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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
Rhea Fernandes Leatherhead Food International v . Technical Team Leader for her valuable editorial contribution. Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged. My gratitude is extended to Dr Peter Wareing. Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product. published by Leatherhead Food International and RSC Publishing. for his scientific input and Victoria Emerton. I also acknowledge and thank Jackie Apps. guides to HACCP. Fish and Seafood. The books are intended as a source of information for microbiologists and food scientists working in the food industry and responsible for food safety. I dedicate this book to my husband . and a detailed list of contacts for various food authorities. which has been the chief factor in ensuring the completion of this book. thank you for all the support. Alison Turner. and growth characteristics for key pathogens associated with the product. and Meat Products. All handbooks also contain a review of the related legislation in Europe and UK. Training Manager. and Ann Pernet for the work put into typesetting and indexing. I thank the contributing authors for their collaboration.Goldwyn.FOREWORD The Microbiology Handbook series includes Dairy Products. both in the UK and elsewhere. potential hazards associated with the consumption of a range of products. The books in the series are designed as easy-to-use guides to the microorganisms found in foods.
2 Initial Microflora 1.6 Pathogens: Growth and Survival 1.7 Published Microbiological Criteria 1. 3. vii .1 Introduction 2.3 Processing and its Effects on the Microflora 4.1 Definitions 3.1 Definitions 1.1 Definitions 4.8 References 1.5 Spoilage 2.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2. 4.5 Pathogens: Growth and Survival 3.7 References 4.3 Initial Microflora 2.7 Published Microbiological Criteria 2.8 Further Reading CRUSTACEAN SHELLFISH 4.4 Spoilage 4.4 Processing and its Effects on the Microflora 2. CHILLED AND FROZEN RAW FISH 1.6 Published Microbiological Criteria 3.8 References MOLLUSCAN SHELLFISH 3.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.5 Pathogens: Growth and Survival 4.3 Processing and its Effects on the Microflora 3.6 Pathogens: Growth and Survival 2.7 References 3.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.4 Spoilage 1.2 Definitions 2.2 Initial Microflora 4.2 Initial Microflora 3.3 Processing and its Effects on the Microflora 1.5 Factors Affecting Fresh Fish Spoilage 1.4 Spoilage 3.6 Published Microbiological Criteria 4.
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 6.4 Amnesic Shellfish Poisoning (ASP) Toxins 7. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.1 Introduction 8.1 Definitions 5.3 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 212 9.2 Initial Microflora 6.6 References 6. 9.4 Regulation (EC) No.3 Stages of a HACCP Study 8. 2006 No.8 Ciguatoxins 7.8 Guidance 222 9. 7.10 Further Reading 223 viii .5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.1 Introduction 7.3 Processing and its Effects on the Microflora 6. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.3 Processing and its Effects on the Microflora 5.5 Regulation (EC) No.9 Azaspiracids 7. 8.2 Legislative Structure 190 9.4 Implementation and Review of the HACCP Plan 8.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.6 Regulation (EC) No.9 Other Relevant Legislation 223 9.7 Further Reading FISH AND SHELLFISH TOXINS 7.7 Food Hygiene (England) Regulations 2006.1 Definitions 6.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.10 Cyclic Imines 7. EU FOOD HYGIENE LEGISLATION 189 9.I.7 References 5.5 Pathogens: Growth and Survival 6.2 Definitions 8.11 Conclusion 7.5. S.8 Further Reading FERMENTED FISH 6.3 Tetrodotoxin 7. CURED.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.4 Spoilage 5.5 Pathogens: Growth and Survival 5. 852/2004 on the General Hygiene of Foodstuffs 191 9.6 Published Microbiological Criteria 5.2 Initial Microflora 5.6 Lipophilic Shellfish Toxins (LST) 7.1 Introduction 189 9. SMOKED AND DRIED FISH 5. 14 (Hygiene requirements specific to the UK) 221 9.4 Spoilage 6.
8 Vibrio cholerae 10.10 Vibrio vulnificus 10.5 Plesiomonas 10.4 Listeria spp.3 Clostridium perfringens 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .6 Salmonella spp.7 Staphylococcus aureus 10. 10.1 Aeromonas spp. 10. 10.9 Vibrio parahaemolyticus 10.2 Clostridium botulinum 10.10 PATHOGEN PROFILES 10.
. Saebo A. Catches can be gathered from seas. Geographical region.H. Seafood and Vegetables. Meat. 94767. Often contamination is from human and animal sources. fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins. the consumption of fish and seafood has increased. 2007. smoked and dried fish.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish. Nollet L. 2006. Eds. and.. fermented fish. Cross N. for fish.B.. cured. Safety and Processing of Wild and Farmed Fish.. Luten J. Further Reading General Fish/Seafood Microbiology Cross N. chilled and frozen prepared fish. Hui Y.. Jacobsen C. Seafood Research from Fish to Dish: Quality. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs. Clark S. 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.INTRODUCTION Fish and seafood are a main source of animal protein in the diet. The second edition of this handbook is a review of the entire book for currency of information.M. Wageningen.. rivers and lakes whose water can range from pristine to contaminated. thus. season. Seafood processing: basic sanitation practices. Bekaert K. in Handbook of Food Products Manufacturing: Health. Because of their health advantages over red meats. and fish and shellfish toxins. xi . molluscan shellfish.A. Poultry. Wageningen Academic Publishers.. The Microbiology Handbook. Hoboken. Wiley. crustacean shellfish. Chandan R. Oehlenschlager J. Milk..
Biogenic amines in fish and shellfish. Seafood: processing. Blackwell Publishing.International Commission on Microbiological Specifications for Foods. Heinitz M. Ashie I. 2002. Seafood processing: basic sanitation practices.P. in Seafood from Producer to Consumer: Integrated Approach to Quality. Journal of Food Protection. 245-66 Bremner H. 91-8. Eds.. New York. Hafsteinsson H. Gram L.A. Nip W-K. The fish industry. London. 1997... 8 (2). Dalgaard P. Noordwijkerhout. Food Control.L. Blech Z.B. Blackie. 2003. Gram L. Taylor S. Johnson J. Predictive microbiological modelling and seafood quality. Martin R. 200-23. Blech Z. Ohlsson T. Simpson B. 167-77. Oxford. in Microorganisms in Foods 6: Microbial Ecology of Food Commodities. Microbiology of fish and fish products. Ed. Cambridge. 60 (4). in Food Processing: Principles and Applications.L. 318-23. Noordwijkerhout. Cambridge. Bruinsma B. CRC Critical Reviews in Food Science and Nutrition..M. 459-72. 87121.H. in Fish Processing Technology. 1998. Elsevier. Amsterdam. Huss H. in Food Plant Sanitation.E.Y. Salmonella spp.J.D. 413-30. Gorham J. Vo1ume 39.B.. Gudmundsson M.. International Commission on Microbiological Specifications for Foods.H.K. in Minimal Processing Technologies in the Food Industry.. 2002. International Journal of Food Microbiology.L. 2005. basic sanitation practices.M.R. in Kosher Food Production.. Ed. Woodhead Publishing Ltd. Dalgaard P. Control of indigenous pathogenic bacteria in seafood. Smith J. 1996. 36 (1-2).. Stanfield P. Luten J. Fish and fish products (Microorganisms in fish products). 1997. McCarthy S. 329-65. Davies A.. Stanfield P. in Advances in Food and Nutrition Research. Huss H. Woodhead Publishing Ltd.Y. November 1995. Safety and Quality Issues in Fish Processing.. Marcel Dekker. Proceedings of the International Seafood Conference. Minimal processing in practice: seafood.. November 1995.R.N. 33 (1). 2004. 372-6. 1996.A. Ed. Luten J.A. Hui Y. Hall G. 121-37.H. Ventresca P. Amsterdam. Hui Y. London. Microbiological spoilage of fish and fish products. Huss H. Ed.S.. Ed. 1997.H. 431-43. Eds. New York. Academic Press. Rawles D.H. 2004. and Clostridium botulinum in smoked fish and shellfish. 543-61. Flick G. Eds. Oxford. Bengtsson N. Ed. Elsevier. 61 (3). Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. 1996. Blackwell Publishing. The incidence of Listeria spp. Smith J. Journal of Food Protection. Modified-atmosphere packaging of fish and fish products. Spoilage and shelf-life extension of fresh fish and shellfish. Tong P.. in Seafood from Producer to Consumer: Integrated Approach to Quality.. Kluwer Academic / Plenum Publishers... 174-249. 1997. Proceedings of the International Seafood Conference. xii .S. 1997..
R..D. 46 (1). 1992.. 305-17. Chapman and Hall. August 1991. International Journal of Food Microbiology. guidelines. McMeekin T.D. 2257. 1991. 26 (3). Sterling Publications Ltd. Liston J.. Eds. International Journal of Food Microbiology.M. 166-73. 1994. Liston J. in Quality Assurance in Seafood Processing: A Practical Guide. Evaluation of the bacteriological quality of seafood. A new look at microbiological standards.J. Amsterdam. Microbiological quality of fishery products.H. Dalgaard P. 25-39.A. Amsterdam. Qualitative and quantitative characterization of spoilage bacteria from packed fish. London. Ward D. Hobbs G. Poultry and Fish Products. Application of predictive microbiology to assure the quality and safety of fish and fish products. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Bacterial spoilage of seafood.R. Blackie. Eds. 319-33. Ed. Twigg B. The microbiology of seafood products. Ed. in Public Health Aspects of Seafood-borne Zoonotic Diseases. Liston J. Jakobsen M. Jakobsen M.. in Quality Attributes and Their Measurement in Meat. Martin A. Gram L. 1994. Amsterdam. 269-82. 1996. Ross T. Chapman & Hall. Garrett E. Elsevier. 11 (2). Ward D. Microbial hazards of seafood consumption. 1994. Eds. poultry and fish. 118-20..R. 1990. Pearson A. 1990. in Food and Beverage Technology International: USA. Ed. Fish: microbiological spoilage and safety.H.. Bonnell A. Microbiological indicators and seafood processing. Liston J.. 93-105.S. Modelling of microbial activity and prediction of shelf-life for packed fish. Blackie. Microbial growth and its control in meat. 90-106. 1994. 1995. International Commission on Microbiological Specifications for Foods. Fish spoilage and safety. Volume 5: Microbiological Specifications of Food Pathogens. London. 1989. Liston J.A. 54-61. 121-8. in Microorganisms in Foods. Seafood toxins of microbiological origin. 359-403. PHLS Microbiology Digest. Dalgaard P. xiii . Bacterial pathogens transmitted by seafood. Ed. 16 (1). 5 (3). Huss H. Food Technology. 1995. Bonnell A. Eds. Institut fur Veterinarmedizin. Copenhagen. 1994. Bernoth E-M.M. and specifications for seafood products. Activities Report of the R&D Associates. 265-79. 1992. Elsevier. London. Berlin. August 1991. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. 26 (3). International Journal of Food Microbiology. Gilbert R.. in Fisheries Processing: Biotechnological Applications.. 1992. Elsevier.N. Evaluation of the bacteriological quality of seafood.H.M. Copenhagen. Glasgow. 44 (12). 5662. Dutson T. August 1991. Ed. Gram L. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. 1-17. London.International Commission on Microbiological Specifications for Foods. Gibson D. Olley J. Huss H. Sofos J. 1992. Huss H. Copenhagen. 459-78. Food Science and Technology Today. Jakobsen M.
R. Lamprecht E.Pedrosa-Menabrito A. 1987. 11 (2). Ed. Volume 1.. 104-9. guidelines. Hobbs G. Dicharry A. 42 (3). Hobbs G.a review.spoilage of fish. Simmonds C. London. Microbiological hazards associated with fishery products. in Developments in Food Microbiology. 117-27. Part I . Microbiology of frozen fish and related products.K. xiv . Hodgkiss W. 1986. 8-16. Eyles M. 199-226.J..C. Food Technology. Chichester. Davies R. Ed. Shelf-life extension of fresh fish . 1988. Ed.K. 1985. Microbiology of fish. 169-208. London. 103. Norris J. 46 (1). Wiley. in Microbiology of Frozen Foods. 1982. Applied Science Publishers. Elsevier Applied Science Publishers. Regenstein J.. Food Technology. 71-117. The bacteriology of fish handling and processing. Journal of Food Quality.M. Garrett E.. 42 (3). Seafood-borne bacterial pathogens of marine origin. Microbiological standards.S. CSIRO Food Research Quarterly. 90-3. Robinson R.R. and specifications and inspection of seafood products. in Essays in Agricultural and Food Microbiology. 1988. 1988. Hackney C.
The lipid content. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. However.1. most fish contain omega 3-fatty acids and other essential nutrients. 1 . Muscle composition and structure of fish also differ from those found in other meat.2 . Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. which can be up to 67%. “fish” will be used for fresh and seawater finfish. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. lean fish are those fish that store most of their fat in the liver. Fish fillet composition can vary significantly within the same species due to feed intake. The connective tissue content of fish is also lower than that found in meat.20%. Protein content in fish fillet varies typically from 16 . while fatty fish have fat cells distributed along their bodies. and excludes molluscs and Crustacea. Fish fillets are a poor source of carbohydrates. and other forms of aquatic animal as part of their ‘fish’ definition.5% (1). fins and a streamline body. ‘fish’ also refers to the flesh of such animals used as food. respectively (1). CHILLED AND FROZEN RAW FISH Associate Prof. crustaceans. In this chapter.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. migratory patterns. Fish flesh is dominated by the abundance of white muscle in relatively short segments. it shows a typical season pattern especially in migratory species such as herring or mackerel. but typically present a low fat percent when compared to other meats.21%. and is mostly interspersed between the muscle fibres. Fish are an important part of a healthy diet since they contain high quality protein. In addition. giving it its characteristically flaky structure. Fish can be divided into fatty and lean fish. and spawning season. In addition. typically 3 and 15% of total weight. The lipid fraction is the component showing the greatest variation. offering less than 0. typically fluctuates between 0. This super class of vertebrates includes all the bony and cartilaginous finfish.
and maintained at temperatures just below the freezing point. transportation and sale. firmness. Superchilled fish is fish that has been cooled to. transportation and sale. Quality refers to palatability and organoleptic characteristics such as tenderness. during storage. Spoilage describes changes that render fish objectionable to consumers. spoilage microflora describes an association of microorganisms that. juiciness. odour and texture. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident.FISH AND SEAFOOD Chilled fish is fish that has been cooled to. and maintained at or below -2 °C (normally below -12 °C) during storage. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. Controlled-atmosphere packaging (CAP) refers to packaging in an atmosphere where the composition of the gases is continuously controlled during storage. and flavour based on the maturity. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. Evisceration is the removal of the viscera from a fish. Fresh fish is raw fish that has not been processed. and maintained at or below 7 °C. colour. Organoleptic refers to qualities such as appearance. frozen or preserved. and texture of the fish. but no further control is exercised during storage. through their development on fish. renders that fish objectionable to consumers. Frozen fish is fish that has been cooled to. at -2 to -4 °C. This technique is primarily used for bulk storage. DMA is dimethyl amine. but not below 3 °C during storage. colour. nitrogen (N2) and oxygen (O2). transportation and sale. The proportion of each component is fixed when the mixture is introduced. usually carbon dioxide (CO2). 2 . marbling. hence. Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases.
some species (most Vibrios) require sodium chloride for growth. However. healthy fish is considered sterile and should not present any bacteria or other microorganisms. It is difficult to estimate how many microorganisms are typically associated with fish. In fact.2 Initial Microflora The subsurface flesh of live.I). The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. Clostridium. Gram-positive organisms such as Bacillus. 3 . Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters. Micrococcus. It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish. It must be noted that mesophilic bacteria can rapidly overgrow psychrotropic organisms. as with other vertebrates. potential human pathogens may be present in both scenarios. although these are influenced by water quality and feed. molecular-based methods have not yet been used to address this issue. More accurate. Lactobacillus and coryneforms can also be found in varying proportions (1). Shewanella and Pseudomonas. On the contrary. 1. processing or storage (Table 1. Gastrointestinal tracts and gills typically yield high bacteria numbers. Human pathogenic bacteria can be part of the initial microflora of fish. Flavobacterium. and typical of the fish flora. the season and the method of harvest. since they heavily depend on the type of sample analysed and the protocol used for isolation. whenever possible. In general. are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest.) are also common aquatic bacteria. For example.CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed. several culture media containing sodium chloride and more than one incubation temperature should be used. Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp. and those that although not autochthonous to the aquatic environment. Moraxella. posing a concern for seafoodborne illnesses (2). microorganisms colonise the skin.II) (3). These pathogens can be divided into two groups: organisms naturally present on fish (Table 1. The number and diversity of microbes associated with fish depend on the geographical location. standard culture-dependent methods can only recover between 1 to 10% of total bacteria present in any given sample. the natural fish microflora tends to reflect the microbial communities of the surrounding waters. gills and the gastrointestinal tract of fish.
103 High (107 . and mud Ubiquitous.1 .106 cells/g (exception: V. Escherichia coli Yersinia enterocolitica 101 .26 °C 10 . Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp.19 µg/kg bodyweight from < 102 .II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.109 cells/g) * Some sources considered L. 4 . and risk associated with the consumption of seafood is low (2.102 101 . numbers of pathogenic bacteria in raw fish tend to be low. vulnificus) 5 .14 . In addition. However.37 °C Unknown Unknown TABLE 1. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. 105 . stream water. Plesiomonas shigelloides 3 . sewage.0. 4).35 °C 8 . during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria. birds.FISH AND SEAFOOD TABLE 1. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . 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.37 °C High for most species.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00.106 cells/g. Toxin levels 0.>106 Salmonella spp. estuarine environments.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.
in which the bacterial load 5 . offering better control of fish health prior to. human hands and clothing. boxes and/or baskets. mackerel or herring. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria. and spoilage. cortisone levels increase during prolonged stress and can alter the fillet quality. chilled brine or refrigerated seawater at -2 °C. with very little control over the condition of the fish at the time of death or the duration of the killing process.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. with fish such as tuna and cod being more commonly eviscerated than sardines. After capture.3 1. 1. The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea. hook and line. However.a practice that may have both advantages and disadvantages. and conditions of storage. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. in which the health of each animal can be assessed prior to slaughter. Physical damage such as loss of scales. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating. in recent decades. ropes. finfish will almost invariably come into contact with nets. In eviscerated fish. and their re-use will increase the crosscontamination of other fish with such microorganisms. handling and processing Wild finfish are usually caught by net. decks. has an effect on the amount of stress and physical damage that the fish will suffer during capture. however. icing or freezing. The length of time that set nets have been in the water or the time trawlers’ nets are towed. and the killing process is designed to minimise stress. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. During capture and storage. or traps. This contrasts greatly with the meat industry. and especially when fish is stored on board for longer periods. The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside. digestion and off-flavours in uneviscerated fish. Fish may be eviscerated prior to storage at sea . In addition. potential contaminants added during handling and processing. If evisceration is carried out at sea.3. with the production of off-odours. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms. aquaculture practices have been expanding worldwide. and during harvest. birds and soil. Increasingly.1 Processing and its Effects on the Microflora Capture. the cuttings provide areas of exposed flesh that are open to microbial attack. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating.
potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). and of the quality of the water used for ice and to wash fish. As with all foods. fish have a higher post mortem pH. 7). These containers are also used for displaying the fish during auction at the quayside. Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. for example. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. if there is some oxygen demand from the product. MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation.3.FISH AND SEAFOOD can be substantial.2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. 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. Physical barriers not only protect from physical damage. and minimising handling will all reduce microbial cross-contamination. Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. a reduction in the oxygen tension is achieved which. partial fermentation of sugars occur leading to lower pH. in time. However. In the absence of oxygen. use of effective detergents and disinfectants. and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO). By vacuum packing foods. but if applied properly should extend the life of the product.). Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. These changes can be deleterious. not part of the normal flora of the meat animals (8). from water activity (aw) through to physical damage. What is more. allowing more growth of spoilage organisms. they are usually process contaminants. Anaerobic atmospheres have less effect on fresh fish shelf life. Packaging changes the intrinsic and extrinsic parameters affecting a product. but isolate the food in an environment different from the bulk atmosphere. however. 1. and to limit the loss of quality (5). ferric ion (Fe3+)). In MAP. will result in fully anoxic conditions. The atmospheric conditions surrounding a product may be passively or actively altered. Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. cleaning of the work environment. nitrogen. often in the absence of adequate refrigeration. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. oxygen etc. by actively altering the composition of the 6 .
Modified-atmosphere preservation of fish was first reported in the 1930s. the pathogens that may be able to grow before spoilage occurs. and the problems of muscle structure damage by the modified-atmosphere (11). Micrococcus and Moraxella). In the United States. The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). including salt. The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. phosphates. may not be considered ‘fresh’ fish. although less information on their effectiveness is available. Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). gutted or filleted fish. VP is one of the oldest forms of altering the interior gaseous environment of a pack. but only in recent years has it seen a marked expansion in use and market share. The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. and increase in-pack drip (12). without overt signs of spoilage. a modified-atmosphere may contain any gas necessary for the desired effect. an inert and odourless gas. The products. which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains. High concentrations of carbon dioxide have the most pronounced microbial effect. botulinum. does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. which are able to grow rapidly at low temperature. Flavobacterium. but can dissolve into fish liquids and deform packages. the inhibition of common 'spoilage' bacteria (Pseudomonas. Gases used in MAP of fish most commonly include carbon dioxide and nitrogen. Many additives have been tried in conjunction with changed atmosphere. the higher pH and reduction potential (Eh) of the fish muscle. This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. and the promotion of a predominantly Gram-positive. the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . slower-growing flora (11). nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. after additive application. Replacement of oxygen with nitrogen. discolour pigmented fish (11).3 °C and produce toxins. as they are able to grow at temperatures as low as 3. Sulphur dioxide.CHILLED AND FROZEN RAW FISH surrounding gas. The microflora of meat is not the same as that of whole.
g. however. adequate shelf life and cooking requirements. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. 5. use of high-quality raw fish. The problem of water loss in MAP fish is common. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1. Although all fish species need to be treated separately. at all times. as already stated. greater advantages are seen. Only the highest-quality fish. may result in unacceptable colour and textural change and hence little shelf life extension (11). 2. Subsequent storage must be at or below 3 °C. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). 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). trimethylamine and sulfides). For other less fatty marine fish. adequate product labelling in respect of storage temperature. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. should be used. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. For fatty fish. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused.g. 3.3 °C at all points from packaging onwards. grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. owing to the dissolution of cellular structures. The replacement gas should contain at least 40% carbon dioxide. TMAO) that can be reduced to produce 8 . The growth of these bacteria is reduced by high carbon dioxide concentrations but. and the use of a HACCP plan. 4. this problem occurs to a lesser extent with meat. however. but is recognised as a drawback of carbon dioxide application. In the United Kingdom. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). The addition of high levels of carbon dioxide to these fish. from both a microbiological and a chemical standpoint. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. botulinum. 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. These conditions include storage of the product at ≤ 3. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13).
C (C=reject) Significantly. Over time. to brown and slimy with an intense off-odour. and metabolic activities of microorganisms present in the fish. as traditional spoilage patterns of the external organs are typically very distinct. so lower carbon dioxide concentrations may have better preservative effects.CHILLED AND FROZEN RAW FISH off-odours. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids).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. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 .60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). these are summarised in Table 1. and the flesh turns from bright and elastic to dull and soft (1). 17. or for that matter evaluation of the taste of the fish. Loss of freshness and spoilage cannot be separated as processes.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). 1. The eyes turn from convex and clear to concave and opaque.III.4. the skin turns from iridescent to dull and bleached with bacterial slime. A. TABLE 1. B. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. with no smell. Degradation of whole fresh fish stored in ice generally follows a set pattern. 18) and are probably the most widely used. the gills from pink and shiny. When working with fillets.4 1. smell and 9 . Various methods for whole fish freshness evaluation by a trained panel have been proposed. Taste panel evaluation of cooked fish may be used to score or grade colour. reactions caused by activities of the fish’s own enzymes. texture. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. and this pattern is the basis of freshness grading schemes.
The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical. After death. During the initial storage. As the fish degrades. the Eh of fish muscle remains relatively high (20). rigor mortis is the first noticeable change in the fish. %). the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). The pH of the muscle will drop after death depending upon the amount of residual glucose or glycogen that is reduced anaerobically to lactate with the coproduction of ATP. From being flaccid. 19). is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. the muscles relax again and the fish returns to a flaccid state. spoiled and putrid. TMAO permits a high Eh to remain in the muscle tissue.5 (18. chemical.g. a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. sea-weedy taste. ii) Neutral taste. biochemical and microbiological actions.FISH AND SEAFOOD taste of cooked fish. an intense odour compound. little flavour. post-rigor values are typically in the range of pH 5. all marine fish use/have TMAO (21). After some time. By measuring the concentrations of the six components. development of sickly sweet. rigor resolves. ammoniacal. 23). Generally. To a varying extent. or simply a waste product present due to bioaccumulation (22). cabbage-like. texture becomes soft and watery or hard and dry. possibly slightly metallic. this will generally correlate with the length and severity of rigor. HxR and Hx) gives a quantity (K-value.8 to 6. the conductance generally increases. the pH drop of the muscle is correspondingly less. 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. reduction in ATP and increase 10 . However. AMP. thus. iv) Enhancement of the spoilage characteristic of phase iii. an anti-freeze. or its remaining shelf life (1). iii) Traces of sour. which increases from 0% towards 100% with time. ADP. RT Freshmeter or Fishtester). as little endogenous reduction of TMAO occurs (18). the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. 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. sulphurous and/or rancid smells. bacterial reduction of TMAO to TMA. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. Because fish tend to have relatively little residual glycogen compared with mammals. an osmoregulator. The process of ATP breakdown is used as an indicator of fish freshness (24). After death. These processes are strongly influenced by the physical conditions of storage. fruity and/or bitter off-flavours. the Torrymeter. IMP.
therefore. This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. However. During spoilage. 7). not to describe freshness. does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection. and are especially important in the spoilage of frozen fish. 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. using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. and a segregated weak acid to absorb them. the importance of microbial action increases with time (5.CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). which may also be formed by bacteria (25). or a specific fraction of the volatile bases. 18). however. 31). however. Lipid oxidation and other oxidative changes lead to oxidative rancidity. as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. For some fish species. Quality indices based upon the products of microbial metabolism do not explain changes in quality until microbial growth produces measurable changes in the fish. Comparing results for different fish species. as microbial spoilage is limited by the low storage temperature (1. Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. a correlation can be made between the spoilage of the fresh fish and the production of TVB-N. The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. is later significantly reduced by bacterial action. Chemical degradation continues after the initial post mortem phase. Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. colour changes. Oxidative rancidity is known to reduce the quality of fatty fish in particular (19. 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). these measurements are usually used to quantify the amount of spoilage. hypoxanthine. 27) produced by aerobic respiration and other forms of metal ion reduction (28). Evaluation of Total Volatile Base-Nitrogen (TVB-N. 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. also termed TVN). 11 . Neither does the change in pH during storage correlate well with the production of TVB-N. the residual acid is then titrated. so the measurement is effective only during the initial loss of freshness. 26). for example the TMA fraction.
biochemical and microbiological reactions taking place within and on the fish.FISH AND SEAFOOD 1. The factors are interactive and cannot be completely isolated. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH.2 Fish-spoilage bacteria The parameters affecting the multiplication of microorganisms in foods have been categorised into two general groups: intrinsic (inherent qualities of the food) and extrinsic (qualities of the food environment) (35).5.1 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. 1. but also crucial for the assurance of consumer safety. yoghurt spoilage by yeasts and clostridial spoilage of cheese (5) are examples where it has been recognised for many years that a particular minority of the microbial flora present in the product is responsible for its spoilage. For example. careful temperature control during storage is not only important in terms of quality loss. For fresh fish. by altering the gas atmosphere (extrinsic). 34). 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. the results of altering one parameter may have far-reaching consequences on others. a minority are often associated with the majority of the spoilage. Fish spoil as a result of the chemical.5 1. For fresh fish iced immediately after capture and continually stored in ice. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent.and temperate-water fish spoilage rates may be many. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. the slower the spoilage processes proceed (within limits). Also. the lower the temperature of storage.5. 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. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 . The concept of specific spoilage organisms (SSOs) is not new. 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 reasons for the differences between tropical.
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. frozen) fish products. The importance of Sh. 35). Sh.1 Pseudomonas spp. 35.10% of the total flora of fresh fish from temperate marine waters. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. plus other characteristic compounds of the species responsible for spoilage.5. They are generally characterised as Gram-negative rods.3. catalase-positive. it is often isolated as about 1 . 13 . therefore. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. motile with polar flagella. ‘oniony’ and ‘faecal’ odours from the production of ketones. or faster than the rest of the flora of ice-stored fresh marine fish (9).e. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. Pseudomonas spp. 1. These organisms are described as potential spoilers. It is thus an important spoilage organism of gadoid fish such as cod. putrefaciens growth may be used as indicators of spoilage (30). 37). although only since the late 1960s has it been realised that specific metabolites of Sh. the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. 36). The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp. obligate respiratory bacteria. its ability to reduce TMAO to TMA. As previously noted. putrefaciens spoilage of fish is due to its biochemical action on muscle. produce hydrogen sulphide (H2S) from cysteine.5. i. It is also present in fresh water.2 Shewanella putrefaciens Sh. oxidase-positive. more especially.CHILLED AND FROZEN RAW FISH started in the 1940s.g. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s. which produced the characteristic spoilage compounds of the fish (30. It is able to grow as fast as. Members of the genus are able to produce pigments. and may play some role in the spoilage of freshwater fish. aldehydes. During the 1970s. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms.3. the spoilage of a product may be strongly influenced by the conditions under which the product is held. (9) and they are considered SSO of iced freshwater fish (35). processed (e. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. on fish are diverse and in many cases species-specific. mediated spoilage is characterised by ‘fruity’.
putrefaciens was not present in such numbers (23. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). whereas >108 cfu/g Sh. Shewanella denitrificans. they are heterogeneous. does not show carbon-source chemotaxis. with several new species being described. Shewanella oneidensis. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). but Sh. Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection.how to reduce a molecule that they are not able to take into their cells efficiently. a population of Ph. Ph. The prevalence of the bacterium in so many environments. and Shewanella profunda were described. the bacteria have a significant problem to overcome . Another distinct feature of the physiology of Sh. Shewanella livingstonensis.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). phosphoreum sufficient to cause spoilage was found.3. Shewanella waksmanii. as the ferric iron is insoluble. 1. It is also able to produce hydrogen sulphide and a range of other off-odour compounds. This wide-ranging respiratory capability is thought to be unique (39). mesophilic strains of Sh. additional psychrotrophic species such as Shewanalla baltica. Sh. owing to its cell size and activity. and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. putrefaciens were required. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). 41). was capable of being in a significant numerical minority on the MAP fish but still able to yield the majority of the TMA thought to be responsible for the rejection (23. Sh. it is strictly respiratory. Shewanella gelidimarina. putrefaciens growth has been collected (12. phosphoreum has also been identified as responsible for histamine fish poisoning. In 50% N2/50% CO2 MAP cod. Shewanella olleyana. at the time of rejection. Later. Ph. putrefaciens has been studied. This type of intoxication occurs when bacteria convert the histidine 14 . but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39). The ferric reductase activity of Sh. and recent taxonomic description using modern molecular methods bifurcated this species. 38). phosphoreum were required for 50% of taste panellists to reject a sample. In fact. like all members of the Pseudomonadaceae.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. Research has shown that approximately 107 cfu/g of Ph. Initially. Sh. putrefaciens lies in its chemotaxis: the bacterium. putrefaciens were identified as Shewanella algae. it was proposed that this organism.5. baltica has been identified as the main H2S producer in cod during cold storing (37). although. putrefaciens strains are characterised as microaerophiles or anaerobes. In terms of its taxonomy. 41). Shewanella frigidimarina. unlike many other motile bacteria. Shewanella affinis. and Shewanella aquimarina.
B. types B. a convenient but diverse species of bacterium with a number of different types. However. 6).e. The heterogeneity of the types (A. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. previous exposure and access to supportive treatment. algal toxins. where a particular 15 . a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. B. but with proper cooking. When inoculated into VP corned beef and sliced ham.3.4 Brochothrix thermosphacta and lactic acid bacteria B. Acetate production has been reported as a good indicator of spoilage by this organism. bacteria and higher parasites all present some risk associated with fish consumption (1.1 Clostridium botulinum C. Review of outbreak data suggests fresh fish to be safe. Adult disease is caused by the production of the botulinum toxin. 5). including antisera. E and F include bacteria with minimum growth temperatures of approximately 3. 5. E. Types A. Some Ph. 1. Recent studies have investigated the dominance of B. lightly preserved fish products are associated with botulism. thermosphacta did not produce off-flavours until 2 . B. C2. this is probably due to spoilage occurring before toxin elaboration. Of these.6. (and very rarely F and G) have been reported to cause human disease. they do not produce significant product spoilage). Of 404 intoxication outbreaks of type E C. age. E. D. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. Fewer still are capable of growing on fresh fish. F and G) is well documented (44). There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. these risks are substantially reduced (1. botulinum recorded up to 1963. Growing evidence suggests a role for B.3 °C (Group II) and these are non-proteolytic (i. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. presents a potential risk. especially type E botulinum intoxication.6 Pathogens: Growth and Survival Endogenous chemicals. 75% (with a 35% mortality rate) occurred in Japan. botulinum. 1. Huss (1. 1. human viruses.5. C1.CHILLED AND FROZEN RAW FISH present in fish into histamine. it is bacterial risks that increase after capture of the fish. 5) reports that fresh fish consumption has never been shown to cause human botulism. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42).3 days after having reached 108 cfu/g (42). thermosphacta in the spoilage of some MAP fish.
accounted for the majority of cases (1. Proper chilling.g. Japan. Other lightly or semi-preserved (e. parahaemolyticus cases have increased across the world. parahaemolyticus O3:K6 is responsible for many of the recent V. V. storage and preparation (11). including epidemics in India. Despite the ubiquitous nature of type E C. Currently. which can cause human gastroenteritis. A unique clone of V. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. parahaemolyticus outbreaks. there is no specific guideline that describes a minimum level of V. again. Only about 1% of marine isolates produce a thermostable haemolysin. and when in contact with contaminated water (no CDC reported cases from fish.6. V. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. sushi) (5). which is believed to be required for virulence. whether stored aerobically or in modified-atmospheres.3 °C) will eliminate all risk. This strain has been responsible for 50 to 80% of all V. and North America. 5). is small. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. The first report of the organism being a foodborne agent was with shirasu. In Japan. which was probably contaminated from an uncooked food source or an excreting foodhandler. vulnificus is associated with warm marine and estuarine waters. the risk posed to the consumer of fresh fish. parahaemolyticus is a marine organism. smoked. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. a Japanese boiled and semi-dried sardine dish. 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. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . the majority are associated with products not cooked immediately before consumption. 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.g. Russia. I-sushi.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. the majority of outbreaks are caused by consumption of raw fish products (e. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). 1. and proper cooking (boiling for 1 minute. Since 1996. USA. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. Proper temperature control (<3. France. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. The bacterium causes primary septicaemia. 11). were associated with finfish between 1978 and 1998 (46). 1978 . Southeast Asia.2005). due to the cultural preference for raw fish dishes.2 Vibrio parahaemolyticus and other vibrios V. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). salted or pickled) fish have also been associated with botulism. Generally.
5. are associated with human diarrhoeal illnesses.6. their potential to produce disease-causing exotoxin in food has not been fully examined (48). cholerae O1. Despite this.26 to 0. They are aquatic organisms with an epidemiology that is yet to be fully understood. listeriosis cases have been reportable to the CDC. others are psychrotrophic mesophiles (49). the causative agent of cholera. There is no risk associated with the consumption of properly chilled and cooked fresh fish. which is often fatal (>50%). and especially Aeromonas hydrophila.55 cases per 100. coli on isolation media (50). there is no risk from the consumption of properly handled and cooked fresh fish. V.6. monocytogenes incidence has been between 0. however.20 °C. Some isolates are true psychrophiles. 17). shigelloides are encountered. Chill storage may not eliminate growth. The major virulence factor appears to be the production of toxins with enterotoxic. L. are present on fish and can grow to significant numbers during storage (1.. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. and fish and shellfish are probably the major reservoirs for the organism (50). shigelloides is similar to Aeromonas spp.3 Aeromonas Aeromonas spp. cytotoxic. with minimum growth temperatures around 0 °C and optima of 15 .4 Listeria monocytogenes L. in its habitat. In the US. possibly due to under-reporting because of their likeness to E. sodium channel blocking and haemolytic activity (48). The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis.CHILLED AND FROZEN RAW FISH disorders). P. monocytogenes has been well documented as a foodborne human pathogen. proper chill storage will eliminate any risk associated with this organism. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. Since 2000. is historically associated with faecally contaminated water. infection. and especially estuarine environment. It is known that Aeromonas spp. Again. Similar problems as those encountered for Aeromonas spp. 1. 1. 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. but proper cooking should substantially reduce risk of infection.000 17 . in assessing the risk of fishborne infection by P. and proper cooking will substantially reduce risk. Most cases have been sporadic rather than associated to large outbreaks. but the bacterium is known to survive and grow in the shallow marine. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. Although the psychrophiles are unable to grow at body temperature.
Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). leads to the production of histamine. Like other toxins. Klebsiella pneumoniae and Hafnia alvei. 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. Lightly preserved products. 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. 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. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied.pregnant women. alcoholics. and Lactobacillus spp. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. it is not apparent to the consumer and it cannot be destroyed by cooking. full risk analysis is not possible (52). Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . Being a psychrotroph... Clostridium spp.98 minutes in cod and D60 of 4. 1. some Vibrio spp. D60 of 1.48 minutes in salmon have been reported (53). but more especially by Morganella morganii. 1.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. Photobacterium spp. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. If fish are not properly refrigerated. but proper cooking will reduce any risk. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. neonates. It is regularly isolated from seafoods (51). within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). 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.6.2002 (55)) and is also common worldwide.. at risk groups . Fortunately. The organism poses a serious risk in chilled products not cooked before consumption.FISH AND SEAFOOD persons. foetuses.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. there is a possibility for growth of the organism on chill-stored fish.6. but cooking will significantly reduce the risk of infection. Infections are mostly associated with the consumption of raw or 18 . especially pickled fish.
No.. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. especially in warm-water areas and where fish are to be consumed raw. coli Case 1 4 Plan 3 3 n 5 5 c 3 3 m 5 x 105 11 M 107 500 Additional tests to be carried out when appropriate Salmonella V. area or lot (CAC.7 Published Microbiological Criteria Microbiological criteria for food defines the acceptability of a product or a food lot based on the absence or presence or number of microorganisms. 1997) Huss H. aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration.IV Sampling plans and recommended microbiological limits for fresh and frozen fish (Adapted from ICMSF 1986) Limit per gram or cm2 Test Aerobic plate count E. including parasites and/or quantity of their toxins / metabolites per unit of mass. Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease.1997.. parahaemolyticus (see Table 1. volume. Faecal coliform counts may be used instead of E. Gram L. it may be desirable to test for Salmonella and V. 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. coli counts where this method is preferred. parahaemolyticus Staph. In summary.IV) (57). TABLE1. Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it. Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. long storage under refrigeration or one of the former prior to freezing.H. Rome.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. which in many cases will eliminate the risk. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. For fish from inshore or inland waters of doubtful microbiological quality. FAO. 1. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF). 2003. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). Ababouch L. EC. 444.
. 1992. Jehanno D. Assurance of seafood quality. International Journal of Food Science and Technology. 11.H. 243-60..R. Davies A.H.. Armstrong D.J. Reddy N. Rhodehamel E. 1996. 20 ..K. 6. Food and Agriculture Organisation of the United Nations. 5 (1). Journal of Food Safety. M. Ahmed F. 2002. Control of indigenous pathogenic bacterial in seafood. Critical Reviews in Food Science and Nutrition. 283-94. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). Food Control. Ashie I. 1992. Food Control. Dalgaard P.R.E.. Huss H.. Huss H.J. in FAO Fisheries Technical Paper No. in FAO Fisheries Technical Paper No.. 121-37.H. Food Control. 37. Simpson B. 67-71.. 5. Spoilage and shelf life extension of fresh fish and shell-fish. References Huss H. 91-8. Sivertsvik M. International Journal of Food Microbiology.A. parasites. Rome.J. 8 (2). 12. Microbes and Infection.N. International Journal of Food Microbiology. 3. 1994. 9. 13. 1989. Fletcher R.. Jeksrud W.A. Food and Agriculture Organisation. 2000.H. The role of seafood in bacterial foodborne diseases. 334. 5-7. 19.8 1.H. 12. Ed.. Codex Alimentarius Commission. Italy. 12. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth.P. 14. Huss H. 1651-60. Quality and quality changes in fresh fish... 1995. 10. Smith J. 1997. Nychas G. Microbiological spoilage of fish and fish products.K.. 291-304. Codex Alimentarius Commission.R. 27. Food and Agriculture Organisation. Ed.S. Incidence and growth of psychrotrophic Clostridium botulinum in foods.FISH AND SEAFOOD 1. Gibbs P. International Journal of Food Sciences and Techonology. International Journal of Food Microbiology. Davies A. Feldhusen F. and natural toxins. 33. 107-27. 4. 348. Gram L. 2001. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres. Food and Agriculture Organisation of the United Nations. Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review. Capell C. activities and safety. 2003. 1994. Gram L.. 1996. Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef.. Kirby R. 87121. Huss H.R. 1993. Davey K.A. Rosnes J.E. 36. 24. Incidence of foodborne pathogens in European fish. 7. 2.. Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms. 2. 8. Rome. Kautter D. 87-118.T.
21.J..edu/haccp/compendium/chapt08.E.. Reginstein J. in Advances in Seafood Biochemistry. 1982. Journal of Applied Bacteriology. Flick G.. Microbiological spoilage of fish and fish products. Food and Agricultural Organisation of the United Nations. Edinburgh. Flick G. 25. 30.J. AVI Publishing. 29.spoilage of fish. 22. Shelf life extension of fresh fish . Herbert R. 22-36.E. Chemical composition. Qualitative and quantitative characterisation of spoilage bacteria from packed fish. Saito T. 1995.E. 1.J. Non-enzymatic oxidation of seafood lipids.N. Gill T. http://seafood. Lancaster. Huis in't Veld J..H.G. 1959. Sea Fish Industry Authority. 1985. 1933. 1-18. Conway E. 21 16. 2007. Autolytic changes in fish post mortem. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage. 1971. Huss H. 1995.. 1992. Ed. 419-29. 19. Leblanc P.H. 1983. Børresen T. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout. Gram L. Quality and Quality Changes. Flick G. 17.. 20. Eds.. Food and Agricultural Organisation of the United Nations... 305-17. Eds. Journal of Agricultural and Food Chemistry. 11. Hebard C. in Fresh Fish. International Journal of Food Microbiology. Quality and Quality Changes.J.M. 42-54. Ratnayake W. 121-37. 26. Food Technology. 23. Letters in Applied Microbiology.CHILLED AND FROZEN RAW FISH 15. Gibson D.htm Anon.E.. 33.J. The micro-determination of ammonia. 37. 28. Pedrosa-Menabrito A. An absorption apparatus for the micro-determination of certain volatile substances. Huss H. Hendrie M. Martin R. 36. 41-50. Microbial and biochemical spoilage of foods: an overview.M... March 20. International Journal of Food Microbiology. National Advisory Committee on Microbiological Criteria for Foods. 33.M. Lipid oxidation in seafood. 1415.. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products.A.M. 27. 1992. 354-8. 245-67. 1996. 1995. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish. Hebard C. Byrne A. 27. Rome. 21.S..ucdavis. Schwall D. 18.R. 1996.... 117-27. Surette M.J. Ackman R. Khayat A. Westport. Martin R.E. Slade A. Journal of Food Quality. Yajima T. in Fresh Fish.. Biochemical Journal. 24.E. Shewan J. Bacteria active in the spoilage of certain seafoods. 1995. 1987. Nature. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere. 1988. 130-2. 26. Ed. 19-22. Rome. International Journal of Food Microbiology.H. Part I .J. Gill T. Davies A. 184. 34. National Advisory Committee on Microbiological Criteria for Foods. Arai K.H. Dalgaard P.A.C.a review.J. Changes in purine nucleotide of red lateral muscle of rainbow trout. Adopted. . Huss H. Technomic. in Chemistry and Biology of Marine Food Products. Washington D. Martin R.
Lee J..S.H. 67. and regulation.eu/LexUriServ/LexUriServ. Gram L. and an Achromobacter species. Venkateswaran K.R.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J. 1984. 1985. Fish spoilage bacteria – problems and solutions. Occurrence and importance of proteolytic bacteria as spoilage indicators. Applied and Environmental Microbiology. 40. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches. 174. International Journal of Food Microbiology. 1992. Rome. 4. Nychas G..R. 38. Satomi M.. Environmental Biotechnology. Archiv für Lebensmittelhygiene. Applied Microbiology. Blackwell Scientific Publications.L. Role of proteolytic bacteria in fish spoilage. 61-6. 62-70. 71. Dixon N. 36. Myers J. Sumner J. 1984. 37. 43. Quality and Quality Changes.FISH AND SEAFOOD 31. 26.. Dalgaard P. Journal of Applied Bacteriology. 317.M.. Kell D. Huss H. Cohen D. 323-9. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. The effect of storage temperature on shelf life. Food and Agricultural Organisation of the United Nations. FAO Fisheries Report. Processes in Pathology and Microbiology. Vogel B. 328-9. in Fresh Fish. Pseudomonas fluorescens. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish. Iron and manganese in anaerobic respiration: environmental significance..H. II. Magno-Orejana F.B. 1973.. Taussig M. 72-80.J.. 3429-38. 48.europa. Jørgensen B. Nealson K.. Drosinos E. Huss H. Growth and activity of Shewanella putrefaciens isolated from spoiling fish. 1994. Scanlan R. Saffarini D. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere. Brochothrix thermosphacta. Gorczyca E. Ed. Miller A.. 109-36. Italian Journal of Food Science.M. 33.A. 42.. 18-21. 22 .E. Libbey L. 39.. Oxford. Annual Review of Microbiology. 311-43.. 2002. Karnop G. Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens. 1982.H. Dalgaard P.M. 1995. Myers C. 35.J. 44.H.F. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens. 13.. 9. 1996. 6689-97. 33. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia. 41. 1989.. physiology. Brady P. Rome. 34. 36. 2005. Gram L. The inhibition by CO2 of the growth and metabolism of microorganisms. 32. 262-6. 334. Journal of Bacteriology.. 51-62. 1989.
54. Panicker G. 23 . Braden C. Daniels N. Genigeorgis C. 1993. Embarek B. International Journal of Food Microbiology.. Bishop R. 1997. 172-249. 1987. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures. Griffin P. Smith J. 1986. Ljungh A. 58. Food Technology. 47.. 55 (SS10).. 2005. 1-37.. Hammond R. Surveillance for Foodborne-Disease Outbreaks . E and F in salmon fillets stored under modified atmospheres at low and abused temperatures. Kluwer Academic/Plenum Publishers. Aeromonas and Plesiomonas as food. The International Commission on Microbiological Specifications for Foods..H. 1-34. Golden D..... Wilson S. Modern Food Microbiology. 1991. Huss H. 1661-6. Woodruff R. Embarek B. Myers M. The International Commission on Microbiological Specifications for Foods. Whiting R. The International Commission on Microbiological Specifications for Foods. 57. Lynch M. Ray B. 2006.N. 56. Fish and Fish Products (Microorganisms in Fish Products).CHILLED AND FROZEN RAW FISH 45. Altekruse S. 50. 2003. International Journal of Food Microbiology. in Microorganisms in Foods 6. 181-8. 2000. Morbidity and Mortality Weekly Report..W.. Garcia G. 2194200.M. 2005. Majeed K.. 1998-2002. 20.K. Bean N. Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B. 303-12. 1973-1998. Sampling for Microbiological Analysis: Principles and Specific Application. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico. 23. 51.P. Microbial Ecology of Food Commodities. Jay J. Mac Rae I. 330-6. 48. Bej A.A. Lindroth S.K.C. 49... Ed.M. Springer. Miller A..M. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets. Journal of Food Protection. Painter J. Use of risk assessment to reduce Listeriosis incidence. Thompson S. USA. Wadström T.K.. 53... New York.. 1991..and waterborne pathogens. 1994. 51 (4). Journal of Infectious Diseases.P. 100-3. Lehane L. 17-34. Presence.H.L.. International Journal of Food Microbiology. Slutsker L. Blackwell Scientific Publications.C.J. 55.L. 2000. 12. Oxford. Histamine fish poisoning revisited.. International Journal of Food Microbiology. 69.United States.. 181.. Olley J. detection and growth of Listeria monocytogenes in seafoods: a review. 85-95. 12. MacKinnon L. International Journal of Food Microbiology.J.A. 181-96. Microorganisms in Foods 2. 50. Applied and Environmental Microbiology. 52. Vibrio parahaemolyticus Infections in the United States. Loessner M. 46..
H. Sumner J.M.D.. Smith G. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. The Microbiology of Seawater Fish.M.. 13-32.D. Eds.. Hobbs G. Gibson D. Cann D. 1. 65-72. Bulletin of the Faculty of Fisheries (Hokkaido University)..1983. Ravn Jørgensen B... Fisheries and Food. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches.. Technomic Publishers. Huss H.L. International Journal of Food Microbiology. Ed. Magno-Orejana F. Gibson D..FISH AND SEAFOOD 1. 1989. in FAO Fisheries Report No. in Torry Advisory Note No 93.P.. Aberdeen. Shahidi F. International Journal of Food Microbiology. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Lancaster. Torry Research Station of the Ministry of Agriculture..1 Further Reading Processing Anon. Handling and processing of scad. 271-85.. Ed. 1986..M. 127-34. Rome. Simard R.G. Food and Agriculture Organisation of the United Nations. 1984. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. 15. Estimation of the bacteriological quality of fish by automated conductance measurements. 35. 1992. Cho Y.H. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage.9..A. 1992. Kitts D. Shinano H. 1984.. Jones Y. 24 . Villemure G. Journal of Food Protection. McMeekin T. Dalgaard P. 1988. Journal of Food Science. Gram L.). Taoukis P.C. International Journal of Food Microbiology. Borgstrom G.9 1. Labuza T. Florida. 1961. Olley J. Fisheries and Food. 741-50. Ogden I. 51. 317-21.... Food and Agriculture Organisation. Houston N.9. Changes in microflora and chemical compounds in mackerel stored by partial freezing. 1997. 1.. Academic Press. 73-89. 1.. Trolle G. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. 1989. 1985. in Fish as Food. 317 (Suppl. Huss H. Akiba M. Application of predictive microbiology to assure the quality and safety of fish and fish products. Picard G. International Journal of Food Microbiology. Huss H. in Seafood Science and Technology. 6. Torry Research Station of the Ministry of Agriculture.H.. 295-307.S. Microbiological quality and shelf life prediction of chilled fish. Torry Research Station. Aberdeen. 4.2 Spoilage Shewan J.E. 55. Ross T. 62-70. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere.
.H.europa.. London. 1.R. Trolle G.H. 59. 1997. Liston J. Blackie Academic & Professional.J. 1. 137-46. in Fish processing technology. 20023. Applied and Environmental Microbiology. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. Klausen N.. 5. Otero A.. Modified atmosphere packaging of fish and fish products. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish.. Huss H. Elsevier Science. 1987. 322-6. in Quality Assurance in the Fish Industry. Garcia M. International Journal of Food Microbiology..L. 65-72. Jacobsen M. 1992. 4. Hall G. Gram L. Ed.9. Huss H.eu/LexUriServ/LexUriServ. http://eur-lex.H. 25 . Effect of culture age.3 Pathogens Condon S. 1993. Amsterdam.K.CHILLED AND FROZEN RAW FISH Einarsson H.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. International Journal of Food Microbiology..9. 1987.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. Gram L. 72. Journal of Applied Bacteriology. Sala F. 479-88.M. 1992. Eds. 2197-203.. Huss H. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. A rapid method for detection of histamine producing bacteria.
although recent developments in improved supply chains. clarity of eyes. or could be disguised by sauces. due to the early cessation of spoilage. the exceptions being Sushimi (high quality raw fish).g. products of lesser quality. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr. 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. colour of gills.1 Introduction It was often thought that whilst the highest quality seafood products are processed and sold as whole fish and/ or fresh fillets (e. smoked (hot) mackerel. This is no longer the case with many processed products now available in stores. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2. Sushimi tuna). The same used to be said for the difference between fresh and frozen products.).g. smoked (cold) salmon. Even though heating prior to consumption will eliminate pathogenic microbes. smell etc. The majority of seafood products produced require a cooking stage before consumption. but still acceptable and meeting all safety criteria.2. 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. seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. Few seafood products are sold ‘ready-to-eat’. 27 . and crustacea such as shrimp and crabs. it does not necessarily apply to the toxins that may have been produced. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e. and on-board freezing mean that products frozen at the point of capture are now perceived by the consumer as being of a better quality than a fresh product. emphasising the high quality of the seafood and the gourmet nature of the prepared products. colourings etc.
from both wild capture and aquaculture sources. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish.. Doubts have been raised over the microbiological status and handling practices of reef-caught fish. the processing industry has developed a wide range of value added products. They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP).. Microbiological hazards include Salmonella. At the same time. Listeria spp. prawns. and are well known for their diet consisting of waste materials. shellfish and molluscs.FISH AND SEAFOOD Recent trends in the market for food and seafood products. and these are increasingly being presented with an ever-expanding range of accompaniments. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein. and this has produced a greater availability and diversity of products in the marketplace. Prepared natural fillets are fillets of fish without any additional coating. etc. This includes all categories of seafood including trout and other freshwater species. and the consumer only has to heat (cook) the product. where the preparation has already been carried out. Frozen fillets may have a surface layer of 28 . they are usually supplied filleted with head on. which can be farmed. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. often using the simplicity of a microwave oven.2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. have shown the increasing importance of convenience foods. Nevertheless. They are typically flown into export markets. including freshwater fish. Shelf life on ice. The expected shelf life attainable for this format is usually up to 8 days. and pathogenic Vibrio spp. in particular. but should be equivalent to cold-water products. molluscs. 2. Seafood is the term used descriptively to include all categories of product. as they tend to lose colour and sheen after defrosting. rather than frozen. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted. There is also a limited market for filleted material. or in MAP depends on catch conditions and treatment. they are usually of a better standard than species such as tilapia. but these are now finding favour as products in their own right. and it is worth noting the additional hazard to fish handlers from Vibrio vulnificus from injuries received during handling and filleting of spiny tropical fish.
and the product then shaped and heat-processed to cook the product before being chilled. The expected shelf life of these products is up to 8 days (or longer for some chilled. swordfish steaks). They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes.g. They are sold from wet fish counters.g. Cold smoking is essentially a flavouring process. The result is fish with a typical residual salt content of 1. natural fillets of fish such as herring. Added colours can be included with the brine solution to meet specific requirements for products e. a protective glaze may be added to the frozen steak. which may. Fresh fillets are sold from wet fish counters. resulting in a product that is ready-to-eat without any further cooking. annatto with smoked haddock. The shelf life for MAP fillets is typically up to 5 days. salmon. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. or pre-packed frozen with added glaze. but may be increased (up to 1 month) by packaging methods like MAP or VP. Some products may be salted for flavour only (e.5%. which is approximately 30% (or 1 . A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. cod).g.g.2. sawdust with other woods adjusted for flavour profile and burn duration). 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. Pâtés are prepared with raw fish. tuna steaks) or band-sawn from a frozen block (e. Hot-smoked fish are prepared from brined. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. haddock. VP products) because of the high salt content acting as a barrier to spoilage. or pre-packed in MAP or vacuum packaging (VP). Again. mackerel. etc.. in VP or through mail order. the ingredients premixed. with a minimum residual salt content of 3.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product. Fish steaks can be cut from a natural fillet or loin (e. This process in effect cooks the product and reduces or eliminates microbial contaminants. be flavoured with herb dips or peppercorns. 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.5% in the aqueous phase.5 . in addition. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. mackerel.2 days) longer than that of non-prepacked fish. and salmon. 29 . 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.
and seafood mixes. This ‘catch-all’ category includes seafood salads. Other ingredients may be natural or reconstituted. and heat-fixed. Natural prawns from cold or warm waters can include those peeled. ‘fingers’ before being coated in batter and/ or breadcrumbs. 30 . flavours. There is an increasing market for high value fresh or thawed fish (e. The majority are products made from various types of whitefish (cod. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process.or a five-stage process. The cakes are usually coated using a three. Prawns. mixed whitefish. taramasalata.g. or cut from a preformed frozen fish blocks into shapes. jellied eels. cephalopods.) Raw material is typically fish fillet. and may also include additional ingredients such as fresh herbs and spices. fish nuggets/bites with dips and seafood-based snack products. long-life seafood mixes using sorbate and benzoate as preservatives. which may include molluscs. Band-saw dust and fish trimmings from fishblock preparation materials may be used.g. pasta or potato. It is often used as a protein supplement.FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate.. coated in batter or breadcrumbs. Other. salmon) coated and sold as chilled products. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. tuna. Surimi is the insoluble protein fraction of fish after washing and processing. rice or pasta.g. etc. and in the shell. it can be used as a seafood analogue. with an expected shelf life of around 7 days. rice. e. haddock. addition of ingredients such as colours. Breaded fillets/steaks and battered fillets are either natural or shaped fillets. Fish cakes are products produced by mixing fish with potato and herbs. They are pasteurised products and treated as high risk raw materials. e. binders. Seafood ready meals (frozen or chilled) differ from seafood recipe meal centres in that they contain a source of carbohydrate such as a potato topping. The protein source can be fish fillet or blocks. Scampi are tails of Dublin Bay prawn or langoustine. and formed into round cake shapes. With further processing.
Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. including seasonal variation. which may become a focus of contamination deep within the muscle.3.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. Only after death. storage. and intestines of the fish. 6). In summary. For a summary of background information on the hazards associated with fisheries products and ingredients. product handlers. Dalgaard et al. or visible damage from gaff marks (4). searing or bar marking). After capture and during any subsequent processing. Grading is based on size of fish (>25 kg). 2). and processing conditions such as dehydration or poor filleting practices. it may be used for other products or markets depending on the product specification. and ingredients added to the final product. The edible tissue of some species of fish are more susceptible to gaping than others. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. Contamination will. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). and pathogenic flora associated with seafood.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage.I. and have emphasised the origin and sustainability of the fish as key marketing points. i.and turtlefriendly long lining to the less discriminatory netting.e. remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. fat content. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. see Table 2. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. skin. as a result there is an increase in spoilage rate thereby reducing the product shelf life. is not used for sushi or other high risk (and high value) products. species (Bluefin>Bigeye>Yellowfin>Albacore). the microbial flora of seafood is associated with the gills. 31 . Recent public opinion has favoured the use of dolphin. handling or preparation stage. Tuna that does not meet the highest of grades based on colour. A primary role in fish spoilage has been assigned to this organism.3 2. will the saprophytic flora start to invade the surface flesh (1. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. sensory assessment of freshness etc. whilst gaping is also dependent on a range of other factors that effect the muscle composition. on the whole. and the potentially toxic by-products of histamine-producing bacteria (5.
coryneforms. Pseudomonas.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. cereus Staphylococcus aureus. and so are the potential microbial contaminants associated with them. intermediate processing. Penicillium. shelf life determination. yeasts Pathogenic microflora/toxins Salmonella. Shigella. streptococci. moulds Bacillus Bacillus. B. moulds. Clostridium botulinum.4 2. and. Escherichia coli. histamine Cryptosporidium. for chilled products.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. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter.4. Bacillus subtilis B. Staphylococcus. protozoa and viruses B. staphylococcal toxin. be considered at an early stage during new product development. toxin. 32 . Table 2. moulds Bacillus. Clostridium Lactobacilli. Staph. pseudomonads. TABLE 2. however.I provides an overview of common raw materials. Aspergillus. Aeromonas. spore-formers. coliforms. anaerobic spore-formers. Listeria. 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. Fusarium.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. Vibrio cholerae. raw materials. product assembly. Clostridium perfringens. Bacillus cereus. 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. by the HACCP team or by a hazard and risk assessment programme. cream. and associated spoilage and pathogenic microorganisms. cereus B. yeasts Lactobacilli.3. and milk Cheese Water/Ice Bacillus. cereus Bacillus licheniformis.FISH AND SEAFOOD 2. Moulds Enterobacteriaceae Salmonella. enteric viruses 2. In addition. Enterobacteriaceae. nuts and fruit Associated microflora Lactobacilli. mushrooms. moulds Bacillus.
the core temperature. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. Figures 2. but it is assumed that any band sawing or dice process will introduce heat from blade friction.1 . defined holding time at a chill temperature (0 – 3 °C). Block-sawn dust that is required for use in fish cakes is swiftly transferred to a chiller to avoid water loss and spoilage from process contaminants.4. Examples of tempering techniques include the use of heated air. up to -5 °C so as to improve the cutting and handling properties during processing. Generic examples have been used to illustrate only the major points. but will also need to ensure that the optimum shelf life can be safely obtained. For some components.2. but it also poses a risk of cross-contaminating other products undergoing tempering. and potential opportunities for growth of foodpoisoning bacteria. this may be an incidental effect. 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. All have the same intention.4.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. 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. Band-sawn materials or fish dice are normally promptly further processed. depending on the size of the fillet 33 .CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. as the full complexity of the processes used for specific products cannot be covered in a general text. 2. Effects on the microflora are minimal during tempering (7). to evenly raise the temperature of the entire frozen product from -20 °C or lower. and contamination from equipment to clean fish muscle. 2.3. Rapid controlled cooling of heat-processed components will help to minimise spoilage. as a heat process is often required for functional or organoleptic reasons.1 Coated products Seafood products are coated with a cereal-based batter or crumb with up to 30 50 % of the final product weight accounted for by the batter or crumb. It is recommended that the principles of HACCP are applied to novel processes. Where the raw materials are frozen. or the use of microwave technology.4. there are a number of distinct methods used.
2. chilled or sold chilled on defrost with an expected shelf life of 6 .1. are placed into plastic boxes and covered in ice to preserve their condition.1A . On arrival at the processing plant. It is then size-graded.FISH AND SEAFOOD processed. 2.4.Fish Cakes). 2. This material is transferred to a processing facility with the ice replaced as necessary to ensure that the product is maintained at the temperature of melting ice. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. The “blown” meat is graded.4. The processing of three examples types are described in Figure 2.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). by mincing and mixing with a binding agent (as described in section 2. frozen in a nitrogen blast freezer and weighed into sacks for later processing.3. after landing. Fig.1B .Shaped Fillets. with the coating heat-fixed. may not rise above 0 ºC during processing.1.2.2). The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.1.1 (2.Scampi. Frozen cores are usually batter-enrobed. drained and then refrozen for storage until required for further processing. the scampi is washed in potable water to remove foreign bodies.3. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. quickfrozen. Raw material preparation stages for chilled or frozen coated fish products 34 .1C .7 days.
ii) Fish-filleting waste .3. and water is added to rehydrate the components. those that use frozen blocks of whole fillets. The mix is shaped and formed ready for packing or for enrobing. 2. or sold as a chilled product after thawing. 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.off-cuts or belly flaps. because of its nature. or promptly processed to avoid further quality loss. 35 . but.2 Shaped products This type of product falls into two categories. from potato flake. The fish component of the cake will. quickfrozen. chilled.g. more seriously. fish fingers). with an expected shelf life of 6 . pathogens such as Salmonella and Listeria. spoil more rapidly than the dry ingredients. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. It is for this reason that all by-products from primary processing. that are to be reused for other foods. once the dry components have been rehydrated.3.4. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish.1. The remaining dried ingredients are mixed with the fish. additional spoilage organisms may be introduced.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet.. This is especially the case with block-sawn dust. Waste material produced from this type of processing is often reused in the third category (fish cakes). Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. or those where the trimmings. Fish cake cores may be batter-enrobed. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . such as aerobic spore formers and Staphylococcus spp. with the coating heat-fixed. Therefore. material will need to be transferred swiftly to a chiller. are handled and stored as any other food product. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. as the physical structure of the material has been destroyed. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. iii) Fresh/fillet/minced fish block.7 days. unless it is poorly handled.4.block-sawn dust.1.3 Fish cakes Fish cake preparation is used as an example of the production of fish products with fresh or dried ingredients added during the process.
Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. IQF/pre-formed fillets c. a. vibrios and other fishassociated spoilage organisms. The function of this is to improve the adhesion of subsequent coatings. such as Listeria and Vibrio spp. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. Spore-forming anaerobes such as C. The first stage. starch. raising agents. 2. potentially. as well as ensuring that there is minimal contamination from human processors.. perfringens could. and Enterobacteriaceae from flour. any excess is removed with an air knife.e. which can occasionally be isolated from protein-handling processes. Batter contains water. i. Fish products. a characteristic flora will develop from the batter components.2 illustrates the typical stages of enrobing or breading. Bacillus spp. e. is the addition of a pre-dust. Initially. or sold as a chilled product after thawing. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. absorb any excess moisture on the surface of the fish portion. rusk or crumb. with the coating heat-fixed. spices. e. egg. Breaded or battered fish fillets are fully heat-processed before consumption. 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. with an expected shelf life of 6 . A large variety of product formats and types can be batter-enrobed. which typically consists of flour.4 Battering/enrobing process Figure 2. from flour. because of the intensive handling required during processing. If a crumb coating is required.or fivestage) frozen or chilled. and may include milk powder. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process. which is product-dependent and not always required. Products are preformed and presented to the process (either three.g. some of this initial flora 36 .7 days. and possibly Staphylococcus spp. milk powder and spices. quickfrozen.FISH AND SEAFOOD In all of the above examples. rapidly multiply if strict temperature control was not maintained. Tempered band-sawn materials b. Natural fillets d. and if required. fish cakes Portions or fillets may be batter-enrobed.g. flour. and to carry flavourings where required. chilled. lactobacilli from the milk powder and flour. Over time.4.
replaced in part by process contaminants and bacterial flora from seafood. dry batter mixes are all potential sources. cereus can proliferate under certain conditions. It is advisable that some control mechanism is employed to prevent toxinforming organisms from multiplying to dangerous levels. This can be achieved by frequent clean-downs and adequate hygiene. Flow diagram outline of a typical three. and.2. and both of these can produce heat-stable toxins that can survive a batter heat-fix process. The origin of contamination for the two organisms is often unclear. but it is believed that poor hygiene regarding product and food handlers. time-limitation or production run temperature control. this is not of great significance to the microbiological standard of the finished product. contaminants such as Enterobacteriacae can proliferate and develop during the process. aureus and B.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.CHILLED AND FROZEN PREPARED FISH PRODUCTS associated with the ingredients will be diluted out. but because batter is heat-set. cereus. With inadequate temperature control of batter systems.2. Staph. in the case of B. Fig. The FDA has issued clear guidelines on acceptable 37 .
pasta or potato. A temperature no greater than 10 ºC for 12 hours cumulatively or 21. and must not show evidence of disease. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products.2. as this could introduce off-flavours and reduce shelf life. For example. The protein source can be fish fillet or block.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe.g. Fig.1 ºC for no more than 3 hours is recommended (1). Figure 2.3. Flow diagram outline for recipe dish ingredients preparation 2. but they are supplied without a source of carbohydrate e.5. to produce a specific product.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). The vegetables must withstand processing.4. There are two main categories.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers.4. 2. 38 . and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. canned fish may be used in their preparation owing to it having wider availability and low wastage. rice.
Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. and may include molluscs or prawns that may or may not have undergone a minimal heat process. and E. frozen block or canned fish. In an attempt to improve washing and reduce chlorine taint.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. Blanching of vegetables can be an effective means of reducing or removing surface contamination. will disperse the saprophytic flora of vegetables . colour and water will occur. treated as a highrisk ingredient.3 log reduction in bacterial numbers on the surface of vegetables. with the accompanying release of additional nutrients by processing. coliforms and pseudomonads throughout the mix. coryneforms. they should be stored under temperature-controlled conditions. and protected from any post-processing contamination. all-year-round fresh vegetables are preferred. However. any processing stage.lactobacilli. For example. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. rice or pasta. encourage more rapid decay and loss of structural integrity. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. 39 . Any assumptions regarding the application of good agricultural practice to produce should be made with caution. Salmonella typhi. Once vegetables have been blanched. i. Washing with a biocide will at best achieve a 2 . if the process is not appropriately controlled. but also as components in recipe meals. not just as accompaniments. coli O157. 8) will. A number of examples of direct faecal contamination of vegetables exist in the literature (8). However.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. For premium products. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. loss of quality. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components. The presentation of vegetables has a significant effect on the product quality. Products must be washed thoroughly to remove pests and soil. The majority are products made from various types of whitefish (cod. for example. pastry. spore-formers. Although the processes are claimed to be superior to using unmodified hypochlorite for washing. and may include sauce accompaniments. Raw material is typically fish fillet. and mixed whitefish).5. For example. control is more onerous. and reduce spoilage organisms and potential pathogens. haddock.e. even those with pH-modified dressings. The opportunity for growth of L. with cases of foodborne disease traced to Listeria monocytogenes. chopping and mixing of material. 2. (7. the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. or breadcrumbs.4.
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. and then cooled slowly allowing germination and re-growth.4. On hydration.g. 40 . where composite meals are heat-treated. Placing such products in a standard chill store to cool down could result in condensation. cereus).6.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. but temperature-abused pasta can also spoil rapidly by mould growth. Alternatively.) to the product (12. cereus. Sauces can be multi-component preparations or more simple preparations. if subjected to temperatures between 20 and 40°C. B. with the range always expanding as new tastes and cuisines are developed. 11). spore-formers can proliferate if adequate control of temperature is not maintained. An incident involving Staph. rely on the use of microbiologically sound ingredients. Products or dried ingredients contaminated with either spores of aerobic bacteria (e. aureus toxin is well documented (10. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose. Appropriate handling facilities or equipment will be required to protect the product. a heat process in excess of 73 ºC will be required to rehydrate this component fully. especially in those products that have a vegetable component or rice accompaniments.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. cereus are well known and are still a common issue in the UK (9). If the sauce contains modified starches for freeze/chill stability. 2. such as butter garnish with black pepper. can germinate and rapidly multiply. and an increased risk of contamination from L. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. Typically. Potato toppings made from flake or powder will normally have little associated spoilage flora.4. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. This is a particular issue especially where products are heat treated to eliminate vegetative cells. For this reason all products. 13). Sauces or marinades are added to fish to broaden appeal or to accompany components.
Marinade-type products are becoming more popular. The format uses flavour compounds . the sauce component will spoil rapidly and the possibility of vegetative pathogens from post-process contamination or raw components growing to significant numbers cannot be ruled out.herbs. for instance. 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. suspended in an acidified sauce with an oil component. 13). flour products. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit.4.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. etc. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. 41 .5 illustrate the recipe dish ingredients preparation and assembly and packaging. spices. and handling of processed sauce can be less rigorous than.2. 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. that of a dairy-based sauce. Without barriers. Figures 2. respectively.4 and 2.
2 Modified-atmosphere packaging Originally developed for the red meat industry. as it dissolves in the surface moisture to form a weak acid.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. and tuna loins are packed by placing the product in a plastic pouch and removing all the air. The oxygen-free conditions will however allow other anaerobic bacteria such as C. the method replaces air in the sealed packaging with a defined mixture of gases.4. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. 42 . The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues.FISH AND SEAFOOD Fig. unless the temperature is strictly controlled to below 5 °C throughout the supply chain. These include: 184.108.40.206.5.7. botulinum to grow. 2. generally carbon dioxide and nitrogen. which may restrict either microbial growth or enzyme activity.2. Flow diagram outline assembly and packaging of recipe dish 2.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).
but is not limited to. However. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). as this organism is a common contaminant of freshwater fish species (19). phosphoreum. Not only will these organisms have a numerical advantage over other proposed spoilage organisms. initially. but they will also be readily adapted to the nutrient source and conditions. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. 2. Shewanella putrefaciens and Ph.1 Chilled storage For chilled products or chilled components of frozen products. and Yersinia spp.10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. Vibrionaceae will succeed (17 ). It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage.. At refrigeration temperatures (5 . Aeromonas spp. in numerical terms. Nonetheless. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15). and finally flavour loss. both organisms are capable of growth at chill temperatures.5 Spoilage During processing. a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood. The case for growth and contamination of products with Aeromonas is stronger. where the protein component is nearly always heat-processed.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’. The fish component of recipe dishes. 43 . This is considered to be the one major difference between fish and meat products. is typically uncooked or at best lightly processed to retain eating characteristics. by a loss of texture. The literature has good information on spoilage of individual components of recipe meals. Beyond this. and in influencing the final eating quality of the product. at slightly higher temperatures (10 ºC). (20).5. This is also true of chill storage and the distribution chain. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18). Spoilage will be most readily detected in proteinaceous components of ready meals. the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. followed by moisture loss. temperature control is important in preserving chilled integrity of ingredients. but little on in-pack spoilage. except for some crustacea. Spoilage of this material is not expected to differ greatly from the processes described elsewhere.
released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). and Aerobacter spp. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution.. These have been proved to impart off-flavours. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. freezing also has a protective role in improving the safety of raw materials by.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. Serratia spp. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. eliminating nematodes in products that do not have a heat treatment step. The conditions and freezing process most favourable for good-quality products will. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24). Still. As a note of interest. within dairy products are well known. producing characteristic off-flavours. 44 . This reinforces the need for prompt processing and temperature control of ingredients and finished products. 2.5. where the variety of components and ingredients used in manufacture is varied and complex. Products that are developed with an extended shelf life beyond the 6 . botulinum. this should also take into account shelf life of product components used in the assembly of the final product. for example. By retaining the intrinsic spoilage flora of seafood. The type and nature of the enzymes and their activity will depend on the bacterial species. which would warn the consumer of the potentially unsafe nature of the product.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. For example. also be those that have the least effect on the viability of microorganisms (21).g. Bacillus spp. 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.. auto-oxidation of fats/oils). however. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. However.FISH AND SEAFOOD 2. enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. the shelf life of products is limited by the natural spoilage process described earlier.5. Pseudomonas spp. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e..
any risk of C.104/ g should be treated with some concern (26). 2.3 Clostridium perfringens A study examining C. are dealt with elsewhere in this book. air. 2. botulinum when held in favourable conditions for growth (28) i. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. A number of other Bacillus spp. 13. and Campylobacter are recorded in filter feeding molluscs such as mussels.6. 2. 27). Vibrio spp. Counts of between 103 . farmed or free-living crustacea (30). perfringens contaminating seafood will depend on the source of raw material. Additional hazards from marine bio-toxins. with pathogenic microorganisms or viruses. and this issue may be of greater consequence for warmwater. botulinum type E and Vibrio parahaemolyticus (25). Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. Hazards attributable to the non-seafood components are listed below and this information. water. Incidents in such diverse materials as cream. and some effects related to product type and time to toxin production have been noted. especially molluscan filter feeders. cereus.6. histamine intoxication. low oxygen or anaerobic conditions. Therefore.e. 45 .6. typhi. along with previous discussion.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. These are C.. spices and herbs and vegetable shoots have been reported (12. agricultural run-off or sewage can contaminate in-shore species. may be of benefit within any risk assessment for chilled and frozen prepared fish products. Cases quoting contamination of seafood with organisms such as S. etc. permit rapid growth of B. Inappropriate holding times and temperatures will.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. in some cases. perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). Germination of spores requires favourable conditions.1 Bacillus cereus This is a common contaminant of dust. potato.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. In addition. or a heat shock followed by holding at a temperature suitable for growth. and many raw or processed foods (26). have been linked with food poisoning and should also be considered. soil.
46 . It is unclear.6. if any other potentially pathogenic organisms. Shigella spp.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. Chilling may help Shigella to survive. 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). infected food handlers. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. but under the appropriate conditions.5 Shigella spp. aureus (35). Rates of contamination with smoked seafood are much higher than those with unsmoked products.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). 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces.6. and this is believed to be due to a tolerance to brine used in the smoking process (32). 2. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). and inadequate cooking or holding within the temperature growth range (27). and will therefore cause “food poisoning” even in a product that is cooked before consumption. monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. These conditions will allow the production of the toxin. monocytogenes as a contaminant in foods (31). L. An acceptance has now developed in the UK of the presence at low levels of L. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). growth may occur in foods (26). but. indigenous or other. However.6. will colonise factory environments in a similar fashion except for possibly Vibrio spp. the restrictions on growth rate seen with seafood will not apply. at the present time. 2. which is not affected by any subsequent cooking process. Sources of contamination are many because of the widespread distribution of this organism in the environment. These include inadequate refrigeration and preparing foods far in advance of planned service.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. L. when introduced or inoculated into sterile components of chilled prepared recipe dishes. 2. aureus enterotoxins in foods.
2. are not mandatory. For instance.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts. and as such the monitoring is for verification purposes rather than control. such as batter mixes. unless the criteria is for a pathogenic organisms where only non detection is acceptable.III based on IFST Guidelines (39)) provide example guidelines for seafood products. as published by for example. (36). cereus emetic toxin are often confused owing to similar symptoms. although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”.CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph. 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. Microbiological specifications do not necessarily relate to product safety. the fresh product is often consumed before the results are obtained. and their association with similar materials. with respect to the number and types of ingredients used. and not to final products. it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur.7 Published Microbiological Criteria Due to the complexity of many processed food products. and are a recognised means of providing data on manufacturing standards. Microbial guidelines. It should be remembered that 47 . etc. aureus enterotoxins and B.II and 2. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40). although due to long analysis times required. Microbial levels at the end of manufacturing (m) will be set much lower than those at the end of shelf life (M) which in turn should still be safe to consume. but these may be eliminated by a subsequent heat-treatment stage.6. but provide the industry with an indication of the microbial level at which action is required to ensure product safety. pastry. 2. . Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems. 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. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). The following tables (Table 2.7 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. pasta.
g. coli GMP* <104 <102 <10 Maximum Product-dependent 104 103 GMP* Not detected in 25 g Not detected in 25 g <102 <20 Not detected in 25 g <50 ppm (<5 mg/100 g) Maximum Not detected in 25 g 103 104 103 102 <50 ppm (<5 mg/100 g) *GMP = Values found immediately after production under good manufacturing conditions. Maximum= Levels at end of stated shelf life 48 .fish products. cereus Staph. 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. monocytogenes B. parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E. aureus V.FISH AND SEAFOOD the microbial criteria for other products (rice.II IFST Guidelines for part-cooked foods – e. L. coated fish. pasta. coli GMP* <102 Maximum 104 TABLE 2. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp. batter etc) should also be consulted when establishing the specifications for raw materials and final product. TABLE 2. fish fingers.III IFST Guidelines for processed foods .
Collette R. Hobbs G.H.C. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. References US Food and Drug Administration.R. 5 (3). Microorganisms in Foods 6: Microbial Ecology of Food Commodities. 47. 3.G. Ohgiya S. 11. Letters in Applied Microbiology.. Huss H.R. 67-73.. 1997. 167-93..L. Murrel H.J. England and Wales Quarterly Report. Incidence and levels of Bacillus cereus in processed spices. 373-8. 24 (5). 1997. Beuchat L. Yumoto I. 1986.M. Microbiology of processed spices. Journal of Food Science.T.. London. 1962.. Food Science and Technology Today. Tu A. Ishizaki K. 38 (11). Sakamoto T. in Fish Inspection Quality Control and HACCP: A Global Focus. 96. 166-73.W.R.A. Importance of Photobacterium phosphoreum in relation to spoilage. 2001. Matsuyama H. Taylor and Francis. 204-16. 2. Recent Advances in Food Science. Latt T. 2005. 215. 14. Mejholm O. Letters in Applied Microbiology.. Lawyer R. Animals and Spoilage. Communicable Disease Report. Slavin J. Ward D. 1991. 1997. 1993.C. Fish and Fisheries Products: Hazards and Controls Guidance.. Bartlett C. Woolaway M.. Proper processing and handling of raw tuna. Journal of Milk Food Technology. Ltd. Wienke A. 1991. International Commission on Microbiological Specifications for Foods. 663-87. 1976. 8. Pvt. Journal of Food Protection. 15.. Van Nostrand Reinhold. Kluwer Academic. Journal of Milk Food Technology.. Oxford. The bacteriology of fresh and spoiling fish and some related chemical changes. Powers E. 6. 7 (24). . Dalgaard P. 1982. Fish: microbiological spoilage and safety. 1976. Powers E.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). Bartram P. Kumeta H.M.K. 1311-7.. Pathogenic microorganisms associated with fresh produce. Microbiology of Marine Food Products. 1997.E. Cambridge.. 1. 10. 39 (10).. 233-7... 7. Christiansen T.L. Washington D. Masuoka Y. Aureli P. Hoshino T.. 49 4. London. Gilbert R. Shewan J.R. Communicable Disease Report. 9.. Journal of Hygiene. 13. 70-2.L. Behling A.R. Brown T.C.J. 5. Eds.8 1. IBH Publishing Co.M.. Outbreaks of foodborne illness in humans. 59 (2)... Martin R. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. Bacterial histamine production as a function of temperature and time of incubation. Hackney C.. New York. Toxin Related Diseases: Poisons Originating from Plants. 2. 25 (1). 668 -70.. 12. Taylor S. Food and Drug Administration.
Microbiology in fishery science. Anon. Speck M. Liston J. 1995. Golden D. Connel J.. 25. Journal of Food Protection.. 28 (4). in The Microbiological Safety and Quality of Food.. Glasgow. Andersson A.J. Her Majesty’s Stationery Office. in Anaerobic Bacteria in Habitats Other than Man. Food Microbiology. Blackie Academic Press. SAB Symposium Series No. Food Control. Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants.. Gram L. 1996. Mead G. Lund B. Anaerobes in relation to foods of plant origin. Ed. Fishing News Books Ltd. 1993. Clostridium perfringens in the environment. 9. Huss H.. Aeromonas and Verotoxigenic E. Ray B. 1986. 28..K. 1987.W. Huss H. 18. 11..C.R. Gould G. 26. 22. Blackwell Scientific Publications. Halligan A. 29. Loessner M. 1979.V. 1994.H. 2000. 1992. International Commission on Microbiological Specifications for Foods. Leatherhead. 351-72.L. Oxford. Barnes E.. Effects of freezing and storage on microorganisms in frozen foods: a review. 51-158 Saxby M. The Emerging pathogens . Springer. Trolle G. Leistner L. Brooks J. Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens.J. 28. 34-40. 1992. 27. 50 17. coli (VTEC) . 333-6. 13. Food Research International.. 655-60. Advisory Committee on the Microbiological Safety of Foods..Yersinia. a foodborne pathogen? Review of the clinical data.J...A Literature Survey. 59-72. Baird-Parker T.E. Granum P. 4 (1). 19. 31. 23. 65. Food preservation by combined methods. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service. Gaithersburg. 1988. 274-86. Wood L. 4. New York. International Journal of Food Microbiology. London. Aspen Publishers.M. PHLS Microbiology Digest.M. What problems does the food industry have with spore forming pathogens B. Survey of the incidence of Aeromonas and Yersinia species in retail foods. 24. Curran D. Ronner U. Applied Microbiology. . 40 (5).C.J. Eds. Matches J. 1990. Eds. 2005. 309-16.H. Liston J. in Advances in Fish Science and Technology. Food Taints and Off-flavours. Report on Vacuum Packing and Associated Processes. Gram L. Leatherhead Food Publications.J. Keusch A.... Fresh and processed fish and shellfish . 30. U. Morgan D.R. Walker S. Lund B. 13 (1).FISH AND SEAFOOD 16. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures. London. Jemmi T. 41-3. 1977. perfringens? International Journal of Food Microbiology. 32. 155-9. 472-506. Journal of Food Safety. Blackie Academic and Professional.A. 1974. Is Aeromonas spp. Jay M. 25 (2). 20.C. 21. cereus and C. Modern Food Microbiology.. 1996. 1996.
A. Principles and Practices for the Safe Processing of Foods. 1998. 60 (4).chilledfood. 50 (9).L.pdf Shapton D. 36. U. Microbiological Criteria.F. Chilled Food Association. 1987.. 38. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. Shapton N.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology.uk/cday/posters2008/npd6. http://eur-lex. 788-801. 41. 40. 34. 2006. Psychrotrophic Bacteria in Foods: Disease and Spoilage. Kraft A. Institute of Food Science and Technology.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. 1997. London.2.H. Development and Use of Microbiological Criteria for Foods.europa. Campden & Chorleywood Food Research Association. CRC Press. Varnam A. 1999. Smith J.A.. London. Wolfe Publishing. 35. Journal of Food Protection.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33. 51 . 37. Chilled Food Association.K. Evans M. 1992. British Retail Consortium. 372-6.K. http://www. McCarthy S. U.campden. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs. Cambridge. Foodborne Pathogens an Illustrated Text. http://www.G.co. Gloucestershire.eu/LexUriServ/ LexUriServ. 1991. Shigella as a foodborne pathogen. 39. Journal of Food Protection.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. Phil Voysey.A. Woodhead Publishing Ltd.
oysters and clams can process about 3. completely encloses the body. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. the adductors. which includes oysters.6 litres. and 2. can hold the shells tightly shut against enemies (1). mussels. chemical contaminants and other impurities on the mucus of the gills. while 53 . mussels. Many microorganisms ingested by shellfish survive the digestive process (3. the shell. mussels. Box 3012 Bothell WA 98041-4421 United States of America 3. scallops.S. they also trap bacteria. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. The class having species of economic and public health importance is the Pelecypoda. 4).1 Definitions General features of the mollusc The phylum Mollusca. Pacific Regional Laboratory Northwest U. P. Pelecypoda. The term shellfish in this context is limited to oysters. lying well within the mantle cavity. and scallops.1. S. MOLLUSCAN SHELLFISH Carlos Abeyta. oysters.E. Jr. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. Food and Drug Administration 22201 23rd Dr. octopi. cockles. Pelecypods have little need for locomotion.1 3. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). where it is eventually swallowed and digested. clams. Plankton and algae form the bulk of their diet from seawater. The phylum consists of five classes: Amphineura. Other species of shellfish such as crabs.7 litres of water per hour. whelks. The class Pelecypoda includes all of the bivalves such as clams. cockles. Special tracts of cilia move this mucus toward the mouth. In most species. respectively. chitons. lobster. squid. slugs. cockles and scallops. In the process of filtering water. snails. and shrimp. Strong muscles. Gastropoda. composed of two valves hinged together dorsally. O. clams. and Cephalopoda.3. For example. abalones. In these forms. Scaphopoda. viruses.
Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 .2 International commercial products Globally. and usually at higher concentrations than found in their habitat.South America Pacific – Japan Quahaug (also known as hard clam and hard shell clam) Coquina clam Hen clam Butter clam (also called the Washington clam) Mogal clam Little neck clam (also called rock Cockle Pacific littleneck) Atlantic – North American/Europe Atlantic – Europe/North America Japan Atlantic/PacificNorth America Donax spp. dwelling on the bottom or attached to structures in the water column. The habitat of oysters and mussels are sessile. but maintain contact with the water by means of a siphon tube (5). Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific .North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp.I). are not as vulnerable to contamination through pollution of their beds as the bivalves. 3.I Partial list of bivalve molluscan shellfish products in commercial use internationally (5) Common name Clams Soft (shell) clam Pacific/Atlantic North America Pacific/Atlantic North America Pacific . many species of bivalves are edible (Table 3. Clams are benthic and live burrowed in the mud. Whatever contaminants are in the water will eventually get into the shellfish.1. The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3.FISH AND SEAFOOD commercially valuable.
Pacific – North America/ Australia/New Zealand Pacific – North America Crassostrea gigas Western oyster (also called Olympia oyster) Cockle Cockle Sydney rock oyster Rock oyster Dredged oyster Cockles Common cockle Ostrea lurida South America Japan Australia New Zealand New Zealand Ostrea chilensis Ostrea laperousei Crassostrea commercialis Crassostrea glomerta Ostrea lutaria North Atlantic – Europe/ North Africa Pacific – North America Cardium edule Cardium corbis Cardium aculeatum Cardium tuberculatum Spiny cockle Knotted cockle Mussels Blue mussel Atlantic/Mediterranean Atlantic/Mediterranean North Atlantic – Europe Pacific – New Zealand Pacific – North America Europe Atlantic/Mediterranean Mytilus edulis Common mussel Horse mussel Bearded horse mussel Mytilus californiaus Modiolus modiolus Modiolus barbatus 55 .MOLLUSCAN SHELLFISH Gulf clam Pismo clam Surf clam (also called bar clam) Freshwater clam Oysters Common oyster Portuguese oyster Blue point oyster (also called American oyster) Pacific oyster Europe Europe Atlantic – USA Ostrea edulis Crassostrea angulata Crassostrea virginica Gulf of Mexico Pacific – North America Atlantic – North America Titaria cordata Trivela stuttorum Spisula solidissima Japan Corbicula spp.
Most clams are only a few centimetres in their maximum dimensions. hard-shell or soft-shell clam. or siphons. littleneck. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint. extended above the surface to maintain water currents needed for respiration and feeding. geoduck.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. cherrystone. The muscular foot is used to burrow in mud or sand. and other names. but 56 .1 Clam Clam is the common name of a large group of often edible. Buried clams leave tubes.2. A mantle tissue encloses the body of the clam. Different species are variously called quahog. mostly marine bivalve molluscs. Jacques Iceland scallop Bay scallop Weathervane scallop (also called Alaska scallop) Atlantic North East Atlantic North Atlantic West Atlantic Pacific – North America Pacific – North America Pacific – Japan Common scallop Sea scallop (also called giant smooth) Japan Atlantic – North America Pecten varius Pecten maximus Chlamys islandica Argopecten irradians Pecten caurinus Pecten aequisulcatus Pecten laquaetus Pecten yessoensis Penten magellanicus or Placopecten magellanicus Pecten meridionalis Pecten novaezealandiae Pecten jacobaeus Aequipecten gibbus Chlamys opercularis Chlamys varius Amusium balloti Mediterranean – South East Europe New Zealand Australia Mytilus galloprovincialis Mytilus canaliculus Mytilus planulatus Commercial scallop Australia New Zealand Great scallop Calico scallop Queen scallop Variegated scallop Scallop saucer Atlantic/Mediterranean Atlantic – North American Atlantic Atlantic/Mediterranean Australia 3.1. laterally compressed.
oyster soup. all in cans. sun-dried. uncooked. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. packed in weak brine. skewered with bamboo sticks. Fresh: in shell. Canned: clams are steamed open and the meats removed from the shells. hermetically sealed. washed.2. uncooked (shucked). Frozen: in shell. Canned soups include clam chowder. or shelled meats. trimmed and packed. oyster bisque.1. Dried: Hoshigai (Japan) shelled meat. smoked meats are also canned. Semi-preserved: meats cooked and packed in spiced vinegar. after precooking and brining. Oysters lack a foot or have a reduced foot. then heat-processed. Smoked: meats.2 Oyster Oysters are marine bivalve molluscs. Dried: shelled meats. 57 . Frozen: shelled meats. and no siphon. Soups: oyster stew.MOLLUSCAN SHELLFISH the giant clam of the Indian and Pacific oceans may be nearly 130 cm long and weigh 227 kg. or shelled meats. These molluscs feed on minute plants and animals carried to them by the current. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. clam madrilène and clam liquor. Live: in shell Fresh: in shell or shelled meats. Live: in shells. Canned: meats removed from the shell by steaming. having rough and irregularly shaped shells that are closed by a single adductor muscle. uncooked meats packed unprocessed. 3. either whole or minced. in cans with either brine or clam liquor. the meats are smoked and packed in oil in cans or jars. tidal waters or in brackish water in the mouths of rivers.
and held apart. flattened ear or wing at the hinge area.2. They have filibranch gills.2. and the anterior end smaller and pointed. The familiar marine mussel is asymmetrical. boiled. with the posterior ends of the shells (valves) broad and rounded. by interlocking tufts of hair-like cilia. vinegar solution.FISH AND SEAFOOD 3. Smoked: meats may be canned in edible oil. in which the individual branches. 3. Unlike other sessile bivalve molluscs. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves. or filaments. The shells are fluted and round with a broad. The adductor muscle is the only edible part of the animal. Salted: cooked meats bottled in brine. or packed with jelly. mussel paste. smouldered and afterwards dried (Japan). 58 . are united. sauces.) Dried: peeled.1. cooked meats packed in dry salt for transport. Bait: used live extensively for baiting fish hooks for line fishing. scallops can swim or leap by a series of rapid closings and openings of the valves. gutted. brine. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. The posterior halves of the valves contain a large adductor muscle that holds the valves closed.1. Internally.3 Mussel Mussels are marine bivalve molluscs that live either partially buried in the sea bottom or attached to rocky surfaces by means of byssus threads. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). Fresh: shelled meats (often only the adductor muscle and the roe are eaten. mussel in butter sauce. Live: whole with shells. Fresh: cooked meats.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. also with other ingredients. in mussel liquor. In some species. scallops possess a single large adductor muscle. Semi-preserves: cooked meats with vinegar-acidified brine and spices. The common scallop is about 5 cm long. Canned: cooked meats.
which affects the temperature. Because molluscan shellfish are filter feeders. Fresh: meats removed from the shell by boiling. Cockles range from about 1 to 15 cm in length and have two valves of equal size. mostly in oysters. For example sessile and benthic shellfish dwelling on the bottom or burrowed in marine sediment are especially vulnerable to pollutants (sewage and/or wastewater) introduced into the shellfish water beds.2. 3. 3. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding.MOLLUSCAN SHELLFISH Frozen: shelled meats. Live: in shell. also bottled in brine. salinity. Salted: meats either lightly or heavily dry salted. Vinegar cured: bottled in malt vinegar after brining.1. nutrient concentration and pollutants of the water column. clams and mussels. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. The microflora are dependent upon a number of factors such as season. The following pathogenic microorganisms causing illness in humans are characteristically associated with molluscan shellfish: Vibrio parahaemolyticus Vibrio cholerae O1 Vibrio fluvialis Vibrio hollisae Vibrio vulnificus Vibrio cholerae non-O1 Vibrio mimicus Vibrio furnissii 59 . Canned: in brine. and the environment. they accumulate pathogenic microorganisms from polluted waters. Canned: in own juice. depending on length of journey. and butter. are vectors of disease transmission (8). pH.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves. Raw and partially cooked shellfish. Whatever contaminants are in the water will eventually get into the shellfish (7). in sauce.
rakes.1. and yeast cells of Rhodotorula rubra and Trichosporon spp. 15).. The largest groups found are the Pseudomonads. 12.3 3. Campylobacter coli Bacillus cereus Hepatitis non-A. They are termed viable non-culturable microorganisms. and cockles The processing of bivalves takes two forms. clams. Temperature.3. Other microorganisms found in lesser numbers are the Achromobacter spp. that have never been cultured on laboratory media.2.. Pseudomonas spp. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. shellstock (live) which is shellfish in the shell. mechanical harvesters. whole or in part. and Aeromonads.. 10.. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. spirochetes of the genus Cristispira and Saprospira. and shucked shellfish that is shellfish. 11...1 Shellstock 3. but have never been cultured (14.1 and 3. nutrients. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp. Acinetobacter spp.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs.. (9.1. responding to the environment in which they are harvested. 13). salinity.1.000 /g. Micrococcus spp.. There are microorganisms. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. Bacillus spp. dredges. 3. Flavobacterium spp.3.1 Processing and its Effects on the Microflora Oysters. or by 60 . from which one or both shells have been removed. In summary. Water temperatures affect the concentrations of microflora in the water. the initial flora composition and concentration of bivalves are highly variable. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100.3. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. 3.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. Vibrios. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. Enterococci spp. observed microscopically. For example. Moraxella spp. Alcaligenes spp. mussels. At harvest..
Thirdly. usually in open-mesh containers and/or burlap sacks. or from other sources approved by the State Shellfish Control Agency (SSCA).1. If by boat. or immediately placed in refrigerated trucks. Secondly. Water used for shellstock washing is from approved growing areas. by truck or boat. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale.MOLLUSCAN SHELLFISH hand (Figure 3. after harvesting. 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.3. The shellstock is transported dry. Shellstock are placed in drycooler storage containers in the facility. Generic flow diagram of live shellfish from harvester to point of sale 61 . There are three primary reasons for wet storage: first. Shellstock may be wet-stored at the processing facility in tanks of clean seawater. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters.1). for transportation to the point of sale. Fig. in containers from which they can be retrieved easily. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. or in bay-approved shellfish-growing waters prior to sale or processing. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock.
The washing of sediment and detritus soon after harvesting will prevent further contamination. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating.2 ºC (45 ºF) or below (19). 22). Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. 26). involving 691 cases was attributed to oysters contaminated in a wet storage area (27). with the exception of A. 17). Generally. and the sanitary practices applied during harvesting and shipping. and Citrobacter species) and Vibrios. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. and temperature storage of shellstock to point of sale. At this temperature the multiplication of faecal coliforms (E. 21. synthetic seawater for the purpose of de-sanding and salt uptake. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. bivalves tightly close their shells thus trapping water and microflora. 24. coli. are prevented. including V. 16. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). The most important factor in controlling bacterial growth in shellstock is temperature. for a considerable length of time. and thus contaminating the shucked meat at the point of sale. however. the bacterial counts in the meats will increase. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. At harvest. handling. Harvest conditions are another factor that influences the growth of bacteria in shellstock.1. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. with prolonged storage the microbiological quality of the bivalve decreases. hydrophila (18). some pathogenic organisms will survive and multiply in shellfish (20. Excessive sediment on the shells and dead shellfish may increase bacterial 62 .1. 23. there is evidence that in the absence of adequate refrigeration. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. Furthermore. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. 25).FISH AND SEAFOOD 3. Over a period. If long delays occur before reaching the processing facility and/or refrigeration. vulnificus.3. Enterobacter. Generally. Water used for shellstock washing should be of good sanitary quality. shellstock are not refrigerated on the harvest boat. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. to avoid possible contamination of the shellstock. without adequate refrigeration. shellstock are placed in refrigerated trucks for transport or placed in wet storage. Klebsiella. when it reaches the processing facility. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. An outbreak of infectious hepatitis in Sweden in 1956.
washing. In addition. are applied to the shucking operation of shellfish. the tank is opened through a drain gate and chute. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station. Due to the nature of the shucking operation.1. Shuckers routinely use either cotton or rubber gloves during the shucking operation. clothing becomes very soiled. Thus.3. 3. If shellfish are not reasonably clean at the time of processing. Studies have shown the use of rubber gloves reduces the contamination level. and fish. and packing shellfish into containers (Figure 3. removal of shellfish (shucking).MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage.2 Shucked shellfish 3. or subjected to splash from foot traffic. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. To remove the shellfish attached to the remaining valve. whereas cotton become wet and 63 . thus contributing to high bacterial counts in the final product (28). lobsters. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. the finished product may become contaminated.1. 3. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. Containers are placed in packing boxes for shipping at refrigerated temperatures. 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. In the blower-washer.1.1 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. through which the washed meats are discharged for rinsing and then packed in containers.2.2).2. Under these conditions if the shuckers enter the packing room.2 Effects of microflora during processing The same considerations as previously discussed. 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).3. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported. After washing of meats. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank. shellstock is subjected to contamination when stored on the floor in standing water. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). washing and culling shellfish prior to storage is essential. for the effects of microflora during the processing of shellstock.3.
Separate rooms or lockers for storage of clothing. Shucking containers are rinsed clean with running water.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). aprons. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. 3. There is a tendency to store such articles on the shucking benches or in packing rooms.2. and gloves help prevent the contamination of shellfish. which may lead to contamination of product. and sanitised before each filling. Shucked meats are thoroughly drained. 64 . cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. 2.2 ºC or less. Fig 3. 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. 4. within one hour.
34. 65 . As described previously. For example. a carbohydrate. Lactobacilli seem to be the predominant microflora in shellfish. Alcaligenes. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. are an excellent medium for growth of bacteria. where there is a breakdown of temperature control or prolonged storage of shellfish. particularly shucked meats. whereas bacterium such as Pseudomonas. streptococci. glycogen levels range from 0. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. Pseudomonas. 32).2 ºC. Micrococcus. at the turn of the century. 33. The spoilage process in shellfish may be divided into three stages: increase of acidity. Achromobacter and Flavobacterium levels decrease during storage. skimmers. Much of the work has been done with oysters and clams. clean gloves. shucking pails. 36). and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. in oysters. and yeasts. abundant gas production. Shellfish. 31.47 to 6. particularly oysters stored at 7 ºC. Flavobacterium. Earlier studies.2 ºC or less. blowers and other utensils that have cracked. The initial flora found in shellfish is the major factor of spoilage in bivalves. In summary.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. these include lactobacilli. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. All equipment coming in contact with the shellfish such as colanders. and proteolysis (10). is stored in the bivalve as a source of energy. This process changes the bacterial profile to microorganisms favouring low pH.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. rough. Proteus.8% (37). anywhere during the process of shellfish from harvesting to the point of sale. and packing the shellfish in a timely manner. Controls that should be applied include using clean packing equipment. spoilage will occur. 3. Glycogen. Temperature abuse will result in high bacterial counts. Shucked shellfish are held and transported at temperatures of 7. The environment has an effect on the bacterial population found in shellfish. 35. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. 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. Glycogen is hydrolysed by the initial flora found in shellfish. thus producing acid and gas (38). These organisms may play a significant role in the spoilage processes. showed that organisms such as Achromobacter. Shucked shellfish are an excellent medium for the growth of bacteria.
5 cases/100. Diseases caused by Vibrios are enteric in nature. headache and fever. nausea. and harvesting of shellfish in the cooler months. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. This bacterium is present in all the major processing steps of a shellfish plant.5.48 hours with abdominal pain.FISH AND SEAFOOD 3. 44). of V. prevention of cross-contamination. during processing.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. Sporadic cases are frequent along the coast of the United States. coastal areas). however. and very common in Japan. 66 . V. approximately 23 cfu/g.1 Vibrio parahaemolyticus V. widespread in both polluted and unpolluted waters. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). One of 250 cases reported involved shucked products with low levels. 41).000 population/year (46). The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. parahaemolyticus causes gastroenteritis lasting 24 . ranging from epidemic cholera to sporadic cases of diarrhoea.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. Between May and September 1997. parahaemolyticus. Fortunately. and estimates based from a hospital study show less than 0. from harvesting to finished product (i. It is a common marine bacterium.1. These organisms can be present in the shellfish from the point of harvesting. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. diarrhoea. there is a recent report of low levels of V. V. 3. shellstock and packing of shellfish in jars) (29). Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. They can be introduced to the shellfish from the environment or added during processing. lot and retail outlet. Major outbreaks are associated with the warmer months. primarily in brackish or saltwater. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). although non-hemolytic strains have been associated with illness (43. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. where outbreaks occur regularly. The incidence of V. only a small percentage of strains in the marine environment are potentially pathogenic (45). there were over 250 cases of illness on the West Coast of North American extending from California to Canada. parahaemolyticus possibly producing illnesses (40). parahaemolyticus from shellfish is low. Products involved were from the same processor. and to the point of sale.5.e. 3.
cholerae. cholerae non-01 has been associated with septicaemia. The occurrence of V.48 hours of onset. vulnificus.000 population/year (8. and nausea within 24 . parahaemolyticus the estimated annual incidence rate is very low. Control of V. abdominal cramps. 46. 23 of 24 major USA-West Coast estuaries were positive for V. It causes diarrhoea. chills. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. cirrhosis. with a fatality rate of 50% for individuals with septicaemia. cholerae non-01 is ubiquitous in the estuarine environment. the organism has been found in the Gulf of Mexico. 3. It is one of the most severe foodborne infectious diseases. gastroenteritis. parahaemolyticus. Primarily. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. leukaemia. 54. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. like V. 48). Hazards from this bacterium can be controlled by thorough cooking of shellfish. parahaemolyticus growth in shellfish meats is temperature dependent. In one study. Illnesses are usually associated with the consumption of raw oysters as described in Table 3.MOLLUSCAN SHELLFISH Control of V. in the range of 0.5. 53. Oysters are the most hazardous food associated with V. cholerae non-01 (51). V. nausea. V.5.2 Vibrio vulnificus The symptoms of V.1. fever. 57. diabetes. They are sensitive to cold. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). and rapidly become inactive at low storage temperatures. which have been implicated in shellfish-related disease outbreaks (31). vulnificus are fever. 56). V. The bacterium is an estuarine species that causes wound infections. however. The organism attaches to the intestine and release of a toxin is suspected. are particularly susceptible to primary septicaemia. Inadequate refrigeration will accelerate the growth of V.8 case/100. Again. those who have liver disease. 3.II (8. or primary septicaemia.4 . Healthy individuals are susceptible to gastroenteritis. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). and preventing cross contamination once the product is cooked.0. Control of oyster-associated 67 . and vomiting.7 days. vulnificus is by rapid refrigeration during warm weather months. Death has been reported to occur within 36 hours from onset. 55. 46. or immunosuppression.3 Vibrio cholerae non-01 Illness caused by V. 58). 49). lasting 6 . High-risk individuals. Onset of symptoms occurs in 48 hours and can be severe. these individuals are advised not to eat raw shellfish. cholerae non-01 is generally a less severe gastroenteritis than that caused by V.1. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. All isolates are considered pathogenic.
is the same as previously discussed for the non-01 strains. 61. parahaemolyticus and V. Onset of disease is 6 hours to 5 days. In general. salinity.48 hours with abdominal pain. Control of infection from these organisms is no different from the other Vibrios. There are several other Vibrio spp. nausea. fever. and southern Asia including India and Bangladesh.S. 3. cholerae 01. and the transmission of the disease by consumption of raw.FISH AND SEAFOOD V. or cross-contaminated shellfish (57. They are common marine bacteria. Onset of illness is from 3 to 5 days after ingestion of contaminated food. abdominal pain. 3. 3. vomiting. Illness can include abdominal cramps.5. imported product from a country experiencing an epidemic. It causes diarrhoea.g. and V. and shock.5. cholerae non-01 illnesses is by cooking seafood thoroughly. cholerae 01 establishing in the U.5%) and 2 . 62. e. nausea. dehydration.10% carbon dioxide for optimal growth conditions. There is evidence of strains of V. In developing countries. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. respectively. associated with shellfish-borne illness outbreaks. processors should know the product source. The frequency of infection of these organisms is no different from other Vibrios. however.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. mimicus.1. The control of V. hollisae. and illness lasts 7 . Gulf coast environment. these organisms cause gastroenteritis lasting 24 . V. Illnesses are associated with the warmer months. vulnificus. Death may occur after severe fluid and electrolyte loss. V. 63). the pathogenic severity is lower than V. drying. In addition.1. preventing crosscontamination of cooked seafood.2 Campylobacter jejuni C. including V. particularly in oysters. undercooked. The presence of these strains appears to be in low numbers and non-virulent (59). fluvialis. acidity and heat. headache and fever. low pH. Poor sanitation and contaminated water supplies will spread the disease. and nausea. jejuni requires reduced levels of oxygen (3 . widespread in both polluted and unpolluted waters. 60). cholerae 01 in molluscan shellfish. 68 . diarrhoea. cholerae 01 is transmitted by faecal contamination of food or water.5. (8.10 days. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59).5 Other Vibrios spp. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. V. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American.
however. 83.5. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. Campylobacter appear to survive well in oysters during storage. V.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. V. fluviallis.II Molluscan shellfish associated outbreaks caused by Vibrio spp.500 cells/gram of Campylobacter may cause illness (64. Campylobacter spp. however. 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).3.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. The seasonality of 69 . 3. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. 73). jejuni begins in the harvest of shellstock from approved waters. 81. V. and V. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. V. In one case. 69. of Incidents 4 279 0 1 0 No. although their role as a pathogen is highly controversial and circumstantial. 82. They have been implicated in limited shellfishassociated gastroenteritis cases (8. jejuni were recovered. 65. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. hydrophila group (75.MOLLUSCAN SHELLFISH TABLE 3. parahaemolyticus. 66). 77. have been isolated from shellfish beds throughout the world (67. 68. jejuni does not multiply in shellstock at refrigerated temperatures. from Pacific shellstock oysters purchased from a retail grocery store (74). mimicus. Human feeding studies suggest that about 400 . 80) are ubiquitous in the marine environment. 72. C. hollisae Infective dose varies with host susceptibility and virulence of the strain. infection occurred among participants of a firemen's banquet.5. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). ranging from 0. control of C. 84. 76.4 to 114 cells. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. Recently. 78) and P. shigelloides (79. This finding is significant in that C. they survive well at refrigerated temperatures. 71). and their immediate proper refrigeration. cholerae 01 and non-01.3 Other organisms of concern 3. 70. Although. 85). To reduce the risk of shellfish-borne campylobacteriosis. C. of Cases 4 362 0 1 0 a Includes V.
000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10.0001 negative for the presence3 Under review4 Source (89). dried milk/whey drink powders. Control of these organisms in shellfish products is limited. parahaemolyticus Staph. Caution should be exercised by individuals with haematological malignancies (88). and where indicated are intended to be used for official control purposes.600. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. vulnificus in molluscan shellfish. jejuni V. 87) of the outgrowth of A. 2 Source (90). They are easily destroyed with mild cooking temperatures. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. 3. prepared sandwiches. aureus C. cholerae V. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. being isolated more often during the warmer month. hydrophila group stored at refrigeration temperatures. soy products. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. non-dairy frozen desserts. 70 .000 cfu/g1 <230/100 g2 <1000 per gram. Test Standard Plate Counts E. negative for heat-labile toxin (LT) or heat-stable enterotoxin (ST) 1 Negative for the presence1 Negative for the presence of toxin producing 01 or non-01 organisms 1 <10. FDA/Administrative Guidelines 3 Source (91). after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500.000/gram (29). FDA/ Administrative Guidelines.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide.000 to 3. Note: Although molluscan shellfish are not included in the selected high risk food. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. coli Salmonella V. In one study. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. Similar results were seen by other investigators (86.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. coli Enterotoxigenic E.
Schmidt C. U. 6. 1936.. 755-76. Philadelphia.S. Gill S. Applied and Environmental Microbiology. Bernard F. Effects of harvesting waters and storage conditions on yeast population in shellfish. 7. 3. 64. 2.S. The American Oyster. Hood M.N. Applied and Environmental Microbiology.A. Tubiash H. A typhoid fever epidemic caused by carrier bootlegging oysters. 71-6. 7. 1983. Fish and Wildlife Service. Biology of the oyster in relation to sanitation.. Smith F. USA. 15. in Multilingual Dictionary of Fish and Fish Products. Colwell R. Bacterial flora of the market oyster. 1975. Liston J. Rippey S.C.K. in Microbiology of Marine Food Products. 245-7.E. Microbiology of bivalve molluscan shellfish. Applied Microbiology.. Van Nostrand Reinhold. Organisation for Economic Co-operation and Development. Microbiological characteristics of Dungeness crab (Cancer magister). Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). 13.S. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. 72-8. Veliger.B.. Washington D. Walker W. Galtsoff P. in General Zoology. References The Phylum Mollusca.K. 30. Vanderzant C. Ward D. Nauman R. Ed.S. 44. 231-51. 46.D. 33-6. 11. 104-9.. 36. Ray S. Proceedings of the National Shellfisheries Association. Eds. Pfeifer D. 42. Villee C..C. Cook D. Jr. 1994. 12. 8. Viability and release Salmonella charity and Escherichia coli from oyster faeces.A. Fleet G. American Journal of Public Health. Rowse A. 71 .E. 82-7. 10. Journal of Food Protection. Colwell R. 419-25.H.R. in Fishery Bulletin 1964 No. 1984. 1991. 5.M. Hackney C. Applied Microbiology. 1964. American Journal of Public Health. 30. 1995. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. Old H. 9. Clinical Microbiology Review. Fate of enteric bacteria in estuarine sediments and oyster faeces. Journal of Milk and Food Technology. New York. 1981.R. 1970.MOLLUSCAN SHELLFISH 3.W. 8.L. 1973. Eds. 447-52.W. 544-8. water and sediment from Galveston Bay. 1926..R.R.F.. Lovelace T. Cook D. Jr. 105-8. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). 16. 1960. 1982..A. 33643... Eliot C. 13. Lee J.7 1.R. Galtsoff P. Infectious diseases associated with molluscan shellfish consumption. 29. Saunders Company.J.R. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay.. 26. 4. Tall B. 19-34.. 633-40. 58. 1963. Wiley-Blackwell. 14. 1968. Thompson C. W. American Journal of Hygiene. Journal of the Mississippi Academy of Sciences. Government Printing Office. 1940. 6. 17.
A. Presnell M.H. 26. 31. Survival of enteric organisms in shellfish. American Society for Microbiology.. Journal of Food Science. Abeyta C.. Public Health Report. Liston J. U. in International Symposium Vibrio parahaemolyticus.. 167-71. 1961. Applied and Environmental Microbiology. 30. 343-5. Fujino T. Incidence of Samonellae in clams. 994-1002.B. 40. Food and Drug Administration. 1995. Beck W. 1983. in National Shellfish Sanitation Program Manual of Operations. Applied and Environmental Microbiology. 47 (5). P-1. 19. oysters. 1985. 32.W. 1992. Arcisz W.B. 5..A. Native Littleneck clams (Protothaca staminea). Hoff J. FDA Gulf Coast Technical Services Unit.S. 4. Crassostrea commercialis. Journal of Food Protection. Fraiser M. Washington D. Olympia oysters (Ostrea lurida). 1973.D. LaRock P. Effects of storage on microbial loads of two commercially important shellfish species. Tokyo. Behaviour of pathogenic bacteria in the oyster..J..F.A. Crassostrea virginica and Mercenaria campechiensis. 27.A. 45. Fleet G. Williams L. and storage. Vaughn M. Hepatitis epidemic conveyed by oysters.W.J. Bacteriological studies of commercial shellfish operations on the Gulf Coast.. Special Report U. 1221-8. Public Health Service Technical Report. 24. F61-9.C.. Stott R.W. re-laying. Bacteriological survey of an oyster bed in Tangier Sound.. Influence of U. Maryland.. 21. 1971. 29. Kelly C. Cook D. 1954. Ed.. 22...S. Establishment of critical control points in west coast oyster processing plants. 1989. 324. Trost P. Son N..T. Chesapeake Science. 23. crabs and mullet. Bacteriological changes in live shellfish: Pacific oysters (Crassostrea gigas). Koburger J. Jr..FISH AND SEAFOOD 18.A.. Sadaguchi G. Jones W. 50. 1964. Part II Harvesting. Ericksen T.M. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. Presnell M. 20.E.S. Kelly C. Sakazaki R.H. Ness G. Kaysner C. during depuration. Public Health Service.. 1984.S.W. 1980. 69. 53. Journal of Food Protection.B.G. Rodrick G. and Manillal clams (Venerupis japonica). Food and Drug Administration.W. U. Roos B.. 989-1003.E. 1205-10. 25. Eds. 1490-5. Ltd. 52. U. 123-8.J. 343-9.. Takeda Y. Blake N. 1967. Temporal occurrence of Vibrio species and Aeromonas hydrophila in estuarine sediments.S. Food and Drug Administration. Hood M. seafood-handling procedures on Vibrio parahaemolyticus. 28. 1956... Saikon Publishing Co. The influence of time and temperature on the bacteriological quality of shell oysters during processing and shipping. Svenska Lakartodningen. Timetemperature effects on the bacteriological quality of stored shellfish. Vasconcelos G. Colburn K. 1214. 72 . Applied and Environmental Microbiology.C. Handling and Shipping Shellfish..A. Wekell M. Ruple A. I.
1969.D. Journal of Clinical Microbiology. Parasitenkunde. Journal of Bacteriology.. 2891-3. Abeyta C. 2. DePaola A. In vitro haemolytic characteristics of Vibrio parahaemolyticus: Its close correlation with human pathogenicity. Arcisz W.P. Kato T. 44.N. Joseph M. Geiger J...T.W.C... 1147-9. 53. Washington Report October 14. 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. 353-7. 1916. The bacterial content of market oysters. Crassostrea virginica. Obara Y. Bothell. 1976. 43. Yokohama Medical Bulletin.M. Abbott S.. Takeda Y. 133-47. 1997.E. 2820-2.. 985-93. Academic Press. Kaysner C. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States west coast. 36... Ishibashi M. 45.W. 1989. in Fish as Food. 1989.G.. Journal of Food Science. 160. Volume 1. Stroh E. Wekell M. 1937. Kelly M. 1914. Epidemiology and spectrum of Vibrio infections in a Chesapeake Bay community. Vibrio infections associated with raw oyster consumption in Florida. Applied Environmental Microbiology. 1961. Peeler R. Yamai S. 46. 13-7. Berry F. 47. Israel E. Geographic and monthly variation in composition of oysters. Stott R.S. 27. 100. Bacteriological control of oysters during processing and marketing.M. 1. Watsky D. Kaysner C.M.. 204-5. Sidwell V. Marine Fisheries Review. 97. Sample T. 313-22.. 33. 73 .. 25. Shellfish Specialist. Jacobson M. Akiyama S. Libonati J. Journal of Infectious Disease. Journal of Food Protection. 1958. 27. 38. U. Bacteriological examination of oysters stored at low temperatures.D.. Kelly C. Jr. Hlady W.A. Hoge C. 1997. 42. U.. Powers C. Jr.. Urease-positive. 40. 34. 41. 128-31. 1989. 37. Vibrio parahaemolyticus illnesses... Food and Drug Administration. 1987. 35.MOLLUSCAN SHELLFISH 32.. 60. Serological and hemolytic characteristics of Vibrio parahaemolyticus from marine sources. 41. (2)..A. kanagawa-negative Vibrio parahaemolyticus from patients and the environment in the Pacific Northwest.. Thompson C.. Loomis A..B. 273-80. 41. The bacterial flora of market oysters. Grodner R. Fieger E. Vanderzant C. Joseph S. Journal of Clinical Microbiology. Journal of Infectious Disease. Janda J. 69.. Takizawa K. 107-8. 39. Miyamoto Y. Infectionskrankheiten und Hygien.M. Takikawa I.L. Morris J. West Coast States and British Columbia.. Bacteriological findings in Baltimore oysters...F. Studies on pathogenic halophilic bacteria. 1926.. 1954. Borgstrom G. Public Health Reports. 38. Ward W. Zentralblatt für Bakteriologie. Novak A.A.M. 1979.L.S. Ed. Journal of Bacteriology. Leitch J. Microbiology of shellfish decomposition. Tanikawa E.A. Bulletin of the Johns Hopkins Hospital. New York.G. 1349-51...
91-103.I.R.T.R. Jacobs R.V...... Hebert S. Vibrio vulnificus: Man and the sea... Spite G.L.D. 1982.. 299-312... 1979.O. Annals of Internal Medicine. Black R. 51.M. Non-01 Vibrio cholerae bacteraemia: Case report and review. 1012-7. 1987. Rahman R.. Pons V...M. Jr. Tacket C. Leitch J..P. Journal of the American Medical Association.R..W..A. Applied and Environmental Microbiology.. 8. Jr.. 55.. New England Journal of Medicine... Kaper J.G.B. Blake P. Hug A. Siebeling. Puhr N. New York...... Lockman H.E. McCay C. Davis B.. Plaard R. Non-O group 1 Vibrio cholerae gastroenteritis in the United States: Clinical.. Journal of Food Protection..T. 656-8. Cholera .. Ecology. Applied and Environmental Microbiology.E. Applied and Environmental Microbiology. 54. Caraway C.A. 60. 802-6. Small P.G..M. Illness caused by Vibrio damsela and Vibrio hollisae. Journal of Infectious Disease. Riddle C. Feeley J.M.G.. Wekell M. 5972.. 1... 49.O.N. Roberts N.T. Motes M. Colwell R. Morris J. Gulf Coast..B. serology. Becker R. Allegra D.M. 275-83. 312. Blake P. Review of Infectious Disease. Hollis D. New England Journal of Medicine..A. Safrin S.. Morris J. Applied and Environmental Microbiology..M. 52.T. Twedt R. Presnell M. 10.. Blake P.. Incidence of Vibrio cholerae from estuaries of the United States West coast.W. 57. Weaver R. Duran W. Conte J. 53.M. Joseph S. West P.G. Miller H. 1983.A..R. 56.A..G.J.E. Twedt R. Morris J.J. Maden J. Stott R..W.. 45. Colwell R. Lee J. 1294-7.. Hunt J. 343-50. Clinical features and an epidemiological study of Vibrio vulnificus infections.. Jr.C.T. Kaysner C.. Adams M. 1981. McPhearson R.. Zywno S. 1982.J.G. McFarland L. sediment and waters of the U. Jr. Jr. 53.G. 74 ..A. Wilson T.. Cholera and other vibrioses in the United States. Bradford H. Snyder J. Blake P. in Vibrios in the Environment. Wilson R.G. Roderick G. Microbial Ecology. An epidemiological study of V.S.F.A. Brenner F. Non-01 Vibrio cholerae in shellfish.F. 1344-8. DePaola A. 305. Jr. Francis D. 37.A. 1980. Wathen H..A. John Wiley & Sons Inc. Morris J. 41. 1475-8.A possible endemic focus in the United States.M. 46.K..E.B. Characterisation of Vibrios cholerae isolated from oysters.. Wachsmuth I.. 1984. Bradford H. 1981. Ed. Vibrios in the Louisiana Gulf Coast environment.W.. 61. Abeyta C. 59... 1988.C.. 253... 558-61. Feldman R. Barrett T.. Lancet.FISH AND SEAFOOD 48.D. Hickman F.M. Tacket C. 149.. McFarland L. Kaper J. Johnston J. 1983. 1985. 94. DePaola A. R. and laboratory characteristics of sporadic cases. and enterotoxin production of Vibrio cholerae in Chesapeake Bay. 50. epidemiologic.. 2050-3.F.. Craig J. 305-9.O. Hug M. 58. 1984. Colwell R. Jr. Wazenski T. Becker S. Ecological relationship between Vibrio cholerae and planktonic crustacean copepods. Peeler J. cholerae in Louisiana. 1985..
S. Wilson T..G. Newell D. 6-9 September 1983. Eds.G. Feldman R. 65. 4. 607-9.. 1995.A. 1584. International Journal of Food Microbiology.G. EF-6.M. 1983.R. 257.F. Austin S. 323-5.. Journal of Rapid Methods and Automation in Microbiology. Moore J. Abeyta C. British Medical Journal. New York..M..J. a newly recognised Vibrio species. 1991. 13.. Brenner D. Wekell M.M. Helicobacters and Related Organisms.A. Huq M. coli and lari) from shellfish and the marine environment. in Campylobacters. Shandera W.G.G. Murphy P. Clements M.M. from patients with diarrhoea.A.. Alam A.MOLLUSCAN SHELLFISH 62. Huges T.B. New York.. Morris G. 80. Journal of Food Protection. 69.R. Isolation of vibrio-like group.E. 1988.M. Campylobacter gastroenteritis associated with raw clams. Ketley J. Blaser M.R. Abeyta C.. 1980. Jr... 1983. 1993. 75 ....A. S41. 4. 63. Abeyta C... 72. 282.A.X. Newell D.D. Stott R.R.A. Davis J. Journal of the Medical Society of New Jersey. 169-71. Arumugaswamy R. in Campylobacters... Levine M. Blaser M.M. Experimental Campylobacter jejuni infection in humans. 4. (jejuni. Wilson T.. in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections. 472-9. 66. Hughes T... 1980.A.G..K... Studies of Campylobacter jejuni infection in volunteers.K.M.J. 71. Wilson I. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N.A. 64. 1981. Wareing D. Kaysner C. Disease from infection with Vibrio mimicus. 70. Murphy P.M. 11.J. Proudford R. Infective dose of Campylobacter jejuni in milk.W. 621-4.P. 157.... Annals of Internal Medicine. Moore J. Journal of Infectious Disease. Jr. Ireland. Dalley E.A..M. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp.. 1996.. 73.. Ketley J.L. Feldman R.D. Eds. Microbial Ecology in Health and Disease.I... Wilson I. Plenum Press. 256. Brussels. 51-64. Wekell M. Levine M..G. Plenum Press. 1996. Johnston J.K.E. London. 67. 68.J. Jr.H. 56. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness.. Robinson D. 101-4. Eds.S. Journal of Clinical Microbiology.. Kaysner.. Hunt J. Helicobacters and Related Organisms. Kaysner C.. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster.R. Public Health Laboratory Service. Fitzpatrick M. Deeter F.L. Matsuda M. Jones D. A method for isolation of Campylobacter spp.. 1987. Wareing D.E.P. Skirrows M.. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation. 99.. Griffin M. C. Pearson A.E. Black R.. Blake P. Clements M. Rowe B. Davis B. infections in man. Black R.
. Jr.K. 84.A. Cook D. Annals of Internal Medicine.A. 690-4. Journal of Rapid Methods and Automation in Microbiology. 343-9.. Fla. 48. 979-85.. 1980.C. Kaysner C.M. and treatment of Plesiomonas shigelloides diarrhoea.. Infection and Immunity. Peterson D.. Fliermans C. 1978. 27. 1981. Kelly M. Cooper M. Plesiomonas enteric infections in the United States.. 82.. Bark D. 83. Herrington D. 849-55. Kain K.FISH AND SEAFOOD 74. Blake P. Seidler R. 7-12. 1986.P.J. Singelton F. Esch G. Kaper J. 1987.. Stelma G. Beaman J. Vandepitte J. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. Hunt J. Van Damme L. vibrios and Aeromonas hydrophila. 449-57. Lockman H. Abeyta C. 1997.. 1146-9.J. Wekell M. Hickman-Brenner F.. 1985. Journal of Food Protection... 81.M. 5. In vitro and in vivo pathogenicity of Plesiomonas shigelloides. 52. 731-8.J. 47. Applied and Environmental Microbiology. 75.C. Levine M. 39. Gracey M. Colwell R.L. Abeyta C. GA.. Petersburg....J. 77. Hood M.. 86. Trost P. The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish. Hirsch R. Tenge B. 1989. 79. 1988.W. Sullivan J. 598-601. 76 . Atlanta. 359-77. 52.M. III.. 78. Hazan T.. Journal of Food Protection. Wekell M..H..A. Jr.. Journal of Applied Bacteriology. 55. Clinical features. Abeyta C. Jr.A. Effects of processing and storing oyster meats on concentrations of indicator bacteria. Frequent isolation of Edwardsiella tarda and Plesiomonas shigelloides from healthy Zairese freshwater fish: A possible source of sporadic diarrhoea in the tropics. 1984.B.W. Koburger J.L. 1990. Stott R.M. 80. Applied and Environmental Microbiology.L. Department of Health and Human Services/Public Health Service/Food and Drug Administration. Journal of Clinical Microbiology. Journal of Food Protection.. 1989.. 475-9. Robinson J.R. 47.A.N.D...F. 105. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness. Applied and Environmental Microbiology. 36. Kaysner C...A.. Burke V. Miller M. Robins-Browne R. Holmerg S.M.. Wachsmuth I.. 998-1001.W. 1984. Prevalence and distribution of Aeromonas hydrophila in the United States.A. Report (1/13/84). Incidence of motile aeromonads from United States west coast shellfish growing estuaries. Journal of Food Protection. 53. Baker R. 50... Ruple A.... Kaysner C.. Aeromonas hydrophila: ecology and toxigenicity of isolates from an estuary. Wekell M.A. Plesiomonas shigelloides: An opportunistic food and waterborne pathogen.M.. Tall B.. Journal of Food Protection. Partridge K.R. epidemiology.D. 87. 85. Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species. Tzipori S. 76. Oyster borne disease outbreaks in St.M.T. Farmer J.D. Herrington T. 223-47..
M. mussels. 2000. U. in U.030.12)..S. Inc.1987. 7.E. Lund B. Food and Drug Administration. 89. 3..C. Baird-Parker T..S. 1995. Burdge D.Pathogen monitoring of selected high risk foods (FY 88/89).C.600 (CPG 7119..S. Slavin J. U.S.S. 91. 1991. Government Printing Office . and HACCP: A Global Focus. Food and Drug Administration. Ed. 90.. Ed. Washington D. U. 560. U..L.R. National Academy Press.W.8 Further Reading Ahmed F. Clams. Food and Drug Administration.MOLLUSCAN SHELLFISH 88. Smith J. Food and Drug Administration. Collette R. Aspen Publishers Inc. Chapter 5: Foods. 63-8. Martin R. Food and Drug Administration Compliance Policy Guides. U. Lancaster.C. Food and Drug Administration Compliance Program Guidance Manual.S. Fish Inspection.S.. Food and Drug Administration. Part I Sanitation of Shellfish Growing Areas.H.S. Technomic Publishing Co. Quality Control. The Microbiological Safety and Quality of Food.A. Food and Drug Administration. Washington D. in National Shellfish Sanitation Program Manual of Operations. Sherlock C. 1997.C. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. U. Food and Drug Administration. 77 .W.. Food and Drug Administration. Washington D. 1995. Gould G. 1987. Ed.. oysters. Program 7303. Gaithersburg.E. in U. fresh or frozenadulteration by bacteriological contamination Sec. Seafood Safety. Pittsurgh.
e. and available in 0. packed and frozen in 0.g.Ph. Such products are usually only partially cooked before freezing. lobsters.5 kilo pack (boxes or bags). jointed limbs and a chitinous exoskeleton.4. Crayfish. crayfish and shrimp (1). Peeled and cooked shrimp are ready-to-eat after controlled thawing.5 and 1.. Shrimps.. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). peeled product. and frozen in 1 and 2.0 kg polythene bags. Pandalus borealis (deep-water shrimp). Pandalus montagui and Crangon crangon (inshore shrimp).0 kg polythene bags.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda.. Species eaten from tropical waters include Penaeus spp. crabs. the front pair ends in claws. with shells and heads on or off are usually packed. They possess a segmented body. 79 . Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. CRUSTACEAN SHELLFISH Linda Nicolaides.5 and 1. M. with additional water to form a glaze. and Trachypenaeus spp. and are usually aquatic. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. (deep-water shrimp). Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. Parapeneopsis spp. lobsters and crabs have ten legs. Peeled ready-to-cook shrimp are raw.
The structure of the circulatory system of crabs is not closed. packed into polythene bags. In comparison. Cooked and chilled. it is important that shrimp are handled and iced correctly in order to keep them in as fresh and uncontaminated a state as possible. particularly after it has been cooked. Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. Following capture or harvest. However. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. spoilage bacteria are of little importance as. shrimp die immediately after capture. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. boiled to kill the crustaceans and cook their meat prior to consumption. As with finfish. particularly for members of the genus Vibrio (2). stored and transported as frozen product. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp. e. For example.. or frozen crab. If these products are subjected to temperature abuse.g. e. Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP). the geography of the area they are captured from or are grown in. for crabs and lobsters. the season. storage above the recommended storage temperature. during the peeling of cooked shrimp. with the increasing demand for ready-to-eat/cook crustacean products.g. 4. and the temperature and quality of the waters in which they exist. Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance.FISH AND SEAFOOD Crabs. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors. which include the environment from which they have been captured or harvested. hence. which means that the haemolymph can be a reservoir of bacteria. the potential exists for crosscontamination from both the processor and from the environment. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria. only live carcasses are processed. lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. lobster tails and cooked crabmeat presented decoratively in their shells. so that spoilage may occur during landing and transportation to the processing plant. Shewanella putrefaciens and 80 . Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function. Pasteurised crabmeat is edible meat removed from the exoskeleton by hand. traditionally. tropical waters.2 Initial Microflora As with finfish. their feeding and living habits.
chilled 81 . After capture and transport to the processing plant. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected. with “head on” or “head off” whereby the edible tail is removed from the head. from the time of capture and processing. glazed and frozen (see Figure 4. The claws are usually bound together to prevent the animals fighting and causing damage to each other. Pseudomonas spp. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. They are then taken to market alive in either holding tanks. Alcaligenes.1). or with the shell and central vein removed. A large proportion of shrimps are produced by aquaculture. or cooked. In contrast. and Achromobacter spp. graded according to size. uncooked prawns and shrimp is presented in Figure 4. lobsters and crayfish are harvested from cages or pots containing bait.1.for example. whether GHP are in place. the genera associated with tropical species include Enterobacteriaceae. so they are either sold on the local market or are processed in local processing plants.. Hence. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . In comparison shrimps die quickly after capture. corynebacteria. Micrococcus spp. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. however. Shrimp might be further processed with the shell on.10 days for temperate water species (3).. Shrimps can be processed whole.3 Processing and its Effects on the Microflora Crabs. they are washed. or transported directly to processing plants.3. 4. The rate of spoilage will be a function of time and temperature. These animals can survive out of water if the surrounding environment is kept cool and damp. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. When the shrimp arrive at the processing plant they are washed and then sorted into size. bacterial contamination and spoilage are inevitable.1 Prawns and shrimps An example of the process used for preparing frozen. especially if they are stored in ice or iced water. the rate of deterioration of the crustacean shellfish by these organisms is reduced.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots. chilled sea water. thus. 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. gills and thorax. Vibrio spp.. peeled and cooked. Shrimp will then be glazed and frozen in a raw state. or are found in brackish water around the shores. 4. up to 16 days for tropical species compared with 8 . As with finfish.
Flow diagram . and packed to meet the customer’s specification. 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. 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.frozen uncooked prawns 82 . Good Manufacturing Practice (GMP).1. Fig.FISH AND SEAFOOD frozen. Breaded products introduce an added risk of spore forming bacteria. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour. spices and cooking batter. After cooking.
The shells have a crisp. and have a sweetish. there is no indication of life.4.4 Spoilage The main genera of Gram-negative bacteria responsible for the spoilage of crustacean shellfish are the same as those that cause spoilage in finfish. indicative of protein degradation and the growth of the principal spoilage bacterium Sh.3. If the tail drops. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. bright and dry appearance. since crabs and lobsters are traditionally consumed immediately after cooking. viruses or parasites. putrefaciens. contaminating microflora may include enteric bacteria. Heat is produced if the shrimps decompose in a confined space. 4. The shell will be soft. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. the joint between the tail and carapace is open and the limbs hang loosely. In comparison. for crabs. 4. slightly iodine smell. the meat is either repacked and sold in the shells.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. immediately prior to boiling. Once removed and cleaned. and a strong ammoniacal odour is produced. they include Sh.1 Prawns and shrimps Freshness indicators for prawns and shrimps are for the shells to be crisp and dry and feel sharp and cool to the touch.4. or in hermatically sealed cans that have undergone an appropriate heat treatment. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. or packaged and processed. dull and sticky. with the growing demand for ready-to-eat products from crab and lobster meat. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. If hygienic procedures are not followed. If boiled. there is no associated spoilage problem. bacteria of public health significance. With spoiled crabs and lobsters. 4. 83 . However. whilst boiled crustaceans have a pleasant slightly sweet smell. An offensive odour is also produced. wet and ‘soapy’ or ‘jammy’ to the touch.CRUSTACEAN SHELLFISH 4. putrefaciens and Pseudomonas spp. spoiling prawns and shrimps have shells that are soft.2 Cooked crabs and lobsters As indicated earlier.
The presence of L. Moreover. Although foodborne illness had not been associated with the product. monocytogenes was absent from 40 samples of cooked prawns. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). Hence. 5. The above results prompted the FDA to develop a compliance programme for the pathogen.5% of frozen raw shrimps from different countries (13). monocytogenes in cooked ready-to-eat foods. animal manure. as well as for Salmonella. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). in domestic/imported shrimps (15). and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14.5. Following this.FISH AND SEAFOOD 4. in 24.5. a common pond fertiliser used in aquaculture. Further surveys identified Listeria spp. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . Salmonella cannot be completely removed from raw shrimp during processing. although efficient cooking will quickly eliminate it (6). Aquatic birds and the hands of workers have also been identified as sources of the pathogen. shrimp and shellfish (11). 6. 4.cooked prior to consumption. 16). monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). shrimps and cockles sold (17).2 Listeria spp. and usually for both. even if GMP is followed. fish fingers. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. 7). They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. Salmonella species are more commonly found in crustaceans reared by aquaculture or growing in faecally contaminated brackish water compared with species of marine origin (4.1 Pathogens: Growth and Survival Salmonella spp. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. An import alert was put in place in June 1987. However.5 4. later surveys in 1990 recovered the pathogen from retail raw. cooked and smoked fish. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. nearly 30 years elapsed before the potential hazard of L.
g. Crabs are covered in a chitin exoskeleton. Researchers have demonstrated that inoculated shrimp supported toxin production by C. 4. V. aureus is not a member of the intrinsic flora associated with crustacean shellfish. is widely distributed in aquatic environments. Crustaceans have been linked with food intoxications attributed to Staph. 4. therefore. 24.5. 4. To date.70% of healthy individuals carry toxin-producing strains of Staph. Vibrio cholerae. botulinum type E at 10 °C but not at 4 °C (20). about 50 . cholerae is divided into two groups. which can become colonised by V. cholerae non-O1. pasteurisation and cooking. The former. whilst boils and cuts are also common sources of the pathogen. 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. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. particularly crabmeat (18. The main source of contamination is the hands of processing personnel. cholerae. 22) as well as from undercooked crustacea when harvested from contaminated waters (23. Once the competing microflora has been removed by a heat treatment. 19). so is able to survive heat treatments that eliminate vegetative cells. although contamination by types A and B may occur from the environment. especially in products that have been vacuum packaged.3 Staphylococcus aureus Staph.CRUSTACEAN SHELLFISH old. monocytogenes has been attributed to crustaceans. was associated with seafood consumption. pregnant women and the immuno-compromised. Control should be achieved by promoting GHP and staff training programmes. However. the causative agent of cholera. the pathogen can grow at temperatures as low as 3 °C. botulinum type E is commonly associated with fish and fishery products.5. as part of the natural bacterial flora. 25).5. combined with the degree of handling of these types of product. aureus because of the ubiquity of this organism in the processing environment. e. V. no incidence of foodborne illness caused by L. An epidemic that began in South America in 1991. cholerae O1 and V. aureus on their hands. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. The pathogen has resistant spores.4 Clostridium botulinum C.5 Vibrio spp. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. 85 .
the potential hazard exists as C. vulnificus is an invasive and lethal pathogen associated with wound infections. Quantitative levels were below limits of detection in all cases (<0. However. non-B Hepatitis viruses (31). However.5. Japan.5. whilst 5. hydrophila and A. 4. It is more commonly implicated in countries where crustacean shellfish are eaten raw. 4. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. 31.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. Hepatitis A virus presents the most serious hazard. in a survey carried out in the United States between 1977 and 86 . hydrophila is able to grow well at refrigeration temperatures (0 . although A. Parasitic infections are controlled by either cooking infected material prior to consumption. particularly trematodes (6). a marine halophile. Norovirus. but has the potential to cause fatal foodborne illness (26). coli. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat.7 Aeromonas hydrophila and Aeromonas sobria A. e. Caliciviruses.8% of samples contained C. occurs in crustaceans from warm environments. Astroviruses and non-A. however.5. to date. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. South America. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). V. Of these. 32). been implicated in a case of foodborne infection associated with crustaceans.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. 4. 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.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. A. parahaemolyticus. hydrophila has been isolated from prawns (29). or freezing at a temperature of -23 °C for 7 days (30.30 MPN/g) (27).8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans.g.5. V. 4.6 Campylobacter jejuni and Campylobacter coli C. jejuni has not.
47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). No foodborne incidents have been attributed to this source to date. 4. They can survive low pH levels. Selected parts from this regulation is presented below. consistent with that in the stomach. The risk of such hazards is far higher with fin fish and molluscan shellfish.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. which could be transferred to the meat during unskilled removal from the shell (35).CRUSTACEAN SHELLFISH 1981. from farm to fork (37. Provisions laid down concerning the use of additives would also apply to these types of products. which results in a persistent and apparently permanent loss of short-term memory (36). 4.5.11 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. 39). 38. or chlorine concentrations similar to those found in drinking water. 4. 87 .5. risk-based system based upon the seven principles of Hazard Analysis and Critical Control Point (HACCP) supported by pre-requisite or good practices programmes at all stages of the food chain.6. produced or imported into the European Union (EU). are included in the Commission Regulation (EC) No. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34).1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. A summary of microbiological criteria for crustacean shellfish is listed below. only 0. although it is one of the European Union (EU) legislated allergens. The toxin causes amnesic shellfish poisoning (ASP) when consumed. 2073/2005. 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.
FISH AND SEAFOOD
220.127.116.11 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 18.104.22.168.1 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. 22.214.171.124.2 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. 126.96.36.199 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.
This can be called ‘kench’ curing and is mainly used for non-fatty white fish. CURED. Fish curing is practised worldwide. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. It includes the salting. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. Salting is both a method of preserving fish. and before significant spoilage takes place. fish is immersed for a few hours in a relatively low concentration brine. 93 . split and opened out flat. rather than for preservation itself.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. where the salt replaces moisture in the fish flesh. drying and smoking processes. Wet salting or brining. both to preserve and to create a variety of new products. SMOKED AND DRIED FISH Dr. alternating fish layers with a layer of salt. dependent on the country or the type of product.5. Usually. and marinating processes. The processes take anywhere from a few hours to a few days or even months. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. The fish is headed. and the extracted moisture is allowed to drain away. and salting is done by stacking the fish in layers. Fish can be salted in three different ways or their combination: Dry salting. and a preliminary operation to some smoking. their combinations and/or variations. drying. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. and is sometimes known as ‘pickling’.
spices). Traditionally. is used as a preservative instead of salt. evaporated water is removed by vacuum pump. as concentrated brines are formed. Marinating or pickling. The products are usually consumed raw. such as ‘gravad’ fish. After the freezedrying process. Thus. It has found limited application with fish. are added to the marinade. Vacuum drying. Longer shelf life is achieved in the products when salt is added. usually vinegar. The product is dried by contact with heated surfaces or by radiation. and the fish can be immersed in such brines for long periods. Heat is transferred to fish from heated air or a heated surface. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. Food products can be dried in three different ways: Air or contact drying. Usually. Included in this group are some lightly preserved products where no acid is added to the marinade. without other preservation methods applied after boiling. This is a mixture of a dry and a wet salting process. Fish in contact with refrigerated plates freeze. depending on the type of product. under normal pressure. Smoking is an ancient process for preserving and flavouring food. where an organic acid. as well as other flavourings (sugar. Marinades are acidified brines. As the water is drawn out of the flesh by the salt a brine is formed. It is not commonly used for fish products. or boiling it with dry salt and water. thus delaying spoilage for a few days. the product rehydrates quickly but does not retain reabsorbed moisture on cooking.FISH AND SEAFOOD Dry / wet salting. This is the common method of drying fish. Salted-boiled fish are the products obtained on either boiling fish in a brine. this was done naturally by exposure of the fish to the air. Freeze drying. The original principle of preservation was due to a combination of lowered water activity of the product and the uptake by the product of bactericidal and antioxidant components 94 . Salt. this ‘brine’ can be collected and placed again over the fish several times. prior to or during the boiling process. thus making the product unsuitable for microbial growth. When the process is running in a closed system. the preservative effect is due to the low pH used in the process. and the air movement removes the vapour. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms. The fish is mixed with dry salt but in a watertight container. Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. it can also be named ‘pickling’ or ‘blood pickling’. thus reducing the water activity of the product. the ice sublimes and the vapour is removed from the fish by vacuum pumps. Salt-boiling.
It can be smoked either traditionally. After the salting step. The smoke itself may contain spores of moulds. which impart the flavour of smoke to fish. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. or by means of smoke flavouring processes. SMOKED AND DRIED FISH of smoke.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. by exposure to the wood or sawdust smoke. sporeforming bacteria or osmophilic moulds. The secondary objective is to impart sensory characteristics such as smoke colour and smoke flavour to the product. 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. temperatures below 30 °C are used. Hot-smoking. The different spices used in the immersion brines are also a potential source of microorganisms. Also. Smoked fish is prepared by subjecting the fish to the direct action of smoke from wood or wood sawdust burning. or added to the brine with other flavourings. leaving only elevated levels of carbon monoxide. such as injection or dipping in a solution of liquid smoke. including pathogens. Clear smoke. In this process. 95 . the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. With the exception of some Asian countries. or by means of smoke-flavouring processes with liquid smoke. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke. and filtered to remove tars and particulate matter. Smoked-flavoured fish. It is commonly used in place of traditional smoke. Usually. and the preservative effect is only slight. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. It can be injected into the product. Usually. or liquid smoke. Smoking can be either traditional. The process was designed so that the smoke is filtered to remove all smoke particles. the fish is subjected to heat for a period that does not coagulate protein. the salt used in the processes may contain halophilic bacteria.CURED. However. In this process. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour. Cold-smoking. process temperatures are over 60 °C. by exposure to the wood or sawdust smoke. fish can be smoked by means other than the direct action of smoke. smoking is now used mainly to give the fish a characteristic flavour. 5.
% 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. processes and producers’ private recipes.4 52.6 132.4% salt (Table 5. Different brine strengths are required according to the curing processes and/or the type of product. or salt in the water phase. only the most common processes can be described here.6 264. At 18 °C a 100º brine is produced when the solution contains 26. This should be distinguished from the salt content. When a milder brine.I). Therefore. Salt concentration means the percentage of salt in the water contained in the fish. it causes swelling because the fish absorbs water from the salt solution (1). about 50º (132.4 184. sometimes the salt crystallises onto the surface of the fish. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. a 90º or above brine (237.2 g of salt per litre of brine).0 g of salt per litre of brine). it is impossible to establish standardised processing stages in the curing of fish. the fish is usually salted with a 70 . When working with salting.0 158. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish. TABLE 5. Thus.FISH AND SEAFOOD 5. a 0º brine saturation means pure water.0 .3 Processing and its Effects on the Microflora Owing to the wide variety of products.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. A 100º brine solution means that the solution is saturated and no more salt can be dissolved.8 211.2 105. and salt content. For smoking salmon. etc.8 79.80º brine (184. it is important to understand the concept of salt concentration. Salt in the water phase links the two conditions of moisture and saltiness. all using salting steps.6 g of salt or more per litre of brine) may cause salt burn in this way. The dissolution of salt in water is dependent on the temperature.2 237. is employed. smoking or marinating processes. When a 100º brine is used. The strength of the solution is described as degrees of saturation (brineometer degrees).8 211.
1 Dry salting / Kench curing Usually. it tends to lead to a slight yellowing and softening of the product. if very pure sodium chloride is used for curing purposes.3. when the air humidity is high. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). having a short section at the end of the tail. producing characteristic pink patches. lean white fish is used for this purpose. ‘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. caught in North Atlantic waters. addition of levels of 0. are likely to cause hydroscopicity of the product and. Salting can be done in a number of different ways. See Figure 5. The salt used should be of food-grade quality. When pure salt is used. Usually it is followed by other preservation techniques such as drying and/or smoking. ‘Green cured’ split cod with about 60% of the backbone removed. This condition is known as ‘putty fish’ (4).1 Salting Salting of fish for preservation purposes has been used since pre-history.0. the fish is restacked with fresh salt after about a week.35% and 0. 97 . The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. SMOKED AND DRIED FISH 5. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. respectively. and essentially free from iron and copper (3). The calcium and magnesium impurities. However.15% calcium and magnesium.1 ppm and 10 ppm. and the salt has penetrated throughout the fish (3). mainly in tropical conditions. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. and a ‘putty’ fish product is also obtained (1). A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. On the other hand. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. the fish is mixed with one part of salt per eight parts fish. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. For a light salting. 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.1. can be dried further to produce ‘klipfish’ or ‘bacalão’.CURED. Also. 5. outgrowth of halophilic bacteria may occur. and salt can speed this oxidation process. but the concentration of copper and iron must be kept under 0. are recommended to help make the product whiter. low in calcium and magnesium. for which there is a high demand in Mediterranean countries. however. which include calcium and magnesium. This is because fatty fish turn rancid quickly. respectively (1). and for a heavy salting a ratio of 1:3 salt:fish is used (1).1. The most common fish used in the northern hemisphere is cod (Gadus morhua). and such delays.15 .3.
it appears that water activities much less than 0. Processing of dried salted cod 5. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole.2 Wet salting or brining As referred to above.1. For large fish it is advisable to split it before salting. although toxin production would be unlikely (4). such as Staphylococcus aureus could survive and grow at such water activities in the product. eviscerated and/or split).5.1.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. such as 98 . The greatest barriers to salt penetration are the skin and scales.90 are rarely achieved.3. Where fish are salted in their whole form. Fig. Food-poisoning organisms. the process is mainly for flavouring purposes prior to the use of other preservation processes. so that the risk of spoilage is high. when a light brine is used. Only small fish can be salted whole.
The fish are kept immersed in their own brine near saturation point for several days. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.09 8.95 3.26 3.61 3.3 days.80 5. Fresh sardines. a saturated brine is used. the brine is then replaced over the fish.CURED.23 2.17 4.65 2. thus.44 2.0 4.61 3.25 5.00 3.85 3.44 2.11 3.86 2.76 99 .88 5. and a red burgundy colour will develop in the product.85 3.1.51 5.45 3. the liquid that comes out of the fish forms a brine. limiting the amount of rancidity that can develop.22 6.26 4. TABLE 5.11 4. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.35 4. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .03 4. with a shelf life of about 4 weeks (water activity (aw) about 0. Anchovies are the main fish used for these purposes.II.19 3. herrings and other fatty fish can be used.16 7.41 6.30 2. the fish enzymes play an important role by maturing or ripening the product.90 (1).66 5. but mackerel. after being drained and pressed at 465 kg/m2 for 15 hours.5 10.78 6.99 2.60 2.41 3.76 4.3.5 4.49 4. the fish is dry-salted.3 Dry / wet salting In this process.05 7.46 4.78) (1). This is why it is the preferred method for fatty fish.98 6. if split fish is dry-salted.23 2.98 5. Here.96 1.41 4.70 6. Histamine formation is of importance when using scombroid and some clupeidean species.41 5. sardines.03 2. SMOKED AND DRIED FISH smoking.73 1.78 2. headed and gutted. but as it is placed in containers.26 7.66 5. with pressure being applied to the top of the container to help the brine to drain away.76 4.26 1.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) 2.91 2.33 5.17 5. they can be stored at 28 °C.70 3.54 5.74 9.56 6.23 8.11 9.26 2.06 4.0 3.5 3. There are several variations of this process. For example.64 4. Histamineforming bacteria do not generally grow at water activities below 0. The main objective of the brining method of salting fish is to limit air penetration during processing.25 5.10 1.04 5. See Table 5.09 8.98 6. and can be collected and replaced again over the fish.23 3.51 3.76 4. For preservation purposes.47 6. the product obtained is called gaspé-cured (5).84 1. can be cured in a saturated brine solution for 6 days and.5 3.76 4. 5.76 4.0 2.
The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. and even death. 5. because of the salt content of the product. Another salt-boiled Indonesian product. Other preservation techniques can be used for extending the shelf life of these products. but it has further smoking. and packed (3). the occasional implication of pindang in cases of foodborne illness. drying and salt-boiling steps until a hard translucent block is obtained (4).5% 120grain distilled vinegar. containing 1 kg of salt for every 25 kg of fish. The first process can give a longer shelf life. up to 1 . salt.3.2% acetic acid and 2 . A much firmer product is obtained when the fish is salted prior to acidification.4% of salt) for distribution and sale (1). and 6% salt. ‘ikan kaju’. All these products are ‘semi-preserves’. or a pH <5 and they are stabilised for a limited period.6 days without refrigeration (1). The fish can be soaked in a high concentration of vinegar (about 4 . then packed in barrels with a 70º brine. usually flavoured with onions and hot chillies. As referred to above. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. and lasts for 3 . has a shelf life of several years.3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. they are packed for consumption within 2 weeks at chill temperature (6). and spices) is added to the brine. until the flesh is soft. a Peruvian product. underlines the need for careful control (4). outgrowth of bacterial spores after processing may occur (4). Afterwards. In a second step. Usually. herring or other fatty species are used. even if the product is hermetically sealed. flavouring (sugar.3. However. Once cooled. After this. they can be cut and cured for 3 days in a solution with 3% white distilled vinegar.30 minutes.7%) for as long as 3 weeks. is made of different varieties of fish. which is dipped into a boiling brine for 10 . The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. The lime juice plays the role of the vinegar. it can be repacked in milder brine (1 . Herring can also be cured for several days in an 80 to 90º brine with 2. Another variation uses sour cream instead of the second brine step. giving a cooked appearance to the product.2 months. placed in a marinade of lime juice. whereas the latter method preserves the fish for only a few days. or can be boiled by being placed in a basket. Ceviche. There is epidemiological evidence that these products have been the cause of some 100 . Traditional jellied eels are a product where pieces of fish are cooked in boiling water. They will have a salt content >6% (w/w) in the water phase. usually requiring refrigeration below 10 °C for longer-term storage. only the nonspore-forming organisms are eliminated in the process and.FISH AND SEAFOOD 5.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour.
It takes about 2 days to marinate completely. Shark pectoral fins. Dried shrimps are widely used in Asia. and the stacks are placed in the refrigerator. 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. Stockfish is extensively used in Portugal. In some coastal areas of Portugal. the temperature and the amount of pressure applied (5). after frying. Other products. used as an appetiser. mixed with salt. 5. etc. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. jellyfish. air flows naturally. and a fine product is obtained. while in the latter case there is a provision for fans or other means of mechanically moving the air (1).0) (7). Usually. Matjes are made of fat young herring. Dried octopus is also a very common dish in the interior of the country. depending on the thickness. are likely to survive in these mild-processed products. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. In Scandinavian countries. gutted. Stockfish (dressed. 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). Fresh dill is placed between the layers of fish. Italy and Spain (5).5. gravad fish. Foodborne pathogens. are sundried and. sea cucumbers and seaweeds. a very fine delicacy for Oriental food lovers. Gravad fish is a Scandinavian product made of salmon fillets. It is important to assure the quality of the raw material. split and dried gadoid species) are traditionally made in cold climates. under a light pressure. the fish is dried naturally in a relatively cold environment. Provided the product is kept in dry and fairly cool conditions. it will last for many years (5). such as squid. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). are commonly dried in Asian countries. by using either solar heaters or mechanical dryers. still made without prior salting on a large scale. sugar and spices. In the first case. South America and the Caribbean. some fish species such as moray eel and varieties of dog fish. At pH levels below 4. are generally air-dried at sea. Some bacteria and enzymes are still active in the presence of acid and salt.CURED. and the flesh will break down.4 Air drying This is probably the oldest method of fish preservation.There are several products named marinades (matjes. The most common method used for fish products is the air-drying process. They are an important dried product. the fish is dried after salting or brining.) where no acid is added to the process. 101 . Stockfish products have a golden brown colour and are very hard. but sometimes it is dried without prior salting. such as Listeria monocytogenes and Clostridium botulinum. bacterial toxins and parasites (7). marinated products will keep for several months at a temperature of 4 °C.3. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. which are placed in a mixture of salt.
the product can be held for months at room temperature. When no more water can be removed from the fish.the lower the humidity. In conclusion. or wooden drying rack to dry in the sun. the protein must be set or denatured with low-temperature drying before applying higher temperatures. They must be soaked in water. The product is dried to a final water content of about 25 to 38% (3). 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. France.83 (9)).g. there is an increasing demand from people that have emigrated to industrialised nations for traditional products. botulinum growth might occur in the bulk of the product. that may have contaminated the product during the drying process.FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. Whilst the low aw of the final product will inhibit microbial growth. To prevent this. 102 . and increasing amounts of such products are making their way into the market place. and it is exported to Portugal. Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s. some of which are listed in Table 5. aureus) or spores (e. Spain. It is important to balance the drying rates and quality/safety issues. Thus. Staph. making a water barrier and consequently. The fish must be stored in reasonably dry conditions to avoid moisture absorption. but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). the skin will form a hard case (case-hardening). which leads to mould growth (aflatoxins. Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. the fish must be dried slowly enough to avoid case-hardening. neither it. In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. Iceland and Canada. and has the primary objective of creating a product with specific taste and textural characteristics.75 (1). Italy and South America. the better the moisture transfer. by warm air circulating in a drying chamber. the diffusion of the bulk water to the surface of the fish is blocked. However. dried salted cod (bacalão) is now mainly produced in Norway. skin up. A highly demanded dried fish product in Mediterranean countries is dried salted cod. if the rate of water removal is too high in comparison with the diffusion rate. Dried salted products must have a water activity below 0. more commonly nowadays. After dry salting. C. the quality is more difficult to control. tarpaulin. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain.g. Whilst the production of dried fish products is tightly controlled in industrialised countries. nor subsequent cooking operations will affect the presence of toxins (e. typically in stews and soups. Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources. botulinum).III. The amount of moisture transfer from the fish to the air is dependent on the relative humidity . and the water must be changed several times before cooking. toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0. spoilage and possibly C. the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or.
However. making full description impossible here. Maintaining the temperatures of pyrolysis between 425 and 200 °C. The smoking procedures vary widely. mussels and many others from tropical countries. Aroma and flavour are a blend of smoke components (3). clams. In the mechanical kiln. The quality of the smoke is related to the type of wood used. as referred to above. cod roes. Hardwoods are considered better because of the milder flavour they impart to the food. Top-quality fish must always be used. tuna.salmon. trout. white mangrove and abura has an inhibitory effect on some microorganisms. haddock. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish.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. aureus and Saccharomyces cerevisiae (10). botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. swordfish. eel. sprats. using electrostatic filtration of the smoke. There are many fish species that are used for smoking purposes . Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . It has been reported that the smoke from mahogany. cod. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10). SMOKED AND DRIED FISH TABLE 5. Blends of various woods can be formulated to produce sawdust with specific flavours. It can be used for cold or hot smoking. 5. oysters. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns. herring.CURED. and the use of smoke generated by superheated steam or liquid smoke. Staph. is mainly a process for flavouring fish rather than a preservation technique. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. mackerel. such as Escherichia coli. softwood colours the product quickly.5 Smoking Smoking today.3. aureus (toxin) C.
Drying should be minimal at the start. temperatures must be kept under 30 °C throughout the process. The fish can be dry-salted using a weight of salt 5 . but a ‘safety’ (i. covering the fish surface completely (10). Cod roes. Wet salting or brining is preferable to dry-salting when smoking fillets because. Also. wooden boxes or master cartons (10). Depending on the type of product. as well as many other products prepared in the same way. skin off or on. When frozen/thawed fish is used. the following step is salting. the excess salt is removed with water. the highly desirable cold-smoked salmon. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. If frozen fish is used. it will reduce the salty flavour. are boiled before smoking (12). Some herring products. which is important for the appearance of these products (4). low temperatures will slow the uptake of salt. skin on or off).and hot-smoking is the temperature of the process. spices. The maintenance of temperatures below 30 °C avoids protein coagulation. shrink packaged. The length of the smoking process for such products is much greater than for the hotsmoked fish. and the cold smoke is denser. tuna and swordfish. but it will increase towards the end of the process (10). such as smoked salmon-trout. Thus.FISH AND SEAFOOD water flow system with chlorinated water (25 . gutted or filleted. thus allowing the deposit of smoke on the fish surface. Whether the product is prepared headed. however. whether cold. Sugar can be added together with the salt. but this concentration will again depend on the type of product and the length of the brining stage. The salting period is dependent on the size of the fish and its presentation (whole.1 Cold-smoking The major difference between cold. smoked cod fillets and finnan haddock are usually cooked before consumption.50 ppm) should always be done just before process initiation (3). usually dry-salted and cold-smoked.3. giving the product the desired appearance and flavour (4). Sugar. 104 . pasteurisation) temperature is not achieved in any stage of the process. 5. 70 – 80 º brine is commonly used.18 hours at 4 °C.10% of the weight of the fish. or packed in retortable foil pouches. are not cooked prior to consumption. owing to the elution of soluble proteins. it will take up salt twice as fast as fresh fish (10). In cold-smoking. thawing must be done slowly for 12 . If wet salting is chosen. canned or bottled. they can be vacuum packaged. all cold-smoked products should be cooked before consumption. filleted. This difference is crucial for safety issues because the flesh is not cooked. All the smoked products. Brining at 10 15 °C is recommended (10). permitted colourings or smoke flavours can also be added to the product in the brining step. after being smoked need to be chilled immediately to 4 °C before being packed. it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11).5. However. Thus.or hot-smoked. as it retains more moisture. an attractive glossy pellicle on the fish fillet surface is achieved after draining. or quick-frozen and stored at -18 °C (2).e.
such as Yersinia enterocolitica. secondly. a final cooking period at 80 °C (12). there is no point in the process that can fully assure the absence of L.5. Smoked products can be also packed under controlled-atmosphere packaging. Although other human pathogens that can grow at refrigeration temperature. gravad salmon (also called gravad lax or gravlax). stored at 4 °C and packed. 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. in order to produce toxin. Length of the smoking process at each stage will depend on the species and the type of product. can be isolated from seafood. The exception is vacuum packed fish. and their sensitivity to salt (13). the products are cooled. refrigerated storage and vacuum packaging still allow its growth (12). a drying period at 30 °C.3.2. can grow with salt levels of 10% in the water phase. During smoking.CURED. as. but they should be cooled to a temperature of 10 °C within 3 hours. Pathogenic vibrios. but the proteolytic types A and B. are of concern when smoking fish from tropical waters (13). botulinum type E present an even longer time than before to grow and produce toxin (14). The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. Neither the smoking temperature nor the salt content is enough to kill the organism. further cooled to 3 °C within 12 hours and then packed. botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. A typical temperature set is as follows: firstly. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). and hotsmoked salmon (kippered) (3). Afterwards. if they are taken from a chill room at 3 °C and packed. if present. If the refrigeration chain is maintained.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. typically mesophilic. This temperature should be maintained during storage and distribution (3). which underlines the danger of storage at abuse temperatures of >10 °C (13). Vacuum packing is recommended only for coldsmoked salmon. Hot-smoked products require a longer cooling stage. See Figure 5. a partial cooking period at 50 °C. On the other hand. 5. To avoid growth of non-proteolytic C. thirdly. and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). if a 105 . thus allowing any psychrotrophic C. SMOKED AND DRIED FISH During cold-smoking. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). Furthermore. Smoked salmon fillets are usually vacuum packed whole or after slicing. These products should be vacuum packed at room temperature. monocytogenes.
FISH AND SEAFOOD product is packed too warm without being cooled to room temperature. Fig. 5. it will have a reduced shelf life. and moulds will grow on the product (12). Process of cold-smoked sliced salmon 106 .2.
(Penicillum cyclopium and Penicillium puterellii) (19). Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). cooking in brine. a very expensive and fashionable smoked product in the Netherlands. fatty fish will remain in good condition for 6 days. Aspergillus ostianus. 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). they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). but mechanical drying is used instead of sun drying. are used. Eels. to have no further advantage over other packing methods for other fish (12). SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. In terms of safety. white fish about 8 days. 18). Aspergillus tamarii) and several Penicillium spp.3.CURED. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. Vacuum packaging retards the onset of rancidity when fatty fish is used.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. but seems. The classical liquid smoke is 107 . Katsuobushi. Boiled or steamed clams.6 hours and smoked for 3 . are usually hot-smoked after being immersed in 80º brine or dry-salted (6). similar to masmeen. break down of fats and the moderation of smoke flavour. 5. 23). are usually 80º brined for 5 . The production method is similar to masmeen. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. They will keep well frozen. 17. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. 22. synthetic smoke flavours. 21. then hot smoking and finally sun drying it. including ‘umami’ (16. rewetted and remoulded to give the desired shape and consistency. including Eurotium repens and Eurotium rubrum. and many closely related Aspergilli (Aspergillus ochraceus. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). Hot-smoked products made from white fish will usually keep better than those from fatty fish. The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. Growth of these moulds on the tuna results in the extraction of moisture. Aspergillus oryzae.3½ hours using the typical temperature set for hot-smoking referred to above (12). such as yeast derivatives. 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). When refrigerated at 3 °C. moulding it into long thick sections. Mackerel. chilled or freeze-thawed. resulting in a characteristic flavour (20.5. Natural smoke extracts. They are generally not cooked prior to consumption. apart from the improved appearance. The product is removed from the drying cycle. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. and substances unconnected with smoke but with a smoky flavour or smell.
4 Clear smoke treatments The “Clear” or “Tasteless” smoking process is a processing technique whereby smoke is generated as in a normal food smoking process but is filtered to remove the characteristic smoke flavours before application on to otherwise untreated (uncured) tuna. or smoke extracts in organic solvents. When liquid smoke was used. 5.6 to 2. 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. smoking. The European Scientific Commitee for Food.03 μg/kg of benzopyrene. the salt level could effectively be reduced from 4. In experiments comparing traditionally smoked fish with liquid smoke. Fresh fish spoilage may be measured by 108 . drying. 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. nutritional neutrality and antioxidant activity (27). Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. 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. According to some recent research. spraying is preferable because it is easiest to control flavour and acceptable with this method (3).06 μg/kg of benzoanthracene in the final foodstuff (28). and less than 0. Liquid smoke was considered to provide advantages of safety. 5. It has been shown that liquid smoke can reduce the formation of PAHs (3). so as to impart an unnatural red colouration to the flesh.5.3. 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. 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). injecting it into the fish flesh. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). consistency of flavour and colour.4 Spoilage Brining. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. both of which are considered potential carcinogens. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25).FISH AND SEAFOOD smoke condensate that is dissolved in water or oil.8% and still provide protection for 7 days (25). applying it as a dip after brining. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. or applying it as an atomised spray within a modern automatic smoke kiln.
chemical oxidation. or extended refrigerated storage. Spoilage needs to be carefully defined for each product.3%). but. Once placed into brines.000 cfu/ml of brine may build up. and coryneform bacteria. otherwise. spoilage is due to endogenous microorganisms and enzymes of the fish. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. When heavy brining or salting is important for product shelf life. as the fish are added. which may also be produced to describe the quality attributes and their loss over time for processed fish. Often. a very dry product is required to produce 'shelf-stability'. rock and manufactured salts did not usually contain halophilic bacteria (29). pickling and dry-salting all introduce bacteria to the fish. a product may become undesirable for consumption as a result of changes in its sensory characteristics. Once these changes are defined. contact with the bacteria 109 . and of smoke to add chemical preservatives. these halophilic organisms may grow and cause spoilage. contaminant bacteria from other materials used to produce the product. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). the fish flora are transferred to the brine mix. and high concentrations are usually required to inhibit their growth fully. SMOKED AND DRIED FISH grading schemes or quality indices. Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2. Some brines are re-used over a period of days and a load of 10. however. The use of drying and/or salt to lower the water activity. the consequent reduction in the pH will help to inhibit spoilage organisms. therefore. the fish are cut and splayed to allow greater surface contact with the salt. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). which are found in the salt. the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. for others. a process similar in microbiological effect to that of saturated brining. or their products require refrigerated storage.CURED. and. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime.. limits the types of microorganism capable of growing. Most marinated and smoked fish products are first brined or salt-pickled. When acid marinating or smoking. Micrococcus spp. these bacteria tend to continue to dominate the flora in the liquid. the shelf life may be estimated and investigation made into the causes of the loss of sensory quality.g. In salts. for some. Usually. Stronger and longer brining has an effect like that of pickling. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure). The microflora attached to the fish is relatively unaffected by light brines used before smoking. marination or drying). also. the bulk of the microflora comprises Bacillus spp. Within the limits of safety. both of these will affect the spoilage patterns. For some products. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt. Brining.. the production process will modify the microbial flora present on the product. Even saturated brines allow some halophilic organisms to grow. Upon salt packing.
32. as little if any growth will occur below 5 °C on salted fish (29). may also be found in the salt used in production. This condition is often associated with case-hardening. sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). or after drying if the fish is stored in excessive humidity. niger. sometimes covering the whole of the fish with a whitish. though only the latter two species were considered likely to have grown on the fish (32). ‘Pink’ describes a condition associated with surface growth of the pigmented halophilic bacteria Halococcus spp. and Halobacterium spp. if not too contaminated. including A. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. were also isolated. spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. Unlike ‘pink’. W. E. Aspergillus wentii. (syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. powdery growth. together with a range of Aspergilli. The metabolism of the organisms will produce off-odours. Sporendonema sebi)). flavus. including hydrogen sulphide (26). and still produce a reasonable dried product. 34) have shown that P.g. Only halophilic microbes may grow. yeasts and fungi. lactic acid bacteria (LAB) and moulds. or fat breakdown) caused by enzymes rather than by microbial growth. the best way of controlling the organisms is to refrigerate the fish. B. pisce was the main fungus isolated. and Aspergillus penicilloides. it was found in about 50% of samples examined. which. rubrum are also common isolates from dried fish. The condition describes the growth of the halophilic moulds (Wallemia sebi. Aspergillus sydowii. At the pre-drying stage. A. like the ‘pink’ organisms. although it may occur in other salted fish. protein breakdown. (syn. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage. gaspé). most notably Penicillium citrinum and Penicillium thomii. halotolerant or halophilic micrococci. repens and E. Eurotium amstelodami. as the water activity of saturated brine solutions is low enough to inhibit most bacteria. Several Penicillium spp. The microbiology of heavily salted products is dominated by Gram-positive. ‘dun’ is not characterised by the production of offensive odours. ‘pink’ and ‘dun’. Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration. halophilica was isolated 110 . Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. Studies conducted on dried and salted fish from Indonesia (31.FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. The bacteria form coloured colonies on the surface of salted fish prior to drying. it is possible to wash off the infected parts. spore-forming bacteria. 33). yeasts.. The bacteria are osmosensitive and will lyse in water. A number of spoilage organisms can grow and produce conditions such as ‘sliming’. rancidity due to lipolysis. Basipetospora halophila. 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). Therefore.
LAB. Refrigeration is necessary. so the bacteria spoiling these products will be psychrotrophic or psychrophilic. SMOKED AND DRIED FISH infrequently at low levels. In Vietnam. as already stated. Fish processed by cold-smoking. Marine vibrios were the dominant flora in smoked salmon that was drysalted. it was. around the water activity of saturated sodium chloride. botulinum type E.747. however. where a high internal temperature is reached. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life. flavus. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. The preservative effect of the smoke will inhibit some spoilage microorganisms. has been isolated from dried fish (35). The brining to 3 . which does not create the salty conditions required for spoilage by those halophilic microorganisms. Aspergillus spp. The salt in the water phase must be high enough to inhibit the growth of C. but. Vacuum packing limits chemical oxidation.. not the growth of spoilage organisms.5% sodium chloride in the water phase will inhibit some bacteria. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation.IV. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). For more information on spoilage see Table 5. but this will not prevent the growth of spoilage bacteria. Cold-smoked fish will contain microbial flora representative of that of the raw fish.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). 33. able to grow on dried salted fish. 111 . Pseudomonadaceae. The heat during hot-smoking will kill vegetative microorganisms but not all spores. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. Aspergillus clavatus and Aspergillus niger. Table 5. with a lower aw limit for growth of 0.CURED. and microbial spoilage is more likely. including A. will have a different residual post-smoking flora from 'hot-smoked' fish. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations. It has been reported that the type of salting (dry.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. Smoked fish products tend to have a light brine treatment. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. 36).
IV Spoilage of salted. sebi and B. + to +++ strength of preservative effect 4 Post-process contaminants 112 .FISH AND SEAFOOD TABLE 5.not used/unchanged. halophila Halophilic spoilage organisms Halophilic spoilage organisms. boiled. 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. . and Halobacterium spp. 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.W. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted.variable. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. LAB. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. Dun.
0-9. The presence of salt in cured products has a great effect on the growth of the bacteria. the non-proteolytic type E strains can grow in up to 5% sodium chloride. when present in the raw material. It has been shown that. botulinum type E. but if the fish products are stored at temperatures <10 °C. 9 and 8 of the outbreaks.5-8.9 0. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39).V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. Since neither the smoking nor the drying parts of this curing process are particularly severe. Before. the importance of cold-chain integrity must be emphasised. aureus Yeasts Moulds Lactic acid bacteria pH 5. botulinum type E and production of toxin.5-8. and as botulism is a serious hazard in foods. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. SMOKED AND DRIED FISH TABLE 5. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth.5-10.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. could not grow and produce toxin.5 4. popular demand for healthy foods has led to a change in the tastes preferred by the public and.97 0. with this change. botulinum type E Staph. and C. pressure on the traditional methods of curing. the products were heavily salted.97 0. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. respectively. In the United States. the fermented products being the most dangerous group (7). types A and B are not affected by 3 . However. under optimal conditions.5 5.86 0. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. The concentration required to prevent growth at room temperature can vary from as low as 3. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). although the combination of the two factors.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). smoked and dried. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). salted and pickled products accounted for 10.5 5. sodium 113 . botulinum Staph.5-10 4.5% to 5%.5 0-11 3.CURED.61 Organisms Most Pseudomonadaceae C.8 1.75 0. botulinum.5.8 0. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E.
if recontamination occurs and storage conditions are favourable.5. the risk of infection can be eliminated by proper cooking. as they require more handling. has been reported (42).4 Listeria monocytogenes The psychrotrophic L. gills and guts as well as in several lightly preserved fish 114 .86 and 10% sodium chloride. However. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. As they are poor competitors.5% salt in the water phase. they will grow rapidly (7). more importantly. if the person handling the product is a carrier of Staph. Once the product is contaminated. The FDA established that hot. the organism may contaminate the product.5. An outbreak of Shigella in an Italian cruise ship.94 (7). It is a halotolerant bacterium able to grow at water activities as low as 0. Shigella and E.5. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . Thus.or cold-smoked fish should have at least 3. As with other organisms.0 . 5. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). aureus growth.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. if it contains sodium nitrite ≥100 ppm. they do not grow well in the raw material. the organism can survive most of the light curing processes. 5. are particularly susceptible to contamination and. The organism is mesophilic with a minimum growth temperature of 7 °C. these parameters change as a result of the interference of food components and other limiting factors that can act in an additive or synergistic way to prevent Staph.3 Enterobacteriaceae The pathogens Salmonella. and the minimum pH for growth is 4. 5. Sliced smoked fish products.30% of the population positive for enterotoxin-producing strains (41). Usually. growth and toxin formation may occur. but. related to the consumption of a raw smoked fish meal. if storage temperatures are over 10 °C. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. Infection by Shigella sonnei has increased in the UK. but requires higher temperatures for toxin production (>10 °C). the human nose.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. it should contain not less than 3. In foodstuffs. throat and skin. after processing. but.5. when the bacterial load of the product is lower. coli all occur on fish products as a result of contamination from animal/human reservoirs. aureus.0% salt in the water phase (30).4.
0. Of microbiological concern in the northern hemisphere are two parasites . have been shown to have an anti-listerial activity in many food products. the CT 3162-FAIR program. SMOKED AND DRIED FISH products such as cold-smoked fish. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. Pseudoterranova dicipiens. a short period of freezing either of the raw material or of 115 . in the EU Fisheries R&D-FLAIR program. the USA established ‘zero tolerance’ for the organism. or nematodes. the only concern to public health is when eating raw or uncooked fish products. 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.514. Some LAB. marinades (ceviche) (43) and gravad fish (44). They have complicated life cycles. and other local traditional products (7).8 . a round worm.9.2. This includes matjes herring. The round worms. do not survive 1 min at 60 °C. there were no reported outbreaks related to cured fish. Its limiting conditions for growth are 0 . lightly salted caviar. are considered unsafe.95 (45). The use of LAB in fish preservation is a topic that has been studied in projects (UP. and the tape worm Diphyllobothrium spp. which. most of them are of little concern to public health.45 °C.12%. 5. has also been detected in cod (7).the round worm Anisakis spp. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). from freshwater fish.CURED. such as ‘lightly preserved’ fish products. monocytogenes’ in 25 g of the finished product.5. salt levels of 8 . Two outbreaks in the United States have been reported to be linked to salmon (10).6 and water activities below 0. favours growth of psychrotrophic LAB. and in CT95-1207FAIR). monocytogenes pathogenic for only specific segments at risk. from seawater fish. in combination with curing agents and refrigerated storage. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. As they are heat-sensitive. a number of fish products. which include intermediate hosts. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. monocytogenes in levels at or above 102 per gram (46).92 . Thus. pH 4. Unlike some European countries. cold-smoked fish. gravad fish. which considered L.5 Parasites Despite the presence of parasites in fish being very common. which means that the process must conform to a level of ‘no detectable L. widely used as starter cultures in food processes. Until 1997. both being detected in cold-smoked salmon. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48).
the organism should be identified # N/A denotes not applicable 116 . E. ‡ On occasions some strains may be pathogenic. and the levels or absence of indicator organisms or pathogens.100 >100 N/A Pathogens Salmonella spp. 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.<103 N/A 100 .<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. perfringens B. aureus C.VI Guidelines for the microbiological quality of ready-to-eat smoked fish.<104 104 . smell. † Guidelines for aerobic colony counts may not apply to certain fermented foods. texture. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E.<104 20 .<100 20 .<107 ≥107 N/A <100 <20 100 .6 Published Microbiological Criteria TABLE 5. salami. 5.<100 200 .<100 20 . (not L.<100 >104 >100 N/A N/A <20 20 .<104 100 . cereus and Bacillus subtilis# <20 <20** <20 <10 20 . Acceptability is based on appearance. cholerae Vibrio parahaemolyticus L. If the Bacillus counts exceed 104/g.<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . not detected in 25 g Campylobacter spp. soft cheese. monocytogenes Staph. for example.<100 10 . and unpasteurised yoghurt. These foods fall into category 5.FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). coli (total) Listeria spp. coli O157 & other VTEC Vibrio. monocytogenes) (cfu/g unless stated) <106 106 .
Leatherhead. monocytogenes Salmonella spp. 1990. 7. Moody M. References Regenstein J. 2007. 56. 1994.. FAO Fisheries Technical Paper.. McLay R.M. 9. 82 (revised). where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. except in the viscera of dungeness crab. 117 .M.CURED. Birnie K. 3.. 4. or presence of toxin 0. Preservation of fish by curing (drying. Bannerman A.. 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. Doré I. Van Nostrand Reinhold. 2. 8. * Staph. Torry Advisory Note No. 1997.. Catching. 1982. Flick G. 1984. Smoked. The Smoked and Cured Seafood Guide.F. Handling and Processing Eels. Food spoilage fungi. Regenstein C. in Fish Processing Technology. 6. Torry Advisory Note No. Wareing P.W. in The Seafood Industry. Eds. Marinades. Hall G. Eds. Van Nostrand Reinhold. Wareing P. parahaemolyticus Vibrio vulnificus C. salting and smoking). Fernandes R. Fernandes R. 37(revised). aureus * Vibrio cholerae V. 120-38. SMOKED AND DRIED FISH TABLE 5. Martin R. botulinum * Limit of tolerance 1x 103 ETEC/g. in Micro-Facts: The Working Companion for Food Microbiologists. USA. Horne J. New York. 1979. 3273. 334. 381-406. Huss H.A. Urner Barry Publications Inc. Horner W. London. 1991.E.7 1. Hot Smoking of Fish. Torry Advisory Note No.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. coli (ETEC) L.8 ppm (saxitoxin equivalent) 20 ppm domoic acid.. 1993. 254-82 5. New York.F. cured and dried fish.H. Leatherhead Publishing. Chapman & Hall. Ed. Introduction to Fish Technology. Assurance of Seafood Quality. Flick G.
Druschitz A. 1994. 24. 11. 15.. Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish. Torry Advisory Note No.M..... Biotechnology. 22.... Recommendations for the Preparation of Smoked Salmon. in Fisheries Processing. 843. Eklund M. Bioscience.E.M. Kinoshita Y... Kinoshita Y..P.. Bioscience. Doe P.. Microbiological control for fish smoking operations. Teeny F.. Yamauchi H... 25.. 45 (10). 16. Pelroy G. Perusing Asian soup options. Halpern B. Food Product Design. The discovery of umami.H. Peterson M. Martin A. 19.. 27 (9). Food Reviews International. 1998. Packing Fish in a Modified Atmosphere. Yajima I. Matsui M. 23.M. Doi M. Ed. 935-41. Flavour constituents of dried bonito (Katsuobushi). 1995. Journal of Food Protection. 13. 14.C.B. Biotechnology and Biochemistry. 1992. Lancaster. Chapman & Hall. 108 (3).. 1980. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour. 1984. Hosokawa M.F. Sakakibara H. 18. 22 (revised).. 61 (8). 553-72. Processing Cod Roes. Martin A. 2008. and Biochemistry. 5 (revised). Shuto Y. Shuto Y. Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species. Cann D.. Yokayama S. Shuto Y.. 17. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour. 1989. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor. 1990. 51-81. 118 . Lindemann B. Shiemori Y. 6 (4). 88. 1995. Jeppesen V.A. What’s in a name? Are MSG and umami the same? Chemical Senses. Doi M.FISH AND SEAFOOD 10. 6 (6). Torry Advisory Note No. Biotechnological Applications. Food Control.R. 53 (4). Biotechnology and Biochemistry. Bannerman A. 1981.. 18. Ogiwara Y. Torry Advisory Note No. 21. Doi M. O-methylation of 2. 1997. 1977.. Technomic Publishers. 1051-5. Doi M. Kanayama T. 2002.W. 335-40. 508. 2007. Ninomiya M. Cann D. Paranjpye R. 17 (9). 958-60... London. Horne J. Bannerman A.E. Bioscience. 20. Ninomiya Y. Aoshima H. Botulism and Fishery Products. Control of biological hazards in cold smoked salmon production. 12. Matsui M.C. 1386-7. 1982.. Mitou M. 59 (12)..K. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors. Food Chemistry. Fish Drying and Smoking: Production and Quality. Patel T. Huss H. Agricultural and Biological Chemistry. 56 (6). 2324-5. Dillon R. Torry Advisory Note No. Hayashi K. Embarek P. 840-6.
I.M.. Adebajo L. Fungal flora of South Vietnamese fish and rice. Townsend J.D.D.O. 119 . The control of botulism in hot-smoked trout and mackerel. Press Inc.. 32.. Cann D.I. New York.A. 14. Pitt J. ceviche. 1994. 1995. Luxembourg. SMOKED AND DRIED FISH 26. The microbiology of sea-water fish. 29 (2). Modern Food Microbiology. 5. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. Nahrung. Rockville.. Washington DC. 197-208. 74. Fuchs R. 348.J. 2005. Blackie. Ed. Seafood Safety..V.C. Hansen L. 1996.K. Cox J. 34. 27686. 39.D. 1985. 40. Food Research International. V.M.CURED. Gill T.E. Golden D..H. 1991. Smoke flavourings. 1971. Fungi associated with Indonesian dried fish. 88-90. 30. Studies on salt fish. Hocking A. in Fish as Food. 1993. 74.T. Diyaolu S.A. Smoked and smoke-flavoured fishery products. 351-7. 43. 1994. 41. Food and Drug Administration. Mycotaxon. Simini B. 35..A. Frank M.. Journal of Tropical Medicine and Hygiene. 218-25.R. Commission of the European Communities. 1979. Tour highlights production and uses of smoke-based flavours. Røntved S. Academic Press. Lancet. Scientific Committee for Food. Food Technology. Journal of Applied Bacteriology. Pitt J. Institute of Medicine. Studies on Sporendonema epizoum from ‘dun’ salt fish. Borgstrøm G. 1941.. Pszczola D. Taylor L. 164-9. Sprouse R. New species of fungi from Indonesian dried fish.. National Academic Press. Committee on Evaluation of the Safety of Fishery Products. 37. Importance of autolysis and microbiological activity on quality of cold-smoked salmon. An outbreak of Shigella. 107-11.I. Antimicrobial properties of smoke. 1986.M. Journal of Food Technology.D. Shewan J. 22.F.. Pitt J. US Food and Drug Administration. Sirvas S. 3 (4). 311-7. 27.A. 1961. Hocking A. Jay J. Ahmed F. Loessner M. 742. Hocking A.. in Reports of the Scientific Committee for Food (34th Series). in Fish and Fishery Products Hazards and Controls Guide. Ed. 12. in Smoke in Food Processing. Oxford. 29. Incidence of Listeria monocytogenes in an acidified product.Y.. 49 (1). 38. Food Microbiology.. 1997.S. Lucas F. CEC.E. Ed. London. 70-2. Interactions among xerophilic fungi associated with dried salted fish... 1991. Food and Drug Administration. Wheeler K. Boca Raton. 1995. Letters in Applied Microbiology.1988. Ed. 36. 1996. 31. Maga J.. Journal of the Fisheries Research Board of Canada. 123-9. Wheeler K. Springer Science. 42. 38 (3). Hess E. CRC...B. 28. 98100. Fungi and Food Spoilage.F. 181-8. 74 (2). Huss H. 33. Anggawati A. Hocking A.
37 (1).. in smoked and "gravad" fish.. Beltran A. Pelroy G. Zeitschrift für Lebensmittel-Untersuchung und –Forschung. 26 (4). 45 (9).fda. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon. 863-6. 5 (1).. 1989. Journal of Food Protection. Washington D.. Cann D.A. 1982.. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus. Torry Advisory Note No. Journal of Food Protection.. 120 . http://www. 5... Prevalence of Listeria monocytogenes and other Listeria spp. Eklow A.. Hobbs G. Huss H.C... Gibbs P.. de Louvois J. Nye K.. and smoked fish establishments good manufacturing practices.. 55 (4). 46.D. 1980. Persson I. Food Control. 507-10..M. Pelaez C. 197-209.8 Further Reading Gibson D. 22. Susuki E. 1980. 5-7. Amsterdam. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale. Danielsson-Tham M-L.FISH AND SEAFOOD 44. Ribeiro C. 1992. Hurst A.J. 47. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions. Peterson M. Acta Veterinaria Scandinavia. 1976.L.J. Moral A. Ed. salted. Journal of Clinical Microbiology. Ericsson H. Danielsson-Tham M.. Paranjpye R.W. Pathogenic microorganisms of importance in seafood. Hughes A.W. Appendix 5: FDA and EPA Guidance Levels. An outbreak of listeriosis suspected to have been caused by rainbow trout. Eklund M. US FDA Centre for Food Safety & Applied Nutrition.C. Wooten R.. Unnerstad H. Advances in Food Research. Clostridium botulinum and its importance in fishery products. Food and Drug Administration. Davies A. Communicable Disease and Public Health. 58 (8)... Tham W. Loncarevic S.html Gilbert R. Little C..N. 49-59. Loncarevic S. Journal of the Association of Food and Drug Officials. Cured. 1995. 2000. in Fish and Fishery Products Hazards and Controls Guide. 1994. 188 (3).. 163-7. 232-6. Incidence and growth of psychrotrophic Clostridium botulinum in food. Jacobsen M.A. 135-85. Fletcher R. Garren D. 45.. in Quality Assurance in the Fish Industry.A. 48.. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects. 13-8. Huang Y.M. Roberts D. 3.H.V... Canadian Journal of Microbiology. US FDA Centre for Food Safety & Applied Nutrition. 80... Bolton F.E. 833-41. Round Worms in Fish. 1997... Donovan T. Association of Food and Drug Officials. Mentzing L-O. Richards J. 1996. 35 (11) 2904-7.cfsan.gov/~comm/haccp4x5. 1991. Tham W. Harrison M. Elsevier Science Publishers B. Eds.R. 2001. Liston J..S.
P. Peterson M. Beuchat L.. Letters in Applied Microbiology. 309-16. Hitchins A. Kramer D.CURED.. Beuchat L. Hudson J. Presence. 1994.C. 25. Slade A. 1991. New York. van Spreckens K. Yndestad M. Food Microbiology. 293301.. Ratnam S.. 61 (12). Keusch A.A. Bayliss A.. Interacting effects of pH. 58 (5). 217-36... Lashbrook L. 866-70. Journal of Food Protection.R. solute.J. 1995. Davies A. Letters in Applied Microbiology... Contamination pattern of Listeria monocytogenes and other Listeria spp. 1995. Incidence of Listeria in hot and cold-smoked fish. Poysky F.W. 1994. 11. 42. Rørvik L. 1992.. 1987. 11. Journal of Food Safety. 121 . 1734. 19-27. International Journal of Food Microbiology. SMOKED AND DRIED FISH Sutherland J. 1994. in a salmon slaughterhouse and smoked salmon processing plant. 1992. Liston J. 23 (1). Applied Microbiology. 502-8. Patel T. 4389-95.R. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. 309-18. Caugant D. Combined effects of water activity. 1075-80. Jemmi T. 1995.R. 1973. Taylor S.K. detection and growth of Listeria monocytogenes in seafoods: a review. Dallas H. in Seafood Quality Determination. 27 (6).. pH and sodium chloride. Histamine production by psychrophilic flora. International Journal of Food Microbiology. Nicolaides L... Ben Embarek P. 21 (3)..A..S. Food Technology. Paranjpye R. Fuchs R. Eklund M. and salt concentration on growth and survival of Vibrio parahaemolyticus. 19.N. 394-6. 94-8. and temperature on the growth of Vibrio parahaemolyticus. temperature. Dillon R. 1988. 14. International Journal of Food Microbiology. Pelroy G. Prevalence of Listeria in smoked fish.T. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. Marine toxins of microbial origin. Competition of food bacteria with Listeria monocytogenes during enrichment culture. 1974. Letters in Applied Microbiology.E. 25 (5). Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants.. Applied Microbiology.. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. 1994. Roberts T. Tran T.A. Journal of Food Protection. Poindexter C. 55 (11).178-80. Elsevier. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout.. 844-6. Eds.
added during preparation. For example. 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. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part.1. in economic terms. North American and other markets and are. 6. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. the most important of all fermented fish products. FERMENTED FISH Prof. typically in the range 15 . they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. koji. Fermented fish products are largely confined to east and south-east Asia. The relative importance of these two activities depends on the formulation of the product. though some are produced elsewhere. Those to which only salt is added tend to be mainly autolytic.I.25% and are used mainly as a condiment. the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru.6. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. lactic fermentation can play a significant role. Essentially. there are numerous variants of some common themes and a host of local names used to describe them. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. As with most traditional products that are still produced principally on a cottage industry or domestic scale. whereas in those where a carbohydrate source such as rice or sugar is also added. they are exported to European.1 Definitions As applied to fish products.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. Carbohydrates are added to some fish sauces during their preparation. 123 . The fish sauces are produced on the largest scale.
III. made from fish livers in Greece. where the bacterial production of lactic acid is a major feature.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. salty taste. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans.II. This is well exemplified by some of the fish/salt products of Thailand described in Table 6. jeotkals (shrimp) nam-pla (pa) Budu Patis nam-pla nuoc-mam padec Belachan (shrimps) bagoong kapi mam-ca man-tom (shrimps) Laos Malaysia Philippines Thailand Vietnam Even within the limitations of fish and salt as the sole ingredients. and pissala. to products analogous to other lactic fermented foods such as salami. 6. Fermented fish products are of very minor importance outside Asia.1. may be direct descendants. In Europe. made from whole fish in France. A selection of Asian products is presented in Table 6. and the more widely known Worcestershire sauce a very distant relative. 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. 124 . These have long since fallen from use. who had adopted them from the Greeks.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred.FISH AND SEAFOOD TABLE 6. although garos. a variety of products can be produced within a single country.
FERMENTED FISH TABLE 6. som-pa.g. e. burong-bangus pla-ra. cencalok burong-isda. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps. crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 .II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. tai-suchi phaak. crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 . pa-khem. salt/fish ratio Production period Type of use Hoi-dong Molluscs Whole or without shells Whole - Main dish Kapi (fish or shrimp paste) Nam-pla (fish sauce) Nam-budu Small fish. mam-chao.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. mam-pa-kor. som-fak/ som-fug 125 Malaysia Philippines Thailand .12 months 10% raw sugar added. shrimps. ayu-sushi. funa-suchi. e.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 . mam-seeing sikhae som-kay-pa-eun.3 months Main dish Tai-pla Bowels 1:3 6 . pla-chao. burong-dalag. pa-chao. burong-ayungi.g.8 months Main dish TABLE 6. som-pa-keng pekasam. pla-som.
made from herring. TABLE 6. For example. and a large local freshwater reservoir (2). for example. especially in fish/salt products. Penaeus indicua Many of the fish used in fermentation are uneviscerated. paddy ponds beside rice fields used for collecting fish when the field has dried up. Table 6. relatively mild-tasting product. the most popular varieties of which are rakørret. and old descriptions of their production often refer to the addition of whey.the flooded rice fields. Here. Lactic fermented fish products are often associated with inland areas such as the Central Luzon region of the Philippines and the north-east of Thailand. in Sweden. which has been increasing in popularity over the last 50 years. malt or flour (1). however. and counts ranging from below 104 /g to in excess of 107 have been reported. and the initial microflora is likely to be considerably larger in these cases. which employ lower levels of salt than Asian products. freshwater fish are the usual raw material. fermented trout. rakefisk or surfisk. some fermented fish products remain popular. These are often described as fish/salt products. is a traditional.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. There are also more heavily fermented products. A modern account of their production. 6. states that sugar is often included with the salt. 126 . The diversity of fish used in fermented products (see. This could lead to quite marked variations since the same fish can be obtained from a variety of sources.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. Norwegian gravlaks. it was found that several different sources were used for the fish .FISH AND SEAFOOD In Northern Europe. and their microflora tends to reflect their local environment more than that of marine species.2 Initial Microflora Composition of the initial microflora of fish is described in some detail elsewhere in this Handbook and will not be reiterated here. in Norway and surstrømming. or buried salmon. in a survey of fermented fish products in north-east Thailand.
Endomycopsis and Saccharomyces .4. Mucor. It also demonstrated their significant proteolytic activity during the first month of fermentation. and the anaerobic conditions that prevail in the bulk of the material. (3. Similar results were seen with patis. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. after one month. the Philippine fish sauce. More recently. and studies 127 . It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). in many cases. either directly or as the partially saccharified rice they produce.1 – 6. 4). These preparations contain a mixed microflora of moulds and yeasts. which included nam-pla and patis. Other halophilic species have been identified from fish/salt products. Hansenula. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns.3 6. sufficient to saturate the water phase present and produce an aw of 0. 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.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 some cases. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter.FERMENTED FISH Where other ingredients are used. principally species of the genera Rhizopus. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. however. however. 6. traditional starter culture preparations such as koji (Japan). mean that microorganisms have been generally held to play little or no role in the process that follows. were found to be the predominant isolates. 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. In another microbiological study of four fish sauces. When this is a simple carbohydrate source such as sugar. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. Bacillus spp. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). angkak (Philippines and Thailand) and look-pang (Thailand) are used. they can contribute to the initial microflora. Generally. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. or boiled or roasted rice. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7).3. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). This.74 or below.
6. Belacan (Condiment) Fig.2.1.12). Fig. 6. Patis (Condiment) 128 .FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 .
FERMENTED FISH Fig. thought to be largely the result of autolytic breakdown of the fish tissues. eviscerated fish are sometimes used to slow the autolytic process.4.3. the 129 . and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. however. In fish sauce production. 6.6. Ika-Shiokara (side dish) Fig. 14). Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are. To produce semi-solid fish pastes as opposed to fish sauces.
2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. particularly glutamate.5. Hoi-Malangpu-Dong (side dish.5).3. snack) 130 . Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. filtered and bottled. for example. 6. which is eventually separated from the residual fish tissues.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. 6. Some examples of products and their formulation are presented in Table 6.6 – 6. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. 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. Fig.8.V and Figures 6. Figure 6. volatile fatty acids and nucleotides. In contrast to free-swimming fish.
5-1.4 are also produced using shrimps.6. the prefix 'pla' is replaced by 'kung'.03 Preparation time 6 . N. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus.12 months Main dish 2 Pla-jao C.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. Fig.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0.FERMENTED FISH TABLE 6.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .2-4 Fish (pieces): salt:khaomark (fermented rice). C.7 days 5 . N.12 days 3 .20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. N. C NE. N S 5 . NE Formulation Fish (whole or pieces): salt: roasted rice. north-east. north. NE.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0.6. In these cases. S.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . south of Thailand.12 months Shelf life Use 1 . central. NE 10 . Kung-Chom (staple food. side dish) 131 . C NE. 3:1:0.
(from Arroyo et al. their growth and metabolism are nonetheless slowed. of the two factors contributing to shelf life .VI). 106) Fig. Red burong isda 132 . typically in the range 2 . This is readily explained by the susceptibility of LAB to elevated salt concentrations. Generally.2.7. Phillipines Journal of Food Science and Technology.6. the salt is of far greater importance in determining the product’s long-term stability.10%. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. giving a reduced rate of acid generation. but there is considerable variation in this.1 The use of higher salt levels results in a longer production phase but a better keeping quality product. This can be clearly seen from the data on reported shelf lives of a number of fish salt carbohydrate products and their respective pH ranges and salt contents (Table 6.salt and lactic acid. The LAB isolated from these products are those with more marked salt tolerance but.3. then this will take longer to develop than if lower salt levels are used.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. It also indicates that. If acidity is a desired sensory property of the product. 2.. while they are able to grow at high salt levels. these products are salted at lower levels than the fish sauces and pastes.
6.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).4 .7 .6 3.2 4.FERMENTED FISH (from Arroyo et al.8. 133 . White balao-balao TABLE 6.8 .5 ..9. 2. Philippines Journal of Food Science and Technology.0 . 106) Fig.5.3 .188.8.131.52.5 3.2 4.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.3 years 1 .8 pH 4.1 4.5 .9 .5.0 .17.2 7.0 Shelf life 3 weeks 6 .8 2.3 5.6.9 4.9.8 .1 .5.9 4.0 .12 months 1 .5.
Although the ability to ferment starch is not widespread in the LAB. This results in a more active lactic fermentation and a lower pH. alternatively. such as salt content. the rice may be added in a partially saccharified form such as kao-mark (Thailand). Garlic. Where they are used. 21). a common ingredient in these products. Addition of carbohydrate will. 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. although the final population of LAB may be similar. inulin.FISH AND SEAFOOD 6. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3.4.VII). 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 .2. TABLE 6.0 . they increase the amount of soluble sugars produced and thus the range of LAB that can grow. 19).3. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6.1 .5) despite other factors.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. Rice is the most common carbohydrate source used. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. In several products. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. of course. also serves as a valuable source of the more readily fermented carbohydrate.9) compared with the white product (pH 4. 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).VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. For example. When added as boiled or roasted rice alone. being similar (18. also lower the unit cost of the product and this is probably a significant factor in many cases.3.
135 . which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak.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). Traditional packing techniques used with lactic fermented fish products. reporting the product’s pH to range from 4. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). The growth of yeasts and moulds could. presumably correlating with the salt content. markedly extending the shelf life (25). There is little published information on the factors that limit the shelf life of the low-salt products. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24).2. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. the pH drops from 6. and there is a marked regional variation in the preferred type.17. Clearly. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. Some authors have claimed that the function of the roasted rice added during pla-ra production is to improve texture and slow the fermentation rather than to serve as a substrate for lactic fermentation (22). the Korean fermented fish product. is something of an exception since millet is used along with garlic as the carbohydrate source.FERMENTED FISH Sikhae. but they are likely to behave similarly to lactic fermented products produced from other commodities. the level of salt employed is on the high side for fish/salt/carbohydrate products. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. 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.7 to below 4. in turn. Excessive growth and acid production by the LAB could render products too sour for acceptability. such as wrapping in banana leaves.3 weeks at around 20 °C. 6. which ranged from 7. allowing other spoilage organisms to grow. lead to an increase in pH. Generally.3 days. Results of a product survey were consistent with this.7 up to 6. The mixture is allowed to ferment for 2 .8 . around 12%. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. In a successful fermentation.2 at the end of fermentation. thereafter declining to about 4.5:9:3. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage.5 in the first 2 .9%. will not reliably exclude air. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products.5. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products.
Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. 6. which accounted for 88% of outbreaks in the period 1952 . outbreaks of 136 . The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). In Japan. There is little in the way of documented information associating fermented fish products with foodborne illness. and consumed without cooking. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. As in Norway. fish products are the most common source of botulism. 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 Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. parahaemolyticus has been reported growing down to a pH of 4. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. In the fish sauces and pastes. Because of the severity of the illness it causes. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. typically 30 °C. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. V. The bacterial pathogens most commonly associated with the living environment of fish. under conditions inimical to growth and high ambient storage temperatures. viable numbers will probably decline quite rapidly. After capture. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. although these limits apply when all other factors are optimal (26).FISH AND SEAFOOD Products where a substantial population of moulds and yeasts is deliberately added in the form of saccharified rice or a starter culture would spoil as a result of their continuing activity breaking down the rice and producing unacceptable levels of alcohol in the product. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. 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. are Vibrio parahaemolyticus and Clostridium botulinum. Several measures have been suggested for reducing the risk of botulism. During fermentation. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures.8 and at salt levels as high 10%. however.87. but we are unaware of any reports of this happening.
6. beaver tail and paw.. particularly in south-east Asia and the western Pacific regions (30). With this particular organism. while increasing the acetic acid content of the formulation had little or no effect (32). which is generally consumed raw (29). salt content was found to have the greatest impact on the viability of larvae. Mahidol University. and warm weather can result in rapid growth of C. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. Saisithi P. Rakefisk and Herring in the Norwegian Tradition. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada. Journal of Food Science. Oslo.. 1966. Dollar A. 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). Many of these products are described as fermented.O. and these are often used in the production of fermented fish products. Foodborne trematode infections are emerging as a major public health problem.FERMENTED FISH botulism have become less frequent in recent years. whale meat. though 27% survival was still recorded (33). they could act as an important vehicle of transmission. Yhoung-Aree J. One study with the trematode Haplorchis taichui. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). Raw materials subject to this process include seal flipper. 1990. 2. 1989. 31.. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. botulinum.. giving an alkaline fermentation. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). Novus Press.. botulinum and the production of toxin. Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. Salting levels are often insufficiently low to prevent growth of C. References Riddervold A.M. and salmon heads and eggs. 3. Greenland and Alaska. Liston J. walrus nose. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31).6 1. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. Fermented Fish Products Study (Thailand) Phase 1. Thailand. Where these products are consumed without final cooking. 105-10. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. Microbiology and chemistry of fermented fish. 137 . Institute of Nutrition. affecting an estimated 40 million people. Baseline Study (1987-1988). Lutefisk. although the absence of any source of fermentable carbohydrate means that the process is putrefactive. Puwastien P. Dhanamitta S. which will persist into the product. Kasemsarn B.
M. 3649.. Fukuda H. 185-92. Journal of Food Science. Constantinides S. Reddi P..H. Fermented fish products of south east Asia. Halophiles in Thai fish sauce (Nam pla). Kaneko Y. Sands A.R. Aslam Z. 11. Byun M. 2006. 1957. Journal of Food Technology. Nov.. Thawai C. 5. Production and characterisation of NaCl-activated proteinases from Virgibacillus spp.. Orejana F. Takada K. 43.. Dymsza H. 6338. 2007. 1986. microbiological and chemical properties of salted and fermented squid. Tanasupawat S.. 2007. from fermented shrimp paste in Thailand. Lee S.. in General and Applied Aspects of Halophilic Microorganisms.. 16. 353-63... 8.. Ahn H. Microflora of four fermented fish sauces. Cooke R.H. Dong-Ho K.. 1995. SK33 isolated from fish sauce fermentation. Lim J. Yasir M.. Salinococcus jeotgali spp. Journal of Scientific and Industrial Research (India).. Liston J.R. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries...K. Journal of Fermentation and Bioengineering. A halophilic serine proteinase from Halobacillus spp. Plenum Press. Kudo T. Yongsawatdigul J.. 1395-401. Some chemical changes in fish silage. 29.. 934-9. Tanasupawat S. 1972. 14... Process Biochemistry. 9.A. Tsunemi M..T. Effects of gamma radiation on sensory qualities.R. 3.V.S. New York. International Journal of Systematic and Evolutionary Microbiology. Journal of Food Protection.. 643-7... 37. Tropical Science. Ed. Government of Australia. isolated from jeotgal. Canberra. SR5-3 isolated from fish sauce: purification and characterisation. Bioscience. 57. 1991. 381-8.J. Dhamcharee B. Rodtong S.D. Backhoff P.. Thongthai C.. Velankar N. Im W. Pakdeeto A.. Moonmangmee S. Nov. 1976. Crisan E.. Catheptic activity of fish muscle. 63. Yook H.W. Biotechnology and Biochemistry. 57. Takano M. 141-2. Ono H. Suntinanalert P..FISH AND SEAFOOD 4. 10.T. Namwong S. Saono S. 12.K. Lentibacillus kapialis spp. 124-30. 16A. Kyong-Haeng L. Protein hydrolysate from fish. 17. 11. Rodriguez-Valera F. 2000.. Agents of proteolysis and its inhibition in patis (fish sauce).M. Journal of Food Science and Technology. 1985. Hiraga K. 80. 138 . International Journal of Systematic and Evolutionary Microbiology.. a traditional Korean fermented seafood. 25. 15. Chung Y. 6. 1979. 7. Adams M. 13. Sinsuwan S. 70. Applied Microbiology. 17-30. 106-8. Rattagool P.. Itoh T.... Jae-Hun K. Oda K. 1975.. Olympia M. Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme.. 61-73. Phil. Hull R. 2008.
Rustad T. burong-isda. in Clostridium botulinum: Ecology and Control in Foods. 26.L. International Journal of Food Microbiology. 28 (6). Dodds K.D. 1815. 69-104.. a Thai low-salt fermented fish product. 1990. Sakai H.B.. 25.H. Caldo G. International Commission on Microbiological Specifications for Foods... 104-6. Characterisation of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation.H. 2002. Eds. Valyasevi R.. 274-86. Blackie. Journal of Food Science and Technology.B. 305-22. Gram L. Yamprayoon J. 103. Volume 5: Microbiological Specifications of Food Pathogens. ASEAN Food Journal. Journal of Agricultural Science (Tokyo). Dodds K.FERMENTED FISH 18...H. Marcel Dekker. Ben Embarek P. Owens J. Patrick D. Isaac-Renton J. Canada Communicable Disease Report. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak. 105-12. in Microorganisms in Foods. 29. Marcel Dekker. 30. Wainwright R. New York.. Vibrio parahaemolyticus.. 1983. 24. 28. Saisithi P. The fermented fish food. Erikson U. 46. a Thai low-salt fish product. Kozaki M.D.. Caldo G. Hauschild A. Huss H. 2007. Benjakul S. Gram L. Control of foodborne trematode infections.. 92. Visessanguan W. 27.. Journal of Applied Microbiology. Ostergaard A. Traditional fermented fish products with special reference to Thai products.. Kozaki M.. 138-44. Epidemiology of human foodborne botulism. 22. in the Philippines. 21. 28. International Commission on Microbiological Specifications for Foods. Yeast-flora in red burong-isda a fermented fish food from the Philippines.A. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation. 28.. World Health Organisation Technical Report Series.. Fermentation and spoilage of som fak.W. 19.W. 1996.. Hazards from northern native foods. WHO Study Group. Avurhi J. Paludan-Müller C.M. 1998. Tropical Science.H.W. Fung C.H. Gram L. 38. 1999. 219-29. Martin J. Eds. 139 . 1992. 3-10. 1-4.. 1995. 1992. Moody L. Sakai H. Journal of Agricultural Science (Tokyo).H.. 426-35. Food Chemistry. Tanaka M. Ed.. Paludan-Müller C..K. 1987.. 3.. Wedell-Neergaard C. 20. Riebroy S. Hauschild A.. New York. London. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper.. Two outbreaks of botulism associated with fermented salmon roe – British Columbia. 27. in Clostridium botulinum: Ecology and Control in Foods. 1983. Hauschild A. 23. Huss H.A.L. 307-14. 2002... Dawar M. Huss H.
. Roepstorff A. Sripa B. 117. Wongsawad C.. Miwa M. 29. northeast Thailand. 33.... 2005. 23. Hanai S..R. 1995.R.R. Kittiwatanachot P. Huss H.. 15998. Honjo S. Thai traditional food preparation. Sriplung H... Gaithersburg. Aspen Publishers Inc. Fermented flesh foods.FISH AND SEAFOOD 31. 36. Southeast Asian Journal of Tropical Medicine and Public Health.. Adams M. Fermentation and Food Safety. International Journal of Cancer... Nout M. 6. 32.. Jedpiyawongse A. Bloemsma B. 110-1. Chuboon S.. 2005.7 Further Reading Adams M. Survival of Anisakis larvae in marinated herring fillets.. Srivatanakul P. Todoroki T. Horst K. Nithikathkul C.. 2001. 140 . Uchida K. International Journal of Food Science & Technology. Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom.J. Kikukawa H. Survival of Haplorchis taichui metacercariae in lab-pla. 1986.. 661-70.. Deerasamee S. Ruamsuk A. 854-60. Progress in Industrial Microbiology.H.
7. While some of these toxins only occur in seafood from a restricted geographical region. An emerging technology is the use of immunosensors (1 . a number of new toxins and toxin families have been identified. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. including the well known marine toxins 141 . Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins. Another book dealing with the clinical effects of a variety of poisons and toxins. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity.3) and some practical applications for detecting shellfish toxins have been reported (4. Wright. It remains to be seen if this approach gains momentum in the future. physiology and detection of marine and freshwater toxins has also appeared (8).D. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. Nevertheless. Although most of these antibody-based test kits have yet to receive international regulatory approval.C. Ph. FISH AND SHELLFISH TOXINS Jeffrey L. several scientific reports have described their efficacy and potential. and a second updated edition dealing with the pharmacology. Since the original chapter in this series was published (6). FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7. The manual on harmful marine algae and their products. 5). Scientific and Cultural Organisation) has been updated (7).M. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. Increasingly sophisticated instrumentation (e.g. C.. known and unknown. most notably the azaspiracids and the cyclic imines as well as some additional polyethers.. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world.
g. and sub-tropical waters and habitats (15). An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10).1). Saxitoxin Group instead of the Paralytic Shellfish Poisons).FISH AND SEAFOOD has been produced (9).2. Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. and a tropical species Pyrodinium bahamense (16). PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. Not all the toxins found in shellfish extracts are microalgal products. a great variety of shellfish from all latitudes. 142 . In addition. The Ncarbamoyl sulfates are readily hydrolysed to STX. neoSTX and Gonyautoxins (GTX) 1-4. 7. Gymnodinium catenatum. dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). 7.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. with intoxications occurring in many countries even today (15). and a review on the risk assessment of marine toxins has also been produced (14). some are the result of biotransformation processes by the filter-feeding host. temperate. 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. Consequently. even crustaceans such as lobsters. all of which contain a tetrahydropurine skeleton (Figure 7. Many of these species are extremely common and are found throughout the world in a variety of cold. 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. can become contaminated with these toxins.2 Paralytic Shellfish Poisoning (PSP) Toxins The PSP toxins are arguably the most recognised group of marine toxins and continue to be regarded among the most dangerous. and another. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). All PSPs degrade rapidly in alkaline conditions. Around twenty PSP toxins are known.
Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. PSPs bind to site 1 of sodium channels reversibly blocking sodium conductance in nerve and muscle membranes by binding to a specific receptor site located on the outside surface of the membrane close to the ion channel opening.1. However. and fingers within 5 . the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. and humans seem to be particularly susceptible. tongue. this is never sufficient to render the seafood safe to consume. 143 . Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. 7.2. Human symptoms begin with tingling and numbness of the lips.2 Toxicology PSP toxins affect the central nervous system of mammals.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. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis.30 minutes of consumption. 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).
An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). 17). China. However it remains as the monitoring method of choice in many countries. other derivatives have been reported.2). 15. The mouse bioassay has been in place for decades and the method has been standardised.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. Most intoxication events have occurred in South East Asia. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. Thailand. a criticism has been that such methods are time consuming. Alternative HPLC methods have subsequently reduced this to a single HPLC run. specifically 144 . 24) (Figure 7. 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.2. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins.FISH AND SEAFOOD 7. TTX most commonly occurs in many species of puffer fish. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). and has also been evaluated by a number of EU laboratories. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. a pre-column method has received Association of Analytical Communities (AOAC) approval (18).8 µg/g) of shellfish or seafood tissue. 7. After some development and various trials. and while simple and relatively inexpensive to operate. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21).1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). Taiwan. 23. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. 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 over ten analogs are known (22.3. 7. including Japan. While the chemical detection methods are most promising for the future. and islands of the South Pacific. Since its discovery. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. Two approaches have been taken here. as well as other parts.
2. 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. hypotension. The onset of symptoms usually occurs rapidly. Fig. 26). where puffer fish of the genus Fugu are a delicacy. respiratory failure and coma. A review of TTX detection methods has been published (12.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions.2 Toxicology Like the PSP toxins. Severe poisoning causes paralysis and respiratory failure. nausea and vomiting. gonads and roe (24). 25.3. it is served in speciality restaurants with qualified chefs to properly prepare the fish. raising speculation that TTX is produced by a bacterial source. 23. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. 27). and even newts. preventing depolarisation and propagation of action potentials in nerve cells. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. Since then.FISH AND SHELLFISH TOXINS in the liver. molluscs. and in mild cases results in distal muscle weakness. and blocks diffusion of sodium through the sodium channel.3. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. and more recently by an actinomycete Norcardiopsis dassonvillei (26). and once again the mouse bioassay has been used to detect the presence of the toxin. the blue-ringed octopus. horseshoe crabs. 7.2 mg of purified toxin can be lethal. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. An oral dose of 1 . Typically this has been accomplished using HPLC followed by post 145 . In Japan. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. 7. Structure of tetrodotoxin 7. All of the observed toxicity is secondary to the action potential blockade.
but all of them are considerably less toxic than DA itself (29). 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.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). but 146 . Fig. a polar glutamate agonist. 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.4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA).3 for examples). Currently 12 isomers of DA have been reported (see Figure 7. and more rapid LC-MS methods [(M+H)+ m/z 320] have been reported for the detection and identification of TTX and its various derivatives (27).3. Structure of domoic acid and three naturally occurring photoisomers 7.FISH AND SEAFOOD column oxidation to the fluorescent derivatives. and while promising. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA.4. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. 7. Immunoassay methods have also been investigated. but it is known that such congeners are less toxic than DA itself (30). 7.
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. multiseries. Pseudonitzschia australis. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. seizures. which has been well documented in studies along the west coast of the US. has been reported as a medium-level producer of DA (33). Levels of DA in the other strains are generally <1 pg/cell. Okinawa and more northerly parts of Japan.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. Victims display a variety of symptoms including headache. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample.39). 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. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell).. cormorants and pelicans (34). For example. The oral absorption of DA is around 5 .4. scallops. found in the Pacific around Vietnam.FISH AND SHELLFISH TOXINS only three strains (P. including persistent short term memory deficits (36 . 147 . 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. Indeed. disorientation and coma. and this has led to ASP toxicity in birds. 7. 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. To a great extent. and is mainly distributed in the blood. southern and western portions of Ireland. as does impaired renal function which increases serum concentration and residence time of the toxin. but any impairment of the blood brain barrier results in additional risk. Penetration of the blood brain barrier is poor in healthy subjects.10% of the administered dose. and in locations around New Zealand (32. Another factor is the movement of the toxin through the food chain. another diatom species Nitzschia navisvaringica. nausea. and clams. and review of the toxicologic pathology of DA has been published (36). The original 1987 event was followed some years later by a more widespread episode along the western coast of the US and today toxin-producing strains of Pseudonitzschia have been found in the Gulf coast of the US and along the western shores of the US and Canada. and sea animals such as sea lions and even whales (35). 33). Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. Recently. the Philippines. the Mediterranean.
and this is typically the method of choice. and an immunosensor method has also been reported (4). produce a variety of other 148 . are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. as can LC/MS. Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. 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). australis. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. 41). Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. 7. although in the latter study the probes for Pseudonitzschia pungens. but just over 15 years ago a third toxin.FISH AND SEAFOOD 7. Since then.4).. The benthic Prorocentrum spp. To further complicate matters. California. However.. DA can be detected by the mouse bioassay but the regulatory limit for shellfish is set below this level at 20 µg/g (20 mg/kg) shellfish tissue. the heterotrophic dinoflagellates Dinophysis spp. dinophysistoxin-1 (DTX-1). Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. in Monterey Bay.4. 49). and consequently were often overlooked as the real source of the toxins. and dinophysistoxin-2 (DTX-2) (Figure 7. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. 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. In the context of this chapter. DTX-2 was discovered in contaminated Irish shellfish (46). shellfish tissue can be efficiently extracted with aqueous methanol (1:1). DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). Alternatively. and these methods have been reviewed (40. Dinophysis spp.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. though this method is most commonly used to analyse plankton and seawater (42).5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). 48.
and include Dinophysis acuminata. are recognised as DSP toxin producers. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. although occasionally fatty acyl esters linked through the 7-OH group (referred to as DTX-3 compounds) are also found and are likely a result of bioconversion in shellfish tissue. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). While these esters are not toxic in vitro.5. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms.. DTX-5. and Dinophysis norvegica among others (47. Dinophysis fortii. and Prorocentrum belizeanum.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. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. 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. The parent acids are the only DSP congeners to be found in shellfish tissue. Dinophysis caudata. DSP toxins produced by benthic Prorocentrum spp. Lately these compounds. In comparison. which include the pectenotoxins and the yessotoxins. Structure of okadaic acid and the two related DSP toxins 7. 149 .FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay.4. Fig. In this case Dinophysis was suspected as the culprit organism. 7. are also recognised as a threat to human health. such as Prorocentrum lima. Dinophysis acuta. This was probably not the first DSP episode. Prorocentrum maculosum. 49). and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. Today over ten Dinophysis spp.
it does not provide information on the DSP toxin profile. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. and abdominal pain. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. 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. DSP-Check is a commercially 150 . nausea. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. and DTX-2. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . In recent years. DSP episodes have been reported worldwide and are included in many monitoring programs (47). even with the minimal of clean up steps (48). and a detection limit of 100 µg/kg has been reported (12).3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. Due to the lipophilic nature of the toxins. most progress in DSP toxin detection has been using new or improved LC/MS methods and a detection limit of 10 µg/kg has been reported. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method.5. various European countries. although a comparative study of in vitro phosphatase inhibition with mouse bioassay data has shown that OA is about twice as potent as DTX-2 (51). and while this provides critical seafood safety information. DSP episodes have been reported from Japan. vomiting. or indeed on the presence of other associated LSTs that can occur with DSP toxins and which result in a positive mouse bioassay result (vide infra). 7.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. A regulatory level of 80 . New Zealand and Chile. which can lead to an underestimation of the total DSP content in a sample. 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.3 days. 7. Since the late 1970s. 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.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A.5. Human symptoms include diarrhoea.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). over 20 PTXs are known (56) and more than 30 YTXs have been reported (57.5 and 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). 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. 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. with a claimed detection limit of 20 µg/kg. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. However. 55). Fig.5. 7. Indeed. Within this group. for many years these compounds were included in the DSP toxin complex.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. and the latter group are usually found to be responsible for any reported shellfish toxicity (59).6. with good agreement with the mouse bioassay and LC–based methods (54. 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. In yet another approach. 58). 7. Some representative examples of both groups of toxins are shown in Figures 7. and another rapid antibodybased test kit for DSP toxins has also been reported (53).
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. many parts of Europe and the Mediterranean.6.6. Structure of the two parent yessotoxins. 7. 7. the toxin content and profile may vary among different strains.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). Japan. which may co-occur in blooms with other organisms.FISH AND SEAFOOD Fig. eastern Canada.6. New Zealand and South America (49. While 152 . As is typical for many algal species. 58).1 Origins and distribution It is now well recognised that PTXs are co-produced with DSP toxins by various strains of the dinoflagellate Dinophysis and hence the distribution of both families of toxins in shellfish is similar (49). 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). The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. 57. YTX and homo-YTX 7.
Estimates of acute toxicity of the parent compound following IP injection vary considerably. The YTX group has not been reported to cause human illness. but less so following oral dosing (61). While LC/UV detection is possible for PTXs.7). but various congeners are acutely toxic in the mouse bioassay following IP injection. but a fluorescent derivative has been prepared for others. However.FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. 58). The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63.3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue. The structures and chemical data including the molecular weight of each member have been collated (56). the PTXs show considerably less toxicity when administered orally. chemical analysis is best carried out by LC/MS and several approaches have been described (57. There are no clear cases of human illness resulting from ingestion of PTXs (59). The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds. 7. Many YTX toxins contain a conjugated diene system. Because of the number and complexity of the YTX group. though its cell effects may be linked to modulation of calcium channels (62). and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. which is further compounded by differences in the experimental design. Although there is no evidence that yessotoxins are responsible for human illness.6. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7. it may be compromised by other biological materials in the matrix. 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. For many years it was believed that 153 . 64). The crude extract may be used directly in the mouse bioassay. Some of the PTXs are potent cytotoxins in vitro (60). a regulatory level of 1 mg/kg shellfish tissue has been set. which aids detection by LC/UV. 7. Their toxicology and mechanism of action remain uncertain. the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present.
brevetoxins are found in mussels. In the Gulf of Mexico. namely Chatonella marina. and cockles (65. and Heretosigma akashiwo.7.7. 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. 7. whelks. These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). which are found in Japanese waters (66). Fibrocapsa japonica. 66). but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65.FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida. the same toxins have been reported to be produced by three genera of Raphidophytes. Interestingly. Many of these species are associated with massive fish kills. 154 . They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. Fig.S. was chemically characterised as PbTx-2 (67). and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf.1 Origins and distribution The brevetoxins are produced by the dinoflagellate Karenia brevis (formerly known as Gymondinium breve and Pytochodiscus brevis) a dinoflagellate found in the Gulf of Mexico. and off the coast of New Zealand (65). 66). PbTx-1 and PbTx-2. and one of the brevetoxin metabolites BTX-B2 7. Structure of the two parent brevetoxins. oysters.
In this application. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. 7. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). and mammalian body fluids without pretreatment or dilution (72). Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. vertigo. and in severe cases seizure. malaise. The mean time to onset of symptoms is around 3 hours. 66.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). which can result in a cascade of associated events in a cell (66).7.7. In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. 7. 69). with hot spots in the coral reef areas of the Caribbean and French Polynesia.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. the competitive ELISA method correlated most closely with the mouse bioassay results. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. nausea. this family of toxins has proven to be a challenge both to identify and monitor. The mouse bioassay is used as the primary detection method. shellfish homogenates. The human health implications following exposure to NSPs have been reviewed (69). with a mean duration of symptoms of around 1 day.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. While the mouse bioassay continues to be used. 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. and diarrhoea. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). 7. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). Ingestion of brevetoxins results in a variety of symptoms including parathesia.
CTXs are large. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins.8. 7. but more are suspected to exist. These include species belonging to the genera Ostreopsis and Coolia.8). and only a few structures have been identified. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish.8. Fig. lipid soluble polyethers.FISH AND SEAFOOD transferred through the food chain to the larger carnivores. complex. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red. green and brown seaweeds and also occur in sediments and coral debris. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin. most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7.1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins. a group of closely related polyethers that are considered the source of ciguatoxins. Distribution through the food chain begins with 156 . A detailed comprehensive review of ciguatoxins has recently been published (74). The diversity of benthic phytoplankton that exist on reefs further complicates the picture.
pruritus. Like the brevetoxins. groupers. only trace amounts of CTXs can cause illness following consumption of contaminated fish. weeks or even months (74 . all of which can be linked to the effect of CTX on excitable membranes. and the sensation of loose teeth (77). Other symptoms include a metallic taste. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. arthralgia. Another symptom that can develop over time is extreme fatigue. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. 75. although not all CTXs are equipotent. and in fact CTX poisoning is often only identified after consumption of contaminated fish. and there are suggestions that such incidents are increasing and spreading worldwide (74). they bind to voltage-dependant sodium channels in the cell membrane causing them to open. 7.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. and temperature sensation reversal.3 Detection methods As a consequence of their potency. Even when suspect samples are available. These include paresthesias (numbness and tingling of the extremities). Patients have reported symptoms lasting weeks and even months.8. 7. the neurological symptoms can persist for days. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel.1 µg of ciguatoxin can result in human illness (74. Globally. Local island populations attempt to avoid poisoning by catching only smaller fish. and although the gastronomical problems may dissipate within a day or so. barracudas. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . myalgia. there are neither standards nor an official testing program. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. 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. which can be confused with chronic fatigue syndrome. sharks. which can return following the consumption of alcohol and certain foods such as oily fish (74.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. which in turn are consumed by carnivores such as snappers. The onset of symptoms can occur rapidly within 24 hours. 78).8. 75).76). and moray eels. where cold objects feel hot and vice versa. and it is estimated that ingestion of as little as 0. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs.
minor revisions of a few stereocenters were made. This is then subjected to LCMS/MS analysis. high-throughput screen (74). Recently. almost 20 additional derivatives have been reported (83 . and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. Recent reviews of this new group of toxins have appeared (83. The original structure proposed revealed an entirely new class of shellfish toxin.85). and another dealing with the chemistry and detection methods has been published (85).FISH AND SEAFOOD Lethal Dose (MLD) 0. specific for CTX activity. only a few laboratories are able to undertake such analyses. Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. and the corrected structure was published in 2004 (86) (Figure 7.20%) present in lesser amounts (Figure 7. has been developed.9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland.25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required.g. Since the original discovery and identification of the toxin. another in vitro sodium channel-based assay.9). and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. at C-3 and C-23). In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). Usually the major AZA derivatives found in shellfish are AZA1 (50 . following consumption of contaminated mussels Mytelius edulis (82). 84). and following synthesis of the molecule some time later. The majority of these have been proposed based on LC/MS analytical data. 158 .9).30%) and AZA3 (5 .5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. Consequently. which are based on an LC-MS/MS approach.80%). and only a few structures (AZA 1 . followed by clean-up on silica and aminopropyl cartridges. A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine. with AZA2 (10 . but this only has application in a clinical setting (81). although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). 7. Immunoassays may be an alternative detection method (79). which features characteristic fragment ions corresponding to sequential losses of three molecules of water. Chemical detection methods are severely compromised by the lack of standards. Acetone extracts of fish tissue are subjected to solvent partitioning.
Ensis siliqua. cassipes produces AZAs in culture. oysters (Crassostrea gigas. and to date this has been the only source from which these toxins have been found at toxic levels. clams (Tapes philipinarum. Following the original episode.9. these molluscs were never considered to be the biogenetic source.9. 7.3 days. diarrhoea and stomach cramps. and off the coast of Morocco (83. 84. and to date no long-term effects have been recorded (83. Scandinavia. vomiting. which are ingested by P. AZA toxins do 159 . it has been impossible to establish that P. 7. 88. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). which can persist for 2 . but current data indicates that unlike the DSP compounds. cassipes (83).).9. 89). While this result is not necessarily in debate. However. Mytilus galloprovincialis). The bivalves affected include mussels (Mytilus edulis. scallops (Pecten maximus). There has been considerable debate over the source of azaspiracids. Portugal.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. AZA occurrence has been reported by several European countries including France. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. Ostrea edulis). 90).FISH AND SHELLFISH TOXINS Fig. These symptoms are not unlike those experienced following the consumption of DSP toxins. cockles (Cerastoderma edule). In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. as in other cases of shellfish toxins. and most recently in a crustacean (Cancer pagurus). and some time after the original episode in Ireland. Structure of the three principal azaspiracids found in shellfish 7. Donax spp.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea.
85).12). and ASP. the spirolides. Using the same approach. Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. 7. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . 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. Based on IP minimum lethal dose data. and closely related ptierotoxins (Figures 7. which are particularly amenable to detection by positive ion electrospray mass spectrometry.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. abnormalities in the thymus and spleen. but all contain a spirocyclic or cyclic imine function within their structure.10 – 7. parenchymal cell pyknosis in the pancreas. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. interferences can arise when analysing a matrix as complex as a crude shellfish extract and it is recommended that a simple solid phase extraction (SPE) step be introduced prior to LC/MS analysis (93). and erosion and bleeding in the stomach. the pinnatoxins.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. 91. Once again. PSP. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes.9. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. 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. The mouse bioassay has been used to monitor shellfish extracts. 92). though the possibility exists that they may inhibit other phosphatases.10 Cyclic imines These toxins are represented by several distinct structural types. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). and necrosis of epithelial cells on the microvilli (91). 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. 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. However. 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. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action.
Subsequent taxonomic studies revised this again to a new dinoflagellate K. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. surf clam and New Zealand abalone.. 7. Some time later. 161 .10. cockles. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94.1 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. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). they are sometimes referred to as emerging toxins of uncertain human health concern.10. Structure of the cyclic imine gymnodimine 7. it was proposed that a dinoflagellate identified as Gymnodinium spp.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. 95). two additional congeners. 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. but they are included here for completeness. 95). Fig.FISH AND SHELLFISH TOXINS health.10). 7.1. scallops. At the time of the initial observation of gymnodimine. Pacific oyster. In only two cases (gymnodimines and spirolides) has a planktonic source been identified.10. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 .520 Da (94. selliformis (97).
11). 7. Whether isolated from dinoflagellate cells directly or from shellfish tissue. The time lapse between ingestion and death is between 5 and 15 minutes.FISH AND SEAFOOD 7. together with a lack of evidence of human intoxication. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. They were originally discovered in extracts of mussels (M. There is a rapid onset of symptoms including respiratory distress. abdominal breathing. paralysis of the hind quarters and eventually complete immobility.1. almost a dozen derivatives in total have been identified to date (94. Most cyclic imines lack an extended chromophore. In particular.100).10. and lack of responsiveness to stimuli. Reduction of the imine group eradicated the toxicity. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. 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.3 Detection methods The characteristic “fast-acting. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). A later study using purified toxin established an LD50 dose (Lethal Dose. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. Since their original discovery which resulted in the characterisation of spirolides A-F (99. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode.to10-fold reduction of toxicity (LD50 600 . However. 50%) of 96 µg/kg. and this is the detection method of choice. 95) (Figure 7. 95). finishing with 100% methanol. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction.10. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . Oral administration of the toxin to mice by gavage resulted in about an 8.10.900 µg/kg). 7. Based on this diminished oral toxicity. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. and so spectroscopic detection by UV methods is not favoured. it has been stated that regulation against gymnodimine is not required (98).1.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds.
the western coast of Scotland.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. or by oral ingestion (381 .707 µg/kg. Structure of four spirolide toxins A-D 7. fasted mice). The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . All data reported here are for desmethyl spirolide C.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). 7.2. Interestingly. those spirolides possessing the vicinal dimethyl grouping (see Figure 7.10.2. fasted mice).166 µg/kg. the gulf of Maine. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). 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). these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 .2 Toxicology The spirolides are the most studied group of the cyclic imines. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. 7.10. southern Ireland. Structural differences among the various spirolides result from variations in the number of methyl groups and the sequence of spiroketal rings 6-5-5 in one group and 6-6-5 in the other.8 µg/kg).11. 95). Fig. While oral toxicity is considerably less than by IP injection. by gavage (87 . in fjords in Denmark and Norway. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada.11.
7. Structure of the pinnatoxin group and the closely related pteriatoxins 164 .3 Detection methods As discussed for gymnodimine. consistent with modulation of acetyl cholinergic receptors (103). 102). The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. Fig.2.10. four members of the group have been identified – two of them being the C-34 diastereomers. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C.12). However.12.3 Pinnatoxins and pteriatoxins Pinnatoxins A . 95. Three members of this group have been reported. the mouse bioassay can be used to detect these fast acting toxins.B contain a spiro imine moiety similar to that found in the spirolides. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. and in fact these compounds share a number of other structural features. The compounds are zwitterionic and therefore somewhat more polar than the spirolides.FISH AND SEAFOOD (100). 7. 7. which includes two diastereomers. To date.10. more useful information on the amount and distribution of toxins present in a shellfish extract is obtained by LC-MS analysis usually following a simple clean-up step. A number of methods following this procedure have been described (94. While the mechanism of action remains to be clearly defined.
3 Detection methods No formal detection methods for these compounds have been reported. Coupled with this. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins.FISH AND SHELLFISH TOXINS 7.10. arise from an algal source .10. Despite this progress. have long been suspected of causing human illness after ingestion. There is strong circumstantial evidence to suspect that these toxins. While this is recognised as a real possibility for a wide range of toxins. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105).10. 7.2 Toxicology There is little information describing the toxicology of these compounds (98). With increasing knowledge of the chemistry of toxins. The LD99 (Lethal Dose. there is no compelling evidence to suggest that they have resulted in human toxicity (98).3. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures. while pinnatoxin D was essentially inactive.1 Origins and distribution Shellfish of the genus Pinna.11 Conclusion There has been considerable progress in the detection of known marine toxins. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). However. The LD99 dose for the pteriatoxins is reported to be in the range (8 . This is generally a result of increased monitoring and continually improving detection technologies and methods.3. and more advanced means of detection. 99%) values for pinnatoxins A-C are in the range (22 . but this remains to be conclusively established (106). but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. like the gymnodimines and spirolides. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. which in many cases are now found to be more widespread than previously believed. 7. 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.3. 7. which occupy tropical waters around Okinawa and the south east coast of China.100 µg/kg). to date action or 165 .180 µg/kg). Only the acute toxicity in mice following IP injection has been reported. new toxin groups such as the azaspiracids and cyclic imines have also been reported.most likely a dinoflagellate.
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M. Wanner A. 73. 2002. New fish-killing alga in coastal Delaware produces neurotoxins. Boca Raton.J.G.. Granholm A. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. Buonocore M. 220-6.. Ed. Aerosolised red tide toxins (Brevetoxins) and asthma. 707-23. Gandini C.S.. Kirkpatrick B.E. 77. Naar J. in Seafood and Freshwater Toxins. Manzo L. Chen A. Granade R.A..P. Bienfang P. Pierce R. Whitney P. in Seafood and Freshwater Toxins. Food and Agriculture Organisation. 171 . Prockop L. 74... Steidinger K.. 117-71..H. Poon-King C.. Weisman R..G. 187-94. Environmental Health Perspectives. Abbott J. Butera R. Marine Biotoxins.. Ed... Baden D. West Indian Medical Journal. Fleming L. Poon-King T. Mild ciguatera poisoning: case reports with neurophysiological evaluations.. in Seafood and Freshwater Toxins. 479-516.C. 110..E. Rome. 50. Jacocks H. Poli M.. 110.C.A.. Clark R... Baden D.G. Muscle Nerve...P. Kirkpatrick B. 519-50.E. Baden D. Reich A. Lenzi A.P. Ramsdell J. Squicciarini D. Locatelli C.N. Toxicon. Marine Drugs.G..A. 2007. Flewelling L. Bourdelais A..B.G. Dickey R. Henry M. Bossart G. Tomas C. 2000. 68.S. 2004.... Bourdelais A. Schrank K.. Backer L.M.... 99-115. Botana L. are potent ichthyotoxic neurotoxins that accumulate in fish. 3. Tomas C. shellfish and mammalian body fluid. 66. Naar J. Kubanek J. Bean J. 551-8. Abraham W. Harmful Algae. 2004. 2008. Food and Agriculture Organisation of the United Nations. Bottein M-Y. CRC Press. 2004. 70. Ed. Fleming L. Chest. 76..M.M.. 456-79. 53.. Reich A. Landsberg J... Ciguatera fish poisoning: treatment prevention and management. 131... Naar J. Wang Z.. The molecular and integrative basis to brevetoxin toxicity. Kubanek J.. Neurologic Shellfish Poisoning (NSP). Ramsdell J. and detection. Lancaster J.D. Ayyar R. 465-70.. Abraham W. Botana L...D.. 825-34. Botana L... Dickey R. 2008.. Baden D. 2007.. Literature review of Florida red tide: implications for human health. Wolny J. Baden D. 2002. 23. 2008. Flewelling L. Zaias J. 72.. Cheng Y.. Environmental Health Perspectives.J. 179-85.E.. CRC Press. Fleming L. Dechraoui M-Y. Pierce R.. Backer L.A.... Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2. Freidman M. 50.. Zaias J. 1598-603. 2007... Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater. Backer L.. Cheng Y. Naar J. Food and Agriculture Organisation. Boca Raton.E.. toxicology.M. Detection of brevetoxins in the twenty-first century.. 69. like ciguatoxins.S. Pierce R. 71.S. 75. Toxicon. 6... Ciguatera toxins: Chemistry. Fernandez M...W.. CRC Press. Ed. Gannon D. 67. Johnson D.R. Watkins S. 2008. Brevetoxins. Benson J..FISH AND SHELLFISH TOXINS 65. in FAO Food and Nutrition Paper 80. Boca Raton..
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.
Establish a system to monitor control of the CCP. Validation . quality assurance or control. 5. 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. 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. Conduct a hazard analysis. 2. 8.The application of methods. food chemistry or engineering. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. The team should also include 177 . Establish the critical limit(s).A point. ingredient and packaging buyers. procedure. food microbiology. Each stage is discussed below in detail. 4. Establish documentation concerning all procedures and records appropriate to these principles and their application. Determine the CCPs. Establish procedures for verification to confirm that the HACCP system is working effectively. 8.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. procedures.Obtaining evidence that the elements of the HACCP plan are effective.1 is a flow diagram illustrating this 12-stage logic sequence. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. hygiene managers. tests and other evaluations. 6.1 Assemble HACCP team HACCP requires management commitment of resources to the process. Figure 8. 7. 3. in addition to monitoring to determine compliance with the HACCP plan. operation or stage in the food chain including raw materials.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . from primary production to final consumption. Verification .3.
g. 8. Logic sequence for application of HACCP 178 .FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations.1. it may be obtained from external sources. Fig. biological. The scope of the plan should be determined by defining the extent of the production process to be considered and the categories of hazard to be addressed (e. chemical and/or physical). If expert advice is not available on-site.
which should be described in detail.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora.1. there may be species-specific hazards related to certain fish and shellfish. fermentation. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation.3. such as children or the elderly. naturally present on fish from warm waters (> 15 °C).3. including practices onboard fishing vessels. or in aquaculture operations. However. etc. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments. 8. heat treatment. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. freezing.2 Describe the product It is important to have a complete understanding of the product. physical and chemical structure (including water activity (aw).g.3. marine toxins associated with particular fish and shellfish species. 8. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. pH.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. packaging.3. 8. contaminated fish and shellfish should not be used. processing conditions (e. If the processing does not include a kill step.). such as enteric viruses in shellfish. In addition. aquaculture drug residues and potentially pathogenic Vibrio spp.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.3. Vulnerable groups of the population. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . shelf life. composition. It is also important to identify the consumer target groups.). etc.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. These factors are likely to have a significant effect on some potential hazards. is a cooking process required?).g.3. 8. storage and distribution conditions and instructions for use. The description should include information such as the product name. Expertise on such inherent hazards. may need to be considered specifically.
FISH AND SEAFOOD product. Ideally. 8. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. It is essential that the diagram is accurate. time/temperature data. 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. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. However. features of equipment design. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. and therefore a detailed description of the procedure applied must be available. 8. It may include details of the factory and equipment layout. For example.3.3. cleaning and hygiene procedures and storage conditions.3.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. 180 ● ● . it is essential that the details of each step are fully appreciated and recorded. 8. the qualitative and/or quantitative evaluation of the presence of hazards.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.4. conduct a hazard analysis. or circumstances of any reclaim or rework activity. because the hazard analysis and decisions regarding CCPs are based on these data. it should also include details of CCP steps. survival or multiplication of pathogenic microorganisms. once determined. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. The operation should be observed at all stages. If HACCP studies are applied to proposed new process lines or products. The team should then conduct a hazard analysis to identify which hazards are of such a nature that their elimination or reduction to an acceptable level is essential to the production of safe food. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. 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. ingredient specifications.
Vibrio spp. the number of consumers potentially exposed and their vulnerability.g.). and the potential growth of Listeria monocytogenes on smoked fish. Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. Hazards introduced during processing of fish products depend very much on the characteristics of the process. any food safety objectives or manufacturer’s food safety requirements. and there are an equally varied range of potential hazards associated with them. Pathogens may be part of the resident microflora of the living animal (e. Note: it is important at this stage that no attempt is made to identify CCPs. botulinum. and viruses). modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. For example. 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. or desirable.g. where a hazard is repeated. 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. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual.6. The HACCP team should then determine what control measures exist that can be applied for each hazard. For example.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products.3. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin. The incidence of psychrotrophic. Therefore it is not possible. Salmonella spp. since this may interfere with the analysis. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage. 181 . One control measure may be relevant to several process steps. the hurdle effect. or may originate from polluted water or from post-capture contamination (e. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. 8.
8. it is not possible to generalise on likely CCPs.3.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. it can be said that effective control measures are likely to include the following: • • • • • 8. but for which no control exists at that step or any other.FISH AND SEAFOOD 8. Criteria used to set critical limits must be measurable and may include physical. For example.7. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. However. structured approach to decision making.3. Where possible. 8. Some examples relevant to fish products are: • • • Time and temperature limits for pasteurised and cooked products Temperature limits for chilled fish Brine concentration in cured fish 182 .3. biological or sensory parameters.3.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. 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. However. If a significant hazard has been identified at a step where control is required for safety. 8. it is necessary to specify both time and temperature for a thermal process. critical limits should be specified and validated for each CCP. and the reader is referred to the appropriate product chapter. then the process must be modified to include a control measure. chemical.2) to provide a logical.1 Fish and seafood Again.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. and its use may not always be appropriate. More than one critical limit may be defined for a single step. application of the decision tree should be flexible. given the enormous variety of fish products and processes in use.
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.
Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety. 14-9. International Journal of Food Microbiology. 1992. Development and use of the HACCP concept in fish processing. 1988. Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H.. WHO. 3 (1). Blackwell Scientific Publications. Food Control. 1992. 188 . World Health Organisation. International Commission on Microbiological Specifications for Foods. Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. Oxford. Microorganisms in Foods. 15 (1/2). 1992. Bryan F. Mayes T.L.H. 33-44. Geneva.
and one that plays an important role in most countries’ food legislation.9. including microbiological criteria. 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. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials. 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. A number of standards are provided in food legislation. to personnel requirements. The new legislation was designed to establish conditions. and conditions of raw materials. final product specifications. Hygiene is a general concept that covers a wide subject area. under which food is produced to optimise public health and to prevent. have been replaced by several linked measures on food safety rules and associated animal health controls. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. to sale or supply to the final consumer. transport and delivery vehicles requirements. with the intention of introducing consistency and clarity throughout the food production chain from primary production. 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. the existence of microbiological standards alone cannot protect consumer health.1 Introduction Hygiene is an important aspect of ensuring food safety. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . however. from structural conditions in the factory or process facility. eliminate or acceptably control pathogen contamination of food.
1991. Subsequently. 7. European Communities L 268.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. and sets out what those enforcing the provisions have to do.9. J. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. as last amended by Regulation (EC) No 806/2003. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. 24.1992.15). 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.1). Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation.7.B. p.9. p. A number of more detailed implementing and transitional measures have been adopted at EC level. transportation. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. packaging. as last amended by Regulation (EC) No 806/2003. 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 187.1991. manufacturing. 41).2 Legislative Structure From 1 January 2006. storing. 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. p. 190 • • . processing. 24. handling and offering for sale or supply to the consumer. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. 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. N. 9. distribution.
of stock farming. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. and binding in its entirety on all member states from the date of entry into force. domestic preparation or handling. which will be covered later in this chapter. penalties and powers of entry. Although the regulations have the force of law. is required to give effect to the EU regulations. Wales and Northern Ireland). revocation of existing implementing legislation.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. As regulations. Primary products are defined as products of primary production including products of the soil. setting offences.3 Regulation (EC) No. e. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer.I. There are some exemptions.14. 9.7. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. p. 1 .g. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. etc. Additionally the requirements of EC Regulation 853/2004 must be complied with. with primary production. The Food Hygiene (England) Regulations 2006 (S. 2006 No. 2073/2005. right through to the implementation of procedures based on HACCP principles. 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. Wales and Northern Ireland. 19.I. 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. and equivalent legislation in Scotland. the legislation is directly applicable law.) in England. national legislation in the form of a Statutory Instrument (S. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. 191 . processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. food storage that is for private or domestic consumption. 852/2004. and of hunting and fishing. for example.11) The EU hygiene regulations apply to all stages of food production including primary production.1993. 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.
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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: 184.108.40.206 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.
220.127.116.11 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.
18.104.22.168 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.
22.214.171.124 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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126.96.36.199 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.
188.8.131.52 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.
184.108.40.206 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.
220.127.116.11 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. 18.104.22.168 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.
22.214.171.124 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 126.96.36.199 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). 188.8.131.52 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:
197 . slime or sand. live tunicates and live marine gastropods. where necessary. Mechanically separated fishery product means any product obtained by removing flesh from fishery products using mechanical means resulting in the loss or modification of the flesh structure. to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing. slicing. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. shelling. Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. after preparatory work such as bleeding. skinning. if necessary: chilling or freezing: filleting. gutting and removal of fins and. heading. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. and chopping. where appropriate. reptiles and frogs) whether wild or farmed and including all edible forms. including products packaged under vacuum or in a modifiedatmosphere that have not undergone any treatment to ensure preservation other than chilling. purification centres or dispatch centres in tanks or any other installation containing clean seawater. Conditioning means the storage of live bivalve molluscs coming from class A production areas. heading. filleting. and all mammals. or in natural sites. such as gutting. live echinoderms. slicing. shucking. parts and products of such animals. 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. in particular as a result of feeding on plankton containing toxins. followed by wrapping or packaging. to remove mud. Fresh fishery products means unprocessed fishery products. mincing or processing. whether whole or prepared.
a new mark must be applied to the product. 9.4. For the UK.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. EC. In this case. EY. the mark must show the approval number of the establishment. ES. Extracts of the requirements of Annex II. for other codes.4.1.2 Form of the identification mark (i) The mark must be legible and indelible. 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. (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.FISH AND SEAFOOD 9. EÜ. please refer to Regulation (EC) 853/2004. (iii) Additionally.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. the food business operator may apply the same identification mark to both types of food.2. when a product's packaging and/or wrapping is removed or is further processed in another establishment.1.4. (ii) The mark must indicate the name of the country in which the establishment is located and written out in full or shown as a two-letter code in accordance with the relevant ISO standard.4. EO.2. EB or WE.2 Fishery product requirements 9. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. EG. Section I of Regulation (EC) 853/2004. EK. the new mark must show the approval number of the establishment where this occurred.2. and the characters easily decipherable and be clearly displayed. EK. (iv) If the identification mark is applied in an establishment within the Community. the code would be UK. 198 . Section I of Regulation (EC) 853/2004 are given as follows: 9. the mark must be oval shaped and include the abbreviation CE.
Section VIII of Regulation 853/2004. The requirements of this section accompany those in Regulation (EC) 852/2004. Chapter IV. Chapter III. Therefore. Extracts of the requirements of Annex III. and are intended for further handling. Parts A. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. it applies to those animals when not placed on the market live. This section does not apply to bivalve molluscs. granulate and powdered products of animal origin carried in bulk.4.4. 9. the colourings used must be EC authorised colours. i. and fishery products carried in bulk. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. or be printed on a label affixed to the product. Part A and Chapter V apply to retail. they would need to be obtained following Section VII on live bivalve molluscs. (ii) For products of animal origin that are placed in transport containers or large packages.e. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . a) requirements for establishments. the mark may be applied to the outer surface of the container or packaging. the wrapping or the packaging. C and D.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. echinoderms.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. specifically relating to fishery products are given below. processing. it is sufficient to apply the mark to the exterior of that package only. its wrapping or the packaging. reference should be made to the actual Regulation. wrapping or packaging in another establishment. (v) When the mark is applied directly to products of animal origin. tunicates and marine gastropods when placed on the market live. With the exception of Chapters I and II.LEGISLATION 9. for full requirements.3 Method of marking (i) The mark is to be applied directly to the product. the approval number of the establishment and the abbreviation for the community where appropriate. as amended.1.2. including vessels. (iii) In the case of liquid.
Chapter VII of Regulation 852/2004. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. clean seawater may be used for the handling and washing of fishery products. or to handle or process them after harvesting. (a) primary production covers the farming. i. they also include (i) the transport and storage 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. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 . There is a derogation from point a.) 9. it must be placed somewhere that will not contaminate the water supply. 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.2. Structural and equipment requirements refer to the following points 1. this supplements the requirements of Annex II. 2.1 Chapter I . bleeding. fishing and collection of live fishery products with a view to their being placed on the market. if a vessel has water coming in that is used with fishery products.FISH AND SEAFOOD c) for the water supply.2. gutting. refrigeration and wrapping. removing fins. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material. from the place of production to the first establishment of destination. heading. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking.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.e.4.. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter.
(iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas. tanks must incorporate devices that allow a uniform temperature throughout the tanks. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. A temperature recording device must be present and placed where the temperature in the hold is the highest. Also. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. (ii) Holds must be separated from the engine compartments and from the crew quarters by partitioning that prevent any contamination of the stored fishery products. where necessary. (iii) In vessels equipped for chilling fishery products in cooled clean seawater. 4. and allow the monitoring and. any waste processing unit needs to have a separate hold to store the waste. where necessary. Storage areas for the finished products also need to be sufficiently large and cleanable. Such devices must achieve a chilling rate that ensures that the mix of fish and clean seawater does not exceed 3 °C six hours after loading.LEGISLATION (i) Such vessels need to have holds. recording of temperatures. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. 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. 201 . 3. to ensure that melt water does not stay in contact with the products. and not more than 0 °C after 16 hours. Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C.
e) In cases where whole fish intended for canning is frozen in brine.Requirements during and after landing 9. where fishery products are displayed for sale.2. provided no damage occurs to the flesh. 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. b) Factory vessels on board which crustaceans and molluscs are cooked. chilled and wrapped. and products washed thoroughly.4. B. Chapter II .2. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. the product must be kept at no more than -9 °C. Handlers may use spiked instruments to move large fish or fish that might injure them. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption. c) Fishery products.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. other than those kept alive. landed as soon as possible. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. (ii) In the case of food business operators responsible for auction and wholesale markets etc. 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 . The brine must not be a source of contamination. must be chilled or. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. if this is not possible. Hygiene requirements Hygiene requirements refer to the following: a) Parts of vessels used for storage of fishery products must be cleaned and maintained in a good state with no contamination.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.
9. 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. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. Chapter I.2. They should be wrapped. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed. points 1 and 2. C. B. dispatched.2. prepared or processed immediately after arrival at an establishment on land. Additionally. handling fishery products A. 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.LEGISLATION consumption. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. packaged and chilled as quickly as possible after preparation. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . and where necessary. including vessels.4. (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. 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. (ii) Heading and gutting must be carried out hygienically. part I. Fillets and slices must then be removed from work tables as soon as possible after preparation. they must be stored under ice in appropriate facilities. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice.3 Chapter III . 854/2004. C.
(ii) mackerel. raw materials must have no guts. or a stabilising treatment.2. D. (b) fishery products from the following species. (iii) A document from the manufacturer. 9. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. and whole fish must be gutted and washed beforehand.Requirements for certain processed fishery products A. cooling must continue until a temperature 204 . if the processing is insufficient to destroy nematode larvae.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. Once produced. If no other method of preservation is used. 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. (iii) sprat.4. (iv) (wild) Atlantic and Pacific salmon. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. must accompany fishery products referred to above when placed on the market. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . they must be cooled rapidly.2. and (c) marinated and/or salted fishery products.4 Chapter IV . (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. except when supplied to the final consumer. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. stating the type of process they have undergone.
However. the following conditions need to be met: A. refining and purification steps before being placed on the market for the final consumer. and after this. 2073/2005. 2074/2005.5 Chapter V .Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. B. B. Shelling/shucking is to be done hygienically. Total volatile nitrogen Unprocessed fishery products cannot be placed on the market if chemical tests reveal that TVB-N or TMA-N limits are exceeded (limits are set in Regulation (EC) No. separation. raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. there is an exception: food business operator may refrain from chilling the fishery products when whole fishery products are used directly in the preparation of fish oil for human consumption. 9.2. 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. centrifugation. 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). 2074/2005). (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. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. 205 . Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport.4. 2073/2005).LEGISLATION approaching that of melting ice is reached. pressing.2. and the raw material is processed within 36 hours after loading. processing.
Molidae. The scientific name must accompany the common name on the label.2. it should be kept at no more than -9 °C.6 Chapter VI . in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. 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. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing.4. E. In the case of whole frozen fish in brine intended for the manufacture of canned food. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market.4.FISH AND SEAFOOD D. 9. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice.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. 9.2. Fresh. and when fishery products are wrapped on board fishing vessels.Storage of fishery products (i) Fresh fishery products.2.2. Fishery products contaminated with parasites may not be placed on the market for human consumption. may only be placed on the market in wrapped/packaged form and must be appropriately labeled to provide information to the consumer on preparation/cooking methods and on the risk related to the presence of substances with adverse gastrointestinal effects. 206 . prepared and processed fishery products belonging to the family Gempylidae. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product.7 Chapter VII . Diodontidae and Canthigasteridae. thawed unprocessed fishery products.
9. Helicolenus dactylopterus.2. The temperature of frozen fishery products must not fluctuate above -15°C. provided that the journey is short and the competent authority allows this. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. melt water must not remain in contact with the products.3 Other information: Rules relating to visual inspections for the detection of parasites in fishery products are stated in Annex II (obligations on the competent authorities) of Regulation (EC) No. species belonging to the Merlucciidae family.).4. Note: (i) Frozen fishery products do not need to be kept at a temperature of -18 °C if they are being transported from a cold store to an approved establishment to be thawed on arrival for the purposes of preparation and/or processing. 2074/2005.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them.8 Chapter VIII .Transport of fishery products The above points for Chapter VII apply in relation to transport. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. 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. 9.2. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. Sebastichthys capensis. species belonging to the Gadidae family.2.4. (ii) If fishery products are kept under ice. (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 .
FISH AND SEAFOOD Regulation (EC) No 853/2004. as amended. The sample must consist of about 100 g of flesh. (b) and (c) of Part B(1) of that Chapter. where the raw material complies with points (a). Chapters V.3. (b) In the case of other operations. please refer directly to the regulation.General requirements for the placing on the market of live bivalve molluscs Live bivalve molluscs can only be put on the market for retail sale if done through a dispatch centre. and point 3 of Chapter VII applies to retail. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. Live bivalve mollusc requirements - 9. The information required in the registration document is given in point 4 and includes the health status of the production area. Extracts of the requirements of Annex III. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. 9. shellfish species etc. VI. reference should be made to the actual regulation. they supplement the requirements of Annex I to that Regulation. taken from at least three different points and mixed together by grinding. 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. however. Apart from the provisions on purification. The copy of 208 . Section VII of Regulation 853/2004. this Section also applies to live echinoderms. 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. 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 854/2004.4.3 The regulation details specific hygiene requirements for bivalve molluscs. tunicates and marine gastropods.4. VIII and IX.1 Chapter I . they supplement the requirements of Annex II to that Regulation. specifically relating to bivalve molluscs are given below. for full requirements. Chapter IX applies to pectinidae harvested outside those areas.
(ii) Provided the requirements of chapter V are met.Class A means they can be placed directly on the market . Requirements for relaying live bivalve molluscs Only areas approved by the competent authority can be used for relaying live bivalve molluscs with a requirement that buoys. B.3.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority. B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. Transport needs to allow sufficient drainage for best survival conditions.4. processing or relaying. poles or other fixed means are to identify the boundaries of the sites. collection from: . 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. they need to be sent to a processing establishment and undergo specified treatments such as sterilisation in hermetically sealed containers or specific heat treatments such as immersion in boiled water so the internal temperature of the mollusc flesh reaches at least 90 ºC and this temperature is maintained for at least 90 seconds. 209 .Class C means relaying over a long time before it can be placed on the market For bivalve molluscs not subject to Class B or C treatments.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market . Requirements for production areas (i) The competent authority classes (classed A.Hygiene requirements for the production and harvesting of live bivalve molluscs A. C.2 Chapter II . 9.
Requirements for dispatch centres Handling should not result in contamination to any degree. (iv) yessotoxins. the purification system to allow them to resume /maintain filter-feeding activity. 800 micrograms per kilogram.3. B. for: (i) paralytic shellfish poison (PSP). shells of live bivalve molluscs are to be washed throughout with clean water and the molluscs are to come from one of the following: (i) Class A production area (ii) Relaying area (iii) Purification centre (iv) Another dispatch centre 9.3.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. the levels of marine biotoxins should not exceed the following limits. (ii) amnesic shellfish poison (ASP). Requirements for purification centres There are requirements for live bivalve molluscs to be washed.4.4. 160 micrograms of okadaic acid equivalents per kilogram.3. and there are requirements for tanks and water storage containers.FISH AND SEAFOOD 9. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. 210 . containers and a label to identify purification.3 Chapter III . dinophysistoxins and pectenotoxins together.Hygiene requirements for purification and dispatch centres A.4. 1 milligram of yessotoxin equivalent per kilogram.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc. Before dispatch.4 Chapter IV . 9.5 Chapter V . requirements if there are many batches of live bivalve molluscs.
Identification marking and labelling In addition to general identification marking (contained in 9.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.LEGISLATION (v) azaspiracids.6 Chapter VI . the species of bivalve mollusc and date of packaging (at least the date and month) are required.1).4. Finally. Registration document requirements also apply. Requirements of Chapters III and IV are to be complied with. 9. by way of derogation from directive 2000/13/EC. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. (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. 160 micrograms of azaspiracid equivalents per kilogram.2.3. including the identification mark. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.4.3.4. Also. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’. 9. dispatch centre or processing establishment. they cannot be re-immersed in or sprayed with water.4.4. 211 .Other requirements (i) The temperature at which live bivalve molluscs are stored/ transported in should be at one that does not adversely affect food safety or their viability.3. the label. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9. 9.7 Chapter VII .8 Chapter VIII .3. must be waterproof.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.9 Chapter IX .
1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures. 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. This refers to official controls or production and placing on the market of fishery products. or - - 212 . 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. they derive from: (i) poisonous fish.FISH AND SEAFOOD 9.5. (ii) fishery products not complying with the requirement concerning biotoxins. 9. Section VIII. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. chemical. 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. Chapter V of Regulation 853/2004 apply. Therefore the health standards for fishery products as previously stated in Annex III. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004. Fishery products in particular must comply with the requirements of Annex III of the regulation. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given.5 Regulation (EC) No. or (iii) bivalve molluscs. In this regulation. echinoderms. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation.
600 E. live tunicates and live marine gastropods. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated.LEGISLATION - the competent authority considers that they may constitute a risk to public or animal health or are for any other reason not suitable for human consumption. live bivalve molluscs must not exceed 46.1 Chapter II . chapter V is reinforced. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. coli per 100 g of flesh and intravalvular liquid. as previously stated. 000 E. A summary is given as follows: 9. live echinoderms. - - - - B. Sampling plans and frequency are required. in 90% of the samples. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004.2 This states official controls for the production and placing on the market of live bivalve molluscs. The reference method for this analysis is the five-tube. coli per 100 g of flesh and intravalvular liquid. coli per 100 g of flesh and intravalvular liquid. Also. In the remaining 10 % of samples.5. 4.5. Live bivalve molluscs from Class C areas must not exceed 46. three dilutions MPN test specified in ISO 16649-3.2.Official controls concerning live bivalve molluscs from classified production areas A.000 E. Annex II 9. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. Regulation (EC) 854/2004. Live bivalve molluscs from Class B areas must not exceed. that microbiological criteria and chemical contaminants amongst other requirements are met. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. The reference method for this analysis is the five-tube. Classification of production and relaying areas: This section reinforces that specified in Annex III. 213 . section VII.
E.2.6 Regulation (EC) No. Procedures for re-opening a closed production area are stated D.FISH AND SEAFOOD C. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. boundaries or class of a production area or its closure. 9. 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. 2073/2005. 9. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions.2 Chapter III . 9. which has applied since 1 January 2006. establishes microbiological criteria for a range of foods. 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. opening or closing a production area. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying.5. the Member States may permit the import of bivalve molluscs and fishery products from the countries listed respectively in Annex I and Annex II to regulation (EC) 2076/2005. 214 . the competent authority is to close the production area.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. F. Additional monitoring requirements This concerns the competent authority in relation to monitoring of classified production areas where bivalve mollusc harvesting is forbidden and of relaying and production zones to ensure there is no malpractice etc.5.
volume.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. and the legislation applies to all businesses involved in food production and handling. 9.1 Food safety criteria Chapter 1 of the regulation focuses on food safety criteria that cover foods such as ready to eat foods and fishery products and live bivalve molluscs. ensuring that foods being placed on the market do not pose a risk to human health. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. 215 . presence or number of microorganisms. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. 9. and/or on the quantity of their toxins or metabolites. a batch of foodstuffs or a process.6. per unit(s) of mass.4 referring to fishery products. with chapter 2. area or batch. based on the absence.II. Two kinds of criteria have been established: food safety criteria.I.LEGISLATION The aim of this legislation is to complement food hygiene requirements.6. The relevant criteria are outlined in Table 9. The criteria are outlined in Table 9.
TABLE 9. monocytogenes 1. monocytogenes L. monocytogenes.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.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 1.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life .3 Ready-to-eat foods unable to support the growth of L.2 Ready-to-eat foods able to support the growth of 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. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1. monocytogenes. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1. other than those intended for infants and for special medical purposes(4) (8) L.
17 Live bivalve molluscs and live echinoderms. coli (15) 1.Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1. tunicates and gastropods 9 (18) 2 E.25 Live bivalve molluscs and live echinoderms. 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.
References: 1. Particularly fish species of the families: Scombridae..4 or aw ≤ 0. 79. products with pH ≤ 5. Bouquelet S. biscuits and similar products. A pooled sample comprising a minimum of 10 individual animals. uncut and unprocessed vegetables and fruits. to the satisfaction of the competent authority. — bread. Malle P. products heat treated in their final package). soft drinks. The most recent edition of the standard shall be used.. Dervin C..FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. monocytogenes. shall not apply. Single samples may be taken at retail level. J. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002.1 .25. when he is not able to demonstrate.1.. Assay of biogenic amines involved in fish decomposition. beer. Scombresosidae. live bivalve molluscs. Malle P. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . E. c = number of sample units giving values between m and M For point 1. subject to scientific justification. honey and confectionery. spirits and similar products. coli is used here as an indicator of faecal contamination. Bouquelet S. This criterion shall apply to products before they have left the immediate control of the producing food business operator. m = M. AOAC Internat. Clupeidae. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. that the product will not exceed the limit 100 cfu/g throughout the shelf life. Valle M. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. 43-9. — fresh. Pomatomidae. 5 This criterion shall apply if the manufacturer is able to demonstrate. 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. when recontamination is not possible after this treatment (for example.. 1996.0 and aw ≤ 0. Coryfenidae. cider. 2.92.94. products with a shelf-life of less than five days shall be automatically considered to belong to this category. Other categories of products can also belong to this category. 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. according to which the whole batch is to be deemed unsafe. Products with pH ≤ 4. Duflos G. wine. sugar. Engraulidae. including cocoa and chocolate products. to the satisfaction of the competent authority. — — bottled or packed waters. excluding sprouted seeds.
I The limits given refer to each sample unit tested. if all the values observed indicate the absence of the bacterium. where the limit refers to a pooled sample. if all the values observed indicate the absence of the bacterium. if all the values observed indicate the absence of the bacterium. if all the values observed are ≤ the limit. the mean value observed is ≤ m 2. L. if any of the values are > the limit. if the presence of the bacterium is detected in any of the sample units. monocytogenes in ready-to-eat foods able to support the growth of L. — unsatisfactory. L. monocytogenes before the food has left the immediate control of the producing food business operator when he is not able to demonstrate that the product will not exceed the limit of 100 cfu/g throughout the shelf life: — satisfactory. 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. Histamine in fishery products from fish species associated with a high amount of histidine: — satisfactory. 1999. if the presence of the bacterium is detected in any of the sample units. L.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory. — unsatisfactory. — unsatisfactory. if the following requirements are fulfilled: 1. coli. excluding live bivalve molluscs and live echinoderms. (1) 219 . — unsatisfactory. J. if the presence of the bacterium is detected in any of the sample units. — unsatisfactory. Interpretation of the test results relating to Table 9. AOAC Internat. 82. The test results demonstrate the microbiological quality of the batch tested(1). 1097-101. monocytogenes in other ready-to-eat foods: — satisfactory. a maximum of c/n values observed are between m and M 3. no values observed exceed the limit of M. Salmonella in different food categories: — satisfactory. tunicates and gastropods in relation to testing E. 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.
II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2.TABLE 9.4. 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.
and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. which overrides the 8 °C requirement. if one or more of the values observed are > M or more than c/n values are between m and M. 2006 No. Schedule 4 of the Food Hygiene (England) Regulations 2006. 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.7 Food Hygiene (England) Regulations 2006. 2006 No. For example. tolerance periods for chill-holding of foods and hot-holding variations.II The limits given refer to each sample unit tested. with chill holding tolerance periods.K. if all the values observed are ≤ m.1 In the U. E. 2 Interpretation of the test results relating to Table 9. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. 9. — acceptable.K) Temperature control requirements 9. c = number of sample units giving values between m and M The most recent edition of the standard shall be used. — unsatisfactory. The regulations prescribe a chilled food holding temperature of 8 °C or less.LEGISLATION 1 n = number of units comprising the sample. 14 (Hygiene requirements specific to the U. if one or more of the values observed are > M or more than c/n values are between m and M. 14 details temperature control requirements for foods in general. — unsatisfactory. The regulations provide for defences in relation to upward variations of the 8 °C temperature. had not been 221 . if a maximum of c/n values are between m and M.7.I. the defendant will need to prove that the food was on service or display. if all 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. S. — acceptable. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. The test results demonstrate the microbiological quality of the process tested. and the rest of the values observed are ≤ m. The defendant may be required to produce well-founded scientific proof to support his claims. if a maximum of c/n values are between m and M.I. and the rest of the values observed are ≤ m. 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.8 Guidance In the U. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing. 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. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment. for a limited period consistent with food safety. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety.I.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . 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. 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. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S.gov. 2006 No. and was kept at above 8 °C or the recommended temperature. or. S. Alternatively. 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. 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). 9. For other food.S.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours.food. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. was kept at above 8 °C or the recommended temperature for an unavoidable reason. 2006 No.K. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions.
10 Further Reading Regulation (EC) No. 2074/2005 and Regulation (EC) No. L283. 25.4.6. L226. 3) as amended by Regulation (EC) No. 25. L139. 2074/2005. The revised text of Regulation (EC) No. Regulation (EC) No.6. 2074/2005. L139. 25. 1688/2005. Regulation (EC) No. L226. 2076/2005. 30.2004. 2073/2005 on microbiological criteria for foodstuffs (OJ No. 14. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. 1662/2006.LEGISLATION 9. L338. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 223 . 1663/2006. 62) as amended by Regulation (EC) No. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. 206). 83) as amended by Regulation Nos 882/2004. 1019/2008 and as read with Regulation 2073/2005. 1). Commission Regulation (EC) No. L226. 55). 2075/2005 and Regulation (EC) No.2006. 2076/2005. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. The revised text of 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 (OJ No. as amended. Regulation (EC) No. 854/2004 is now set out in a Corrigendum (OJ No. Regulation (EC) No. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No.12. 1441/2007. laying down common marketing standards for preserved sardines. 22.9 Other Relevant Legislation The following regulations about fish designations need to be considered: • Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products Regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products • Note: Requirements for preserved fish are laid down elsewhere.6. 30.4.2004. Regulation(EC) No. 853/2004 is now set out in a Corrigendum (OJ No. The revised text of Regulation (EC) No. 30. these are not be discussed here. 2076/2005.4.2004. 2074/2005. 852/2004 is now set out in a Corrigendum (OJ No. L139. 1.2005. 9. namely.2004. as read with the corrigenda at OJ No.10.2004.2004. 2076/2005. Regulation (EEC) 2136/89. 22) as amended by Regulation (EC) Nos. Regulation (EC) No.
12. 21. as amended by Commission Regulation (EC) No 1022/2008 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. 6) 224 .2001.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.2005. 184.108.40.2060. 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. 22. (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. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.2005.12.
a 10 to 1.5.2 Oxygen requirements Facultative anaerobe. and 2. Aeromonas has the potential to grow over a pH range of ca 4 .5 pH Under otherwise optimal conditions.0 .4 Heat resistance Aeromonads are sensitive to moderately high temperatures. respectively (1). 10.1. Morphology Gram-negative rods. 225 .5 minutes.5.10 (3).1 Aeromonas spp. and D-values at 45 and 51 °C have been reported as 29. 10.1.1. The optimum growth temperature is 28 °C (2).1 10. under optimal incubation temperatures.1. The minimum pH value for growth is thought to be pH 5.5 µm 10.10.1.3 minutes.0 x 1. At pH 4. 10. PATHOGEN PROFILES 10. At 5 °C.3 Temperature Aeromonas grows at temperatures of 2 . Some death will occur at this pH if the temperature is 4 °C.1.000-fold increase in number is observed after one week in food systems (1).3.45 °C (1).3 . no growth is observed (1). 0.
6). forming at least seven different toxins. or at 90 ºC for a few seconds. but the interior of the food may do so. 10.2 Oxygen requirements Although C.2. botulinum types produce heat labile toxins. 10. 226 . 10. Thus an aerobically packed product may not support the growth of the organism on the surface.30 minutes.5 . F are the types that are responsible for most cases of human botulism (5. 10. botulinum is an obligate anaerobe. 0.4 x 1. being capable of slow growth and toxin production at low temperatures . All C. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C.0 µm.5% NaCl (4). Types A.1. which may be inactivated by heating at 80 ºC for 20 . to a lesser extent. A to G. botulinum are known.2.3 Temperature All strains of C. at 85 ºC for 5 minutes.1 Morphology Gram-positive. and is sensitive to > 4. botulinum is prevented. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth.2.2. but the spores of this organism are more so.even as low as 3 ºC (7).6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 .7 . and groups I and II are those responsible for the botulism cases. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different.2% sodium chloride (NaCl) (1). Group I will not grow at temperatures of 10 ºC or less. botulinum grow reasonably well in the temperature range of 20 45 ºC. botulinum are not particularly heat resistant.22.2. types B and F can be either. spore-forming rods.2 Clostridium botulinum Seven different types of C. All type A strains are proteolytic and type E strains are usually non-proteolytic. B. E and.FISH AND SEAFOOD 10. 10. but Group II strains are psychrotrophic.4 Heat resistance The vegetative cells of C. There are four main groupings of the organism.
97) (10. perfringens . parcooked meals with pH value > 5. botulinum depends on solute. 227 . These values have been established under carefully controlled laboratory conditions. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.2 Oxygen requirements C.3 10. pH and temperature. particularly by cooking. These concentrations correspond to limiting aw of 0.1 Clostridium perfringens Morphology Gram-positive spore-forming rods.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. 10. The organism will grow well in the centre of meat or poultry dishes.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. It will not. Note also that if humectants other than salt are used.like other clostridia . respectively (8. In commercial situations.2. vacuum packed.PATHOGEN PROFILES 10. However.0. the minimum aw for growth may be lower. they may survive mild heat treatments (70 .g. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0.97 for Group II (5).6 and pH 5.7 Characteristics of C. 10. grow on the surface of foods unless they are vacuum or gas-packed.2.2.0 and aw > 0. where oxygen levels are reduced.3.9 x 2. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. safety margins must be introduced to allow for process variability. 10. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. 10.1.21 minutes) (9).94 for Group I and 0.10.0 µm. 9).5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. 11)). botulinum spores The most heat resistant spores of Group I C.is an obligate anaerobe.0 . 0. therefore.3.3 .
FISH AND SEAFOOD 10. Recorded heat 228 . 10.3. perfringens spores The spores of C. lower concentrations may be effective in combination with other factors.3.3. Its optimum temperature for growth is 43 . will survive greater extremes of pH (and aw) (12. 15.3. perfringens in vacuum packed beef (18). 10.50 ºC.3. As in the case of other factors limiting the growth/survival of this organism. The lowest aw recorded to support the growth of C. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. type of solute. Under otherwise ideal conditions.3 Temperature The most significant characteristic of C. 0 . although C. perfringens strains. and their heat resistance is also affected by the heating substrate. The D-value for C.6 Aw/Sodium chloride C.5 minutes (17). At cold temperatures.3.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. pH. perfringens has the potential ability to grow within the temperature range 15 . 18). It grows best at pH values between 6 and 7 (the same pH as most meats).8% inhibit growth of most C. vegetative cells die rapidly (12).10 ºC. 10. perfringens in roast beef at 60 ºC is 14. 10. perfringens can vary quite considerably in their heat resistance. (It is unusual for spores to be formed in foods after the growth of this organism (14)). In addition. the enterotoxin is not heat resistant .97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. perfringens is not tolerant of low water activities.7 Characteristics of C.it is destroyed by heating at 60 ºC for 10 minutes (14. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12.93 and 0. perfringens appears to be 0. perfringens. 16). Salt concentrations of 6 . 13). however.5 pH C. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. Spores.45 ºC. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. While some growth can occur at 50 ºC. depending on strain and other conditions. perfringens is not a tolerant organism with respect to pH. 13). etc. the limits for aw are affected by temperature. Some studies indicate that the presence of 3% NaCl delays growth of C.
The upper temperature limit for the growth of L. monocytogenes in crawfish tail meat at 55. D-values for L. In foods.4 ºC is probably a more likely minimum for growth in foods (19).3 is the lowest value where growth has been recorded.5 . respectively (21).19 minutes.4. 10. using hydrochloric acid as an acidulant).2 has been suggested as the lowest working limit (22).8 minutes for heat sensitive spores (12).4.4. however.0 minutes for heat resistant spores to 1. 229 .4.6 . and where acetic acid is used as an acidulant. or even below 0 ºC.0 µm. 1. it is not a sporeformer.3 .4 Heat resistance L.4 10. being potentially capable of growing . requiring days or weeks to reach maximum numbers. Its optimum growth temperature. 10. -0. 0.64.0. so can be destroyed by pasteurisation. pH < 5. 60 and 65 ºC were reported as 10.2 Oxygen requirements Aerobe or microaerophilic.23. monocytogenes is reported to be 45 ºC (20). 10. monocytogenes is not a particularly heat resistant organism.at refrigeration temperatures down to.albeit slowly . Morphology Gram-positive short rods. However.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. the organism is able to grow at pH values well below pH 5 (pH 4.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic.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.98 and 0. growth at low temperatures can be very slow. 10.5 x 0.1 Listeria spp.4. Under ideal conditions. however.especially at refrigeration temperatures. 10. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.2. is between 30 and 37 ºC. and temperature.2.4 . the lowest limit for growth is likely to be considerably higher .
10.5.7 x 1. 10.5 (24.2 Oxygen requirements Facultative anaerobe.83 (20). 10.0.5. 10. 10.4.1. or even grow.6.3.90. at salt levels found in foods (10 . The bacterium may survive for long periods at aw values as low as 0. 25). the optimum being 30 ºC (23).1 Salmonella spp.5.0 .5 .6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24. monocytogenes is quite tolerant of high NaCl/low water activities.5 pH Plesiomonas has a pH range for growth of pH 4.45 ºC. 10. 0. It is likely to survive. It grows best at aw of ≥ 0.0 x 1 . peritrichous flagella.5.3 µm. 10.6 10.8.8 .12% NaCl or more).3 Temperature Plesiomonas can grow in the temperature range of 8 .0 µm. 0.5 10. but has been shown to be able to grow at an aw level of 0. 25).1 Plesiomonas Morphology Gram-negative.5 .6 Aw/Sodium chloride L.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).97.5.FISH AND SEAFOOD 10. 230 . Morphology Gram-negative short rods.5.
5 . However. The growth of Salmonella is generally inhibited by the presence of 3 . Salmonellae are quite resistant to freezing. Some death will occur at pH values of less than about 4. Salmonella has the potential to grow at aw levels as low as 0. and appropriate heat treatments must be determined experimentally for low-aw foods.9. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments.945. Salmonella senftenberg 775W is about 10 . a heat resistant organism.7.5 pH Salmonella has a pH range for growth of pH 3. therefore. or possibly 0.PATHOGEN PROFILES 10. It is not.6. depending on serotype.20 times more heat resistant than the average strain of Salmonella at high aw (27). some strains are able to grow at temperatures as low as 4 °C (26). However. 10.0.8 .6 Aw/Sodium chloride Where all other conditions are favourable. and with an appropriate acid. substrate. 10. with a z-value of 4 . pasteurisation and equivalent treatments will destroy the organism under normal circumstances. the optimum being 35 .6.5. Furthermore. Salmonellae are quite resistant to drying.4% NaCl. strains vary in their ability to withstand heating. although salt tolerance increases with increasing temperature (28). The optimal pH for Salmonella growth is between 6.6.48 ºC. Growth is slow at temperatures below about 10 ºC. D60 ºC values normally range from about 1 to 10 minutes.5.6. 10. depending on the type of acid and temperature.2 Oxygen requirements Facultative anaerobe.3 Temperature Salmonellae can grow in the temperature range of 7 . under otherwise ideal conditions.6.37 ºC. temperature and pH.93 (as reported in dried meat and dehydrated soup). 231 .5 ºC.4 Heat resistance Salmonella is not a spore-forming organism. 10.
9 µm diameter. aureus can grow within the temperature range 7 – 48.20 minutes.3 (optimum 7. stressed cells can also be less tolerant of heating. the D60 ºC value is 2 . The following relate to limits for growth only. the organism is heat sensitive and will be destroyed by pasteurisation.37 ºC (29).3 Temperature Under otherwise ideal conditions. In meat macerate. but may cause some cell damage (30).7 . NB: The growth of Staph.7. It can survive well at low temperatures. Staph. 10. Under most circumstances.FISH AND SEAFOOD 10.7. however. with an optimum of 35 . aureus than citric acid). etc. aw and temperature (sensitivity to acid increases with temperature).7.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. 0.7. Freezing and thawing have little effect on Staph. depending on aw.1 Staphylococcus aureus Morphology Gram-positive cocci. fat content. 10.2 to 9. aureus is more limited under anaerobic than aerobic conditions.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. Most strains of staphylococci can grow within the pH range 4. under otherwise ideal conditions (29. 232 .0 . 10.5 ºC.).7. 31). As is the case with other bacteria. aw.7 10. The limits for toxin production are also narrower than for growth.5).2 Oxygen requirements Facultative anaerobe.0. 10.7. aureus viability.
2. Staphylococci are more resistant to salt present in foods than other organisms.7.2 . with a temperature range for growth of 10 .8 x 1.9.5) (29.8. It can grow over the aw range 0. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.6 Aw/Sodium chloride Staph.4 . heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. In general. 10.2 Oxygen requirements Facultative anaerobe.PATHOGEN PROFILES 10. 0. 10.8 10.99 aerobically under otherwise optimal conditions. 233 .99 little or no toxin production in anaerobically packed foods. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities. Staph.1 Vibrio cholerae Morphology Gram-negative curved rods. but certain strains can grow in 20% NaCl.0 (optimum 7.8.83 .8. strain.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5.10% NaCl. 31) between 0.6 µm 10.43 ºC (35). atmosphere.5 .7.0. pH. * dependent on temperature.>0.86 is the generally recognised minimum in foods (32). In general. and solute. 10.0 .87 and > 0. However.7 Limits permitting toxin production 10 .7. 34). aureus can grow in 7 . an aw of 0. An effect of increasing salt concentration is to raise the minimum pH of growth.3 Temperature The optimum temperature for growth has been reported as 30 ºC.
4 Heat resistance V. under otherwise favourable conditions.8. but only moderately sensitive to freezing (40).4. with optimum at 7.FISH AND SEAFOOD Also. 10.0% NaCl (35).1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. with an optimum of 37 ºC (39).9 10. but not acid conditions.65 minutes (35).1 . 10. declining in number during storage.2 Oxygen requirements Facultative anaerobe.8.97 .8. and it is sensitive to refrigeration temperatures (0 . 10. respectively. are 5 and 43 ºC. It can grow in the presence of 0.998. cholerae may persist on frozen fish for >180 days (36). cholerae in crab meat homogenate at 60 ºC is 2.9. 10.6 Aw/Sodium chloride V.6 (35. V. The D-value for V.5 ºC). 38).9. it has a pH range for growth of pH 5 . cholerae can grow over the aw range 0.0. there is evidence that V.5 pH This organism is tolerant of high pH values.6. 37. 10. parahaemolyticus is easily destroyed by drying. 10. with an optimum of 0.9. cholerae is sensitive to heat at temperatures of >45 ºC.984 (35).3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.9. 234 .
41). The optimum concentration of NaCl equates to aw of approximately 0.4% (39.8 .PATHOGEN PROFILES 10. V. with a temperature range for growth of 8 .4 Heat resistance V. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0. 10. 235 .1 Morphology Gram-negative curved rods. vulnificus is heat sensitive. 10.5 pH The pH range for growth is 4.6 Aw/Sodium chloride V.6 . parahaemolyticus is unable to grow in the absence of salt. The D-values at 55 ºC for clam and crab homogenate are 0.4 Heat resistance V.02 . 41). parahaemolyticus are not heat resistant.10.3 Temperature The optimum temperature for growth has been reported at 37 ºC.29 minutes.43 ºC (42). and are readily destroyed by cooking.9. 10.5% NaCl.922.10. respectively.10. 10.10.0.9. with an optimum pH of 7.9. respectively (39). but the optimum concentration is 2 .6 (41).5 minutes.98 (35.10 Vibrio vulnificus 10. and 2.11. 10.8. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43). The organism can survive for long periods at freezer temperatures (42). 10.2 Oxygen requirements Facultative anaerobe. parahaemolyticus will grow at aw levels as low as 0. depending on solute.
Lund B. 383-408. New York. New York.8 (42). Eds. Eds. Ed. New York. Great Britain. 1995.H. with an optimum of 2. Gaze J. Stelma G. Juneja V. Gorham J. Blackie. Volume 1. Clostridium botulinum and Clostridium perfringens. 2.. Hui Y. Dodds K.W.W. Gorman J. Smith J. 1-27..R.W. Marcel Dekker.R. 1-19. Montville T. Clostridium botulinum. Diseases Caused by Bacteria.M. 1989.. References International Commission on Microbiological Specifications for Foods.M. Doyle M. 10.L.5. New York. 236 . in Clostridium botulinum: Ecology and Control in Foods.5% (42). Miliotis M.10. 97-131. in Clostridium botulinum: Ecology and Control in Foods. Murrell K. Doyle M. Potential hazards associated with REPFEDS. in Foodborne Disease Handbook. Hauschild A.P. in Foodborne Bacterial Pathogens.D.W.N. Hauschild A.. 1994.L. 5. 329-49.5 pH The pH range for growth is 5 . Marcel Dekker.W. Doyle M.P. Foegeding P.. Cliver D..11 1. New York. Eds. New York.. Volume 1: Diseases Caused by Bacteria. Beuchat L.. 1993. 8.. Johnson E. 4. Palumbo S. Dodds K. with an optimum pH of 7.0% NaCl. Aeromonas hydrophila.M. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’. 1994. in Food Microbiology: Fundamentals and Frontiers. Marcel Dekker.. Principles of control. Kovacek K. 2005. New York. Marcel Dekker... 1989. Hauschild A. Abeyta C. Microbiology and Molecular Biology. London. in Foodborne Bacterial Pathogens. 357-67.D.6 Aw/Sodium chloride V. 279-303. Clostridium botulinum.J..H. Dodds K..N. Faris A.. International Commission on Microbiological Specifications for Foods. Eds. Hui Y. 111-89. Aeromonas. Caister Academic Press. Austin J.10.O. Cliver D. Food Control. vulnificus grows over the range 0. Aeromonas species.D.. 7.L... 10. Ed.E.K. in International Handbook of Foodborne Pathogens.FISH AND SEAFOOD 10. 5-19.O.H. Aeromonas hydrophila group.A.H. Murrell K. 2003. Washington D. 3. Marcel Dekker Inc. Novak J.. Eds. 121-76. Eds.H. 1996. Volume 5: Microbiological Specifications of Food Pathogens.D. 9.L. Notermans S.P.R. in Microorganisms in Foods. Kim J. Marcel Dekker. 11. Betts G. 10. Peck M. 2001.H. Ed. 6. 57-63. 6 (1). Fratamico P. in Foodborne Pathogens. Marcel Dekker. ASM Press... Bier J. Clostridium botulinum... in Foodborne Disease Handbook. Bhunia A.5 .10. Stelma G.C. Eds. 1993.
157-62. in Foodborne Disease Handbook. New York.J. 1996. Clostridium perfringens. Volume 5: Microbiological Specifications of Food Pathogens. Montville T. 20. Lancet.G.PATHOGEN PROFILES 12. 311-25.R.P. in Microorganisms in Foods. 735-42. Doyle M. Eds. Eds.. 26. Diseases Caused by Bacteria. 17. Juneja V. Varnam A. Rinke H. Cliver D. Blackie. Murrell K. 15. 336 (8721).R.H. Volume 5: Microbiological Specifications of Food Pathogens. in Microorganisms in Foods.. ASM Press. 1996. 1994. Plesiomonas shigelloides. 19.P. Cliver D. in Foodborne Pathogens: An Illustrated Text. Ed. Matthews K. Marth E.J. 112-25. Riemann H. Johnson E. 16. London. Washington D.O.K. International Commission on Microbiological Specifications for Foods. 1989. London. Beuchat L.J. Ryser E. Lund B.D.. Varnam A. 129-38. 1990. 22. 1991.. 25. London.. 133-67.P. Koburger J. Ed. 1996.A. 2001. Blackie. ASM Press. New York. Ed. Plesiomonas shigelloides. Doyle M. Evans M. Archer P. 1990. New York. Listeriosis and Food Safety. 2005. International Commission on Microbiological Specifications for Foods.P. Ed. Listeria. 2006. in Foodborne Bacterial Pathogens. 18. Labbe R..T. 13.M. 137-64. Nagaraja K. London.C. Wrigley D. International Commission on Microbiological Specifications for Foods. McClane B. Clostridium perfringens. 23.G. in Food Microbiology: Fundamentals and Frontiers.. Elsevier.. Volume 1. Clostridium perfringens.. London.V. Eds. Walker S. Ed. International Commission on Microbiological Specifications for Foods. 237 . International Commission on Microbiological Specifications for Foods.M. 351-82. Evans M. Halvorson D. in Foodborne Diseases. 229-40. Ed.O.. Marcel Dekker.. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs. Foodborne disease due to Bacillus and Clostridium species. Avian Disease. 24. Clostridium perfringens gastroenteritis.H. Cliver D. Plesiomonas. 201-8. 14.. Marcel Dekker. Wolfe Publishing Ltd.A. Volume 5: Microbiological Specifications of Food Pathogens.O. Gorham J. Clostridium perfringens. in Foodborne Bacterial Pathogens.A. Kim C. 1989. Washington D. 2007. Growth of Listeria monocytogenes at refrigeration temperatures. 1990. 141-82. Journal of Applied Bacteriology. 33. Blackie. 982-6. Clostridium perfringens food poisoning.G. in Food Microbiology . Hui Y. Montville T.. Marcel Dekker.An Introduction... in Foodborne Infection and Intoxication.A. Listeria monocytogenes.. Labbe R. 191-243. in Microorganisms in Foods.A. Academic Press.. International Commission on Microbiological Specifications for Foods. 1989. Eds. Eds.C.. Doyle M. Listeria monocytogenes.H. Marcel Dekker. 68 (2).H. Emery D. 21. New York. London. 208-13.J.R. Banks J.
in Foodborne Bacterial Pathogens.C. Marcel Dekker.O.S. Doyle M. Riemann H. Loessner M. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. 1989.L. Ed. Golden D. 31. 642. Fratamico P. 1996. in Microorganisms in Foods. Volume 5. Springer Science. Ed. Lee-Wong A. Wilkinson B. 1990. Crowcroft N. Blackie.P. Jay J. Cambridge.K.M. Microbiological Specifications of Food Pathogens. Dairy. Academic Press. Cliver D. 2005. 1930. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). in Understanding Pathogen Behaviour Virulence. R15764. 1993. 36.A. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems. 28. 238 . 45 (3). 35. Caister Academic Press.O. Wymondham. New York..J. New York. 1994. London. 1989. New York..62. Microbiological Specifications of Food Pathogens.J. Griffiths M. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. Salmonellae. in Modern Food Microbiology. 327-445. 1996. 37. Bergdoll M. Cleland D. London.M. Golden D... 543-68. Ed. Cholera: current epidemiology. 1983. 2005.FISH AND SEAFOOD 27.. 414-25. Bhunia A. 33. 29. Stewart G. D’Aoust J-Y. 545-66. Ed. Vibrio cholerae.. 32. Volume 5: Microbiological Specifications of Food Pathogens. in Foodborne Disease. Ed.A. London. Vibrio parahaemolyticus. Academic Press. Marcel Dekker. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures. Staphylococcal gastroenteritis. Tohyama Y.A. Food and Environmental Sanitation. 8. 217-64. London. 1996.M. 327-52. Jay J. Staphylococcus aureus. 34. Loessner M.P. International Commission on Microbiological Specifications for Foods.H. 39. in Microorganisms in Foods. 85-106. Warner R. Gustafson J. 40.S. Japanese Journal of Experimental Medicine. International Commission on Microbiological Specifications for Foods. CDR Review. 273-84. 523. 38. Oliver J. Ed. Staphylococcal Food Poisoning. London. 426-35. Eds. International Commission on Microbiological Specifications for Foods. Twedt R. International Commission on Microbiological Specifications for Foods.. 2005..R.. 13 (11).S. International Commission on Microbiological Specifications for Foods. Volume 5. Woodhead Publishing Ltd. in Microorganisms in Foods.C. Smith J. Eds. Eds.M. 985-98.. Applied and Environmental Microbiology. in Foodborne Pathogens. in Foodborne Infections and Intoxications. Stress Response and Resistance. Staphylococcal intoxications. in Foodborne Pathogens: Microbiology and Molecular Biology. 2005.D.P. Blackie. 331-57. Blackie. International Commission on Microbiological Specifications for Foods. Vibrio parahaemolyticus. Ed. Reed G. 30. Cliver D. Salmonella. Doyle M. 4 (13). Bergdoll M.
Blackie. in Microorganisms in Foods. Loessner M. 436-9. Loessner M.W. International Commission on Microbiological Specifications for Foods. London.. Springer Science. International Commission on Microbiological Specifications for Foods.M..D.. Ed. 1996. 985-9. Journal of Food Protection. Vibrio vulnificus. Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters.A. 55 (12).J. in Modern Food Microbiology. Jay J. 2005. Yersinia. Jay J. 239 . 42. 1992. Volume 5. New York. Golden D. and Campylobacter species. Foodborne gastroenteritis caused by Vibrio. Golden D. Cook D.. Microbiological Specifications of Food Pathogens.A.PATHOGEN PROFILES 41. 657-78.M..J. Ruple A. 43.
FISH AND SEAFOOD 240 .
gsi.uk/Topics/Fisheries Seafish Industry Authority 18 Logie Mill Logie Green Road Edinburgh EH7 4HS United Kingdom Tel: + 44 (0)131 5583331 Fax: + 44 (0)131 5581442 Web site: www.org/ .uk Prepared Fish Products Association (PFPA) Federation House 6 Catherine Street London WC2B 5JJ United Kingdom Tel: + 44 (0) 208 78362460 The Scottish Government Fisheries Group Environment and Rural Affairs Department Pentland House 47 Robb's Loan Edinburgh EH14 1TY United Kingdom Tel: + 44 (0) 8457 741741 or + 44 (0)131 5568400 Fax: + 44 (0)1397 795001 Email: email@example.com or firstname.lastname@example.org Web site: www.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: email@example.com Web site: www.seafish.gov.defra.gov.ifm.scotland.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 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: email@example.com Web site: www.
ie/templates/homepage.suite. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.com/ Danish Institute for Fisheries Technology and Aquaculture (DIFTA) illemoesvej The North Sea Centre DK-9850 Hirtshals Denmark Tel: + 45 98 944300 Fax: + 45 98 942226 Email: firstname.lastname@example.org/ 242 .13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: email@example.com/fishery/ European Association of Fish Producers’ Organisations H.bim.dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 . Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo. 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 Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.gov.dk Web site: www.difta.gov.asp Food and Agriculture Organization of the United Nations Fisheries and Aquaculture Department Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57053152 Email: FAO-HQ@fao.C.is Web site: www.no Web site: www.uk Web site: www.eapo.FISH AND SEAFOOD Department of Agriculture and Rural Development.fiskehandlerne.com Web site: www.ifpea@eircom. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne. Andersens Boulevard 37.dk Danmarks Fiskehandlere H.org Web site: www.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: email@example.com 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.dk Web site: www.uk/index/fisheriesfarming-and-food.dardni.fao.fiskeriforskning.matis.
co.hpa.au 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 CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: email@example.com Web site: www.csiro.org.dh.uk/en/index.fdf.au Web site: www.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.com/ 243 .co.gov.uk Web site: www.org.org.nz Web site: www.nz/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.seafood.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: firstname.lastname@example.org.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 Web site: www.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: email@example.com 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: firstname.lastname@example.org.
nhs. Web site: www.euro.who.uk/index.codexalimentarius.who.se/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: email@example.com Web site: www.who.asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.int Web site: http://www.co.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: firstname.lastname@example.org Web site: www.co.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.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.nhs.scot.sfam.uk/scieh.The Swedish Institute for Food and Biotechnology Box 5401 SE-402 29 Göteborg Sweden Tel: + 46 31 3355600 Fax: + 46 31 833782 Email: email@example.com. jsp Food Safety 10 .who.int.org.hps.org Web site: www.net/web/indexen.de 244 .se Web site: www.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: firstname.lastname@example.org/foodsafety World Health Organization (WHO) Headquarters CH .who.int/ SIK .sofht.uk Web site: www.sik.php Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: email@example.com Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: info@who.
gov/foodsafety/ Food Safety Authority of Ireland http://www.int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa. Beverage and Consumer Products Companies 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.org/ The Association of Food.ie/ Food Science Australia http://www.riph.who.gov/ Institute of Food Technologists 525 W.uk/ Society of Food Hygiene and Technology (UK) http://www.org/ Royal Institute of Public Health (UK) http://www.org/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.org.htm Centre for Disease Control and Prevention (CDC) (US) http://www.org/ Institute of Food Technologists http://www.fsai.ift.foodscience.fpa-food.au/ International Association for Food Protection http://www.foodprotection.FoodSafety.co.cdc.ift.fda.sofht.csiro.CONTACTS US FDA .gov/ World Health Organization (WHO): Food safety programmes and projects http://www.org Web site: http://www.eu/pol/food/index_en.cfsan.uk/ 245 . Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: info@ift.
FISH AND SEAFOOD 246 .
growth and survival in crustacean shellfish 86 Aeromonas sobria. to test microbiological criteria 19 Aeromonadaceae. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. causing spoilage of dried fish 110. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. 102 Bacillus spp. in crustacean shellfish 87 Alteromonas spp.INDEX Achromobacter spp. 111 Astroviruses. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. pathogenic microorganisms in molluscan shellfish 59-60 as contaminant in enrobing process 37 as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45 growth in vegetable and rice 247 . as autochthonous bacterial flora of fish 3 Aeromonas spp. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. 69-70 Aeromonas hydrophila. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. source of PSP toxins 142 Alexandrium ostenfeldii. planktonic source of spirolides in shellfish 163 Allergens. preserved by dry/wet salting 99 Anisakis spp. in crustacean shellfish 86-7 ATP. 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.
136.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. of recipe fish meals 43 Cholera. 226 Breading process 36-8 Brevetoxins. 98-9 Brochothrix thermosphacta. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. fish sauce 123 Basipetospora halophila. 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. for PSP toxins 144 Chemical indicators. as autochthonous bacterial flora of fish 3 Clostridium botulinum. preserved by marinating 100 Chemical detection methods. caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. in crustacean shellfish 86-7 Campylobacter coli. associated with consumption of fish/fish products 16. 113. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. of fish – effects on microflora 5-6 Carbohydrate. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. 131 Carbon dioxide. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni. as cause of spoilage of MAP fish 15 Calciviruses. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang. in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage. 137. 68-9 Capture and handling. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. smoked and dried fish 113-6 hazard in mild-processed marinated products 101 hazard in sun-dried products 102 non-proteolytic – risk of growth in fish recipe meals 44 248 . of fresh fish spoilage 11 Chemical pollutants. importance in lactic acid fermentation in fish products 130.
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 autochthonous bacterial flora of fish 3 Crabs. of fish – different processes 94 Dun. 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. source of azaspiracids 159 source of DSP toxins 148-9. 106 Contacts 241-5 Conway method. growth and survival in crabmeat 84 growth and survival in cured. 149-50. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. 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. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA).INDEX pathogen profile 226-7 Clostridium perfringens. for detection of DA 148 ELISA kit (DSP-Check). to detect NSP toxins 155 ELISA kit (Biosense). of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. adverse condition on poorly preserved fish 110 EC grading scheme. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. as contaminants in scampi 36. of prepared fish products 44 Escherichia coli. hot-smoking 107 ELISA assay. processing 334 Cockles. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. to determine fresh fish spoilage 11 Eels. smoked and dried fish 114 Enzymic spoilage. 152 Diphyllobothrium. 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 determine fresh fish spoilage 11 Corynebacteria.
spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. processing 35 Fish component. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes. of fish at sea 5 Fatty fish. of fresh fish spoilage 911 Evisceration. causing pink condition on fish surface 110 in fish sauce 127 Handling. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. mould causing spoilage of dried fish 110 Evaluation. to measure fish skin resistance 10 Flavobacterium. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. concerning crustacean shellfish 87-8 European Grading Scheme. non-B) viruses. member of cyclic imine group 161 Gymnodinium catenatum. as source of ciguatoxins 156 Garlic. fish sauce 123 HACCP. source of PSP toxins 142 Gyoshoyu. 130-5 Fish-spoilage bacteria 12 Fishtester. 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. 102 Gymnodimines.FISH AND SEAFOOD EU legislation. as source of inulin in lactic acid fermentation in fish products 134 Garos. preserved by marinating 101 Gravlaks. fermented fish product 126 Green cured split cod 97. for evaluation of fish quality 9 Eurotium. specific EU hygiene rules 196-224 Fresh fish. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . 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. of prepared fish products 44 Gambiertoxins. of fish – effects on microflora 5-6 Hepatitis (non-A.
outbreak caused by gravid 115 Lobster. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. for microbial inactivation in fish products 127. preserved by marinating 101 Microbiological criteria. 100-1 Masmeen 107 Matjes. as source of botulism 136 International legislation. in fish fillet 1 Lipid oxidation. for fresh and frozen fish (Table) 20 Micrococcus spp. smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. preserved by salt-boiling 100 Ionising radiation. definition 1 Legislation.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 . causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149. enabled by carbohydrate source in fish products 130. preserved by salt-boiling 100 Karenia brevis. 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. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. as source of yessotoxins 152 Lipid content. food hygiene 189-224 Lingulodinium polyedrum. to reduce rate of fish spoilage 12 Ikan kaju. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured. concerning crustacean shellfis 88 Jellied eels. as autochthonous bacterial flora of fish 3 Lean fish. 135 I-zushi. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured.
associated with consumption of fermented fish products 137 Oxidative rancidity. reduction in smoking process 108 Norcardiopsis dassonvillei. smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. smoked and dried fish – effects of processing 96-108 of fermented fish – effects of processing 127-35 of fish – effects of processing 5-9 of ingredients of chilled/frozen prepared fish products – effects of processing 32-42 of molluscan shellfish – effects of processing 60-5 Modified-atmosphere packaging 6-9 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee guidance 7 of fresh chilled products 42 Molluscan shellfish 53-77 commercial products 54-9 definitions 53-9 Moraxella. of crustacean shellfish – effects of processing 81-3 of cured. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. shigelloides. in crustacean shellfish 867 Norwalk virus. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. for detection of azaspiracids 160 for detection of ciguatoxins 157 for detection of DA 148 for detection of DSP toxins 150. 153 for detection of LST toxins 151 for detection of NSP toxins 155 for detection of pinnatoxins and pteriatoxins 165 for detection of PSP toxins 144 for detection of spirolides 164 for detection of tetrotoxin 145 Mussels. as TTX producer 145 Norovirus. caused by lipid oxidation 11 Oysters.
preserved by salt-boiling 100 Pink. detection methods 165 origins and distribution 165 toxicology 165 Pissala. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation. 100-1 Pindang. as source of toxins 159. as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. effects on microflora of fish 5-9 Professional bodies. as source of yessotoxins 152 Protoperidinium cassipes. contact details 241-2 Prorocentrum spp. 225-39 Pathogens. 81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 . causing spoilage of dried fish 110-1 Photobacterium phosphoreum. as component for seafood ready meals 40 Prawns. 69-70 Polycyclic aromatic hydrocarbons. fermented fish product 124 Plankton. 108 Polypaecilum pisce. causing spoilage in vacuum-packed dried fish 111 Pseudomonads. growth and survival in chilled and frozen raw fish 19 growth and survival in crustacean shellfish 86 growth and survival in cured. in molluscan shellfish 60 Pseudomonas spp. as source of DSP toxins 148-9 Protein content. 161. as cause of histamine fish poisoning 15 as spoilage organism in chilled products 43 as spoilage organism in fresh fish 14-5 Pickling 94. smoked and dried fish 115 Pasta.INDEX Parasites. spoilage organism in oysters 65 Protoceratium reticulatum. 163. 152-3 Penicillium spp. as component for seafood ready meals 40 Pathogen profiles. growth and survival in chilled and frozen prepared fish products 46-7 growth and survival in chilled and frozen raw fish 15-9 growth and survival in crustacean shellfish 84-7 growth and survival in cured. smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. pathogen profile 230 in molluscan shellfish 5960. adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164. mould causing spoilage of dried fish 110 Potato. for detection of PSP toxins 144. planktonic source of azaspiracids 159 Pseudomonadaceae. reduction in smoking process 103. 145-6 definitions 28-30 non-seafood ingredients 32 Processing. in fish fillet 1 Proteus. 197 Plesiomonas.
preserved in saturated brine 99 Sauce. fermented fish product 126 Raphidophytes. processing 35 Shellstock. 36 Seafood. for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops.) 41 Recipe fish products 38-40 Rhodotorula rubra. flow diagram (Fig. resulting from poorly salted cod 110 Pyrodinium. smoking and marinating of fish 96 Salt-boiling 94. as carbohydrate source in lactic acid fermentation in fish products 134 as component for seafood ready meals 40 RIDASCREEN. chilled and frozen 1-25 Recipe dish. producers of brevetoxins 154 Raw fish. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. processing 34. for evaluation of fish quality 9 Radiation.) 41 ingredients preparation. 135 Rakefisk. for microbial inactivation in fish products 127. as source of tetrotoxin 144-5 Putty fish. vacuum packing 105 Salmonella spp. description and uses 58-9 Scampi. flow diagram (Fig. importance in lactic acid fermentation in fish products 130 importance in salting. growth and survival in chilled and frozen raw fish 19 Pseudoterranova dicipiens.FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries. to measure fish skin resistance 10 Salmon. antibody-based kit to detect STX 144 RT Freshmeter. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. assembly and packaging. detection methods 165 member of cyclic imine group 164 origins and distribution 165 toxicology 165 Puffer fish. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 growth and survival in cured. hazard in salting of fish 97 Salt concentration. 100 Salting 97-9 definitions 93-4 Sardines. fermented fish product 126 as source of botulism 136 Rakørret. as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. source of PSP toxins 142 Quality Index Method. detected in cod 115 Pteriatoxins. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova. in molluscan shellfish 60 Rice. 83 254 . processing and effects on microflora 60-1 Shewanella.
13. as indicator of fresh fish spoilage 11 Toxins. 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. in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. concerning crustacean shellfish 89 Vacuum packaging. processing and effects on microflora 63-4 Skin resistance. for evaluation of fish quality 9 Torrymeter. as measure of fish degradation 10 Sliming. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. of fish after capture 5 Surstrømming. contact details 241-2 Trematode infections foodborne 137 Trichosporon. role in fish degradation 10. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. 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. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 . to measure fish skin resistance 10 Total Volatile Base-Nitrogen. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO. of crustacean shellfish 80.INDEX Shigella spp. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. grading of 31 hot-smoking 107 US legislation. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru. 101 Storage. 14 Torry Scale. 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. fish and shellfish 141-74 Trade associations. fermented fish product 126 Taste panel evaluation. fish sauce 123 Shrimps.
growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 67 pathogen profile 235-6 Vibrionaceae. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses. 62 Yersinia spp. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. 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. processing for seafood recipe meal centres 39 Vibrio spp.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. 152-3 256 . of shellstock 61. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. 68 Vibrio cholerae non-01.
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