subjected to heat for a period that does not coagulate protein. The smoke itself may contain spores of moulds. The process was designed so that the smoke is filtered to remove all smoke particles. 95 . 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. After the salting step. In this process. Usually. Also. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. The secondary objective is to impart sensory characteristics such as smoke colour and smoke flavour to the product. smoking is now used mainly to give the fish a characteristic flavour. and filtered to remove tars and particulate matter. 5. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. SMOKED AND DRIED FISH of smoke.
0 158.3 Processing and its Effects on the Microflora Owing to the wide variety of products. When a 100º brine is used.4 184. the fish is usually salted with a 70 .0 g of salt per litre of brine).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. it is important to state the temperature. At 18 °C a 100º brine is produced when the solution contains 26.2 g of salt per litre of brine). a 90º or above brine (237. Thus.0 . or salt in the water phase.6 132. Salt in the water phase links the two conditions of moisture and saltiness. Different brine strengths are required according to the curing processes and/or the type of product. processes and producers’ private recipes. When working with salting.6 g of salt or more per litre of brine) may cause salt burn in this way. is employed. The dissolution of salt in water is dependent on the temperature. % 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.8 211.8 79. about 50º (132.FISH AND SEAFOOD 5. This should be distinguished from the salt content. Therefore.2 105.8 211. When a milder brine. only the most common processes can be described here. it is impossible to establish standardised processing stages in the curing of fish. For smoking salmon. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. Salt concentration means the percentage of salt in the water contained in the fish. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. and salt content. it is important to understand the concept of salt concentration. etc.80º brine (184.6 264.4% salt (Table 5. it causes swelling because the fish absorbs water from the salt solution (1).4 52. The strength of the solution is described as degrees of saturation (brineometer degrees). TABLE 5. all using salting steps.2 237.I). smoking or marinating processes. a 0º brine saturation means pure water. sometimes the salt crystallises onto the surface of the fish.
CURED. and for a heavy salting a ratio of 1:3 salt:fish is used (1).0. addition of levels of 0. the fish is mixed with one part of salt per eight parts fish. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. it tends to lead to a slight yellowing and softening of the product. Usually it is followed by other preservation techniques such as drying and/or smoking.1 ppm and 10 ppm. producing characteristic pink patches. 5. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. when the air humidity is high. and such delays. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. if very pure sodium chloride is used for curing purposes.3. However. and salt can speed this oxidation process. The calcium and magnesium impurities. On the other hand. ‘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. but the concentration of copper and iron must be kept under 0. The most common fish used in the northern hemisphere is cod (Gadus morhua). are recommended to help make the product whiter.3. and essentially free from iron and copper (3). Salting can be done in a number of different ways. This is because fatty fish turn rancid quickly. respectively (1).1.35% and 0. The salt used should be of food-grade quality. 97 . respectively. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. mainly in tropical conditions. SMOKED AND DRIED FISH 5. 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.15% calcium and magnesium. lean white fish is used for this purpose. having a short section at the end of the tail. and the salt has penetrated throughout the fish (3). which include calcium and magnesium.1 Dry salting / Kench curing Usually. When pure salt is used. low in calcium and magnesium. outgrowth of halophilic bacteria may occur. the fish is restacked with fresh salt after about a week. ‘Green cured’ split cod with about 60% of the backbone removed. caught in North Atlantic waters. can be dried further to produce ‘klipfish’ or ‘bacalão’. for which there is a high demand in Mediterranean countries.1. This condition is known as ‘putty fish’ (4). Also. and a ‘putty’ fish product is also obtained (1).1 Salting Salting of fish for preservation purposes has been used since pre-history. For a light salting. however. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts.15 . are likely to cause hydroscopicity of the product and. See Figure 5. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage.
the process is mainly for flavouring purposes prior to the use of other preservation processes. Processing of dried salted cod 5.3. such as Staphylococcus aureus could survive and grow at such water activities in the product.1. The greatest barriers to salt penetration are the skin and scales. so that the risk of spoilage is high. it appears that water activities much less than 0.2 Wet salting or brining As referred to above. eviscerated and/or split). such as 98 . For large fish it is advisable to split it before salting. although toxin production would be unlikely (4). Fig.5.1. Only small fish can be salted whole. Food-poisoning organisms.90 are rarely achieved. Where fish are salted in their whole form. when a light brine is used. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat.
70 6.23 2.76 4.16 7.1.51 5.41 3.60 2. thus.66 5. the liquid that comes out of the fish forms a brine. the fish enzymes play an important role by maturing or ripening the product.46 4.78 6.35 4.85 3.45 3.5 3.0 4.26 1. Histamine formation is of importance when using scombroid and some clupeidean species.78) (1).61 3.56 6.41 6.22 6.5 3. For preservation purposes.26 3. 5. the brine is then replaced over the fish.84 1. TABLE 5. if split fish is dry-salted.88 5.23 3. sardines.00 3.5 4.76 4.76 4.51 3.80 5. Fresh sardines. with pressure being applied to the top of the container to help the brine to drain away.26 2.99 2.17 5.74 9. a saturated brine is used.04 5. with a shelf life of about 4 weeks (water activity (aw) about 0.41 4. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.11 9.03 4.54 5.95 3. and a red burgundy colour will develop in the product.11 3.65 2.49 4.II. This is why it is the preferred method for fatty fish.61 3. The main objective of the brining method of salting fish is to limit air penetration during processing.17 4.86 2. the fish is dry-salted.09 8.10 1.70 3.5 10. can be cured in a saturated brine solution for 6 days and.78 2.23 8. but as it is placed in containers. The fish are kept immersed in their own brine near saturation point for several days.II Salt in the water phase (after Ian Doré (5)) Moisture (%) 40 45 50 55 60 65 70 75 80 85 90 Percentage salt in the flesh (by weight) 2.85 3. There are several variations of this process. the product obtained is called gaspé-cured (5).91 2.25 5.47 6.09 8.0 2.33 5.26 4.90 (1).44 2. Anchovies are the main fish used for these purposes. Histamineforming bacteria do not generally grow at water activities below 0. See Table 5.06 4.0 3.76 99 .3 Dry / wet salting In this process.98 6.23 2. but mackerel. after being drained and pressed at 465 kg/m2 for 15 hours.73 1.25 5.66 5.64 4.30 2. headed and gutted.44 2. SMOKED AND DRIED FISH smoking. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .26 7.03 2. Here.41 5.3 days.98 5.CURED. and can be collected and replaced again over the fish. limiting the amount of rancidity that can develop.96 1.11 4.76 4.3. herrings and other fatty fish can be used. they can be stored at 28 °C. For example. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.76 4.98 6.19 3.05 7.
Traditional jellied eels are a product where pieces of fish are cooked in boiling water. Other preservation techniques can be used for extending the shelf life of these products. After this. salt. There is epidemiological evidence that these products have been the cause of some 100 . even if the product is hermetically sealed.3. Herring can also be cured for several days in an 80 to 90º brine with 2. usually flavoured with onions and hot chillies.30 minutes.6 days without refrigeration (1). a Peruvian product. and 6% salt. and even death. or a pH <5 and they are stabilised for a limited period.3. only the nonspore-forming organisms are eliminated in the process and. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. 5. and spices) is added to the brine. and packed (3). and lasts for 3 .2 months. Ceviche. giving a cooked appearance to the product. is made of different varieties of fish. it can be repacked in milder brine (1 . they can be cut and cured for 3 days in a solution with 3% white distilled vinegar.7%) for as long as 3 weeks. Another salt-boiled Indonesian product. The fish can be soaked in a high concentration of vinegar (about 4 . outgrowth of bacterial spores after processing may occur (4). All these products are ‘semi-preserves’. or can be boiled by being placed in a basket.FISH AND SEAFOOD 5. The first process can give a longer shelf life. As referred to above. flavouring (sugar. Once cooled. drying and salt-boiling steps until a hard translucent block is obtained (4). The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. However. A much firmer product is obtained when the fish is salted prior to acidification. underlines the need for careful control (4). usually requiring refrigeration below 10 °C for longer-term storage. Another variation uses sour cream instead of the second brine step. then packed in barrels with a 70º brine.5% 120grain distilled vinegar. but it has further smoking. which is dipped into a boiling brine for 10 . until the flesh is soft. Usually. whereas the latter method preserves the fish for only a few days. up to 1 . The lime juice plays the role of the vinegar.4% of salt) for distribution and sale (1). ‘ikan kaju’. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. They will have a salt content >6% (w/w) in the water phase. placed in a marinade of lime juice. because of the salt content of the product. In a second step. containing 1 kg of salt for every 25 kg of fish. the occasional implication of pindang in cases of foodborne illness. they are packed for consumption within 2 weeks at chill temperature (6). Afterwards. herring or other fatty species are used.3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt.2% acetic acid and 2 . has a shelf life of several years.
bacterial toxins and parasites (7). etc. used as an appetiser. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. such as squid. split and dried gadoid species) are traditionally made in cold climates. gravad fish. Stockfish (dressed. 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. by using either solar heaters or mechanical dryers. Gravad fish is a Scandinavian product made of salmon fillets. It takes about 2 days to marinate completely. gutted. 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). some fish species such as moray eel and varieties of dog fish. It is important to assure the quality of the raw material. Stockfish is extensively used in Portugal. the fish is dried after salting or brining. Fresh dill is placed between the layers of fish. under a light pressure. the temperature and the amount of pressure applied (5). Foodborne pathogens. are generally air-dried at sea. Stockfish products have a golden brown colour and are very hard. mixed with salt. Italy and Spain (5).3. Matjes are made of fat young herring. Dried octopus is also a very common dish in the interior of the country. South America and the Caribbean. In Scandinavian countries. the fish is dried naturally in a relatively cold environment. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. still made without prior salting on a large scale. 5.4 Air drying This is probably the oldest method of fish preservation. 101 .) where no acid is added to the process. and the flesh will break down. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). They are an important dried product. Other products. depending on the thickness. Usually. are likely to survive in these mild-processed products. sea cucumbers and seaweeds. air flows naturally. which are placed in a mixture of salt. Provided the product is kept in dry and fairly cool conditions. such as Listeria monocytogenes and Clostridium botulinum. a very fine delicacy for Oriental food lovers. The most common method used for fish products is the air-drying process. jellyfish. and the stacks are placed in the refrigerator. At pH levels below 4. Some bacteria and enzymes are still active in the presence of acid and salt. but sometimes it is dried without prior salting. sugar and spices. are commonly dried in Asian countries. In some coastal areas of Portugal.There are several products named marinades (matjes. are sundried and. In the first case. then mild-cured with special seasonings and packed in barrels with 80 º brine (5).5. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. Dried shrimps are widely used in Asia. it will last for many years (5).0) (7). and a fine product is obtained. after frying.CURED. marinated products will keep for several months at a temperature of 4 °C. Shark pectoral fins.
Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s. Thus. Whilst the production of dried fish products is tightly controlled in industrialised countries. Dried salted products must have a water activity below 0. Staph. the skin will form a hard case (case-hardening). 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. It is important to balance the drying rates and quality/safety issues. and has the primary objective of creating a product with specific taste and textural characteristics. botulinum growth might occur in the bulk of the product. making a water barrier and consequently. spoilage and possibly C.g. Spain. the diffusion of the bulk water to the surface of the fish is blocked. To prevent this. Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources. C. but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). and increasing amounts of such products are making their way into the market place. botulinum).75 (1). A highly demanded dried fish product in Mediterranean countries is dried salted cod. which leads to mould growth (aflatoxins. 102 . the quality is more difficult to control. some of which are listed in Table 5. The amount of moisture transfer from the fish to the air is dependent on the relative humidity . aureus) or spores (e.FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. Iceland and Canada.g. and the water must be changed several times before cooking. the fish must be dried slowly enough to avoid case-hardening. the product can be held for months at room temperature. neither it. France. In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or. by warm air circulating in a drying chamber. that may have contaminated the product during the drying process. In conclusion. tarpaulin. toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0. After dry salting. and it is exported to Portugal. skin up. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain. nor subsequent cooking operations will affect the presence of toxins (e. dried salted cod (bacalão) is now mainly produced in Norway. Whilst the low aw of the final product will inhibit microbial growth. or wooden drying rack to dry in the sun. the protein must be set or denatured with low-temperature drying before applying higher temperatures. more commonly nowadays. there is an increasing demand from people that have emigrated to industrialised nations for traditional products. the better the moisture transfer. The product is dried to a final water content of about 25 to 38% (3). Italy and South America. if the rate of water removal is too high in comparison with the diffusion rate.III. When no more water can be removed from the fish.83 (9)). However. The fish must be stored in reasonably dry conditions to avoid moisture absorption.the lower the humidity. They must be soaked in water. typically in stews and soups.
SMOKED AND DRIED FISH TABLE 5. using electrostatic filtration of the smoke. aureus (toxin) C. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10). cod roes. 5. making full description impossible here. The quality of the smoke is related to the type of wood used. trout. is mainly a process for flavouring fish rather than a preservation technique. Hardwoods are considered better because of the milder flavour they impart to the food. Staph. clams. mussels and many others from tropical countries. Blends of various woods can be formulated to produce sawdust with specific flavours. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. Aroma and flavour are a blend of smoke components (3). and the use of smoke generated by superheated steam or liquid smoke. as referred to above.3. Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . such as Escherichia coli. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns. It can be used for cold or hot smoking. mackerel. Maintaining the temperatures of pyrolysis between 425 and 200 °C.salmon. softwood colours the product quickly.CURED. cod. oysters. white mangrove and abura has an inhibitory effect on some microorganisms. There are many fish species that are used for smoking purposes . temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish. The smoking procedures vary widely. sprats.5 Smoking Smoking today. Top-quality fish must always be used. In the mechanical kiln. It has been reported that the smoke from mahogany. However. herring. aureus and Saccharomyces cerevisiae (10). tuna. eel.III Hazards and sources of contamination associated with air drying Hazard Enterobacteria Salmonella Source Sewage Reptile Birds Humans Humans Environment Airborne Applied to combat insect infestation Environment Human Vector Human Insect Staph. swordfish. botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. haddock.
Drying should be minimal at the start. whether cold. tuna and swordfish. low temperatures will slow the uptake of salt. however. it will reduce the salty flavour.or hot-smoked.3. This difference is crucial for safety issues because the flesh is not cooked. Wet salting or brining is preferable to dry-salting when smoking fillets because. are boiled before smoking (12). Depending on the type of product. all cold-smoked products should be cooked before consumption. covering the fish surface completely (10). The length of the smoking process for such products is much greater than for the hotsmoked fish. All the smoked products. Sugar can be added together with the salt. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. thawing must be done slowly for 12 . Also. smoked cod fillets and finnan haddock are usually cooked before consumption. as it retains more moisture.e. such as smoked salmon-trout. but a ‘safety’ (i. after being smoked need to be chilled immediately to 4 °C before being packed. 5. are not cooked prior to consumption. it will take up salt twice as fast as fresh fish (10). they can be vacuum packaged. or quick-frozen and stored at -18 °C (2). Thus. gutted or filleted. which is important for the appearance of these products (4). If wet salting is chosen. The salting period is dependent on the size of the fish and its presentation (whole. Whether the product is prepared headed. but this concentration will again depend on the type of product and the length of the brining stage. filleted. In cold-smoking. However. The fish can be dry-salted using a weight of salt 5 .FISH AND SEAFOOD water flow system with chlorinated water (25 .10% of the weight of the fish. canned or bottled. Cod roes. 104 . owing to the elution of soluble proteins. temperatures must be kept under 30 °C throughout the process.18 hours at 4 °C. thus allowing the deposit of smoke on the fish surface. the highly desirable cold-smoked salmon. pasteurisation) temperature is not achieved in any stage of the process. usually dry-salted and cold-smoked.1 Cold-smoking The major difference between cold. wooden boxes or master cartons (10).5. shrink packaged. If frozen fish is used. and the cold smoke is denser. but it will increase towards the end of the process (10). the following step is salting.50 ppm) should always be done just before process initiation (3). 70 – 80 º brine is commonly used. permitted colourings or smoke flavours can also be added to the product in the brining step. or packed in retortable foil pouches. spices. giving the product the desired appearance and flavour (4). it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11). Thus. The maintenance of temperatures below 30 °C avoids protein coagulation. as well as many other products prepared in the same way. When frozen/thawed fish is used. skin on or off). skin off or on. the excess salt is removed with water.and hot-smoking is the temperature of the process. an attractive glossy pellicle on the fish fillet surface is achieved after draining. Sugar. Some herring products. Brining at 10 15 °C is recommended (10).
5. During smoking.CURED. there is no point in the process that can fully assure the absence of L. 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. condensation will occur inside the pack. If the refrigeration chain is maintained. monocytogenes. Hot-smoked products require a longer cooling stage. gravad salmon (also called gravad lax or gravlax). but the proteolytic types A and B. a drying period at 30 °C. See Figure 5. the products are cooled. The exception is vacuum packed fish.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. botulinum type E present an even longer time than before to grow and produce toxin (14). and their sensitivity to salt (13). botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. which underlines the danger of storage at abuse temperatures of >10 °C (13). refrigerated storage and vacuum packaging still allow its growth (12). an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). These products should be vacuum packed at room temperature. The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening.5. Smoked salmon fillets are usually vacuum packed whole or after slicing. thirdly. are of concern when smoking fish from tropical waters (13). can be isolated from seafood. Although other human pathogens that can grow at refrigeration temperature. SMOKED AND DRIED FISH During cold-smoking. and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). thus allowing any psychrotrophic C. a partial cooking period at 50 °C. A typical temperature set is as follows: firstly. Furthermore. Afterwards. Pathogenic vibrios. but they should be cooled to a temperature of 10 °C within 3 hours. Neither the smoking temperature nor the salt content is enough to kill the organism. To avoid growth of non-proteolytic C. as. Length of the smoking process at each stage will depend on the species and the type of product. can grow with salt levels of 10% in the water phase. On the other hand. Vacuum packing is recommended only for coldsmoked salmon. Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. further cooled to 3 °C within 12 hours and then packed. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). and hotsmoked salmon (kippered) (3). This temperature should be maintained during storage and distribution (3). if a 105 . a final cooking period at 80 °C (12). in order to produce toxin. such as Yersinia enterocolitica.3. if present. if they are taken from a chill room at 3 °C and packed. Smoked products can be also packed under controlled-atmosphere packaging.2. typically mesophilic. secondly. stored at 4 °C and packed.
it will have a reduced shelf life.2. 5. Process of cold-smoked sliced salmon 106 .FISH AND SEAFOOD product is packed too warm without being cooled to room temperature. Fig. and moulds will grow on the product (12).
Aspergillus tamarii) and several Penicillium spp. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. They are generally not cooked prior to consumption.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. Eels. The product is removed from the drying cycle. white fish about 8 days. synthetic smoke flavours. 23). 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). The production method is similar to masmeen. They will keep well frozen. but mechanical drying is used instead of sun drying. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). rewetted and remoulded to give the desired shape and consistency. and many closely related Aspergilli (Aspergillus ochraceus. 21. Aspergillus ostianus. then hot smoking and finally sun drying it. are usually 80º brined for 5 .6 hours and smoked for 3 . 22. including Eurotium repens and Eurotium rubrum. The classical liquid smoke is 107 . resulting in a characteristic flavour (20. to have no further advantage over other packing methods for other fish (12). apart from the improved appearance.5. 18). such as yeast derivatives. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). but seems. cooking in brine. Boiled or steamed clams. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. including ‘umami’ (16. Aspergillus oryzae. are used. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). Hot-smoked products made from white fish will usually keep better than those from fatty fish. and substances unconnected with smoke but with a smoky flavour or smell. moulding it into long thick sections. 5. (Penicillum cyclopium and Penicillium puterellii) (19). SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. similar to masmeen. In terms of safety. 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). chilled or freeze-thawed.CURED. break down of fats and the moderation of smoke flavour. Vacuum packaging retards the onset of rancidity when fatty fish is used. a very expensive and fashionable smoked product in the Netherlands. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish.3. 17. Growth of these moulds on the tuna results in the extraction of moisture. Natural smoke extracts. The composition of commercial liquid smoke can vary widely depending on the method of manufacture.3½ hours using the typical temperature set for hot-smoking referred to above (12). fatty fish will remain in good condition for 6 days. Mackerel. When refrigerated at 3 °C. Katsuobushi.
Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. 5. When liquid smoke was used.5. injecting it into the fish flesh. 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. applying it as a dip after brining. and less than 0. In experiments comparing traditionally smoked fish with liquid smoke.06 μg/kg of benzoanthracene in the final foodstuff (28). The European Scientific Commitee for Food. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines.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). It has been shown that liquid smoke can reduce the formation of PAHs (3). 5. According to some recent research.4 Clear smoke treatments The “Clear” or “Tasteless” smoking process is a processing technique whereby smoke is generated as in a normal food smoking process but is filtered to remove the characteristic smoke flavours before application on to otherwise untreated (uncured) tuna.6 to 2. nutritional neutrality and antioxidant activity (27). 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. so as to impart an unnatural red colouration to the flesh. the salt level could effectively be reduced from 4.03 μg/kg of benzopyrene. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). or applying it as an atomised spray within a modern automatic smoke kiln. 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. drying.3. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25).8% and still provide protection for 7 days (25).4 Spoilage Brining. Liquid smoke was considered to provide advantages of safety. both of which are considered potential carcinogens. consistency of flavour and colour. or smoke extracts in organic solvents. 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). smoking. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. Fresh fish spoilage may be measured by 108 .
and. Once these changes are defined. Most marinated and smoked fish products are first brined or salt-pickled. Usually. and of smoke to add chemical preservatives. In salts. SMOKED AND DRIED FISH grading schemes or quality indices. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). spoilage is due to endogenous microorganisms and enzymes of the fish. also. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. or extended refrigerated storage. therefore. but. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime. pickling and dry-salting all introduce bacteria to the fish. both of these will affect the spoilage patterns.CURED. Micrococcus spp. the bulk of the microflora comprises Bacillus spp. and high concentrations are usually required to inhibit their growth fully.g. the shelf life may be estimated and investigation made into the causes of the loss of sensory quality. or their products require refrigerated storage. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure). Often. Brining. which are found in the salt. Even saturated brines allow some halophilic organisms to grow.3%). however. Upon salt packing. the production process will modify the microbial flora present on the product. contaminant bacteria from other materials used to produce the product. chemical oxidation. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). Stronger and longer brining has an effect like that of pickling. limits the types of microorganism capable of growing. Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2. a process similar in microbiological effect to that of saturated brining. Some brines are re-used over a period of days and a load of 10. for some.. marination or drying). contact with the bacteria 109 . Once placed into brines. as the fish are added. a very dry product is required to produce 'shelf-stability'. otherwise. which may also be produced to describe the quality attributes and their loss over time for processed fish. the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. these bacteria tend to continue to dominate the flora in the liquid. the fish flora are transferred to the brine mix. The use of drying and/or salt to lower the water activity. The microflora attached to the fish is relatively unaffected by light brines used before smoking. When heavy brining or salting is important for product shelf life.000 cfu/ml of brine may build up. Spoilage needs to be carefully defined for each product. a product may become undesirable for consumption as a result of changes in its sensory characteristics. rock and manufactured salts did not usually contain halophilic bacteria (29). the consequent reduction in the pH will help to inhibit spoilage organisms. When acid marinating or smoking. these halophilic organisms may grow and cause spoilage. For some products. and coryneform bacteria.. Within the limits of safety. for others. the fish are cut and splayed to allow greater surface contact with the salt.
(syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. halophilica was isolated 110 . most notably Penicillium citrinum and Penicillium thomii. sometimes covering the whole of the fish with a whitish. although it may occur in other salted fish.FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. yeasts. rancidity due to lipolysis. B. it is possible to wash off the infected parts. niger. 32. 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). sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). E. Studies conducted on dried and salted fish from Indonesia (31. halotolerant or halophilic micrococci. if not too contaminated. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. may also be found in the salt used in production. spore-forming bacteria. yeasts and fungi. ‘pink’ and ‘dun’. ‘dun’ is not characterised by the production of offensive odours. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage. powdery growth. gaspé). Basipetospora halophila. and still produce a reasonable dried product. including hydrogen sulphide (26). (syn. W. Only halophilic microbes may grow. pisce was the main fungus isolated. flavus. The condition describes the growth of the halophilic moulds (Wallemia sebi. and Aspergillus penicilloides. ‘Pink’ describes a condition associated with surface growth of the pigmented halophilic bacteria Halococcus spp. lactic acid bacteria (LAB) and moulds. The microbiology of heavily salted products is dominated by Gram-positive. The bacteria form coloured colonies on the surface of salted fish prior to drying. A. The metabolism of the organisms will produce off-odours. 34) have shown that P. repens and E.. including A. rubrum are also common isolates from dried fish. though only the latter two species were considered likely to have grown on the fish (32). 33). or fat breakdown) caused by enzymes rather than by microbial growth. like the ‘pink’ organisms. protein breakdown. Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration. together with a range of Aspergilli. A number of spoilage organisms can grow and produce conditions such as ‘sliming’. The bacteria are osmosensitive and will lyse in water. which. spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. This condition is often associated with case-hardening. Eurotium amstelodami. Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. as little if any growth will occur below 5 °C on salted fish (29).g. as the water activity of saturated brine solutions is low enough to inhibit most bacteria. Aspergillus sydowii. Therefore. and Halobacterium spp. At the pre-drying stage. or after drying if the fish is stored in excessive humidity. Sporendonema sebi)). were also isolated. Aspergillus wentii. Several Penicillium spp. it was found in about 50% of samples examined. the best way of controlling the organisms is to refrigerate the fish. Unlike ‘pink’.
Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. The brining to 3 . able to grow on dried salted fish. Cold-smoked fish will contain microbial flora representative of that of the raw fish. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life. Fish processed by cold-smoking. It has been reported that the type of salting (dry. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. around the water activity of saturated sodium chloride. 36). 33. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. which does not create the salty conditions required for spoilage by those halophilic microorganisms. Aspergillus spp. SMOKED AND DRIED FISH infrequently at low levels. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. with a lower aw limit for growth of 0. botulinum type E. as already stated.747.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). The salt in the water phase must be high enough to inhibit the growth of C. however. Marine vibrios were the dominant flora in smoked salmon that was drysalted. including A. In Vietnam. so the bacteria spoiling these products will be psychrotrophic or psychrophilic.. Pseudomonadaceae. and microbial spoilage is more likely. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations.5% sodium chloride in the water phase will inhibit some bacteria.CURED. not the growth of spoilage organisms. Refrigeration is necessary. but. Table 5. will have a different residual post-smoking flora from 'hot-smoked' fish.IV. 111 . flavus. Aspergillus clavatus and Aspergillus niger. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. where a high internal temperature is reached. The preservative effect of the smoke will inhibit some spoilage microorganisms. LAB. but this will not prevent the growth of spoilage bacteria. Smoked fish products tend to have a light brine treatment. Vacuum packing limits chemical oxidation. has been isolated from dried fish (35).or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. The heat during hot-smoking will kill vegetative microorganisms but not all spores. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. For more information on spoilage see Table 5. it was.
Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. sebi and B.IV Spoilage of salted. and Halobacterium spp. . Dun.W. 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. 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. LAB. + to +++ strength of preservative effect 4 Post-process contaminants 112 .variable.FISH AND SEAFOOD TABLE 5. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted. halophila Halophilic spoilage organisms Halophilic spoilage organisms. boiled.
but if the fish products are stored at temperatures <10 °C. with this change.75 0.5-10. Before. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13).5% to 5%.5 5. the fermented products being the most dangerous group (7).9 0. smoked and dried. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. botulinum type E and production of toxin. botulinum Staph. In the United States.5. under optimal conditions.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). It has been shown that.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39).5 4. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14).61 Organisms Most Pseudomonadaceae C. and as botulism is a serious hazard in foods. 9 and 8 of the outbreaks. The concentration required to prevent growth at room temperature can vary from as low as 3. salted and pickled products accounted for 10.CURED.97 0. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. botulinum. aureus Yeasts Moulds Lactic acid bacteria pH 5.8 1. respectively.5 5. The presence of salt in cured products has a great effect on the growth of the bacteria. the products were heavily salted. when present in the raw material. pressure on the traditional methods of curing. However.5-8. and C. the non-proteolytic type E strains can grow in up to 5% sodium chloride. botulinum type E. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth.5-8. Since neither the smoking nor the drying parts of this curing process are particularly severe. SMOKED AND DRIED FISH TABLE 5.5-10 4. botulinum type E Staph. although the combination of the two factors. sodium 113 .86 0. popular demand for healthy foods has led to a change in the tastes preferred by the public and.8 0. the importance of cold-chain integrity must be emphasised. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. types A and B are not affected by 3 .97 0. could not grow and produce toxin.5 0-11 3.0-9.
0 . An outbreak of Shigella in an Italian cruise ship. if the person handling the product is a carrier of Staph. However.94 (7). but. aureus growth. the human nose. Sliced smoked fish products. gills and guts as well as in several lightly preserved fish 114 . the organism may contaminate the product. when the bacterial load of the product is lower. The organism is mesophilic with a minimum growth temperature of 7 °C. As with other organisms.5. In foodstuffs. but.5.4. It is a halotolerant bacterium able to grow at water activities as low as 0. they will grow rapidly (7). if recontamination occurs and storage conditions are favourable. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. 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. throat and skin. growth and toxin formation may occur. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. As they are poor competitors.0% salt in the water phase (30).30% of the population positive for enterotoxin-producing strains (41). The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . Once the product is contaminated. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13).5. the risk of infection can be eliminated by proper cooking.FISH AND SEAFOOD nitrite can be added to the brine but this is not permitted in EU countries because nitrites might lead to formation of the carcinogenic nitrosamines. 5.86 and 10% sodium chloride. are particularly susceptible to contamination and.5% salt in the water phase. if storage temperatures are over 10 °C. has been reported (42).4 Listeria monocytogenes The psychrotrophic L. coli all occur on fish products as a result of contamination from animal/human reservoirs. related to the consumption of a raw smoked fish meal. 5. Infection by Shigella sonnei has increased in the UK. and the minimum pH for growth is 4. they do not grow well in the raw material. Thus. it should contain not less than 3. more importantly. if it contains sodium nitrite ≥100 ppm. as they require more handling.or cold-smoked fish should have at least 3. but requires higher temperatures for toxin production (>10 °C). aureus. The FDA established that hot. Shigella and E.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. Usually. 5. the organism can survive most of the light curing processes. after processing.5.3 Enterobacteriaceae The pathogens Salmonella. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin.
45 °C. the USA established ‘zero tolerance’ for the organism. As they are heat-sensitive. Of microbiological concern in the northern hemisphere are two parasites .9. Thus. Its limiting conditions for growth are 0 . which. do not survive 1 min at 60 °C. a number of fish products. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. marinades (ceviche) (43) and gravad fish (44). monocytogenes pathogenic for only specific segments at risk. favours growth of psychrotrophic LAB.CURED. are considered unsafe. which include intermediate hosts. a short period of freezing either of the raw material or of 115 . SMOKED AND DRIED FISH products such as cold-smoked fish. Two outbreaks in the United States have been reported to be linked to salmon (10).92 .12%. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47).514. monocytogenes in levels at or above 102 per gram (46). lightly salted caviar. the only concern to public health is when eating raw or uncooked fish products. most of them are of little concern to public health. cold-smoked fish.5. a round worm. salt levels of 8 . Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. from freshwater fish. such as ‘lightly preserved’ fish products. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. from seawater fish. there were no reported outbreaks related to cured fish. Pseudoterranova dicipiens. This includes matjes herring. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). the CT 3162-FAIR program. which considered L.2.95 (45). gravad fish. widely used as starter cultures in food processes. both being detected in cold-smoked salmon. pH 4. and the tape worm Diphyllobothrium spp.the round worm Anisakis spp. Some LAB.5 Parasites Despite the presence of parasites in fish being very common. 5.8 . Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere.0. They have complicated life cycles. or nematodes. The use of LAB in fish preservation is a topic that has been studied in projects (UP. monocytogenes’ in 25 g of the finished product.6 and water activities below 0. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. have been shown to have an anti-listerial activity in many food products. The round worms. has also been detected in cod (7). and in CT95-1207FAIR). in the EU Fisheries R&D-FLAIR program. in combination with curing agents and refrigerated storage. 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. and other local traditional products (7). Unlike some European countries. which means that the process must conform to a level of ‘no detectable L. Until 1997.
E. perfringens B. and unpasteurised yoghurt.100 >100 N/A Pathogens Salmonella spp. texture. coli O157 & other VTEC Vibrio.<100 10 . monocytogenes Staph.<104 20 .<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 .<107 ≥107 N/A <100 <20 100 . the organism should be identified # N/A denotes not applicable 116 . salami.6 Published Microbiological Criteria TABLE 5. 5. If the Bacillus counts exceed 104/g. These foods fall into category 5.<100 20 .<100 200 . not detected in 25 g Campylobacter spp.VI Guidelines for the microbiological quality of ready-to-eat smoked fish. monocytogenes) (cfu/g unless stated) <106 106 . 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E.<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.<104 104 . soft cheese. (not L. cereus and Bacillus subtilis# <20 <20** <20 <10 20 . 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. Acceptability is based on appearance. for example.FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). smell.<100 20 . and the levels or absence of indicator organisms or pathogens. cholerae Vibrio parahaemolyticus L.<103 N/A 100 . ‡ On occasions some strains may be pathogenic. † Guidelines for aerobic colony counts may not apply to certain fermented foods. coli (total) Listeria spp.<104 100 . aureus C.
monocytogenes Salmonella spp. Torry Advisory Note No. 1984. 37(revised). where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. salting and smoking).. McLay R. Regenstein C. 3.A.F. 82 (revised).H. Horne J. coli (ETEC) L. Wareing P. Urner Barry Publications Inc. in Micro-Facts: The Working Companion for Food Microbiologists. Assurance of Seafood Quality. Chapman & Hall. Ed. 1993. USA. 117 . Food spoilage fungi. or presence of toxin 0. New York.. 8. Fernandes R. Catching. Smoked.7 1. Huss H. Eds. Martin R.M. Introduction to Fish Technology. New York. aureus * Vibrio cholerae V. in The Seafood Industry. 9. 1979. Van Nostrand Reinhold. Fernandes R. 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. 6. Bannerman A. 381-406.. 1994. 1991. References Regenstein J.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. parahaemolyticus Vibrio vulnificus C. 1997.CURED. except in the viscera of dungeness crab. 7.. Birnie K.. cured and dried fish. FAO Fisheries Technical Paper. Marinades. 56. botulinum * Limit of tolerance 1x 103 ETEC/g. Van Nostrand Reinhold. Leatherhead Publishing. Leatherhead. 1990. The Smoked and Cured Seafood Guide. Moody M. Torry Advisory Note No. Hall G. 3273. 2. 2007. Wareing P. 254-82 5. Flick G. Eds. 334. London. in Fish Processing Technology. 120-38.E.F. Flick G. SMOKED AND DRIED FISH TABLE 5. Hot Smoking of Fish. Horner W. 1982.M.. Doré I. Torry Advisory Note No. 4. Handling and Processing Eels. * Staph.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. Preservation of fish by curing (drying.
and Biochemistry. Bannerman A.. Doi M. 1989. The discovery of umami. Doi M..C.. Fish Drying and Smoking: Production and Quality. Doe P. 1995.F. 17 (9). Torry Advisory Note No. Horne J..K. Microbiological control for fish smoking operations. Hosokawa M. Packing Fish in a Modified Atmosphere. Dillon R. Eklund M.W. 6 (6). 1994. 18. Teeny F.A. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour. 22 (revised). Biotechnology and Biochemistry. 53 (4). 935-41. Ogiwara Y. Bioscience. Huss H. 27 (9).. Lindemann B. 108 (3). Matsui M. What’s in a name? Are MSG and umami the same? Chemical Senses.P. 118 . 56 (6).M. Pelroy G.. Kinoshita Y. 11. Shuto Y... Biotechnology. 20. Biotechnology and Biochemistry.. Ed. Food Control. Technomic Publishers. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors. Matsui M.. 13. in Fisheries Processing. 16.. 843. 335-40. Paranjpye R. Mitou M. Yokayama S..M. Perusing Asian soup options. 1977. 24. Doi M. Yamauchi H.M.. 1386-7.C. 12. Druschitz A. Processing Cod Roes. Kinoshita Y. 1992. Kanayama T. 23. Shuto Y. Torry Advisory Note No. Bannerman A. 1997.E.FISH AND SEAFOOD 10. 1981.. 22. Food Product Design. Ninomiya M. 45 (10). Biotechnological Applications. 840-6. 15. Hayashi K. Jeppesen V. Yajima I... Ninomiya Y. 14. Botulism and Fishery Products.R. 18. 1984. Embarek P. Food Chemistry. Doi M. 25. 2008. Cann D. Cann D... Torry Advisory Note No. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor. Martin A. 1051-5... Torry Advisory Note No. Peterson M. 1995. Food Reviews International. 958-60. Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish. 5 (revised). Bioscience. Sakakibara H. 508. 553-72. 88. Shiemori Y.. Martin A. 1990. 19.. Agricultural and Biological Chemistry. Flavour constituents of dried bonito (Katsuobushi).B. Halpern B... Chapman & Hall. O-methylation of 2. 2002. Aoshima H. Shuto Y.. Bioscience. Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species. 1998..E. Journal of Food Protection. 51-81. 59 (12).. Recommendations for the Preparation of Smoked Salmon. Control of biological hazards in cold smoked salmon production. 6 (4). 2007. 1980.. London. 21. 17. Patel T. Lancaster. 2324-5. 61 (8). 1982.H. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour.
Mycotaxon.R. 1995. Fuchs R. Fungal flora of South Vietnamese fish and rice. Boca Raton. 164-9. 119 . Røntved S. Golden D. 27686. 348. 3 (4). Pszczola D. 12.D.J. 1995. Wheeler K. Incidence of Listeria monocytogenes in an acidified product. 39. Cann D.A. 29 (2).I. 22. 1986.C. Townsend J. Institute of Medicine. Smoked and smoke-flavoured fishery products. 43.M. Studies on salt fish.. Frank M. 74. Taylor L. Journal of Tropical Medicine and Hygiene. Seafood Safety.. 28. 1994. in Smoke in Food Processing. Anggawati A. Food Technology. CRC. in Fish as Food. 14. 107-11. Wheeler K. 1993.CURED. 311-7. Journal of Food Technology.E. Food and Drug Administration. 1996. 218-25. 1971. 42. 38.I.M. Ahmed F. 1941. Blackie. Hocking A. Scientific Committee for Food. The microbiology of sea-water fish. 27. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. Committee on Evaluation of the Safety of Fishery Products. 29. Importance of autolysis and microbiological activity on quality of cold-smoked salmon. Sirvas S. Diyaolu S.H. 123-9. 742. US Food and Drug Administration.K.M. Ed. V. 1991. Pitt J.O. Nahrung. Loessner M. Jay J. Hocking A. Commission of the European Communities. Pitt J. 36.I. Antimicrobial properties of smoke. Ed. 1961. 1997... Oxford. 2005.. Press Inc. Luxembourg. Springer Science. Rockville.. London. The control of botulism in hot-smoked trout and mackerel. Smoke flavourings. 5.F.. 351-7. in Fish and Fishery Products Hazards and Controls Guide. Simini B. Food and Drug Administration. New York. Food Research International.. ceviche.F. Sprouse R.A.. 74 (2).. Lancet.. Food Microbiology.T. 34. Hess E.. National Academic Press. Gill T..B..D. Fungi associated with Indonesian dried fish. 37. 74. Lucas F. Journal of Applied Bacteriology. in Reports of the Scientific Committee for Food (34th Series). Ed. Interactions among xerophilic fungi associated with dried salted fish. CEC. Journal of the Fisheries Research Board of Canada.E. 35.D. Ed. Maga J.. 88-90. Letters in Applied Microbiology. 40..D..V. Washington DC. 33. 1994. SMOKED AND DRIED FISH 26. Cox J. 181-8. 31. 1985. New species of fungi from Indonesian dried fish. 70-2. 41. Modern Food Microbiology. 38 (3). Hansen L. Fungi and Food Spoilage. An outbreak of Shigella. 98100. Pitt J. 197-208.. 1979. Studies on Sporendonema epizoum from ‘dun’ salt fish.A.1988. Huss H.S. Hocking A. 30.A. Tour highlights production and uses of smoke-based flavours. Shewan J. Borgstrøm G. 32. Academic Press..Y. 49 (1). Hocking A. 1996. 1991. Adebajo L.
Loncarevic S. 1980. An outbreak of listeriosis suspected to have been caused by rainbow trout.. Ericsson H. 13-8. Bolton F. Eds... Communicable Disease and Public Health.M.8 Further Reading Gibson D. Nye K. Persson I. 507-10. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale. 1994. Eklow A. 1982. Amsterdam. Paranjpye R. 1980.. Little C. in Fish and Fishery Products Hazards and Controls Guide..H.. in Quality Assurance in the Fish Industry. Garren D...C.. Roberts D. 26 (4).gov/~comm/haccp4x5. Danielsson-Tham M-L. Harrison M. 120 . Prevalence of Listeria monocytogenes and other Listeria spp.. 46. Gibbs P. Loncarevic S. Davies A. Ed. Jacobsen M. Unnerstad H.J. 833-41. Huss H. 2000.M. Acta Veterinaria Scandinavia..C. Appendix 5: FDA and EPA Guidance Levels.. 1991..cfsan. 197-209.A. Advances in Food Research.. Hughes A. Hobbs G. 163-7.D. Pelroy G. Ribeiro C. Peterson M.J. Donovan T. 80. Beltran A. 37 (1). 47..N. Association of Food and Drug Officials. 55 (4). Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions. Tham W. Tham W.W.. Journal of Clinical Microbiology... 48. 135-85.. 1996. 1989.E. Canadian Journal of Microbiology.. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus. Incidence and growth of psychrotrophic Clostridium botulinum in food. Pathogenic microorganisms of importance in seafood. 5. 1992. US FDA Centre for Food Safety & Applied Nutrition... Hurst A.fda.. US FDA Centre for Food Safety & Applied Nutrition.. Pelaez C. 49-59. Wooten R. and smoked fish establishments good manufacturing practices. 863-6.. Huang Y. Washington D.A. Clostridium botulinum and its importance in fishery products. Journal of the Association of Food and Drug Officials. http://www. 1997.. Liston J. Round Worms in Fish.. Journal of Food Protection. Fletcher R. 58 (8). Food Control. Food and Drug Administration.. Richards J.A. 5-7. 5 (1).L. 232-6. 35 (11) 2904-7.W. 45. Cann D.. 2001. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon. in smoked and "gravad" fish. salted. 1976. Journal of Food Protection.html Gilbert R. de Louvois J. Torry Advisory Note No. Susuki E. Moral A. 22. 45 (9).S. Danielsson-Tham M.R. Elsevier Science Publishers B. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects.FISH AND SEAFOOD 44. 188 (3). 3. Eklund M.V. Mentzing L-O. 1995. Zeitschrift für Lebensmittel-Untersuchung und –Forschung.. Cured.
. Marine toxins of microbial origin. Interacting effects of pH. 1991. Dillon R. Combined effects of water activity.C. and salt concentration on growth and survival of Vibrio parahaemolyticus. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. in Seafood Quality Determination. Beuchat L. 11.J. Hitchins A.. 309-16. Paranjpye R..R. 1992. Letters in Applied Microbiology.A. Dallas H. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants..... 1974. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. and temperature on the growth of Vibrio parahaemolyticus. Poindexter C. Davies A. Journal of Food Safety. Presence. 1994. 1995. detection and growth of Listeria monocytogenes in seafoods: a review. Tran T. 1995.. solute. Ben Embarek P.. 11. Keusch A. Patel T. 121 .N. Elsevier. Letters in Applied Microbiology. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. New York. 1995. 58 (5). Food Technology. Letters in Applied Microbiology. Nicolaides L. van Spreckens K.R.. Pelroy G. 1992. 866-70. 21 (3).. 23 (1). 309-18.. in a salmon slaughterhouse and smoked salmon processing plant. 19-27. International Journal of Food Microbiology. Fuchs R.P. Caugant D. 14. 1988. Incidence of Listeria in hot and cold-smoked fish.A. 61 (12).178-80. temperature. 1973. 217-36.R. 19. 1994. SMOKED AND DRIED FISH Sutherland J.E. Applied Microbiology. Rørvik L. 293301. Beuchat L. 42. 844-6.. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. 55 (11). Ratnam S. Hudson J. Competition of food bacteria with Listeria monocytogenes during enrichment culture. 502-8. Applied Microbiology. 1994. 1075-80. Food Microbiology.T. Poysky F. 394-6. Jemmi T. Journal of Food Protection. International Journal of Food Microbiology. Journal of Food Protection. Taylor S. Histamine production by psychrophilic flora.W. International Journal of Food Microbiology..S.CURED. 25. 1734. Bayliss A.A. Prevalence of Listeria in smoked fish. Roberts T. Slade A. Eds. Eklund M. Kramer D.K.. Peterson M.. Liston J.. Yndestad M. 25 (5). Contamination pattern of Listeria monocytogenes and other Listeria spp. 4389-95. 1987. 27 (6). pH and sodium chloride. 94-8. 1994.. Lashbrook L.
1 Definitions As applied to fish products. Essentially. they are exported to European. Fermented fish products are largely confined to east and south-east Asia.I. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. Those to which only salt is added tend to be mainly autolytic. typically in the range 15 . though some are produced elsewhere. North American and other markets and are.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. added during preparation. 6. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. there are numerous variants of some common themes and a host of local names used to describe them. FERMENTED FISH Prof. they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. Carbohydrates are added to some fish sauces during their preparation. For example. koji. 123 . whereas in those where a carbohydrate source such as rice or sugar is also added. producing the fish sauces and pastes. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6. the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. The fish sauces are produced on the largest scale.1. in economic terms.6.25% and are used mainly as a condiment. The relative importance of these two activities depends on the formulation of the product. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. lactic fermentation can play a significant role. As with most traditional products that are still produced principally on a cottage industry or domestic scale. the most important of all fermented fish products.
who had adopted them from the Greeks. 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. although garos. This is well exemplified by some of the fish/salt products of Thailand described in Table 6. Fermented fish products are of very minor importance outside Asia. 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.1. made from whole fish in France. These have long since fallen from use. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. where the bacterial production of lactic acid is a major feature.II. to products analogous to other lactic fermented foods such as salami. a variety of products can be produced within a single country. A selection of Asian products is presented in Table 6. 124 . In Europe.FISH AND SEAFOOD TABLE 6.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. and pissala. made from fish livers in Greece. salty taste.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred.III. 6. may be direct descendants.
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. pla-som. shrimps. burong-dalag. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 . e. cencalok burong-isda.3 months Main dish Tai-pla Bowels 1:3 6 . ayu-sushi. som-fak/ som-fug 125 Malaysia Philippines Thailand . tai-suchi phaak.g.g. crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 . pa-khem. pa-chao.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. burong-bangus pla-ra. funa-suchi. crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 . e. mam-seeing sikhae som-kay-pa-eun. burong-ayungi. mam-chao. pla-chao. mam-pa-kor.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. som-pa.12 months 10% raw sugar added.8 months Main dish TABLE 6.FERMENTED FISH TABLE 6. som-pa-keng pekasam.
in a survey of fermented fish products in north-east Thailand. There are also more heavily fermented products.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. The diversity of fish used in fermented products (see. A modern account of their production. it was found that several different sources were used for the fish . the most popular varieties of which are rakørret.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. made from herring. Penaeus indicua Many of the fish used in fermentation are uneviscerated. 6. which has been increasing in popularity over the last 50 years. freshwater fish are the usual raw material. Norwegian gravlaks. Table 6. malt or flour (1).FISH AND SEAFOOD In Northern Europe. is a traditional. some fermented fish products remain popular. relatively mild-tasting product. and their microflora tends to reflect their local environment more than that of marine species. in Sweden. in Norway and surstrømming. fermented trout.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. however. and the initial microflora is likely to be considerably larger in these cases. or buried salmon. especially in fish/salt products. 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. rakefisk or surfisk. TABLE 6. and old descriptions of their production often refer to the addition of whey. and a large local freshwater reservoir (2). paddy ponds beside rice fields used for collecting fish when the field has dried up. For example. states that sugar is often included with the salt. Here. These are often described as fish/salt products. 126 . which employ lower levels of salt than Asian products.the flooded rice fields. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. for example. and counts ranging from below 104 /g to in excess of 107 have been reported.
FERMENTED FISH Where other ingredients are used. Hansenula. or boiled or roasted rice. 4).3 6. either directly or as the partially saccharified rice they produce. Bacillus spp.4. Endomycopsis and Saccharomyces .3. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. sufficient to saturate the water phase present and produce an aw of 0. Other halophilic species have been identified from fish/salt products. after one month. Similar results were seen with patis. Mucor. principally species of the genera Rhizopus. they can contribute to the initial microflora. the Philippine fish sauce. were found to be the predominant isolates. mean that microorganisms have been generally held to play little or no role in the process that follows. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). and the anaerobic conditions that prevail in the bulk of the material. angkak (Philippines and Thailand) and look-pang (Thailand) are used. In another microbiological study of four fish sauces.74 or below. When this is a simple carbohydrate source such as sugar. Generally. In some cases. It also demonstrated their significant proteolytic activity during the first month of fermentation. These preparations contain a mixed microflora of moulds and yeasts. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). traditional starter culture preparations such as koji (Japan). (3.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. More recently. 6. 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. 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. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. however. This. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp.1 Processing and its Effects on the Microflora Fish sauces and pastes Detailed flow charts for several fish/salt fermented products are presented as Figures 6. in many cases.1 – 6. however. and studies 127 . its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. which included nam-pla and patis. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp.
1.12).FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 . Fig.2. Belacan (Condiment) Fig. Patis (Condiment) 128 . 6. 6.
and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. In fish sauce production.3.4.FERMENTED FISH Fig. eviscerated fish are sometimes used to slow the autolytic process. however. 14). thought to be largely the result of autolytic breakdown of the fish tissues. 6. Ika-Shiokara (side dish) Fig. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. To produce semi-solid fish pastes as opposed to fish sauces.6. the 129 . Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are.
Hoi-Malangpu-Dong (side dish. Figure 6.6 – 6.V and Figures 6. particularly glutamate.5. 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. 6.3. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. for example. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. filtered and bottled. Fig. In contrast to free-swimming fish. snack) 130 . 6.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. Some examples of products and their formulation are presented in Table 6. which is eventually separated from the residual fish tissues. volatile fatty acids and nucleotides.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.8.5).
central.6.7 days 5 . N. C.03 Preparation time 6 .FERMENTED FISH TABLE 6.12 months Shelf life Use 1 .4 are also produced using shrimps. NE 10 . N. C NE. NE Formulation Fish (whole or pieces): salt: roasted rice. C NE. N.2-4 Fish (pieces): salt:khaomark (fermented rice).12 days 3 . Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. N S 5 .5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. Fig.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. S. In these cases. north-east.12 months Main dish 2 Pla-jao C. Kung-Chom (staple food. north. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C. NE.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . side dish) 131 .5-1. the prefix 'pla' is replaced by 'kung'.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .6. 3:1:0. south of Thailand.
Red burong isda 132 . typically in the range 2 . Generally.7. giving a reduced rate of acid generation.salt and lactic acid. but there is considerable variation in this.2. their growth and metabolism are nonetheless slowed.. of the two factors contributing to shelf life . Phillipines Journal of Food Science and Technology.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. 2.6.3. This is readily explained by the susceptibility of LAB to elevated salt concentrations. (from Arroyo et al. If acidity is a desired sensory property of the product. 106) Fig. the salt is of far greater importance in determining the product’s long-term stability. these products are salted at lower levels than the fish sauces and pastes.1 The use of higher salt levels results in a longer production phase but a better keeping quality product. then this will take longer to develop than if lower salt levels are used. while they are able to grow at high salt levels. This can be clearly seen from the data on reported shelf lives of a number of fish salt carbohydrate products and their respective pH ranges and salt contents (Table 6. The LAB isolated from these products are those with more marked salt tolerance but.VI). A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. It also indicates that.10%.
4 .220.127.116.11 months 1 . Philippines Journal of Food Science and Technology.9.6 3.5 3.8 pH 4.5.6. 133 .9 .FERMENTED FISH (from Arroyo et al.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).9 4.0 .VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 18.104.22.168 years 1 .3 5.6..8 .0 .2 4.5 .8 .4.4.0 Shelf life 3 weeks 6 .8 2. 2.9 4.7 .2 7.2 4.0 . White balao-balao TABLE 6.6.1 .1 4. 106) Fig.5.5.5 .3 .
alternatively. Addition of carbohydrate will.VII). also serves as a valuable source of the more readily fermented carbohydrate. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. Where they are used.0 . although the final population of LAB may be similar. the rice may be added in a partially saccharified form such as kao-mark (Thailand).2.FISH AND SEAFOOD 6.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. Garlic.3. Rice is the most common carbohydrate source used. 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 (20.3. When added as boiled or roasted rice alone. also lower the unit cost of the product and this is probably a significant factor in many cases. of course. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. For example. This results in a more active lactic fermentation and a lower pH.9) compared with the white product (pH 4. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3.1 . such as salt content.5) despite other factors. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. 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 . a common ingredient in these products. 21). Although the ability to ferment starch is not widespread in the LAB. 19). being similar (18.2 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. TABLE 6. 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).4. inulin. In several products.
the Korean fermented fish product. the level of salt employed is on the high side for fish/salt/carbohydrate products.5:9:3. allowing other spoilage organisms to grow. such as wrapping in banana leaves.5.7 up to 6.2 at the end of fermentation.FERMENTED FISH Sikhae. The growth of yeasts and moulds could. reporting the product’s pH to range from 4. lead to an increase in pH. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. There is little published information on the factors that limit the shelf life of the low-salt products. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases.3 weeks at around 20 °C. Results of a product survey were consistent with this. Clearly. 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). in turn. 135 . It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. Traditional packing techniques used with lactic fermented fish products. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. thereafter declining to about 4. will not reliably exclude air. the pH drops from 6.7 to below 4. presumably correlating with the salt content. but they are likely to behave similarly to lactic fermented products produced from other commodities. markedly extending the shelf life (25). and there is a marked regional variation in the preferred type.17. The same product can range from a brown turbid liquid where the fish has been subject to extensive autolysis to brown partly dried pieces of fish.8 .9%. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products. is something of an exception since millet is used along with garlic as the carbohydrate source. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. which ranged from 7. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. 6. The mixture is allowed to ferment for 2 .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). Generally.2. In a successful fermentation. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. Excessive growth and acid production by the LAB could render products too sour for acceptability.5 in the first 2 . red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. around 12%. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage.3 days. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24).
8 and at salt levels as high 10%.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. As in Norway. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. are Vibrio parahaemolyticus and Clostridium botulinum. Because of the severity of the illness it causes. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. although these limits apply when all other factors are optimal (26). but we are unaware of any reports of this happening. There is little in the way of documented information associating fermented fish products with foodborne illness. fish products are the most common source of botulism. After capture. 6. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. 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. typically 30 °C. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. 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). The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90.FISH 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. outbreaks of 136 .87. 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. The bacterial pathogens most commonly associated with the living environment of fish. and consumed without cooking. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. V. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. however. parahaemolyticus has been reported growing down to a pH of 4. 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. In the fish sauces and pastes. During fermentation. viable numbers will probably decline quite rapidly. which accounted for 88% of outbreaks in the period 1952 . In Japan. under conditions inimical to growth and high ambient storage temperatures.
. 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). Mahidol University. they could act as an important vehicle of transmission. References Riddervold A. With this particular organism. Liston J. 3. 1989. 31. Kasemsarn B.. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). Lutefisk. 2. 1966. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O.O. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. 1990. Rakefisk and Herring in the Norwegian Tradition. which will persist into the product. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. Fermented Fish Products Study (Thailand) Phase 1.6 1. whale meat. 105-10. botulinum. and warm weather can result in rapid growth of C. Many of these products are described as fermented. although the absence of any source of fermentable carbohydrate means that the process is putrefactive. giving an alkaline fermentation. Puwastien P. Microbiology and chemistry of fermented fish. Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. salt content was found to have the greatest impact on the viability of larvae. though 27% survival was still recorded (33). One study with the trematode Haplorchis taichui. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada. affecting an estimated 40 million people. particularly in south-east Asia and the western Pacific regions (30). beaver tail and paw.. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. 6. botulinum and the production of toxin. and these are often used in the production of fermented fish products. Raw materials subject to this process include seal flipper. Journal of Food Science. Greenland and Alaska. Yhoung-Aree J. Saisithi P. Salting levels are often insufficiently low to prevent growth of C. Oslo. 137 . and salmon heads and eggs. walrus nose. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. Dollar A..M. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31). while increasing the acetic acid content of the formulation had little or no effect (32). Institute of Nutrition. Foodborne trematode infections are emerging as a major public health problem. Baseline Study (1987-1988). Novus Press.. Dhanamitta S. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28).FERMENTED FISH botulism have become less frequent in recent years. Thailand. which is generally consumed raw (29). Where these products are consumed without final cooking.
Bioscience. Applied Microbiology. 3.K. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries. 124-30. Nov. Namwong S.J. 8. 57.M. Tanasupawat S. Constantinides S.. isolated from jeotgal. a traditional Korean fermented seafood. 2008. Production and characterisation of NaCl-activated proteinases from Virgibacillus spp. Government of Australia. Phil.T. 29. 141-2. 10..M. Rattagool P. Journal of Food Technology. Fermented fish products of south east Asia.. Tsunemi M. Dong-Ho K. 138 . Effects of gamma radiation on sensory qualities. 13. 353-63. Moonmangmee S.. Thawai C. 11.. Process Biochemistry. 2007. Plenum Press. Itoh T. Yasir M....R. Biotechnology and Biochemistry. Tropical Science. Halophiles in Thai fish sauce (Nam pla).. Kyong-Haeng L.K. 15. Dhamcharee B.. 6.. Orejana F. SK33 isolated from fish sauce fermentation. 1979. 17-30..R..V. 2006.. New York. 185-92. 1991. International Journal of Systematic and Evolutionary Microbiology. Yongsawatdigul J..W.A.T. Aslam Z. 17.H. Lim J. Adams M. Velankar N. 6338. Reddi P. Canberra. SR5-3 isolated from fish sauce: purification and characterisation. Byun M. Rodriguez-Valera F... Thongthai C. Kaneko Y.D. Journal of Food Protection. 11. Saono S. Protein hydrolysate from fish. 37. 1995..FISH AND SEAFOOD 4. 5. International Journal of Systematic and Evolutionary Microbiology. Tanasupawat S... Yook H. Liston J...R. Takano M. Sinsuwan S. 9. 934-9. 80... 643-7. Catheptic activity of fish muscle. 7. 16. Microflora of four fermented fish sauces. Kudo T. Hiraga K... Olympia M.. Lentibacillus kapialis spp. Cooke R. 3649. Journal of Fermentation and Bioengineering. 1986.. Fukuda H. 12.S. Lee S. 1985.. 70. Suntinanalert P. Chung Y.. Backhoff P. 14. Journal of Food Science and Technology.H. Agents of proteolysis and its inhibition in patis (fish sauce).. Nov. 1395-401. 1972. microbiological and chemical properties of salted and fermented squid. Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme. Journal of Food Science. 25. Sands A. 1957. Ed. Im W. Some chemical changes in fish silage. A halophilic serine proteinase from Halobacillus spp... 2007. 63. from fermented shrimp paste in Thailand. Oda K.. Journal of Scientific and Industrial Research (India). 16A. 61-73. 106-8. 43. Hull R. 381-8. Crisan E. 2000. 1975. Ono H. 1976. Pakdeeto A. Jae-Hun K. 57. Ahn H. Dymsza H.. Rodtong S. Salinococcus jeotgali spp. Takada K.. in General and Applied Aspects of Halophilic Microorganisms..
1815. 1992. 27. in Microorganisms in Foods.. Martin J... Sakai H. Owens J.. Control of foodborne trematode infections.W. 28. 103. Characterisation of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation. 2007. 1992. Rustad T. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation. 138-44. in Clostridium botulinum: Ecology and Control in Foods. 25. Canada Communicable Disease Report. Dodds K. Journal of Food Science and Technology. Caldo G. 1990. Moody L. Traditional fermented fish products with special reference to Thai products. 21. 30. Tanaka M. Huss H.L. 23.B. a Thai low-salt fish product.. Hauschild A. Journal of Agricultural Science (Tokyo). 69-104. Marcel Dekker. Erikson U. 1998. 46. Gram L. Epidemiology of human foodborne botulism. Wedell-Neergaard C.. 24. 1983. 1983. Dodds K..K. Yamprayoon J.. Yeast-flora in red burong-isda a fermented fish food from the Philippines. in the Philippines. Sakai H. Gram L.B.FERMENTED FISH 18.D. Journal of Agricultural Science (Tokyo).H. Patrick D.M. Kozaki M. Volume 5: Microbiological Specifications of Food Pathogens.. 307-14. 28. New York. Tropical Science. Fermentation and spoilage of som fak. New York. Paludan-Müller C.. 274-86. Riebroy S. 19.A. 1995. 104-6.W. Avurhi J.H. 28 (6). 426-35. Caldo G. in Clostridium botulinum: Ecology and Control in Foods. 139 . Visessanguan W.W. Wainwright R. Journal of Applied Microbiology.. 305-22. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak. 1-4. 28. Benjakul S. Ostergaard A... 20. 38. ASEAN Food Journal.L.. Food Chemistry. 105-12. Valyasevi R. 26. 1999. 92. 2002.H. a Thai low-salt fermented fish product. Gram L. 219-29. Ben Embarek P. Dawar M.. Blackie. 3. Hazards from northern native foods. Fung C. International Commission on Microbiological Specifications for Foods.A. Two outbreaks of botulism associated with fermented salmon roe – British Columbia. Eds. 1987. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper....H.. The fermented fish food. International Journal of Food Microbiology. Vibrio parahaemolyticus. Huss H. Eds. Ed.. Saisithi P. Hauschild A.H.. International Commission on Microbiological Specifications for Foods. WHO Study Group. Isaac-Renton J. 2002. 3-10... World Health Organisation Technical Report Series. Marcel Dekker.. London.D. Kozaki M. Huss H.. Paludan-Müller C. 29.. 22. burong-isda. 1996. Hauschild A. 27.H..
. 29. Gaithersburg. Fermented flesh foods. 1995. 23.. Fermentation and Food Safety. 661-70. Progress in Industrial Microbiology. International Journal of Cancer.. Srivatanakul P. Wongsawad C. Ruamsuk A.. Uchida K. Todoroki T. Miwa M. Hanai S. 2001. 1986. Survival of Anisakis larvae in marinated herring fillets... Nithikathkul C. 140 .R.R. 2005. Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom. Bloemsma B... Sripa B. Huss H. 6. Adams M.7 Further Reading Adams M. Kittiwatanachot P. 854-60.. Honjo S. Sriplung H.J. Southeast Asian Journal of Tropical Medicine and Public Health. 117. 15998. Thai traditional food preparation.H.. Kikukawa H. Chuboon S.. Deerasamee S. northeast Thailand. Horst K.FISH AND SEAFOOD 31. 2005... Jedpiyawongse A... Roepstorff A.R... Aspen Publishers Inc. International Journal of Food Science & Technology. 32. Nout M. 33. 36.. Survival of Haplorchis taichui metacercariae in lab-pla. 110-1.
Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. Increasingly sophisticated instrumentation (e.D. The manual on harmful marine algae and their products. Another book dealing with the clinical effects of a variety of poisons and toxins. several scientific reports have described their efficacy and potential.. most notably the azaspiracids and the cyclic imines as well as some additional polyethers. It remains to be seen if this approach gains momentum in the future.7. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7. While some of these toxins only occur in seafood from a restricted geographical region. and a second updated edition dealing with the pharmacology.3) and some practical applications for detecting shellfish toxins have been reported (4. Although most of these antibody-based test kits have yet to receive international regulatory approval. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. Wright.g.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. C.M. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. including the well known marine toxins 141 . An emerging technology is the use of immunosensors (1 . known and unknown. physiology and detection of marine and freshwater toxins has also appeared (8).. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins. Nevertheless.C. Ph. Since the original chapter in this series was published (6). 5). a number of new toxins and toxin families have been identified. Scientific and Cultural Organisation) has been updated (7). the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. FISH AND SHELLFISH TOXINS Jeffrey L.
Around twenty PSP toxins are known. temperate. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). Saxitoxin Group instead of the Paralytic Shellfish Poisons). and a review on the risk assessment of marine toxins has also been produced (14). with intoxications occurring in many countries even today (15). Consequently.2. Not all the toxins found in shellfish extracts are microalgal products.1). 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. The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). The Ncarbamoyl sulfates are readily hydrolysed to STX. all of which contain a tetrahydropurine skeleton (Figure 7. a great variety of shellfish from all latitudes. 7.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. Many of these species are extremely common and are found throughout the world in a variety of cold. and sub-tropical waters and habitats (15). and a tropical species Pyrodinium bahamense (16). Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. 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. 142 .FISH AND SEAFOOD has been produced (9). can become contaminated with these toxins. Gymnodinium catenatum. In addition. 7. neoSTX and Gonyautoxins (GTX) 1-4.2 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. dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). even crustaceans such as lobsters.g. and another. All PSPs degrade rapidly in alkaline conditions. some are the result of biotransformation processes by the filter-feeding host.
7. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins.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. 143 .2 Toxicology PSP toxins affect the central nervous system of mammals.1. tongue. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. this is never sufficient to render the seafood safe to consume. Human symptoms begin with tingling and numbness of the lips. and fingers within 5 . and humans seem to be particularly susceptible.30 minutes of consumption.2. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. 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). 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.
Since its discovery. a pre-column method has received Association of Analytical Communities (AOAC) approval (18). and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. After some development and various trials. specifically 144 . 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. 17). and has also been evaluated by a number of EU laboratories. TTX most commonly occurs in many species of puffer fish. While the chemical detection methods are most promising for the future.2. 24) (Figure 7. 7.3. a criticism has been that such methods are time consuming. 15. The mouse bioassay has been in place for decades and the method has been standardised. Alternative HPLC methods have subsequently reduced this to a single HPLC run. 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).2). 23. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. China. Thailand. Taiwan.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. 7. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. and while simple and relatively inexpensive to operate. other derivatives have been reported.FISH AND SEAFOOD 7. and islands of the South Pacific.8 µg/g) of shellfish or seafood tissue. including Japan. However it remains as the monitoring method of choice in many countries. and over ten analogs are known (22. The latter test is accepted by the United States Food and Drug Administration (US FDA) for use in the United States National Shellfish Sanitation Program. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). Two approaches have been taken here. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. as well as other parts. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21).3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. Most intoxication events have occurred in South East Asia.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised.
and once again the mouse bioassay has been used to detect the presence of the toxin.2. Structure of tetrodotoxin 7. 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. it is served in speciality restaurants with qualified chefs to properly prepare the fish. The onset of symptoms usually occurs rapidly.FISH AND SHELLFISH TOXINS in the liver. and even newts. gonads and roe (24). raising speculation that TTX is produced by a bacterial source.2 Toxicology Like the PSP toxins.2 mg of purified toxin can be lethal. 25.3.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. Fig. preventing depolarisation and propagation of action potentials in nerve cells. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. 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. All of the observed toxicity is secondary to the action potential blockade. and more recently by an actinomycete Norcardiopsis dassonvillei (26). respiratory failure and coma. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. and blocks diffusion of sodium through the sodium channel. horseshoe crabs. 7. 26). hypotension. In Japan. An oral dose of 1 . A review of TTX detection methods has been published (12. 27). 7. 23. molluscs. the blue-ringed octopus.3. Typically this has been accomplished using HPLC followed by post 145 . Since then. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. nausea and vomiting. Severe poisoning causes paralysis and respiratory failure. and in mild cases results in distal muscle weakness.
4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA).3. Currently 12 isomers of DA have been reported (see Figure 7. but all of them are considerably less toxic than DA itself (29). Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28). However this method has been criticised for being complex. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. a polar glutamate agonist. Structure of domoic acid and three naturally occurring photoisomers 7. and while promising. 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). 7. Immunoassay methods have also been investigated. but it is known that such congeners are less toxic than DA itself (30). 7.3 for examples).1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31).FISH AND SEAFOOD column oxidation to the fluorescent derivatives.4. need further development (24). These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. but 146 . Fig.
and clams. another diatom species Nitzschia navisvaringica. scallops. which has been well documented in studies along the west coast of the US. cormorants and pelicans (34).2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. found in the Pacific around Vietnam. including persistent short term memory deficits (36 . 7. southern and western portions of Ireland. seizures. Penetration of the blood brain barrier is poor in healthy subjects.. it has been reported that cooking or steaming contaminated mussels merely has the effect of spreading the toxin to other tissues of the shellfish as a result of disruption of the digestive gland. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. Indeed. and review of the toxicologic pathology of DA has been published (36). and is mainly distributed in the blood. To a great extent. 147 . 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. Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. the Philippines. multiseries. Victims display a variety of symptoms including headache. 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.4. Another factor is the movement of the toxin through the food chain. Recently. but any impairment of the blood brain barrier results in additional risk. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). 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. 33). has been reported as a medium-level producer of DA (33). nausea. disorientation and coma. Pseudonitzschia australis. Okinawa and more northerly parts of Japan.FISH AND SHELLFISH TOXINS only three strains (P. and sea animals such as sea lions and even whales (35). and in locations around New Zealand (32. For example. the Mediterranean.39). herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. The oral absorption of DA is around 5 . and this has led to ASP toxicity in birds.10% of the administered dose. 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. as does impaired renal function which increases serum concentration and residence time of the toxin.
4). California. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). as can LC/MS. 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. are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. DA can be detected by the mouse bioassay but the regulatory limit for shellfish is set below this level at 20 µg/g (20 mg/kg) shellfish tissue. Alternatively. and these methods have been reviewed (40.. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. and this is typically the method of choice. the heterotrophic dinoflagellates Dinophysis spp. Since then. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. 7. To further complicate matters.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. DTX-2 was discovered in contaminated Irish shellfish (46). Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. though this method is most commonly used to analyse plankton and seawater (42). but just over 15 years ago a third toxin. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. and dinophysistoxin-2 (DTX-2) (Figure 7.4. 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). dinophysistoxin-1 (DTX-1).5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). produce a variety of other 148 .30 ng/g) with a rapid clean-up step using a strong anion exchange resin. 49). In the context of this chapter.FISH AND SEAFOOD 7. australis. in Monterey Bay. and consequently were often overlooked as the real source of the toxins. 48.. although in the latter study the probes for Pseudonitzschia pungens. Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). Dinophysis spp. However. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . and an immunosensor method has also been reported (4). 41).
are recognised as DSP toxin producers. 149 . 7. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48).. Today over ten Dinophysis spp. In comparison. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. and include Dinophysis acuminata. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. and Prorocentrum belizeanum. While these esters are not toxic in vitro.4. are also recognised as a threat to human health. Prorocentrum maculosum. Lately these compounds. This was probably not the first DSP episode. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. and Dinophysis norvegica among others (47. The parent acids are the only DSP congeners to be found in shellfish tissue. such as Prorocentrum lima. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. Dinophysis fortii. DSP toxins produced by benthic Prorocentrum spp. In this case Dinophysis was suspected as the culprit organism. Dinophysis acuta. 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.5.1 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. DTX-5. have been referred to as lipophilic shellfish poisoning toxins (LSPs or LSTs) (49) because they are found in the same lipophilic fractions as the DSP toxins and are described in more detail later in this chapter.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. Fig. Structure of okadaic acid and the two related DSP toxins 7. 49). but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. which include the pectenotoxins and the yessotoxins.
nausea. vomiting. which can lead to an underestimation of the total DSP content in a sample. New Zealand and Chile. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1.3 days. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. DSP episodes have been reported from Japan. DSP episodes have been reported worldwide and are included in many monitoring programs (47). This regulatory level includes the possibility of LSPs in the DSP fraction being tested. DSP-Check is a commercially 150 . 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). even with the minimal of clean up steps (48).3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. In recent years. Human symptoms include diarrhoea. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. and abdominal pain.5. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 .5. and DTX-2. and while this provides critical seafood safety information. 7. various European countries.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48).2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A. 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. 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). Due to the lipophilic nature of the toxins. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. Since the late 1970s. 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. and a detection limit of 100 µg/kg has been reported (12). it does not provide information on the DSP toxin profile. both critical enzymes involved in many key metabolic pathways in mammalian cells (50) and consequently affect a host of other biological processes in a cell including tumor promotion. A regulatory level of 80 . All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. 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. 7.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries.
As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). 7. Within this group.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness.5 and 7. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . Some representative examples of both groups of toxins are shown in Figures 7.6 Lipophilic Shellfish Toxins (LST) Lipophilic extraction of toxic shellfish for DSP toxins often results in the simultaneous extraction of other lipophilic polyethers such as the pectenotoxins (PTXs) and yessotoxins (YTXs). for many years these compounds were included in the DSP toxin complex. Some doubt remains as to the significance of these compounds as marine or shellfish toxins with respect to human health since the PTXs for example are co-extracted with the DSP toxins. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). and another rapid antibodybased test kit for DSP toxins has also been reported (53). However. with a claimed detection limit of 20 µg/kg. 7.5. If a European collaborative study of the fluorometric protein phosphatase inhibition method is successful it may eventually be approved for regulatory purposes in the EU. 55). 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. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. Indeed. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. Fig.6. with good agreement with the mouse bioassay and LC–based methods (54. In yet another approach. 58).
7. the toxin content and profile may vary among different strains.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). 58).FISH AND SEAFOOD Fig. 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. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. Japan. many parts of Europe and the Mediterranean.6. which may co-occur in blooms with other organisms.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). As is typical for many algal species. YTX and homo-YTX 7. eastern Canada.6. While 152 . The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. 7. 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. Structure of the two parent yessotoxins. New Zealand and South America (49. 57.
The structures and chemical data including the molecular weight of each member have been collated (56). 64).7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. The crude extract may be used directly in the mouse bioassay. but various congeners are acutely toxic in the mouse bioassay following IP injection. The regulatory level of 160 µg/kg shellfish tissue has been established and the mouse bioassay is often the primary detection method although LC/MS methods are preferred and this requires some simple clean-up of the crude extract prior to chemical analysis. some of which are found as glycosides and sulfates. However. Although there is no evidence that yessotoxins are responsible for human illness. though its cell effects may be linked to modulation of calcium channels (62). the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present.3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue. There are no clear cases of human illness resulting from ingestion of PTXs (59). 7. While LC/UV detection is possible for PTXs. but less so following oral dosing (61). Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. which aids detection by LC/UV. For many years it was believed that 153 . a regulatory level of 1 mg/kg shellfish tissue has been set.6. which is further compounded by differences in the experimental design. and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. chemical analysis is best carried out by LC/MS and several approaches have been described (57. 7. The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. Because of the number and complexity of the YTX group. but a fluorescent derivative has been prepared for others. The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds. Estimates of acute toxicity of the parent compound following IP injection vary considerably.7). The YTX group has not been reported to cause human illness. the PTXs show considerably less toxicity when administered orally. it may be compromised by other biological materials in the matrix.FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7. Some of the PTXs are potent cytotoxins in vitro (60). 58). Their toxicology and mechanism of action remain uncertain. Many YTX toxins contain a conjugated diene system.
Interestingly. Structure of the two parent brevetoxins. Fig. but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65. which are found in Japanese waters (66). the same toxins have been reported to be produced by three genera of Raphidophytes. namely Chatonella marina. 154 .1 Origins and distribution The brevetoxins are produced by the dinoflagellate Karenia brevis (formerly known as Gymondinium breve and Pytochodiscus brevis) a dinoflagellate found in the Gulf of Mexico.7. They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf. 7. oysters. 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”. verruculosa off the mid Atlantic coast of the U. and cockles (65. was chemically characterised as PbTx-2 (67). 66). 66). In the Gulf of Mexico. Many of these species are associated with massive fish kills. These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). whelks. and one of the brevetoxin metabolites BTX-B2 7. and off the coast of New Zealand (65). brevetoxins are found in mussels. PbTx-1 and PbTx-2.FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida.7. Fibrocapsa japonica.S. and Heretosigma akashiwo.
The human health implications following exposure to NSPs have been reviewed (69). and diarrhoea. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. 69). with a mean duration of symptoms of around 1 day. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). 66. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. Ingestion of brevetoxins results in a variety of symptoms including parathesia.7. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. this family of toxins has proven to be a challenge both to identify and monitor.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. malaise. vertigo.7. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . 7. nausea. with hot spots in the coral reef areas of the Caribbean and French Polynesia. and in severe cases seizure.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. In this application. The mouse bioassay is used as the primary detection method. While the mouse bioassay continues to be used.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71).FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. which can result in a cascade of associated events in a cell (66). 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. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). and mammalian body fluids without pretreatment or dilution (72). shellfish homogenates. 7. The mean time to onset of symptoms is around 3 hours. the competitive ELISA method correlated most closely with the mouse bioassay results. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). 7.
Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red.8. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7.8. complex. a group of closely related polyethers that are considered the source of ciguatoxins. Distribution through the food chain begins with 156 . but more are suspected to exist. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. CTXs are large. These include species belonging to the genera Ostreopsis and Coolia. Fig. The diversity of benthic phytoplankton that exist on reefs further complicates the picture. and only a few structures have been identified.FISH AND SEAFOOD transferred through the food chain to the larger carnivores. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin. lipid soluble polyethers. 7. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. 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. green and brown seaweeds and also occur in sediments and coral debris.8). Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7.1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins.
These include paresthesias (numbness and tingling of the extremities).1 µg of ciguatoxin can result in human illness (74. Even when suspect samples are available. 75).8. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. arthralgia. The recommended method to extract fish tissue has been described in detail (74) and the initial warm acetone extract is partitioned against hexane and diethyl ether where the CTXs are concentrated. and in fact CTX poisoning is often only identified after consumption of contaminated fish. and temperature sensation reversal. and the sensation of loose teeth (77). Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. which can return following the consumption of alcohol and certain foods such as oily fish (74. myalgia. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . they bind to voltage-dependant sodium channels in the cell membrane causing them to open. The onset of symptoms can occur rapidly within 24 hours. pruritus. and moray eels. which in turn are consumed by carnivores such as snappers. only trace amounts of CTXs can cause illness following consumption of contaminated fish. which can be confused with chronic fatigue syndrome. and although the gastronomical problems may dissipate within a day or so. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. 7. Globally. Other symptoms include a metallic taste. Patients have reported symptoms lasting weeks and even months.8. weeks or even months (74 . Nevertheless CTX poisoning continues to be a threat to local and visiting populations. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. Another symptom that can develop over time is extreme fatigue. 78). sharks. all of which can be linked to the effect of CTX on excitable membranes. although not all CTXs are equipotent. there are neither standards nor an official testing program.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish.3 Detection methods As a consequence of their potency. Local island populations attempt to avoid poisoning by catching only smaller fish. the neurological symptoms can persist for days. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. barracudas. and there are suggestions that such incidents are increasing and spreading worldwide (74).76). where cold objects feel hot and vice versa. and it is estimated that ingestion of as little as 0. 75. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. groupers. 7. Like the brevetoxins.
but this only has application in a clinical setting (81). Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. has been developed. A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine. and the corrected structure was published in 2004 (86) (Figure 7. and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. with AZA2 (10 . and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. minor revisions of a few stereocenters were made. Chemical detection methods are severely compromised by the lack of standards. Recently.25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. and following synthesis of the molecule some time later. at C-3 and C-23). which are based on an LC-MS/MS approach. This is then subjected to LCMS/MS analysis. Usually the major AZA derivatives found in shellfish are AZA1 (50 .g.30%) and AZA3 (5 . The majority of these have been proposed based on LC/MS analytical data. 7. another in vitro sodium channel-based assay. Immunoassays may be an alternative detection method (79).9). Acetone extracts of fish tissue are subjected to solvent partitioning. 84). The original structure proposed revealed an entirely new class of shellfish toxin.80%). following consumption of contaminated mussels Mytelius edulis (82). specific for CTX activity. followed by clean-up on silica and aminopropyl cartridges.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis.9). almost 20 additional derivatives have been reported (83 . high-throughput screen (74).9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. Recent reviews of this new group of toxins have appeared (83. and another dealing with the chemistry and detection methods has been published (85).20%) present in lesser amounts (Figure 7. Since the original discovery and identification of the toxin. which features characteristic fragment ions corresponding to sequential losses of three molecules of water.FISH AND SEAFOOD Lethal Dose (MLD) 0.85). although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). Consequently. In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). 158 . and only a few structures (AZA 1 . only a few laboratories are able to undertake such analyses.
which are ingested by P. diarrhoea and stomach cramps. 7. and some time after the original episode in Ireland. Mytilus galloprovincialis). Scandinavia. 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. 90). oysters (Crassostrea gigas. Following the original episode. While this result is not necessarily in debate.). There has been considerable debate over the source of azaspiracids. which can persist for 2 . as in other cases of shellfish toxins. However. cockles (Cerastoderma edule). these molluscs were never considered to be the biogenetic source. 7. and to date this has been the only source from which these toxins have been found at toxic levels.9. The bivalves affected include mussels (Mytilus edulis. vomiting. scallops (Pecten maximus). AZA toxins do 159 . and most recently in a crustacean (Cancer pagurus). 88.9. clams (Tapes philipinarum. Ostrea edulis).2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). cassipes produces AZAs in culture.FISH AND SHELLFISH TOXINS Fig. Portugal. In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. it has been impossible to establish that P. cassipes (83).1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. 89). Structure of the three principal azaspiracids found in shellfish 7. and off the coast of Morocco (83. AZA occurrence has been reported by several European countries including France. These symptoms are not unlike those experienced following the consumption of DSP toxins. but current data indicates that unlike the DSP compounds. Donax spp.3 days. 84.9. Ensis siliqua.
Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. 85). 92). However. parenchymal cell pyknosis in the pancreas. 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). Based on IP minimum lethal dose data.9. PSP. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. and closely related ptierotoxins (Figures 7. it is recommended that the entire shellfish be used for AZA analysis since it has been shown that these toxins can migrate to other tissues. 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 Cyclic imines These toxins are represented by several distinct structural types. and erosion and bleeding in the stomach. 7. Once again. The mouse bioassay has been used to monitor shellfish extracts. Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. AZAs also display cytotoxicity towards various mammalian cell lines (83. Using the same approach. Oral administration of AZAs at substantially higher doses than those used in IP injection studies did not result in any mouse mortalities after 24 hours. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. which are particularly amenable to detection by positive ion electrospray mass spectrometry. and ASP. abnormalities in the thymus and spleen.12). 91.10 – 7. and necrosis of epithelial cells on the microvilli (91). but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. Importantly these pathological outcomes were reported as being different from those caused by DSP. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. 7. the pinnatoxins. but all contain a spirocyclic or cyclic imine function within their structure.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. though the possibility exists that they may inhibit other phosphatases. the spirolides.
7. 7.1..10. it was proposed that a dinoflagellate identified as Gymnodinium spp. 161 .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. In only two cases (gymnodimines and spirolides) has a planktonic source been identified.1 Origins and distribution The original observation of toxicity in oysters led to a wider survey of other shellfish species in New Zealand when it was discovered that the toxin was present in a variety of bivalve molluscs including mussels. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94. 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. they are sometimes referred to as emerging toxins of uncertain human health concern. 95). Structure of the cyclic imine gymnodimine 7. two additional congeners.10. 95). selliformis (97). Pacific oyster. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. Fig. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. Subsequent taxonomic studies revised this again to a new dinoflagellate K. Some time later. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis).520 Da (94. surf clam and New Zealand abalone. cockles.FISH AND SHELLFISH TOXINS health. scallops. but they are included here for completeness.10).10. At the time of the initial observation of gymnodimine.
finishing with 100% methanol.1. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. 7.900 µg/kg). Based on this diminished oral toxicity. paralysis of the hind quarters and eventually complete immobility. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. There is a rapid onset of symptoms including respiratory distress.1.to10-fold reduction of toxicity (LD50 600 . 7.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. abdominal breathing. A later study using purified toxin established an LD50 dose (Lethal Dose. The time lapse between ingestion and death is between 5 and 15 minutes. 95). They were originally discovered in extracts of mussels (M. it has been stated that regulation against gymnodimine is not required (98).10.3 Detection methods The characteristic “fast-acting. Oral administration of the toxin to mice by gavage resulted in about an 8. Most cyclic imines lack an extended chromophore. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. 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 original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. Reduction of the imine group eradicated the toxicity.11).FISH AND SEAFOOD 7. Whether isolated from dinoflagellate cells directly or from shellfish tissue. almost a dozen derivatives in total have been identified to date (94. and lack of responsiveness to stimuli.100). In particular. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. 95) (Figure 7. and this is the detection method of choice. Since their original discovery which resulted in the characterisation of spirolides A-F (99. However. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 .10. and so spectroscopic detection by UV methods is not favoured. 50%) of 96 µg/kg. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). together with a lack of evidence of human intoxication.10.
11.10.8 µg/kg). the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). those spirolides possessing the vicinal dimethyl grouping (see Figure 7. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. 7. 7. Fig. fasted mice).1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. fasted mice).2.166 µg/kg. by gavage (87 . While oral toxicity is considerably less than by IP injection. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . the gulf of Maine. or by oral ingestion (381 . the western coast of Scotland.10. southern Ireland.2 Toxicology The spirolides are the most studied group of the cyclic imines. Interestingly.2. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. 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. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). R1 = CH3) of the cyclic imine ring are the most toxic of the group (98).11. Structure of four spirolide toxins A-D 7. All data reported here are for desmethyl spirolide C.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . 95). in fjords in Denmark and Norway.707 µg/kg.
which includes two diastereomers. 7.FISH AND SEAFOOD (100). Fig. However. 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. four members of the group have been identified – two of them being the C-34 diastereomers.B contain a spiro imine moiety similar to that found in the spirolides.12). The compounds are zwitterionic and therefore somewhat more polar than the spirolides.10. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C.2. 95. 102).10. consistent with modulation of acetyl cholinergic receptors (103). Three members of this group have been reported.3 Pinnatoxins and pteriatoxins Pinnatoxins A . 7. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. To date.3 Detection methods As discussed for gymnodimine. 7. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . While the mechanism of action remains to be clearly defined. the mouse bioassay can be used to detect these fast acting toxins.12. and in fact these compounds share a number of other structural features. A number of methods following this procedure have been described (94.
there is no compelling evidence to suggest that they have resulted in human toxicity (98). However.3.10.3. have long been suspected of causing human illness after ingestion. to date action or 165 . 7. 7. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. The LD99 (Lethal Dose. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. This is generally a result of increased monitoring and continually improving detection technologies and methods.3. new toxin groups such as the azaspiracids and cyclic imines have also been reported. While this is recognised as a real possibility for a wide range of toxins.FISH AND SHELLFISH TOXINS 7.1 Origins and distribution Shellfish of the genus Pinna.3 Detection methods No formal detection methods for these compounds have been reported. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). like the gymnodimines and spirolides.10. There is strong circumstantial evidence to suspect that these toxins. while pinnatoxin D was essentially inactive.11 Conclusion There has been considerable progress in the detection of known marine toxins. which in many cases are now found to be more widespread than previously believed. 7. and more advanced means of detection.2 Toxicology There is little information describing the toxicology of these compounds (98).180 µg/kg). which occupy tropical waters around Okinawa and the south east coast of China. Despite this progress. but this remains to be conclusively established (106). Coupled with this.10. The LD99 dose for the pteriatoxins is reported to be in the range (8 .100 µg/kg). arise from an algal source . has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. 99%) values for pinnatoxins A-C are in the range (22 . Only the acute toxicity in mice following IP injection has been reported. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other.most likely a dinoflagellate. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). 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. With increasing knowledge of the chemistry of toxins.
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Dickey R. Ed.G. Bourdelais A.S. Cheng Y... Manzo L.S... Ayyar R. 23.. Pierce R. Ciguatera toxins: Chemistry. Muscle Nerve. Wolny J.E.E... Botana L..D. Environmental Health Perspectives.. 456-79.P. Ramsdell J. 67. Backer L. Granade R.D. Reich A.C. Jacocks H. Butera R. Boca Raton. Prockop L. Pierce R. 2007.H. 68. 72. Neurologic Shellfish Poisoning (NSP).. Backer L..N. Benson J. Dechraoui M-Y. Environmental Health Perspectives.. 479-516... Johnson D. Pierce R. Landsberg J.M. 2004.. Freidman M. Flewelling L.P. 50. like ciguatoxins. Clark R... New fish-killing alga in coastal Delaware produces neurotoxins.... 2007. 551-8...M... 50. 77. 66. Weisman R. Abraham W.G.. 465-70. 2004. Gandini C.P. Zaias J.. 220-6..W.. Rome. are potent ichthyotoxic neurotoxins that accumulate in fish. toxicology. Bean J.. Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater. Naar J.. Botana L.A... Fernandez M. Baden D... 825-34... Reich A. Baden D. Toxicon.J. Ed. Kirkpatrick B. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. Cheng Y. in Seafood and Freshwater Toxins.G. Harmful Algae. 110.. in Seafood and Freshwater Toxins. Marine Biotoxins. Gannon D. in Seafood and Freshwater Toxins. 110.. The molecular and integrative basis to brevetoxin toxicity. 70. and detection. Steidinger K.. Naar J.A. Bienfang P..S. Zaias J. Boca Raton. Food and Agriculture Organisation of the United Nations. Fleming L.R. 171 .. 2002. 76.C. in FAO Food and Nutrition Paper 80. 2008. Toxicon. 74.. Ed. 2008. Watkins S.E.G.. 2008. 707-23.. Poon-King C. 179-85.. 2000. 117-71. Granholm A. 2008. Wanner A. Dickey R. Bossart G. Squicciarini D. Abbott J. 131. CRC Press.. 53. Locatelli C... Detection of brevetoxins in the twenty-first century.A... Tomas C..A. 2007. Lenzi A. Wang Z. 3. Ed. 73. Chen A. Naar J... 75. Naar J. Kirkpatrick B. Kubanek J. Mild ciguatera poisoning: case reports with neurophysiological evaluations. Botana L. Brevetoxins. CRC Press. Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2.....E.M. 187-94.. Marine Drugs. Abraham W. Aerosolised red tide toxins (Brevetoxins) and asthma. Bottein M-Y. Food and Agriculture Organisation. Boca Raton.E. Schrank K. Ramsdell J. shellfish and mammalian body fluid. CRC Press. Whitney P.. Kubanek J. Poon-King T. Baden D... Henry M. Poli M. 2004. 2002. Fleming L.. Fleming L.FISH AND SHELLFISH TOXINS 65. Literature review of Florida red tide: implications for human health.B... 71. Backer L. 519-50. Baden D.G.M.S. Food and Agriculture Organisation.. Buonocore M. 69. Tomas C.J. Flewelling L. 1598-603. Baden D.. West Indian Medical Journal. 6.M. Chest. Ciguatera fish poisoning: treatment prevention and management. 99-115.. Bourdelais A. Lancaster J.
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.
FISH AND SHELLFISH TOXINS 92. Twiner M.J., Hess P., Bottein-Dechraoui M-Y., McMahon T., Samons M.S., Satake M., Yasumoto T., Ramsdell J.S., Doucette G.J. Cytotoxic and cytoskeletal effects of azaspiracids-1 on mammalian cell lines. Toxicon, 2005, 45, 891-900. Moroney C., Lehane M., Brana-Magdalena A., Furey A., James K.J. Comparison of solid phase extraction methods for the determination of azaspiracids in shellfish using electrospray liquid chromatography-mass spectrometry. Journal of Chromatography, 2002, 963, 353-61. Molgo J., Girard E., Benoit E. Cyclic imines: An insight into this emerging group of bioactive marine metabolites. Phycotoxins, 2007, 319-35. Cembella A., Krock B. Cyclic imine toxins: Chemistry, biogeography, biosynthesis and pharmacology, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 561-80. Seki T., Satake M., MacKenzie L., Kaspar H., Yasumoto T. Gymnodimine, a new marine toxin of unprecedented structure isolated from New Zealand oysters and the dinoflagellate Gymnodinium sp. Tetrahedron Letters, 1995, 36, 295-9. Haywood A.J., Steidinger K.A., Truby E.W., Bergquist P.I., Adamson J., Mackenzie L. Comparative morphology and molecular phylogenetic analysis of three new species of the genus Karenia (Dinophyceae) from New Zealand. Journal of Phycology, 2008, 40, 165-79. Munday R. Toxicology of cyclic imines: Gymnodimine, spirolides, pinnatoxins, pteriatoxins, prorocentrolides, spiro-centrimine, and symbioimines, in Seafood and Freshwater Toxins. Ed. Botana L.M. Boca Raton, CRC Press. 2008, 581-94. Hu T., Curtis J.M., Oshima Y., Quilliam M.A., Walter J.A., Watson-Wright W., Wright J.L.C. Spirolides B and D, two novel macrocycles isolated from the digestive glands of shellfish. Journal of the Chemical Society, Chemical Communications, 1995, 2159-61.
100. Hu T., Burton I.W., Cembella A.D., Curtis J.M., Quilliam M.A., Walter J.A., Wright J.L.C. Characterisation of spirolides A, C, and 13-desmethyl C, new marine toxins isolated from toxic plankton and contaminated shellfish. Journal of Natural Products, 2001, 64, 308-12. 101. Cembella A.D., Lewis N.J., Quilliam M.A. The marine dinoflagellate Alexandrium ostenfeldii (Dinophyceae) as the causative organism of spirolide shellfish toxins. Phycologia, 2000, 39, 67-74. 102. Ciminiello P., Dell’Aversano C., Fattorusso E., Magno S., Tartaglione L., Cangini M., Pompei M., Guerrini F., Boni L., Pistocchi R. Toxin profile of Alexandrium ostenfeldii (Dinophyceae) from the northern Adriatic Sea as determined by liquid chromatography-mass spectrometry. Toxicon, 2006, 47, 597-604. 103. Gill S., Murphy M., Clausen J., Richard D., Quilliam M., MacKinnon S., LeBLanc P., Mueller R., Pulido O. Neural injury biomarkers of novel shellfish toxins, spirolides: a pilot study using immunochemical and transcriptional analysis. Neurotoxicology, 2003, 24, 593-604. 104. Uemura D., Chou T., Haino T., Nagatsu A., Fukuzawa S., Zheng S.Z., Chen H. Pinnatoxin A: a toxic amphoteric macrocycle from the Okinawan bivalve Pinna muricata. Journal of the American Chemical Society, 1995, 117, 1155-6. 173
FISH AND SEAFOOD 105. Takeda N., Uemura N., Suenaga K., Uemura D. Structural determination of pteriatoxins A, B, C, extremely potent toxins from the bivalve Pteria penguin. Tetrahedron Letters, 2001, 42, 3495-7. 106. Kuramoto M., Arimoto H., Uemura D. Bioactive alkaloids from the sea. Marine Drugs, 2004, 1, 39-54.
8. HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
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
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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.
Conduct a hazard analysis. Establish a system to monitor control of the CCP. 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. tests and other evaluations.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. The team should also include 177 . in addition to monitoring to determine compliance with the HACCP plan. 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. procedure. Determine the CCPs.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . Validation . 4.A point. procedures. quality assurance or control. ingredient and packaging buyers.The application of methods. 6. Verification . 3. operation or stage in the food chain including raw materials. Figure 8. hygiene managers. Establish procedures for verification to confirm that the HACCP system is working effectively. 7. Establish the critical limit(s).1 is a flow diagram illustrating this 12-stage logic sequence.1 Assemble HACCP team HACCP requires management commitment of resources to the process. food chemistry or engineering. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. 2. from primary production to final consumption. Establish documentation concerning all procedures and records appropriate to these principles and their application. 8.3. 5. 8. Each stage is discussed below in detail.Obtaining evidence that the elements of the HACCP plan are effective. food microbiology. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available.
g.1. Fig. Logic sequence for application of HACCP 178 . 8. chemical and/or physical). The scope of the plan should be determined by defining the extent of the production process to be considered and the categories of hazard to be addressed (e. If expert advice is not available on-site. biological. it may be obtained from external sources.FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations.
may need to be considered specifically. 8. 8.g. including practices onboard fishing vessels. Expertise on such inherent hazards. fermentation.1.3. etc. composition. storage and distribution conditions and instructions for use. It is also important to identify the consumer target groups. 8. such as enteric viruses in shellfish. packaging. contaminated fish and shellfish should not be used. freezing. processing conditions (e. These factors are likely to have a significant effect on some potential hazards. etc. If the processing does not include a kill step.3.2 Describe the product It is important to have a complete understanding of the product. shelf life. 8.g. there may be species-specific hazards related to certain fish and shellfish. It should provide an accurate representation of all steps in the production process from raw materials to the end179 .).3. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments.3. naturally present on fish from warm waters (> 15 °C). pH. marine toxins associated with particular fish and shellfish species.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. which should be described in detail. aquaculture drug residues and potentially pathogenic Vibrio spp.).3. heat treatment. However. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. such as children or the elderly.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. is a cooking process required?).3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. Vulnerable groups of the population. or in aquaculture operations. In addition. The description should include information such as the product name. physical and chemical structure (including water activity (aw).1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora.3.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries.
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. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. and therefore a detailed description of the procedure applied must be available. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly.6 List all potential hazards associated with each step. time/temperature data. it should also include details of CCP steps. ingredient specifications.4.FISH AND SEAFOOD product. For example.3. conduct a hazard analysis. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. because the hazard analysis and decisions regarding CCPs are based on these data. it is essential that the details of each step are fully appreciated and recorded. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. Ideally.3. It is essential that the diagram is accurate. cleaning and hygiene procedures and storage conditions. survival or multiplication of pathogenic microorganisms. 8. once determined.3. If HACCP studies are applied to proposed new process lines or products. The operation should be observed at all stages. or circumstances of any reclaim or rework activity. However.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. 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. 8. The team should then conduct a hazard analysis to identify which hazards are of such a nature that their elimination or reduction to an acceptable level is essential to the production of safe food. the qualitative and/or quantitative evaluation of the presence of hazards. 8. It may include details of the factory and equipment layout. 180 ● ● . features of equipment design.
3. the hurdle effect. The HACCP team should then determine what control measures exist that can be applied for each hazard. 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. Salmonella spp. Note: it is important at this stage that no attempt is made to identify CCPs. The incidence of psychrotrophic. For example. botulinum. since this may interfere with the analysis. For example. and there are an equally varied range of potential hazards associated with them. Vibrio spp. 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. and viruses). Therefore it is not possible. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards.g.). there are particular hazards associated with contamination of shellfish by microorganisms from human sewage.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products. Hazards introduced during processing of fish products depend very much on the characteristics of the process.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual. any food safety objectives or manufacturer’s food safety requirements. Pathogens may be part of the resident microflora of the living animal (e. 181 . Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. or may originate from polluted water or from post-capture contamination (e.6.g. or desirable. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. 8. and the potential growth of Listeria monocytogenes on smoked fish. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin. the number of consumers potentially exposed and their vulnerability. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. One control measure may be relevant to several process steps. where a hazard is repeated.
7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. biological or sensory parameters.3. However.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. Where possible. but for which no control exists at that step or any other. 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. Criteria used to set critical limits must be measurable and may include physical.1 Fish and seafood Again. critical limits should be specified and validated for each CCP. 8. If a significant hazard has been identified at a step where control is required for safety. chemical. Some examples relevant to fish products are: • • • Time and temperature limits for pasteurised and cooked products Temperature limits for chilled fish Brine concentration in cured fish 182 .8 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. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively.3.7. it is not possible to generalise on likely CCPs.2) to provide a logical. then the process must be modified to include a control measure.8. More than one critical limit may be defined for a single step. and its use may not always be appropriate. However.3.FISH AND SEAFOOD 8. structured approach to decision making. it is necessary to specify both time and temperature for a thermal process. For example. application of the decision tree should be flexible. and the reader is referred to the appropriate product chapter. 8.3. it can be said that effective control measures are likely to include the following: • • • • • 8.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
• • • 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
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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.
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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
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considered and documented, and adjustments made to the procedures as necessary. Triggered reviews/audits should occur as a result of changes, whereas a scheduled review/audit should be annual, as a minimum. 8.5 References
Wareing P.W., Carnell A.C. HACCP – A Toolkit for Implementation. Leatherhead, Leatherhead Food International. 2007. Bernard D., Scott V. Hazard Analysis and Critical Control Point System: use in controlling microbiological hazards, in Food Microbiology: Fundamentals and Frontiers. Eds. Doyle M.P., Beuchat L.R. Washington D.C., ASM Press. 2007, 971-86. Su Y-C., Daeschel M.A. Sanitation and HACCP, in Surimi and Surimi Seafood. Ed. Park J.W. Boca Raton, CRC Press. 2005, 141-62. Huss H.H. Considerations in the application of the HACCP principles to seafood production, in Assessment and Management of Seafood Safety and Quality. Eds. Food and Agricultural Organisation, Huss H.H., Ababouch L., Gram L. Rome, FAO. 2004, 153-77. Chen H-C., Chang P.C-M. Frozen seafood safety and HACCP, in Handbook of Frozen Foods. Ed. Hui Y.H., Cornillon P., Legaretta I.G., Lim M.H., Murrell K.D., Nip WK. New York, Marcel Dekker. 2004, 339-63. Mortimore S., Mayes T. The effective implementation of HACCP systems in food processing, in Foodborne Pathogens: Hazards, Risk Analysis and Control. Eds. Blackburn C.deW., McClure P.J. Cambridge, Woodhead Publishing Ltd. 2002, 229-56. Mayes T., Mortimore C.A. Making the Most of HACCP: Learning from Other’s Experience. Cambridge, Woodhead Publishing. 2001. Mortimore S.E., Wallace C., Cassianos C. HACCP (Executive Briefing). London, Blackwell Science Ltd. 2001. Chartered Institute of Environmental Health. HACCP in Practice. London, Chadwick House Group Ltd. 2000. Jouve J.L. Good manufacturing practice, HACCP, and quality systems, in The Microbiological Safety and Quality of Food, Volume 2. Eds. Lund B.M., BairdParker T.C., Gould G.W. Gaithersburg, Aspen Publishers. 2000, 1627-55. Stevenson K.E., Bernard D.T. HACCP: A Systematic Approach to Food Safety. Washington D.C., Food Processors Institute. 1999. National Seafood HACCP Alliance. IQF cooked shrimp processing HACCP model, in HACCP User's Manual. Ed. Corlett D.A. Gaithersburg, Aspen Publishers. 1998, 454-73. Mortimore S., Wallace C. HACCP: A Practical Approach. Gaithersburg, Aspen Publishers. 1998.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Khandke S.S., Mayes T. HACCP implementation: a practical guide to the implementation of the HACCP plan. Food Control, 1998, 9 (2-3), 103-9. Forsythe S.J., Hayes P.R. Food Hygiene, Microbiology and HACCP. Gaithersburg. Aspen Publishers. 1998. Food and Agriculture Organisation. Food Quality and Safety Systems: A Training Manual on Food Hygiene and the Hazard Analysis and Critical Control Point (HACCP) System. Rome, FAO. 1998. US Food and Drug Administration. Fish and Fishery Products Hazards and Controls Guide. Rockville, Food Drug Administration. 1998. National Advisory Committee on Microbiological Criteria for Foods. Hazard Analysis and Critical Control Point Principles and Application Guidelines. 1997. Gardner L.A. Testing to fulfil HACCP (Hazard Analysis Critical Control Points) requirements: principles and examples. Journal of Dairy Science, 1997, 80 (12), 3453-7. Martin R.E., Collette R.L, Slavin J.W. Fish Inspection, Quality Control and HACCP: A Global Focus. Proceedings of a Conference, Arlington, May 1996. Lancaster P.A., Technomic Publishers. 1997. Codex Alimentarius Commission. Hazard Analysis Critical Control Point (HACCP) System and guidelines for its application, in Food Hygiene: Basic Texts. Ed. Codex Alimentarius Commission. Rome, FAO. 2003, 21-31. Garrett E.S., Jahncke M.l., Tennyson J.M. Microbiological hazards and emerging food safety issues associated with seafoods. Journal of Food Protection, 1997, 60 (11), 1409-15. National Seafood HACCP Alliance. HACCP: Hazard Analysis and Critical Control Point Training Curriculum. Raleigh, North Carolina State University. 1997. Dillon M., McEachern V. HACCP and quality assurance of seafood, in Fish Processing Technology. Ed. Hall G.M. London, Blackie. 1997, 224-48. Savage R.A. Hazard Analysis Critical Control Point: a review. Food Reviews International, 1995, 11 (4), 575-95. Pearson A.M., Dutson T.R. HACCP in Meat, Poultry and Fish Processing. Glasgow, Blackie. 1995. Huss H.H., Food and Agriculture Organisation. Assurance of Seafood Quality. FAO Fisheries Technical Paper No. 334. Rome, FAO. 1994. Bonnell A.D. Process control, in Quality Assurance in Seafood Processing: A Practical Guide. Ed. Bonnell A.D. London, Chapman & Hall. 1994, 39-53. Pierson M.D., Corlett D.A., Institute of Food Technologists. HACCP: Principles and Applications. New York, Van Nostrand Reinhold. 1992. Hudak-Roos M., Garrett E.S. Monitoring, corrective actions and record keeping in HACCP, in Quality Assurance in the Fish Industry: Proceedings of an International Conference, Copenhagen, August 1991. Eds. Huss H.H., Jakobsen M., Liston J. Amsterdam, Elsevier. 1992, 521-31.
Oxford. 14-9. 15 (1/2). International Journal of Food Microbiology. 1988.L. Food Control. International Commission on Microbiological Specifications for Foods.. Bryan F. 1992. Mayes T. Blackwell Scientific Publications. 188 . Microorganisms in Foods. Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality. Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H. Development and use of the HACCP concept in fish processing. 1992. Geneva. 1992. Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. World Health Organisation. 33-44. WHO.H. 3 (1).
Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . final product specifications. 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. however. and conditions of raw materials.9. including microbiological criteria. have been replaced by several linked measures on food safety rules and associated animal health controls. transport and delivery vehicles requirements. A number of standards are provided in food legislation. Hygiene is a general concept that covers a wide subject area. from structural conditions in the factory or process facility. 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. with the intention of introducing consistency and clarity throughout the food production chain from primary production. EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials.1 Introduction Hygiene is an important aspect of ensuring food safety. to personnel requirements. under which food is produced to optimise public health and to prevent. eliminate or acceptably control pathogen contamination of food. and one that plays an important role in most countries’ food legislation. The new legislation was designed to establish conditions. to sale or supply to the final consumer. the existence of microbiological standards alone cannot protect consumer health.
Regulation 854/2004 relates to the organisation of official controls on products of animal origin.1992. 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.9. storing.15). N. A number of more detailed implementing and transitional measures have been adopted at EC level. and sets out what those enforcing the provisions have to do. transportation. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. Subsequently.1991. processing. distribution. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. 190 • • . p. 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. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No.B. European Communities L 268. 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. p.1). 41).7.9. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. 24. existing hygiene Directives including those below were repealed: • Council Directive 91/492/EEC of 15 July 1991 laying down the health conditions for the production and the placing on the market of live bivalve molluscs (Off. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. as last amended by Regulation (EC) No 806/2003.1991. manufacturing. packaging. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. handling and offering for sale or supply to the consumer.2 Legislative Structure From 1 January 2006. 24. as last amended by Regulation (EC) No 806/2003. p. J. 7.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation.
right through to the implementation of procedures based on HACCP principles. is required to give effect to the EU regulations. penalties and powers of entry. for example. 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. and binding in its entirety on all member states from the date of entry into force. Wales and Northern Ireland. The Food Hygiene (England) Regulations 2006 (S. etc. 2006 No. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. Additionally the requirements of EC Regulation 853/2004 must be complied with. There are some exemptions. national legislation in the form of a Statutory Instrument (S. food storage that is for private or domestic consumption. which will be covered later in this chapter. revocation of existing implementing legislation.g.7.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. 2073/2005. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. 191 . setting offences. of stock farming. As regulations. Wales and Northern Ireland). p. 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.I. 852/2004.11) The EU hygiene regulations apply to all stages of food production including primary production. and of hunting and fishing. Although the regulations have the force of law.3 Regulation (EC) No. Primary products are defined as products of primary production including products of the soil. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. 19. 1 . e. the legislation is directly applicable law. Although EU food hygiene regulations are directly applicable in the individual Member States there are some aspects where Member States are required or allowed to adopt certain provisions into their national laws. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No.) in England.I. with primary production. 9.14. domestic preparation or handling. and equivalent legislation in Scotland.1993.
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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: 22.214.171.124 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.
126.96.36.199 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.
188.8.131.52 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.
184.108.40.206 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.
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220.127.116.11 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.
18.104.22.168 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.
22.214.171.124 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.
126.96.36.199 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. 188.8.131.52 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.
184.108.40.206 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 220.127.116.11 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). 18.104.22.168 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.
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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:
shelling. or in natural sites. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. skinning. Conditioning means the storage of live bivalve molluscs coming from class A production areas. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness. reptiles and frogs) whether wild or farmed and including all edible forms. whether whole or prepared. Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. gutting and removal of fins and. followed by wrapping or packaging. purification centres or dispatch centres in tanks or any other installation containing clean seawater. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. where necessary. to remove mud. 197 . to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing. in particular as a result of feeding on plankton containing toxins. and all mammals. parts and products of such animals. slicing. heading. such as gutting. including products packaged under vacuum or in a modifiedatmosphere that have not undergone any treatment to ensure preservation other than chilling. live tunicates and live marine gastropods. 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. mincing or processing. 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. slime or sand. where appropriate. if necessary: chilling or freezing: filleting. and chopping. after preparatory work such as bleeding. slicing.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. filleting. live echinoderms. shucking. heading. Fresh fishery products means unprocessed fishery products.
Section I of Regulation (EC) 853/2004 are given as follows: 9. EÜ.2. EK. Section I of Regulation (EC) 853/2004. the food business operator may apply the same identification mark to both types of food. (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. when a product's packaging and/or wrapping is removed or is further processed in another establishment. EK. 9. EC.2. For the UK.4. ES. the new mark must show the approval number of the establishment where this occurred. (iii) Additionally.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production.4. 198 . the mark must show the approval number of the establishment. EF. 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. EY. Extracts of the requirements of Annex II. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not.2 Fishery product requirements 9. the code would be UK.FISH AND SEAFOOD 9. and the characters easily decipherable and be clearly displayed.1. (iv) If the identification mark is applied in an establishment within the Community.1. the mark must be oval shaped and include the abbreviation CE. EO.2 Form of the identification mark (i) The mark must be legible and indelible.4. for other codes.2. a new mark must be applied to the product.4.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. EG. (ii) However. (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. please refer to Regulation (EC) 853/2004. In this case. EB or WE.
2. (ii) For products of animal origin that are placed in transport containers or large packages. (v) When the mark is applied directly to products of animal origin.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. The requirements of this section accompany those in Regulation (EC) 852/2004. the colourings used must be EC authorised colours. (iii) In the case of liquid. This section does not apply to bivalve molluscs. reference should be made to the actual Regulation. and are intended for further handling. tunicates and marine gastropods when placed on the market live. C and D. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . echinoderms.4. Chapter III.1. for full requirements. they would need to be obtained following Section VII on live bivalve molluscs. it is sufficient to apply the mark to the exterior of that package only. a) requirements for establishments. or be printed on a label affixed to the product.LEGISLATION 9. 9. granulate and powdered products of animal origin carried in bulk. including vessels. and fishery products carried in bulk. the wrapping or the packaging. i.e. Therefore.3 Method of marking (i) The mark is to be applied directly to the product.2. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. it applies to those animals when not placed on the market live. Part A and Chapter V apply to retail. Section VIII of Regulation 853/2004. its wrapping or the packaging. the mark may be applied to the outer surface of the container or packaging. With the exception of Chapters I and II. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. Extracts of the requirements of Annex III. Parts A. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer.4. specifically relating to fishery products are given below. processing. Chapter IV. as amended. wrapping or packaging in another establishment. the approval number of the establishment and the abbreviation for the community where appropriate.
2. (a) primary production covers the farming.e. gutting. this supplements the requirements of Annex II. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products. fishing and collection of live fishery products with a view to their being placed on the market. There is a derogation from point a.) 9.2.4. Chapter VII of Regulation 852/2004. if a vessel has water coming in that is used with fishery products. from the place of production to the first establishment of destination. 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. A. it must be placed somewhere that will not contaminate the water supply. refrigeration and wrapping. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products.. The definition of fishery products covers primary production as well as its associated operations. bleeding.FISH AND SEAFOOD c) for the water supply. or to handle or process them after harvesting. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 . heading.1 Chapter I .Requirements for vessels Food business operators are required to ensure that they meet the structural and equipment requirements as well as hygiene requirements required for vessels used to harvest fishery products from their natural environment. Structural and equipment requirements refer to the following points 1. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. removing fins. i.2. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material. clean seawater may be used for the handling and washing of fishery products. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter.
recording of temperatures. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas. (iii) In vessels equipped for chilling fishery products in cooled clean seawater. Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C. and allow the monitoring and. 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. tanks must incorporate devices that allow a uniform temperature throughout the tanks. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. 201 .LEGISLATION (i) Such vessels need to have holds. and not more than 0 °C after 16 hours. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. 4. 3. Storage areas for the finished products also need to be sufficiently large and cleanable. Also. (ii) Holds must be separated from the engine compartments and from the crew quarters by partitioning that prevent any contamination of the stored fishery products. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. any waste processing unit needs to have a separate hold to store the waste. A temperature recording device must be present and placed where the temperature in the hold is the highest. where necessary. Such devices must achieve a chilling rate that ensures that the mix of fish and clean seawater does not exceed 3 °C six hours after loading. to ensure that melt water does not stay in contact with the products. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. where necessary.
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.Requirements during and after landing 9. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption. Chapter II .2. e) In cases where whole fish intended for canning is frozen in brine.2. 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. must be chilled or. 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 . do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. provided no damage occurs to the flesh.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. b) Factory vessels on board which crustaceans and molluscs are cooked.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. B. if this is not possible. Handlers may use spiked instruments to move large fish or fish that might injure them. chilled and wrapped. where fishery products are displayed for sale. and products washed thoroughly. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. c) Fishery products. (ii) In the case of food business operators responsible for auction and wholesale markets etc. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. other than those kept alive. The brine must not be a source of contamination. 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. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above.4. landed as soon as possible.
2. 9. 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. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 .LEGISLATION consumption. packaged and chilled as quickly as possible after preparation. B. Fillets and slices must then be removed from work tables as soon as possible after preparation. (ii) Heading and gutting must be carried out hygienically. dispatched. 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. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels.3 Chapter III . C. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. including vessels. C. part I. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed. prepared or processed immediately after arrival at an establishment on land. 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. They should be wrapped. and where necessary. 854/2004. Additionally. they must be stored under ice in appropriate facilities. handling fishery products A. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. points 1 and 2.2.4. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. (v) Containers for dispatching and storing unpackaged prepared fresh fishery products kept under ice should ensure that melt water does not remain in contact with the products.Requirements for establishments. Chapter I.
must accompany fishery products referred to above when placed on the market. (iii) sprat. (iii) A document from the manufacturer.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products.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.2. except when supplied to the final consumer. and whole fish must be gutted and washed beforehand. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . (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. cooling must continue until a temperature 204 . (iv) (wild) Atlantic and Pacific salmon. 9. they must be cooled rapidly. and (c) marinated and/or salted fishery products. raw materials must have no guts.4. 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. (ii) mackerel. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing.2. stating the type of process they have undergone. (b) fishery products from the following species. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. If no other method of preservation is used. if the processing is insufficient to destroy nematode larvae. Once produced. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. or a stabilising treatment.4 Chapter IV .Requirements for certain processed fishery products A. D.
2073/2005). raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. 9. the following conditions need to be met: A. B. 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). refining and purification steps before being placed on the market for the final consumer. B. However. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. 205 . 2074/2005. 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.4.LEGISLATION approaching that of melting ice is reached.5 Chapter V . (ii) The production process for fish oil must ensure that all raw material intended for the production of crude fish oil is subject to a treatment including (where relevant) heating. centrifugation. pressing.2. and the raw material is processed within 36 hours after loading. separation. Shelling/shucking is to be done hygienically. C. 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. 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. 2073/2005. 2074/2005). and after this. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No.2. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. processing.
Fresh. and when fishery products are wrapped on board fishing vessels.Storage of fishery products (i) Fresh fishery products. thawed unprocessed fishery products. Diodontidae and Canthigasteridae. Molidae. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market.4.6 Chapter VI . 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.2. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing.2. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption.2. In the case of whole frozen fish in brine intended for the manufacture of canned food.4.7 Chapter VII . prepared and processed fishery products belonging to the family Gempylidae. Fishery products contaminated with parasites may not be placed on the market for human consumption.FISH AND SEAFOOD D. 206 . E. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. it should be kept at no more than -9 °C.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. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. 9. 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. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. 9. The scientific name must accompany the common name on the label.2.
2074/2005.2. Helicolenus dactylopterus. provided that the journey is short and the competent authority allows this.2. species belonging to the Gadidae family.4.4. Sebastichthys capensis.Transport of fishery products The above points for Chapter VII apply in relation to transport. 9.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. 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.3 Other information: Rules relating to visual inspections for the detection of parasites in fishery products are stated in Annex II (obligations on the competent authorities) of Regulation (EC) No. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. The following in relation to TVB-N limits is extracted from the regulation: Unprocessed fishery products shall be regarded as unfit for human consumption where organoleptic assessment has raised doubts as to their freshness and chemical checks reveal that the following TVB-N limits are exceeded: (a) 25 mg of nitrogen/100 g of flesh for the species: Sebastes spp. The temperature of frozen fishery products must not fluctuate above -15°C. (ii) If fishery products are kept under ice. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them.2. species belonging to the Merlucciidae family.). melt water must not remain in contact with the products. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated.8 Chapter VIII . 9. (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 .
1 Chapter I . (b) In the case of other operations. The information required in the registration document is given in point 4 and includes the health status of the production area. The reference method to be used for checking the TVB-N limits involves distilling an extract deproteinised by perchloric acid as set out in Chapter III For distillation apparatus and routine methods which may be used to check the TVB-N limits. Chapter IX applies to pectinidae harvested outside those areas. taken from at least three different points and mixed together by grinding. 9. and point 3 of Chapter VII applies to retail. Apart from the provisions on purification. 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. VI. however. Chapters V.3 The regulation details specific hygiene requirements for bivalve molluscs. The copy of 208 . specifically relating to bivalve molluscs are given below. The sample must consist of about 100 g of flesh. they supplement the requirements of Annex II to that Regulation. this Section also applies to live echinoderms. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. Extracts of the requirements of Annex III. tunicates and marine gastropods. they supplement the requirements of Annex I to that Regulation. as amended. Live bivalve mollusc requirements - 9. Section VII of Regulation 853/2004.FISH AND SEAFOOD Regulation (EC) No 853/2004.4. shellfish species etc. for full requirements. where the raw material complies with points (a).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.3. please refer directly to the regulation. (b) and (c) of Part B(1) of that Chapter. VIII and IX.4. 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. reference should be made to the actual regulation.
3.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. 9. poles or other fixed means are to identify the boundaries of the sites.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority.2 Chapter II . 209 . (ii) Provided the requirements of chapter V are met. Requirements for relaying live bivalve molluscs Only areas approved by the competent authority can be used for relaying live bivalve molluscs with a requirement that buoys.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market . they need to be sent to a processing establishment and undergo specified treatments such as sterilisation in hermetically sealed containers or specific heat treatments such as immersion in boiled water so the internal temperature of the mollusc flesh reaches at least 90 ºC and this temperature is maintained for at least 90 seconds. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. B. Transport needs to allow sufficient drainage for best survival conditions.Class A means they can be placed directly on the market . 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. collection from: .4.Hygiene requirements for the production and harvesting of live bivalve molluscs A. Requirements for production areas (i) The competent authority classes (classed A. processing or relaying. C. B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with.
3. requirements if there are many batches of live bivalve molluscs. 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. the purification system to allow them to resume /maintain filter-feeding activity.4 Chapter IV . for: (i) paralytic shellfish poison (PSP). dinophysistoxins and pectenotoxins together.3 Chapter III . 20 milligrams of domoic acid per kilogram (iii) okadaic acid.3. (ii) amnesic shellfish poison (ASP). Requirements for purification centres There are requirements for live bivalve molluscs to be washed. B. 800 micrograms per kilogram.5 Chapter V . 9. and there are requirements for tanks and water storage containers. 1 milligram of yessotoxin equivalent per kilogram. 210 .3.4.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. 160 micrograms of okadaic acid equivalents per kilogram. the levels of marine biotoxins should not exceed the following limits.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. Before dispatch. containers and a label to identify purification. (iv) yessotoxins.4. Requirements for dispatch centres Handling should not result in contamination to any degree.FISH AND SEAFOOD 9.4.Hygiene requirements for purification and dispatch centres A.
Finally. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’. by way of derogation from directive 2000/13/EC. must be waterproof. Requirements of Chapters III and IV are to be complied with. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9. including the identification mark. 9. the species of bivalve mollusc and date of packaging (at least the date and month) are required. dispatch centre or processing establishment.4. 9.4. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.LEGISLATION (v) azaspiracids.8 Chapter VIII . 9.3.1). (ii) Individual consumer-size packages of live bivalve molluscs must be closed and remain closed after leaving the dispatch centre and until presented for sale to the final consumer.6 Chapter VI .4.Other requirements (i) The temperature at which live bivalve molluscs are stored/ transported in should be at one that does not adversely affect food safety or their viability.3.3. Also.9 Chapter IX . 211 . the label. 160 micrograms of azaspiracid equivalents per kilogram. Registration document requirements also apply.3.7 Chapter VII .2. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre.4.Specific requirements for pectinidae harvested outside classified production areas Requirements for harvesting and handling following harvesting and health standards for live bivalve molluscs must be met.Identification marking and labelling In addition to general identification marking (contained in 9. they cannot be re-immersed in or sprayed with water.4.
or (iii) bivalve molluscs. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. 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. (ii) fishery products not complying with the requirement concerning biotoxins. 9. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. It is a requirement that food business operators give assistance to ensure that official controls carried out by the competent authority can be done properly. or - - 212 . In this regulation. Section VIII. they derive from: (i) poisonous fish. This refers to official controls or production and placing on the market of fishery products.FISH AND SEAFOOD 9.5 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.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. echinoderms.5. chemical. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given. Therefore the health standards for fishery products as previously stated in Annex III. Fishery products in particular must comply with the requirements of Annex III of the regulation. Chapter V of Regulation 853/2004 apply. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004.
The reference method for this analysis is the five-tube. Also. coli per 100 g of flesh and intravalvular liquid. Annex II 9.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. The reference method for this analysis is the five-tube. A summary is given as follows: 9.2. 4. Regulation (EC) 854/2004. 213 . Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. Live bivalve molluscs from Class C areas must not exceed 46. coli per 100 g of flesh and intravalvular liquid. Live bivalve molluscs from Class B areas must not exceed. as previously stated. coli per 100 g of flesh and intravalvular liquid. section VII. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated.5.000 E.600 E.5. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. chapter V is reinforced. live bivalve molluscs must not exceed 46. live echinoderms. Sampling plans and frequency are required. live tunicates and live marine gastropods. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. 000 E.2 This states official controls for the production and placing on the market of live bivalve molluscs.1 Chapter II . three dilutions MPN test specified in ISO 16649-3. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. that microbiological criteria and chemical contaminants amongst other requirements are met. - - - - B. in 90% of the samples. Classification of production and relaying areas: This section reinforces that specified in Annex III. In the remaining 10 % of samples.Official controls concerning live bivalve molluscs from classified production areas A.
2073/2005. 9. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. 9. boundaries or class of a production area or its closure. 214 . 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.6 Regulation (EC) No. establishes microbiological criteria for a range of foods.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. which has applied since 1 January 2006. Decisions after monitoring There are specific requirements whereby if sampling results indicate that health standards for molluscs are exceeded or there is a risk to human health. the Member States may permit the import of bivalve molluscs and fishery products from the countries listed respectively in Annex I and Annex II to regulation (EC) 2076/2005.FISH AND SEAFOOD C. E. 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. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. F. opening or closing a production area. Procedures for re-opening a closed production area are stated D.2 Chapter III .5.5. 9. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. the competent authority is to close the production area.2.
ensuring that foods being placed on the market do not pose a risk to human health.6.I. a batch of foodstuffs or a process. per unit(s) of mass. presence or number of microorganisms. with chapter 2.1 Food safety criteria Chapter 1 of the regulation focuses on food safety criteria that cover foods such as ready to eat foods and fishery products and live bivalve molluscs. The relevant criteria are outlined in Table 9. Two kinds of criteria have been established: food safety criteria. based on the absence. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. area or batch.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria.4 referring to fishery products. 215 . and the legislation applies to all businesses involved in food production and handling.LEGISLATION The aim of this legislation is to complement food hygiene requirements. and/or on the quantity of their toxins or metabolites.II. 9.6. 9. volume. The criteria are outlined in Table 9.
who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1.TABLE 9. monocytogenes. other than those intended for infants and for special medical purposes(4) (8) L.2 Ready-to-eat foods able to support the growth of L. monocytogenes L. monocytogenes.I Food safety criteria Sampling plan(1) n 10 5 Absence in 25 g EN/ISO 11290-1 c m M Products placed on the market during their shelf life Limit(2) Analytical reference method(3) Stage where the criterion applies Food category Microorganisms 1.3 Ready-to-eat foods unable to support the growth of L. monocytogenes 1. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life .1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L. 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.
25 Live bivalve molluscs and live echinoderms.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1. manufactured from fish species associated with a high amount of histidine (17) Histamine .27 Fishery products which have undergone enzyme maturation treatment in brine. 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. tunicates and gastropods 9 (18) 2 E.Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.17 Live bivalve molluscs and live echinoderms. coli (15) 1.
c = number of sample units giving values between m and M For point 1. 1996. 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. to the satisfaction of the competent authority. spirits and similar products. 79. beer. products with a shelf-life of less than five days shall be automatically considered to belong to this category. Engraulidae. A pooled sample comprising a minimum of 10 individual animals. Other categories of products can also belong to this category.1. m = M. that the product will not exceed the limit of 100 cfu/g throughout the shelf life.. Coryfenidae. — fresh. 5 This criterion shall apply if the manufacturer is able to demonstrate. excluding sprouted seeds. Clupeidae. Single samples may be taken at retail level. 43-9. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . subject to scientific justification. This criterion shall apply to products before they have left the immediate control of the producing food business operator. Malle P. Duflos G. Pomatomidae. 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. wine. monocytogenes. when recontamination is not possible after this treatment (for example. J.92. Dervin C. 2.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample.. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002.. biscuits and similar products.0 and aw ≤ 0. when he is not able to demonstrate. products heat treated in their final package). live bivalve molluscs. shall not apply. soft drinks.1 . The most recent edition of the standard shall be used. products with pH ≤ 5. sugar. Bouquelet S.4 or aw ≤ 0.94. Assay of biogenic amines involved in fish decomposition.. Valle M. Malle P. coli is used here as an indicator of faecal contamination. including cocoa and chocolate products. Products with pH ≤ 4. — — bottled or packed waters. Scombresosidae. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. according to which the whole batch is to be deemed unsafe. that the product will not exceed the limit 100 cfu/g throughout the shelf life. uncut and unprocessed vegetables and fruits. Bouquelet S. E. References: 1. — bread.. AOAC Internat. cider.25. to the satisfaction of the competent authority. Particularly fish species of the families: Scombridae. honey and confectionery.
monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory. The test results demonstrate the microbiological quality of the batch tested(1). L. — unsatisfactory. 82. AOAC Internat. (1) 219 . if the presence of the bacterium is detected in any of the sample units.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. monocytogenes in ready-to-eat foods able to support the growth of 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. where the limit refers to a pooled sample. if the following requirements are fulfilled: 1. if the presence of the bacterium is detected in any of the sample units. if all the values observed indicate the absence of the bacterium. — unsatisfactory. monocytogenes in other ready-to-eat foods: — satisfactory. Histamine in fishery products from fish species associated with a high amount of histidine: — satisfactory. L. — unsatisfactory. if the presence of the bacterium is detected in any of the sample units. no values observed exceed the limit of M.I The limits given refer to each sample unit tested. — unsatisfactory. if all the values observed indicate the absence of the bacterium. monocytogenes before the food has left the immediate control of the producing food business operator when he is not able to demonstrate that the product will not exceed the limit of 100 cfu/g throughout the shelf life: — satisfactory. if all the values observed are ≤ the limit. the mean value observed is ≤ m 2. L. tunicates and gastropods in relation to testing E. Interpretation of the test results relating to Table 9. 1097-101. if all the values observed indicate the absence of the bacterium. 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. coli. J. if any of the values are > the limit. a maximum of c/n values observed are between m and M 3. Salmonella in different food categories: — satisfactory. excluding live bivalve molluscs and live echinoderms. — unsatisfactory. 1999.
TABLE 9.4.II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2. coli 5 shucked products of cooked crustaceans and molluscan shellfish 2 100 cfu/g 1000 cfu/g EN/ISO 6888-1 or 2 FISH AND SEAFOOD Coagulasepositive staphylococci 5 Improvements in production hygiene 220 .1 Shelled and E.
7. — acceptable. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. and the rest of the values observed are ≤ m. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health.1 In the U. The regulations provide for defences in relation to upward variations of the 8 °C temperature. 2 Interpretation of the test results relating to Table 9.II The limits given refer to each sample unit tested. which overrides the 8 °C requirement.K. 2006 No. Schedule 4 of the Food Hygiene (England) Regulations 2006. 14 (Hygiene requirements specific to the U. 14 details temperature control requirements for foods in general. c = number of sample units giving values between m and M The most recent edition of the standard shall be used. For example. if one or more of the values observed are > M or more than c/n values are between m and M. Hot-held foods (food having been cooked or reheated and is for service or on display for sale) must not be kept below 63 °C. 2006 No.K) Temperature control requirements 9. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. if one or more of the values observed are > M or more than c/n values are between m and M. the defendant will need to prove that the food was on service or display. The regulations prescribe a chilled food holding temperature of 8 °C or less. S.LEGISLATION 1 n = number of units comprising the sample. if all the values observed are ≤ m. if all the values observed are ≤ m. and the rest of the values observed are ≤ m. The defendant may be required to produce well-founded scientific proof to support his claims. with chill holding tolerance periods. — unsatisfactory. S. had not been 221 . if a maximum of c/n values are between m and M. if a maximum of c/n values are between m and M.I. — acceptable. E. tolerance periods for chill-holding of foods and hot-holding variations. 9.7 Food Hygiene (England) Regulations 2006. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. The test results demonstrate the microbiological quality of the process tested.I. — unsatisfactory.
For other food.S. 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. such as that below. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S.I. for a limited period consistent with food safety.8 Guidance In the U.K. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions. S. 2006 No. 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.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment. 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.I. 3). and was kept at above 8 °C or the recommended temperature. the Food Standards Agency has published guidance notes on the requirements of the EU hygiene and microbiological criteria regulations which can be accessed at the following link: http://www. 9. 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.gov. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. 2006 No. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. Alternatively.food. or. 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.
30. L226. 3) as amended by Regulation (EC) No.4. The revised text of Regulation (EC) No. 2074/2005 and Regulation (EC) No. Regulation (EC) No. Regulation (EC) No. 83) as amended by Regulation Nos 882/2004. The revised text of Regulation (EC) No.6. Regulation (EC) No. 30. 1663/2006. 1688/2005. 2076/2005. 25. 22) as amended by Regulation (EC) Nos. as read with the corrigenda at OJ No. 2075/2005 and Regulation (EC) No.10. 9. 2076/2005.2004. Regulation (EC) No. 206). L226. 1). 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 1019/2008 and as read with Regulation 2073/2005. Commission Regulation (EC) No. Regulation(EC) No. these are not be discussed here. 22. 25. 223 . 2076/2005. 854/2004 is now set out in a Corrigendum (OJ No. 1. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41.2004. Regulation (EEC) 2136/89.6. 62) as amended by Regulation (EC) No. 2074/2005. 2074/2005. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. L139. 14. L226.4. 55). 2073/2005 on microbiological criteria for foodstuffs (OJ No. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito.9 Other Relevant Legislation The following regulations about fish designations need to be considered: • Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products Regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products • Note: Requirements for preserved fish are laid down elsewhere. 2076/2005.2004.2004.2005. 30. 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.10 Further Reading Regulation (EC) No. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No.12. laying down common marketing standards for preserved sardines. L139.2006. 1662/2006. 852/2004 is now set out in a Corrigendum (OJ No. 853/2004 is now set out in a Corrigendum (OJ No. 1441/2007. L139.LEGISLATION 9.6. namely. L283. Regulation (EC) No.2004. as amended. The revised text of Regulation (EC) No. 2074/2005. L338.4.2004. 25.
22.10. 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. 23.FISH AND SEAFOOD Commission Regulation (EC) No 2074/2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC)No 882/2004 of the European Parliament and of the Council. 21.2001. (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004 ( OJ L 338. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. 22) 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 278.1. 6) 224 . 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 22.214.171.1245.2005.2000. 22. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.
1.3.5 µm 10. and 2. Morphology Gram-negative rods.3 Temperature Aeromonas grows at temperatures of 2 .5 pH Under otherwise optimal conditions.2 Oxygen requirements Facultative anaerobe.1. At pH 4. a 10 to 1.5.0 x 1.0 .1. 0. Some death will occur at this pH if the temperature is 4 °C. 10.1. 10.5.1.3 minutes. The minimum pH value for growth is thought to be pH 5. At 5 °C. no growth is observed (1). Aeromonas has the potential to grow over a pH range of ca 4 . under optimal incubation temperatures.000-fold increase in number is observed after one week in food systems (1).1 Aeromonas spp.3 .5 minutes.10 (3).45 °C (1). The optimum growth temperature is 28 °C (2).10. 225 . 10.1 10.1. PATHOGEN PROFILES 10. and D-values at 45 and 51 °C have been reported as 29. respectively (1).4 Heat resistance Aeromonads are sensitive to moderately high temperatures.
5 . botulinum grow reasonably well in the temperature range of 20 45 ºC. 10. A to G. 10. being capable of slow growth and toxin production at low temperatures . botulinum is prevented. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. which may be inactivated by heating at 80 ºC for 20 . or at 90 ºC for a few seconds. F are the types that are responsible for most cases of human botulism (5. All type A strains are proteolytic and type E strains are usually non-proteolytic.1 Morphology Gram-positive.4 Heat resistance The vegetative cells of C.2. forming at least seven different toxins. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C.30 minutes.1. 10.2 Clostridium botulinum Seven different types of C.2. but Group II strains are psychrotrophic. to a lesser extent. Types A.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . B. types B and F can be either. 6). and groups I and II are those responsible for the botulism cases. E and.5% NaCl (4). botulinum are known. 10. Group I will not grow at temperatures of 10 ºC or less.2. 226 .0 µm.2. 10.4 x 1.2% sodium chloride (NaCl) (1). at 85 ºC for 5 minutes.even as low as 3 ºC (7). spore-forming rods.2 Oxygen requirements Although C. botulinum types produce heat labile toxins. botulinum is an obligate anaerobe. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different.7 .3 Temperature All strains of C.2. botulinum are not particularly heat resistant. but the spores of this organism are more so. All C.22.FISH AND SEAFOOD 10. but the interior of the food may do so. and is sensitive to > 4. Thus an aerobically packed product may not support the growth of the organism on the surface. 0. There are four main groupings of the organism.
vacuum packed. particularly by cooking.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4.97 for Group II (5).6 and pH 5.2 Oxygen requirements C.3. The organism will grow well in the centre of meat or poultry dishes.2. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I.0 µm. parcooked meals with pH value > 5. 0.2. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0.like other clostridia .85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. 10. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. In commercial situations.97) (10. 10. pH and temperature.10.9 x 2. safety margins must be introduced to allow for process variability. where oxygen levels are reduced. therefore. 227 . perfringens . botulinum depends on solute.0 . Note also that if humectants other than salt are used.21 minutes) (9).0. grow on the surface of foods unless they are vacuum or gas-packed.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C.0 and aw > 0. they may survive mild heat treatments (70 .94 for Group I and 0.PATHOGEN PROFILES 10. 11)). These concentrations correspond to limiting aw of 0.1 Clostridium perfringens Morphology Gram-positive spore-forming rods. botulinum spores The most heat resistant spores of Group I C.g. 9).7 Characteristics of C. 10.3.3 . It will not. respectively (8.2. the minimum aw for growth may be lower.3 10.1. These values have been established under carefully controlled laboratory conditions.is an obligate anaerobe. 10. However.
13). Salt concentrations of 6 .6 Aw/Sodium chloride C. and their heat resistance is also affected by the heating substrate. pH. perfringens has the potential ability to grow within the temperature range 15 . the limits for aw are affected by temperature. perfringens.it is destroyed by heating at 60 ºC for 10 minutes (14. Spores.5 pH C.8% inhibit growth of most C. While some growth can occur at 50 ºC.5 minutes (17). 13). perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. the enterotoxin is not heat resistant . 10.10 ºC.FISH AND SEAFOOD 10.3.3. At cold temperatures.7 Characteristics of C.3. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. 15. In addition. type of solute. perfringens in vacuum packed beef (18). perfringens is not a tolerant organism with respect to pH. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. It grows best at pH values between 6 and 7 (the same pH as most meats).3 Temperature The most significant characteristic of C. perfringens strains. Its optimum temperature for growth is 43 . Recorded heat 228 . perfringens is not tolerant of low water activities. The D-value for C. perfringens can vary quite considerably in their heat resistance. 10.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. perfringens in roast beef at 60 ºC is 14. 16).93 and 0. although C. Some studies indicate that the presence of 3% NaCl delays growth of C. 0 . (It is unusual for spores to be formed in foods after the growth of this organism (14)). The lowest aw recorded to support the growth of C. perfringens spores The spores of C.50 ºC. depending on strain and other conditions. As in the case of other factors limiting the growth/survival of this organism. vegetative cells die rapidly (12). Under otherwise ideal conditions. 10.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13.45 ºC.3. however. etc. will survive greater extremes of pH (and aw) (12. 18). 10. lower concentrations may be effective in combination with other factors.3.3. perfringens appears to be 0.
1 Listeria spp.5 pH The ability of Listeria to grow at different pH values (as with other bacteria) is markedly affected by the type of acid used.6 .4 Heat resistance L.2 Oxygen requirements Aerobe or microaerophilic. monocytogenes is not a particularly heat resistant organism. Morphology Gram-positive short rods. 10.4.98 and 0.8 minutes for heat sensitive spores (12). 229 . being potentially capable of growing . monocytogenes is reported to be 45 ºC (20). It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.64.albeit slowly .2 has been suggested as the lowest working limit (22). In foods. however. Under ideal conditions. and temperature. monocytogenes in crawfish tail meat at 55. growth at low temperatures can be very slow.3 is the lowest value where growth has been recorded. the organism is able to grow at pH values well below pH 5 (pH 4.0.23. respectively (21).0 µm.4 . However. -0.4.5 . The upper temperature limit for the growth of L. 60 and 65 ºC were reported as 10.4 10. the lowest limit for growth is likely to be considerably higher .4.especially at refrigeration temperatures.4 ºC is probably a more likely minimum for growth in foods (19). 10. or even below 0 ºC. 10. 1.0 minutes for heat resistant spores to 1.4. so can be destroyed by pasteurisation. 10.5 x 0. 10.3 . is between 30 and 37 ºC. using hydrochloric acid as an acidulant). 0.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. D-values for L. however. it is not a sporeformer.2. requiring days or weeks to reach maximum numbers.19 minutes. and where acetic acid is used as an acidulant. pH < 5.4.2.at refrigeration temperatures down to.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. Its optimum growth temperature.
monocytogenes is quite tolerant of high NaCl/low water activities. the optimum being 30 ºC (23).6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.45 ºC. 10.1 Plesiomonas Morphology Gram-negative.97. 10.8. 10.3 µm.5 . 10. 10.5. 10.1.83 (20).1 Salmonella spp.7 x 1. Morphology Gram-negative short rods.5.5. but has been shown to be able to grow at an aw level of 0.0 . 25). 0.0 x 1 .12% NaCl or more).90. 25).5 pH Plesiomonas has a pH range for growth of pH 4.0 µm.5.5.6 10.4. or even grow.5 (24. 10.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).3.2 Oxygen requirements Facultative anaerobe.8 .6 Aw/Sodium chloride L. at salt levels found in foods (10 .5 10. It is likely to survive.5 .0.FISH AND SEAFOOD 10. It grows best at aw of ≥ 0.5. The bacterium may survive for long periods at aw values as low as 0.6. 230 . 0. peritrichous flagella.3 Temperature Plesiomonas can grow in the temperature range of 8 .
10. and with an appropriate acid.5 ºC.5 pH Salmonella has a pH range for growth of pH 3. depending on serotype. the optimum being 35 .5 .6. although salt tolerance increases with increasing temperature (28).20 times more heat resistant than the average strain of Salmonella at high aw (27).93 (as reported in dried meat and dehydrated soup). 10.8 . depending on the type of acid and temperature.48 ºC. It is not. Some death will occur at pH values of less than about 4.6. 231 . Salmonellae are quite resistant to drying.7. Salmonella has the potential to grow at aw levels as low as 0. 10. substrate. and appropriate heat treatments must be determined experimentally for low-aw foods.37 ºC.4 Heat resistance Salmonella is not a spore-forming organism.5. The growth of Salmonella is generally inhibited by the presence of 3 . 10. therefore. D60 ºC values normally range from about 1 to 10 minutes.PATHOGEN PROFILES 10.6 Aw/Sodium chloride Where all other conditions are favourable. temperature and pH.6. However. or possibly 0. pasteurisation and equivalent treatments will destroy the organism under normal circumstances. a heat resistant organism.4% NaCl. Growth is slow at temperatures below about 10 ºC. The optimal pH for Salmonella growth is between 6. Salmonellae are quite resistant to freezing.3 Temperature Salmonellae can grow in the temperature range of 7 . Furthermore. some strains are able to grow at temperatures as low as 4 °C (26).9.5. with a z-value of 4 . high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments.0.6. However. Salmonella senftenberg 775W is about 10 .2 Oxygen requirements Facultative anaerobe. under otherwise ideal conditions.945.6. strains vary in their ability to withstand heating.
the D60 ºC value is 2 .3 (optimum 7. The following relate to limits for growth only. aw.FISH AND SEAFOOD 10. stressed cells can also be less tolerant of heating.37 ºC (29).7. 232 . aureus can grow within the temperature range 7 – 48. under otherwise ideal conditions (29.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. 10. 10. aureus is more limited under anaerobic than aerobic conditions. with an optimum of 35 . As is the case with other bacteria.7. 10.5 pH The pH at which a staphylococcal strain will grow is dependent upon the type of acid (acetic acid is more effective at destroying Staph. fat content.7. 10. 31). 0. Freezing and thawing have little effect on Staph.). however.7 .9 µm diameter. The limits for toxin production are also narrower than for growth.2 Oxygen requirements Facultative anaerobe.7. Staph. aw and temperature (sensitivity to acid increases with temperature).2 to 9. depending on aw.7.5). Most strains of staphylococci can grow within the pH range 4. It can survive well at low temperatures.1 Staphylococcus aureus Morphology Gram-positive cocci.7. aureus viability. aureus than citric acid).5 ºC. the organism is heat sensitive and will be destroyed by pasteurisation. but may cause some cell damage (30). etc.0 .7 10.3 Temperature Under otherwise ideal conditions.20 minutes. In meat macerate. NB: The growth of Staph.0. Under most circumstances.
0.86 is the generally recognised minimum in foods (32).2.4 .8. Staph. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities. but certain strains can grow in 20% NaCl.87 and > 0.83 .8 10.8 x 1. pH.0 (optimum 7. It can grow over the aw range 0.7.>0.9.99 aerobically under otherwise optimal conditions. 31) between 0.2 . In general.1 Vibrio cholerae Morphology Gram-negative curved rods.7. an aw of 0.10% NaCl. 0.43 ºC (35).0 . 233 . with a temperature range for growth of 10 . 34).7. and solute. strain. Staphylococci are more resistant to salt present in foods than other organisms.3 Temperature The optimum temperature for growth has been reported as 30 ºC.7 Limits permitting toxin production 10 . atmosphere.6 µm 10.99 little or no toxin production in anaerobically packed foods.2 Oxygen requirements Facultative anaerobe.5) (29. 10.PATHOGEN PROFILES 10. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant. 10. However. aureus can grow in 7 .8.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5.6 Aw/Sodium chloride Staph. 10. In general.8. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. An effect of increasing salt concentration is to raise the minimum pH of growth.5 . * dependent on temperature.
2 Oxygen requirements Facultative anaerobe. and it is sensitive to refrigeration temperatures (0 .65 minutes (35). 38). The D-value for V. respectively. cholerae may persist on frozen fish for >180 days (36).0% NaCl (35). 10. under otherwise favourable conditions.9.0.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.9. 10. but only moderately sensitive to freezing (40). It can grow in the presence of 0.9.1 .5 ºC). cholerae can grow over the aw range 0. 10. there is evidence that V.6 Aw/Sodium chloride V.1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. it has a pH range for growth of pH 5 .8. with optimum at 7.984 (35). V.6 (35.6.4 Heat resistance V.4. cholerae in crab meat homogenate at 60 ºC is 2. 10. with an optimum of 37 ºC (39).9.FISH AND SEAFOOD Also. parahaemolyticus is easily destroyed by drying.5 pH This organism is tolerant of high pH values. 10. but not acid conditions.97 . 234 .8. are 5 and 43 ºC.8. declining in number during storage.998. with an optimum of 0. cholerae is sensitive to heat at temperatures of >45 ºC. 37. 10.9 10.
The organism can survive for long periods at freezer temperatures (42).6 .8 .9. 41).5 minutes. parahaemolyticus is unable to grow in the absence of salt. The optimum concentration of NaCl equates to aw of approximately 0.98 (35. with a temperature range for growth of 8 .02 . and are readily destroyed by cooking.10. 10.8. 10.4 Heat resistance V.PATHOGEN PROFILES 10.6 Aw/Sodium chloride V.43 ºC (42). 235 .9. 41). 10. respectively.10. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43). depending on solute.9. respectively (39).0.922. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.2 Oxygen requirements Facultative anaerobe.10 Vibrio vulnificus 10.4% (39.3 Temperature The optimum temperature for growth has been reported at 37 ºC. 10. but the optimum concentration is 2 .1 Morphology Gram-negative curved rods. vulnificus is heat sensitive. and 2.6 (41). 10.10.11.4 Heat resistance V. V. The D-values at 55 ºC for clam and crab homogenate are 0.5 pH The pH range for growth is 4. parahaemolyticus will grow at aw levels as low as 0.5% NaCl. with an optimum pH of 7.10. 10. parahaemolyticus are not heat resistant.29 minutes.
in Clostridium botulinum: Ecology and Control in Foods. Aeromonas species. 9..N. Novak J. 6 (1). 2001.11 1..H. 279-303.H. Foegeding P. Great Britain. Eds.A. Hauschild A. Stelma G. in Foodborne Disease Handbook. Doyle M. Bhunia A. in Food Microbiology: Fundamentals and Frontiers. Principles of control. Notermans S.P.K. Clostridium botulinum. Eds. 383-408. Palumbo S.W. 357-67. 329-49. Caister Academic Press.H. Marcel Dekker Inc. Marcel Dekker.R.6 Aw/Sodium chloride V. 7.. Bier J. Peck M.5 pH The pH range for growth is 5 .H.. New York. New York. Marcel Dekker.R. 1-19. Dodds K.L.. Gaze J.M.. Dodds K.D.5. Dodds K. vulnificus grows over the range 0. in Foodborne Disease Handbook. 57-63.. Aeromonas. 1996.10. Miliotis M.P. Ed.. Ed.O.. 11..R. 1994.W. Juneja V.. Ed. 4. Volume 1. Clostridium botulinum and Clostridium perfringens. in Foodborne Bacterial Pathogens. with an optimum pH of 7.. Marcel Dekker.5 . Hauschild A. Microbiology and Molecular Biology.L. New York. Abeyta C. New York.M.W.N. Eds.10.C.E.8 (42).... 10.10. in Foodborne Pathogens. Washington D. 111-89. Marcel Dekker. Marcel Dekker. in Microorganisms in Foods.D. Doyle M.. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’. Eds. Betts G. Eds.J. with an optimum of 2.L.. 1994. 236 .D. Eds. 121-76. Stelma G.H. 6.L. 1989.FISH AND SEAFOOD 10. 10. Smith J. Beuchat L.0% NaCl. Murrell K. Potential hazards associated with REPFEDS. 97-131. 2. Gorham J. References International Commission on Microbiological Specifications for Foods.O... 1989. Marcel Dekker. Aeromonas hydrophila. 3.. 1993. 8. Hui Y... Doyle M.5% (42). in International Handbook of Foodborne Pathogens. Fratamico P. Kovacek K. Murrell K.W. Clostridium botulinum. Johnson E. Cliver D. Cliver D. Gorman J.D. in Foodborne Bacterial Pathogens.. Montville T. Blackie. Aeromonas hydrophila group. Hui Y. 5-19.H. Clostridium botulinum. Lund B. London. Faris A. 1-27. Eds. Volume 1: Diseases Caused by Bacteria.W. New York. 10. 2005.P. New York. Austin J. ASM Press. Diseases Caused by Bacteria. 1995. International Commission on Microbiological Specifications for Foods. 1993. Kim J. 2003. in Clostridium botulinum: Ecology and Control in Foods. Volume 5: Microbiological Specifications of Food Pathogens. New York. Hauschild A. Food Control. 5.M.
P. in Foodborne Disease Handbook. Johnson E.. 33. Doyle M.. 141-82. Halvorson D.A.G. 129-38. Marth E. Wolfe Publishing Ltd. Ed. Cliver D.. ASM Press. Doyle M. London. Matthews K. 1989... London. Banks J. 26. Ed.. Koburger J. 1996. London. in Foodborne Infection and Intoxication..O.P. Ed. New York. Clostridium perfringens food poisoning. 1989. International Commission on Microbiological Specifications for Foods. Ed. Plesiomonas shigelloides. New York. 21. 18.H.K. Volume 5: Microbiological Specifications of Food Pathogens.A. in Microorganisms in Foods. 137-64. 2007. in Foodborne Bacterial Pathogens. Riemann H.. 14.M. 24.C. Academic Press. Listeriosis and Food Safety. Marcel Dekker. 16. McClane B.An Introduction. 157-62.R.. International Commission on Microbiological Specifications for Foods. 68 (2). 1996.. 1989. London. in Foodborne Bacterial Pathogens. Volume 5: Microbiological Specifications of Food Pathogens. 982-6. Marcel Dekker. 2005. Washington D. Hui Y. 237 . Eds. ASM Press. in Foodborne Diseases.H. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs. Labbe R. Blackie. Clostridium perfringens.A. Varnam A. 336 (8721). 735-42.. 2001. Eds.R. Archer P. Ryser E. Labbe R.. Montville T. Walker S. Clostridium perfringens.. Plesiomonas. Murrell K. 1994. Gorham J.H. New York. Marcel Dekker. Blackie. London. in Foodborne Pathogens: An Illustrated Text. Wrigley D... Evans M.J.R. Juneja V. 229-40.G. Nagaraja K. 2006. 25.J. Diseases Caused by Bacteria. Marcel Dekker. 208-13.O.C. Avian Disease. 20. International Commission on Microbiological Specifications for Foods. 19. 351-82. Kim C. Listeria. Washington D. Emery D. Plesiomonas shigelloides. 1990. Eds.H. Varnam A. Eds. Lancet. 133-67. Ed.PATHOGEN PROFILES 12. Volume 5: Microbiological Specifications of Food Pathogens. 1991. Eds. 1990. 112-25. Beuchat L. Rinke H.M. Clostridium perfringens.. Cliver D. Journal of Applied Bacteriology.A. Clostridium perfringens gastroenteritis. International Commission on Microbiological Specifications for Foods. Ed. in Food Microbiology: Fundamentals and Frontiers. 1990. Cliver D. Elsevier. Volume 1.D.P. 201-8.O. Evans M. New York. 191-243. in Microorganisms in Foods.J.P. Blackie.A.T. 22. Listeria monocytogenes. Montville T. 13.G.. 15. Listeria monocytogenes. 311-25. Doyle M. Foodborne disease due to Bacillus and Clostridium species. Clostridium perfringens. London. in Microorganisms in Foods. Growth of Listeria monocytogenes at refrigeration temperatures. International Commission on Microbiological Specifications for Foods.V. International Commission on Microbiological Specifications for Foods.. 1996.J. in Food Microbiology . Lund B. 17. 23.
C. International Commission on Microbiological Specifications for Foods. Vibrio parahaemolyticus.M. Salmonellae. 1983.A. 39. 1989.M. Warner R. Springer Science. Griffiths M.. London. 36. 327-52.. Golden D.O. Blackie. Staphylococcal Food Poisoning. 31. 545-66. Woodhead Publishing Ltd.S. 32.62. 1990. 523. 1994. 38. 414-25. New York. London. Marcel Dekker. Tohyama Y. Volume 5. 642. 1996.. Oliver J. Gustafson J. International Commission on Microbiological Specifications for Foods. in Understanding Pathogen Behaviour Virulence. 1989. Staphylococcal gastroenteritis. Vibrio cholerae. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. London. New York. 34. in Foodborne Pathogens. Doyle M. Japanese Journal of Experimental Medicine. 543-68. New York. 238 . Lee-Wong A. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems.FISH AND SEAFOOD 27. 35.M.O. International Commission on Microbiological Specifications for Foods. 30.S. Cliver D. Blackie.J.A. Cleland D. 33.P. Crowcroft N. 8. Wymondham. 1930. London. D’Aoust J-Y.K. International Commission on Microbiological Specifications for Foods.. CDR Review. 28. Loessner M. Salmonella.P.R. Vibrio parahaemolyticus. Ed... Ed. 273-84. Eds. 985-98. Loessner M. 4 (13). Reed G. in Foodborne Bacterial Pathogens.M.D.C. 1993.J. Microbiological Specifications of Food Pathogens. Eds. 217-64. Stewart G. 1996. 2005. Golden D. Staphylococcus aureus. in Microorganisms in Foods.S. 426-35. in Modern Food Microbiology. 2005. Academic Press. International Commission on Microbiological Specifications for Foods. 327-445. in Foodborne Pathogens: Microbiology and Molecular Biology. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. Volume 5. in Microorganisms in Foods. 29. 45 (3). Staphylococcal intoxications. 2005... 37. Cholera: current epidemiology. Eds. Microbiological Specifications of Food Pathogens. Stress Response and Resistance. Riemann H. 331-57.. Academic Press. Cambridge. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). in Foodborne Infections and Intoxications. R15764. 1996. Volume 5: Microbiological Specifications of Food Pathogens. Marcel Dekker. Ed. Twedt R. Jay J. 85-106. Wilkinson B. International Commission on Microbiological Specifications for Foods. Ed. Bergdoll M. Jay J. Applied and Environmental Microbiology. London. 40. Caister Academic Press.H. Fratamico P.. Ed. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures.L. Smith J. Doyle M. in Microorganisms in Foods. Ed. 13 (11). Ed. Blackie.P. in Foodborne Disease. 2005. Dairy. Food and Environmental Sanitation. Cliver D. Bergdoll M. Bhunia A.A.
43. 55 (12). 42. Jay J. New York.PATHOGEN PROFILES 41. Blackie...M. Volume 5. in Microorganisms in Foods. Jay J. in Modern Food Microbiology.. and Campylobacter species.D. 657-78. Journal of Food Protection. Ed. Ruple A. Golden D. Loessner M. Golden D.A. Cook D.A... 239 .J. Loessner M. International Commission on Microbiological Specifications for Foods. 985-9. Foodborne gastroenteritis caused by Vibrio. London. Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters. Microbiological Specifications of Food Pathogens. International Commission on Microbiological Specifications for Foods. 2005. Yersinia.J. 436-9.M. 1992. Springer Science. 1996.W. Vibrio vulnificus.
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uk Web site: www.uk Web site: www.gov.uk/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: firstname.lastname@example.org/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: email@example.com Web site: www.org.seafish.uk Institute of Fisheries Management (IFM) 22 Rushworth Avenue West Bridgford Nottingham NG2 7LF United Kingdom Tel: + 44 (0)115 9822317 Fax: + 44 (0)115 9826150 Email: info@ifm.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: firstname.lastname@example.org 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: email@example.com/ .co.uk or v.gov.gsi.
dk Web site: www.fiskehandlerne.gov.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: firstname.lastname@example.org/ 242 .dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .dardni. Andersens Boulevard 37.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo.C.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: email@example.com/index/fisheriesfarming-and-food.uk Web site: www. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.no Web site: www.eapo. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne. 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: firstname.lastname@example.org@eircom.com Web site: www.fiskeriforskning.bim.dk Web site: www.org/fishery/ European Association of Fish Producers’ Organisations H.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: email@example.com Web site: www.dk Danmarks Fiskehandlere H.suite.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.fao.gov.org 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.ie/templates/homepage.FISH AND SEAFOOD Department of Agriculture and Rural Development.
csiro.cmar.fdf.com Web site: www.au Web site: www.ifr.org.uk/en/index.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: firstname.lastname@example.org Web site: www.org Web site: www.seafood.bsi-global.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: email@example.com Web site: www.ac.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: firstname.lastname@example.org Web site: www.com/ 243 .org.co.gsi.uk Institute of Food Research (IFR) Norwich Research Park Colney lane Norwich NR4 7UA United Kingdom Tel: + 44 (0) 160 3255000 Fax: + 44 (0) 160 3507723 Web site: www.uk Institute of Food Science and Technology (IFST) 5 Cambridge Court 210 Shepherds Bush Road London W6 7NL United Kingdom TeI: + 44 (0) 207 6036316 Fax: + 44 (0) 207 6029936 Email: email@example.com New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: firstname.lastname@example.org/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: email@example.com 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: firstname.lastname@example.org.
co.se Web site: www.euro.int/foodsafety World Health Organization (WHO) Headquarters CH .int/ SIK .org Web site: www.nhs.who.who.uk/index.euro.sofht.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: email@example.com Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: firstname.lastname@example.org 244 .scot.sik.uk Web site: www.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: email@example.com/web/indexen.int Web site: www.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: firstname.lastname@example.org/ 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: email@example.com Web site: www.uk/scieh.sfam.who.who. Web site: www.asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.The Swedish Institute for Food and Biotechnology Box 5401 SE-402 29 Göteborg Sweden Tel: + 46 31 3355600 Fax: + 46 31 833782 Email: firstname.lastname@example.org Web site: http://www.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. jsp Food Safety 10 .int.scot.nhs.org.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.
CONTACTS US FDA .au/ International Association for Food Protection http://www.org/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa.ift.eu/pol/food/index_en.uk/ 245 .gov/foodsafety/ Food Safety Authority of Ireland http://www.org Web site: http://www.ift.org/ The Association of Food.uk/ Society of Food Hygiene and Technology (UK) http://www.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.gov/ World Health Organization (WHO): Food safety programmes and projects http://www.ie/ Food Science Australia http://www.FoodSafety.fpa-food.who.riph. Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: email@example.com/ Royal Institute of Public Health (UK) http://www.foodprotection. Beverage and Consumer Products Companies http://www.cfsan.gov/ Institute of Food Technologists 525 W.fda.org.co.fsai.sofht.org/ Institute of Food Technologists http://www.csiro.htm Centre for Disease Control and Prevention (CDC) (US) http://www.cdc.
FISH AND SEAFOOD 246 .
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 . breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. causing spoilage of dried fish 110. in crustacean shellfish 86-7 ATP. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. 111 Astroviruses. in crustacean shellfish 87 Alteromonas spp. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. 102 Bacillus spp. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. 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. growth and survival in crustacean shellfish 86 Aeromonas sobria. preserved by dry/wet salting 99 Anisakis spp. to test microbiological criteria 19 Aeromonadaceae.INDEX Achromobacter spp. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. 69-70 Aeromonas hydrophila. source of PSP toxins 142 Alexandrium ostenfeldii. as autochthonous bacterial flora of fish 3 Aeromonas spp. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. planktonic source of spirolides in shellfish 163 Allergens.
description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. fish sauce 123 Basipetospora halophila.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. for PSP toxins 144 Chemical indicators. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. in crustacean shellfish 86-7 Campylobacter coli. as cause of spoilage of MAP fish 15 Calciviruses. caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. 68-9 Capture and handling. 131 Carbon dioxide. in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage. 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. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. 136. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni. 137. as autochthonous bacterial flora of fish 3 Clostridium botulinum. 98-9 Brochothrix thermosphacta. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang. associated with consumption of fish/fish products 16. 226 Breading process 36-8 Brevetoxins. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. preserved by marinating 100 Chemical detection methods. importance in lactic acid fermentation in fish products 130. of recipe fish meals 43 Cholera. of fish – effects on microflora 5-6 Carbohydrate. 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 . 113. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. of fresh fish spoilage 11 Chemical pollutants.
smoked and dried fish 114 Enzymic spoilage. hot-smoking 107 ELISA assay. 149-50. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). 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). smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . of fish – different processes 94 Dun. to determine fresh fish spoilage 11 Corynebacteria. for detection of DA 148 ELISA kit (DSP-Check). detection methods 148 implicated in amnesic shellfish poisoning 146 in crustacean shellfish 87 toxicology 147 Dried fish 93-121 Dry salting 97-8 Dry/wet salting 99 Drying. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. adverse condition on poorly preserved fish 110 EC grading scheme.INDEX pathogen profile 226-7 Clostridium perfringens. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. as contaminants in scampi 36. 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. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. 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. to determine fresh fish spoilage 11 Eels. growth and survival in crabmeat 84 growth and survival in cured. as autochthonous bacterial flora of fish 3 Crabs. 106 Contacts 241-5 Conway method. source of azaspiracids 159 source of DSP toxins 148-9. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. 152 Diphyllobothrium. processing 334 Cockles. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. of prepared fish products 44 Escherichia coli.
fermented fish product 126 Green cured split cod 97. fish sauce 123 HACCP. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. specific EU hygiene rules 196-224 Fresh fish. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. non-B) viruses. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes.FISH AND SEAFOOD EU legislation. as source of inulin in lactic acid fermentation in fish products 134 Garos. causing pink condition on fish surface 110 in fish sauce 127 Handling. 102 Gymnodimines. for evaluation of fish quality 9 Eurotium. 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. preserved by marinating 101 Gravlaks. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . mould causing spoilage of dried fish 110 Evaluation. as source of ciguatoxins 156 Garlic. member of cyclic imine group 161 Gymnodinium catenatum. of prepared fish products 44 Gambiertoxins. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. concerning crustacean shellfish 87-8 European Grading Scheme. 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. processing 35 Fish component. of fresh fish spoilage 911 Evisceration. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. to measure fish skin resistance 10 Flavobacterium. of fish – effects on microflora 5-6 Hepatitis (non-A. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. of fish at sea 5 Fatty fish. 130-5 Fish-spoilage bacteria 12 Fishtester. source of PSP toxins 142 Gyoshoyu.
concerning crustacean shellfis 88 Jellied eels. for fresh and frozen fish (Table) 20 Micrococcus spp. enabled by carbohydrate source in fish products 130. 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. smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. 100-1 Masmeen 107 Matjes. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. in fish fillet 1 Lipid oxidation. for microbial inactivation in fish products 127. preserved by marinating 101 Microbiological criteria. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. as autochthonous bacterial flora of fish 3 Lean fish. food hygiene 189-224 Lingulodinium polyedrum. preserved by salt-boiling 100 Ionising radiation. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 . as source of yessotoxins 152 Lipid content. to reduce rate of fish spoilage 12 Ikan kaju. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. definition 1 Legislation. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. outbreak caused by gravid 115 Lobster. 135 I-zushi. hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149. as source of botulism 136 International legislation. preserved by salt-boiling 100 Karenia brevis.
caused by lipid oxidation 11 Oysters. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . 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. shigelloides. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. as TTX producer 145 Norovirus. associated with consumption of fermented fish products 137 Oxidative rancidity. in crustacean shellfish 867 Norwalk virus.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. of crustacean shellfish – effects of processing 81-3 of cured. 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. smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P. reduction in smoking process 108 Norcardiopsis dassonvillei. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay.
spoilage organism in oysters 65 Protoceratium reticulatum. 161. preserved by salt-boiling 100 Pink. smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. causing spoilage in vacuum-packed dried fish 111 Pseudomonads. 145-6 definitions 28-30 non-seafood ingredients 32 Processing. in molluscan shellfish 60 Pseudomonas spp. planktonic source of azaspiracids 159 Pseudomonadaceae.INDEX Parasites. adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164. as source of toxins 159. 152-3 Penicillium spp. 163. reduction in smoking process 103. 69-70 Polycyclic aromatic hydrocarbons. as component for seafood ready meals 40 Pathogen profiles. in fish fillet 1 Proteus. 197 Plesiomonas. effects on microflora of fish 5-9 Professional bodies. mould causing spoilage of dried fish 110 Potato. detection methods 165 origins and distribution 165 toxicology 165 Pissala. smoked and dried fish 115 Pasta. 225-39 Pathogens. as cause of histamine fish poisoning 15 as spoilage organism in chilled products 43 as spoilage organism in fresh fish 14-5 Pickling 94. fermented fish product 124 Plankton. 81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 . as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. 100-1 Pindang. contact details 241-2 Prorocentrum spp. as source of DSP toxins 148-9 Protein content. as source of yessotoxins 152 Protoperidinium cassipes. as component for seafood ready meals 40 Prawns. causing spoilage of dried fish 110-1 Photobacterium phosphoreum. 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. growth and survival in chilled and frozen raw fish 19 growth and survival in crustacean shellfish 86 growth and survival in cured. 108 Polypaecilum pisce. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation. for detection of PSP toxins 144.
as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. assembly and packaging. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova. smoking and marinating of fish 96 Salt-boiling 94. source of PSP toxins 142 Quality Index Method. resulting from poorly salted cod 110 Pyrodinium. 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. 135 Rakefisk. for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops. flow diagram (Fig. processing 34. 36 Seafood. for evaluation of fish quality 9 Radiation. detected in cod 115 Pteriatoxins. 100 Salting 97-9 definitions 93-4 Sardines. preserved in saturated brine 99 Sauce. importance in lactic acid fermentation in fish products 130 importance in salting.) 41 ingredients preparation. antibody-based kit to detect STX 144 RT Freshmeter. chilled and frozen 1-25 Recipe dish. as source of tetrotoxin 144-5 Putty fish. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 growth and survival in cured. producers of brevetoxins 154 Raw fish. 83 254 . vacuum packing 105 Salmonella spp. processing and effects on microflora 60-1 Shewanella. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products. description and uses 58-9 Scampi. hazard in salting of fish 97 Salt concentration. fermented fish product 126 Raphidophytes.) 41 Recipe fish products 38-40 Rhodotorula rubra. for microbial inactivation in fish products 127. to measure fish skin resistance 10 Salmon. fermented fish product 126 as source of botulism 136 Rakørret. flow diagram (Fig. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. in molluscan shellfish 60 Rice. processing 35 Shellstock. as carbohydrate source in lactic acid fermentation in fish products 134 as component for seafood ready meals 40 RIDASCREEN.FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries.
of crustacean shellfish 80. contact details 241-2 Trematode infections foodborne 137 Trichosporon. in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. 13. processing and effects on microflora 63-4 Skin resistance. concerning crustacean shellfish 89 Vacuum packaging. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. role in fish degradation 10. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. fish and shellfish 141-74 Trade associations. 14 Torry Scale. for evaluation of fish quality 9 Torrymeter. 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. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru. smoked and dried fish 114 pathogen profile 232-3 pathogenic microorganisms in molluscan shellfish 59-60 potential for growth in pasta component of seafood ready meals 40 Stockfish. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO. as indicator of fresh fish spoilage 11 Toxins. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides.INDEX Shigella spp. in molluscan shellfish 60 Tuna. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. fermented fish product 126 Taste panel evaluation. as measure of fish degradation 10 Sliming. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. grading of 31 hot-smoking 107 US legislation. 101 Storage. of fish after capture 5 Surstrømming. fish sauce 123 Shrimps. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 .
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. of shellstock 61. processing for seafood recipe meal centres 39 Vibrio spp. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. 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. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. 62 Yersinia spp. 68 Vibrio cholerae non-01. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. 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 prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. 152-3 256 . growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables.
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