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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
Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged. Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product. and a detailed list of contacts for various food authorities. for his scientific input and Victoria Emerton. I dedicate this book to my husband . The books are intended as a source of information for microbiologists and food scientists working in the food industry and responsible for food safety.Goldwyn. Training Manager. both in the UK and elsewhere. which has been the chief factor in ensuring the completion of this book. Fish and Seafood. thank you for all the support. Technical Team Leader for her valuable editorial contribution. The books in the series are designed as easy-to-use guides to the microorganisms found in foods. guides to HACCP. and growth characteristics for key pathogens associated with the product. potential hazards associated with the consumption of a range of products. My gratitude is extended to Dr Peter Wareing. and Meat Products. Alison Turner. published by Leatherhead Food International and RSC Publishing. I thank the contributing authors for their collaboration. Rhea Fernandes Leatherhead Food International v . I also acknowledge and thank Jackie Apps.FOREWORD The Microbiology Handbook series includes Dairy Products. and Ann Pernet for the work put into typesetting and indexing. All handbooks also contain a review of the related legislation in Europe and UK.
4 Processing and its Effects on the Microflora 2.6 Published Microbiological Criteria 3.6 Pathogens: Growth and Survival 1.5 Factors Affecting Fresh Fish Spoilage 1.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. 4.4 Spoilage 1.2 Initial Microflora 4.8 References MOLLUSCAN SHELLFISH 3.5 Pathogens: Growth and Survival 3.2 Definitions 2.7 References 4.5 Spoilage 2. vii .4 Spoilage 4.7 References 3.1 Definitions 1.2 Initial Microflora 3. 3.4 Spoilage 3.1 Definitions 3.1 Definitions 4.7 Published Microbiological Criteria 2.2 Initial Microflora 1.5 Pathogens: Growth and Survival 4.8 References 1.3 Initial Microflora 2.7 Published Microbiological Criteria 1.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.3 Processing and its Effects on the Microflora 3.6 Published Microbiological Criteria 4.3 Processing and its Effects on the Microflora 1.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.1 Introduction 2.6 Pathogens: Growth and Survival 2.8 Further Reading CRUSTACEAN SHELLFISH 4. CHILLED AND FROZEN RAW FISH 1.3 Processing and its Effects on the Microflora 4.
1 Introduction 7.3 Processing and its Effects on the Microflora 5.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.7 References 5.3 Tetrodotoxin 7.9 Azaspiracids 7.7 Further Reading FISH AND SHELLFISH TOXINS 7.5 References 93 93 95 96 108 113 116 117 120 123 123 126 127 135 136 137 140 141 141 142 144 146 148 151 153 155 158 160 165 166 175 175 176 177 184 186 6.2 Initial Microflora 5.10 Cyclic Imines 7. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9. 8.8 Ciguatoxins 7.1 Definitions 5.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.8 Further Reading FERMENTED FISH 6.5.4 Spoilage 6.9 Other Relevant Legislation 223 9.1 Definitions 6. CURED.2 Initial Microflora 6.1 Introduction 189 9. 2006 No.6 Published Microbiological Criteria 5.8 Guidance 222 9. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.2 Definitions 8.5 Pathogens: Growth and Survival 5.2 Legislative Structure 190 9.4 Regulation (EC) No. SMOKED AND DRIED FISH 5.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.6 References 6.7 Food Hygiene (England) Regulations 2006. 852/2004 on the General Hygiene of Foodstuffs 191 9.6 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.I.6 Lipophilic Shellfish Toxins (LST) 7.11 Conclusion 7.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7. 9. EU FOOD HYGIENE LEGISLATION 189 9.3 Stages of a HACCP Study 8.1 Introduction 8.4 Spoilage 5. S. 14 (Hygiene requirements specific to the UK) 221 9.3 Regulation (EC) No. 7.4 Implementation and Review of the HACCP Plan 8.3 Processing and its Effects on the Microflora 6.5 Pathogens: Growth and Survival 6.5 Regulation (EC) No.10 Further Reading 223 viii .
1 Aeromonas spp.6 Salmonella spp.2 Clostridium botulinum 10.10 Vibrio vulnificus 10.8 Vibrio cholerae 10. 10. 10.7 Staphylococcus aureus 10. 10.5 Plesiomonas 10.9 Vibrio parahaemolyticus 10.4 Listeria spp.10 PATHOGEN PROFILES 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .3 Clostridium perfringens 10.
Seafood and Vegetables. 94767. Safety and Processing of Wild and Farmed Fish.. molluscan shellfish. in Handbook of Food Products Manufacturing: Health... and fish and shellfish toxins.. Chandan R. 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. Poultry. Wiley. Seafood Research from Fish to Dish: Quality. Wageningen Academic Publishers.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish.. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs. Wageningen. the consumption of fish and seafood has increased. Jacobsen C. fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins. Bekaert K. thus. 2006. crustacean shellfish. The second edition of this handbook is a review of the entire book for currency of information. Seafood processing: basic sanitation practices. Meat. fermented fish.B.. Nollet L. Clark S. Oehlenschlager J. 2007. Often contamination is from human and animal sources. Because of their health advantages over red meats. Cross N. Hoboken. Luten J. and.. Hui Y. Geographical region. The Microbiology Handbook.M. Milk.H. rivers and lakes whose water can range from pristine to contaminated.INTRODUCTION Fish and seafood are a main source of animal protein in the diet. Further Reading General Fish/Seafood Microbiology Cross N. Saebo A. chilled and frozen prepared fish. for fish. Catches can be gathered from seas. xi .A.. season. cured. Eds. smoked and dried fish.
Ed. Ed.L.. Ed. New York. London.. Blech Z. Noordwijkerhout.E. Dalgaard P. Cambridge. Control of indigenous pathogenic bacteria in seafood.. Ed.B.M. Rawles D. International Commission on Microbiological Specifications for Foods. 1996. in Microorganisms in Foods 6: Microbial Ecology of Food Commodities. 60 (4). Oxford..A. in Food Plant Sanitation. Blackie. Eds.A. Huss H.. Blackwell Publishing.H. Huss H. London. 543-61. Hafsteinsson H. November 1995.. 2003.H. Microbiology of fish and fish products. Huss H. and Clostridium botulinum in smoked fish and shellfish. 167-77. Simpson B. 1997. Gorham J. 1997. 2002. Luten J.. in Kosher Food Production. xii . Minimal processing in practice: seafood. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. Blech Z. Elsevier. November 1995. 413-30.A.. Stanfield P. 2002. 2005. in Advances in Food and Nutrition Research. Seafood processing: basic sanitation practices. 1997. Food Control. Hall G.S. Taylor S.. Oxford. Vo1ume 39. Eds. Bengtsson N. Gram L. Stanfield P. 8 (2). Journal of Food Protection. 91-8. 431-43. Luten J. Journal of Food Protection. 200-23. 245-66 Bremner H. Cambridge.International Commission on Microbiological Specifications for Foods.R.L.. Flick G. Ed. basic sanitation practices. Heinitz M. Woodhead Publishing Ltd. in Fish Processing Technology.. 1996. in Seafood from Producer to Consumer: Integrated Approach to Quality.Y. New York. Bruinsma B. The fish industry. Spoilage and shelf-life extension of fresh fish and shellfish. CRC Critical Reviews in Food Science and Nutrition. Amsterdam. Kluwer Academic / Plenum Publishers.. Hui Y.H.. Marcel Dekker..R. 174-249. 2004. in Minimal Processing Technologies in the Food Industry. Academic Press. Woodhead Publishing Ltd. 61 (3).J. International Journal of Food Microbiology. Biogenic amines in fish and shellfish. Gram L. Proceedings of the International Seafood Conference. Ventresca P.H.H. Amsterdam. 36 (1-2). Ohlsson T.B. The incidence of Listeria spp.N. Davies A. 87121.Y.. Salmonella spp. in Food Processing: Principles and Applications.. Noordwijkerhout. 459-72. Tong P. 329-65. Hui Y. in Seafood from Producer to Consumer: Integrated Approach to Quality.D.L. Dalgaard P. Martin R. Fish and fish products (Microorganisms in fish products). Elsevier. 318-23. Ashie I. Eds.K. Nip W-K. 33 (1). Microbiological spoilage of fish and fish products. Proceedings of the International Seafood Conference.. Ed. Smith J. Safety and Quality Issues in Fish Processing. 372-6. Gudmundsson M.P. 2004.. Smith J. 1997. Seafood: processing. 1997. 1996. Johnson J.M. Blackwell Publishing. 121-37.S. Predictive microbiological modelling and seafood quality. 1998. McCarthy S. Modified-atmosphere packaging of fish and fish products.
A. International Journal of Food Microbiology. Huss H. Microbial growth and its control in meat.. 1-17. Ed. Huss H. Eds. McMeekin T. Blackie. 305-17. poultry and fish. 1994. Sterling Publications Ltd.. 93-105. guidelines. 44 (12). Chapman and Hall. 25-39. Bacterial pathogens transmitted by seafood. Food Science and Technology Today.H. 46 (1). Amsterdam. International Commission on Microbiological Specifications for Foods. Huss H.A. in Quality Assurance in Seafood Processing: A Practical Guide.R. London. Chapman & Hall. 2257. 1990.D. in Microorganisms in Foods. Liston J.H. Hobbs G.M. Ward D. The microbiology of seafood products. Copenhagen. 1995. 118-20. Ward D. Gram L. Activities Report of the R&D Associates. A new look at microbiological standards. Bernoth E-M.. Liston J. Poultry and Fish Products. International Journal of Food Microbiology. 1990. August 1991...M. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Amsterdam. Dalgaard P. Liston J. London. August 1991. Bonnell A. International Journal of Food Microbiology. Martin A. 1996. Microbiological quality of fishery products. Liston J.. 1989. 1994. 1994. Eds. Gibson D.R. in Quality Attributes and Their Measurement in Meat. 1991. Fish spoilage and safety. Dalgaard P. Jakobsen M. Fish: microbiological spoilage and safety. Blackie. 26 (3). London. PHLS Microbiology Digest. Berlin. 5 (3).H. Jakobsen M. 1992.International Commission on Microbiological Specifications for Foods. Gilbert R. Elsevier. in Food and Beverage Technology International: USA.D. Gram L. Ed. 11 (2). Ed. 54-61. Evaluation of the bacteriological quality of seafood. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. 319-33. Microbial hazards of seafood consumption. Elsevier. Sofos J. 26 (3). Microbiological indicators and seafood processing. 269-82. xiii . 1992. Ed. 1994. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. London. Pearson A. Jakobsen M. Institut fur Veterinarmedizin. 1992. Seafood toxins of microbiological origin.R. Volume 5: Microbiological Specifications of Food Pathogens. in Fisheries Processing: Biotechnological Applications. Amsterdam.S. Qualitative and quantitative characterization of spoilage bacteria from packed fish.. Olley J. Eds. Liston J. Elsevier.J. 1994. August 1991. Garrett E. 90-106. Copenhagen. Ed. Bonnell A.. Evaluation of the bacteriological quality of seafood. 359-403. Twigg B. Food Technology. in Public Health Aspects of Seafood-borne Zoonotic Diseases. 459-78. Bacterial spoilage of seafood. Modelling of microbial activity and prediction of shelf-life for packed fish. and specifications for seafood products. 16 (1). Application of predictive microbiology to assure the quality and safety of fish and fish products. Dutson T.M. 265-79. 166-73.N. Ross T. Glasgow. 1992.. 1995. 5662. Eds. 121-8. Copenhagen.
Volume 1. Applied Science Publishers. 169-208.. London. Elsevier Applied Science Publishers. Hobbs G. 1986. Food Technology. Microbiology of frozen fish and related products.M. 42 (3). Regenstein J. 103. in Essays in Agricultural and Food Microbiology. 1985. 1982. 1988.. Microbiological hazards associated with fishery products. xiv . 90-3. Garrett E. Wiley. guidelines. Part I . Shelf-life extension of fresh fish .J. 104-9. 11 (2). 199-226.K. Microbiological standards.a review.K. in Developments in Food Microbiology. Eyles M. Hodgkiss W. Ed. Ed. CSIRO Food Research Quarterly.R. Lamprecht E. Seafood-borne bacterial pathogens of marine origin. 42 (3). Journal of Food Quality.Pedrosa-Menabrito A.spoilage of fish. Hackney C.R. in Microbiology of Frozen Foods. London. Ed. and specifications and inspection of seafood products. 71-117. Davies R. 1988. 46 (1)... 1988. 8-16. Chichester. Simmonds C. Robinson R.C. Hobbs G.S. 1987. Microbiology of fish. Dicharry A. The bacteriology of fish handling and processing. Food Technology. Norris J. 117-27.
“fish” will be used for fresh and seawater finfish. ‘fish’ also refers to the flesh of such animals used as food. fins and a streamline body. Fish fillets are a poor source of carbohydrates. and is mostly interspersed between the muscle fibres.20%. The lipid content. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. typically fluctuates between 0. CHILLED AND FROZEN RAW FISH Associate Prof. Muscle composition and structure of fish also differ from those found in other meat. crustaceans.5% (1). migratory patterns.1. In addition.2 . Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. The lipid fraction is the component showing the greatest variation. lean fish are those fish that store most of their fat in the liver. giving it its characteristically flaky structure. 1 . which can be up to 67%. Fish fillet composition can vary significantly within the same species due to feed intake. Fish can be divided into fatty and lean fish. Fish are an important part of a healthy diet since they contain high quality protein. while fatty fish have fat cells distributed along their bodies. In addition. This super class of vertebrates includes all the bony and cartilaginous finfish. and excludes molluscs and Crustacea. most fish contain omega 3-fatty acids and other essential nutrients. but typically present a low fat percent when compared to other meats.21%. The connective tissue content of fish is also lower than that found in meat.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. it shows a typical season pattern especially in migratory species such as herring or mackerel. offering less than 0. In this chapter. and other forms of aquatic animal as part of their ‘fish’ definition. and spawning season. Fish flesh is dominated by the abundance of white muscle in relatively short segments. respectively (1). However. typically 3 and 15% of total weight. Protein content in fish fillet varies typically from 16 .
transportation and sale. firmness. nitrogen (N2) and oxygen (O2). but no further control is exercised during storage. Spoilage describes changes that render fish objectionable to consumers. DMA is dimethyl amine. Frozen fish is fish that has been cooled to. This technique is primarily used for bulk storage. Superchilled fish is fish that has been cooled to. Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases. during storage. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. but not below 3 °C during storage. through their development on fish. hence. and flavour based on the maturity. Quality refers to palatability and organoleptic characteristics such as tenderness. and maintained at or below 7 °C. Evisceration is the removal of the viscera from a fish. usually carbon dioxide (CO2).FISH AND SEAFOOD Chilled fish is fish that has been cooled to. colour. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. Fresh fish is raw fish that has not been processed. odour and texture. colour. and maintained at or below -2 °C (normally below -12 °C) during storage. 2 . transportation and sale. Controlled-atmosphere packaging (CAP) refers to packaging in an atmosphere where the composition of the gases is continuously controlled during storage. transportation and sale. and texture of the fish. marbling. and maintained at temperatures just below the freezing point. renders that fish objectionable to consumers. spoilage microflora describes an association of microorganisms that. juiciness. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. at -2 to -4 °C. The proportion of each component is fixed when the mixture is introduced. Organoleptic refers to qualities such as appearance. frozen or preserved.
Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp. several culture media containing sodium chloride and more than one incubation temperature should be used.2 Initial Microflora The subsurface flesh of live. potential human pathogens may be present in both scenarios. However. whenever possible. 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. processing or storage (Table 1. For example. molecular-based methods have not yet been used to address this issue. Moraxella. Flavobacterium. healthy fish is considered sterile and should not present any bacteria or other microorganisms. It must be noted that mesophilic bacteria can rapidly overgrow psychrotropic organisms. Clostridium. and typical of the fish flora. the season and the method of harvest. standard culture-dependent methods can only recover between 1 to 10% of total bacteria present in any given sample. Gastrointestinal tracts and gills typically yield high bacteria numbers. Shewanella and Pseudomonas. Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters.II) (3).) are also common aquatic bacteria. Gram-positive organisms such as Bacillus. posing a concern for seafoodborne illnesses (2). since they heavily depend on the type of sample analysed and the protocol used for isolation. Micrococcus. It is difficult to estimate how many microorganisms are typically associated with fish. and those that although not autochthonous to the aquatic environment. On the contrary.CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed. Lactobacillus and coryneforms can also be found in varying proportions (1). Human pathogenic bacteria can be part of the initial microflora of fish. microorganisms colonise the skin. In fact. It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish. The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest. 1. as with other vertebrates. 3 . some species (most Vibrios) require sodium chloride for growth. In general. gills and the gastrointestinal tract of fish. although these are influenced by water quality and feed.I). More accurate. the natural fish microflora tends to reflect the microbial communities of the surrounding waters.
1 .II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.35 °C 8 . Toxin levels 0. stream water. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. and risk associated with the consumption of seafood is low (2.19 µg/kg bodyweight from < 102 .106 cells/g (exception: V.103 High (107 .>106 Salmonella spp. numbers of pathogenic bacteria in raw fish tend to be low. However.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. monocytogenes as part of the natural aquatic flora It is apparent from the above that there is potentially a very diverse range of organisms present on fish. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . In addition. 4 . 105 . Plesiomonas shigelloides 3 .109 cells/g) * Some sources considered L. vulnificus) 5 . Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp. birds.37 °C High for most species.14 . sewage. during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.0.FISH AND SEAFOOD TABLE 1.37 °C Unknown Unknown TABLE 1. Escherichia coli Yersinia enterocolitica 101 . estuarine environments. and mud Ubiquitous.26 °C 10 .106 cells/g. 4).102 101 .
mackerel or herring.3 1. During capture and storage. handling and processing Wild finfish are usually caught by net. The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration.3. In addition. potential contaminants added during handling and processing. If evisceration is carried out at sea.a practice that may have both advantages and disadvantages. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. with very little control over the condition of the fish at the time of death or the duration of the killing process. ropes. birds and soil.1 Processing and its Effects on the Microflora Capture. hook and line. icing or freezing. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating. and conditions of storage. cortisone levels increase during prolonged stress and can alter the fillet quality. Physical damage such as loss of scales. human hands and clothing. in recent decades. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms. After capture.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. however. aquaculture practices have been expanding worldwide. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria. in which the bacterial load 5 . the cuttings provide areas of exposed flesh that are open to microbial attack. and during harvest. In eviscerated fish. chilled brine or refrigerated seawater at -2 °C. and spoilage. finfish will almost invariably come into contact with nets. has an effect on the amount of stress and physical damage that the fish will suffer during capture. offering better control of fish health prior to. The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea. and their re-use will increase the crosscontamination of other fish with such microorganisms. Fish may be eviscerated prior to storage at sea . Increasingly. digestion and off-flavours in uneviscerated fish. with the production of off-odours. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating. and the killing process is designed to minimise stress. or traps. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside. decks. and especially when fish is stored on board for longer periods. These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans. This contrasts greatly with the meat industry. with fish such as tuna and cod being more commonly eviscerated than sardines. 1. The length of time that set nets have been in the water or the time trawlers’ nets are towed. boxes and/or baskets. However. in which the health of each animal can be assessed prior to slaughter.
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. Anaerobic atmospheres have less effect on fresh fish shelf life. By vacuum packing foods. careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. a reduction in the oxygen tension is achieved which. Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. often in the absence of adequate refrigeration. Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. if there is some oxygen demand from the product. nitrogen. for example. and of the quality of the water used for ice and to wash fish. MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. by actively altering the composition of the 6 . Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. ferric ion (Fe3+)). 1. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). Packaging changes the intrinsic and extrinsic parameters affecting a product. Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. However. they are usually process contaminants.3. allowing more growth of spoilage organisms. As with all foods. and minimising handling will all reduce microbial cross-contamination. use of effective detergents and disinfectants. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. from water activity (aw) through to physical damage. will result in fully anoxic conditions. fish have a higher post mortem pH. however. cleaning of the work environment. oxygen etc. These changes can be deleterious. The atmospheric conditions surrounding a product may be passively or actively altered. 7). Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. and to limit the loss of quality (5). In the absence of oxygen. in time. These containers are also used for displaying the fish during auction at the quayside.2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. but isolate the food in an environment different from the bulk atmosphere. What is more. Physical barriers not only protect from physical damage. In MAP.). but if applied properly should extend the life of the product. and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO). partial fermentation of sugars occur leading to lower pH. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. not part of the normal flora of the meat animals (8).FISH AND SEAFOOD can be substantial.
although less information on their effectiveness is available. Replacement of oxygen with nitrogen. Many additives have been tried in conjunction with changed atmosphere. The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). and the problems of muscle structure damage by the modified-atmosphere (11). and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). The products. sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). Micrococcus and Moraxella). botulinum. Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains. does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. but can dissolve into fish liquids and deform packages. including salt. This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. Flavobacterium. may not be considered ‘fresh’ fish. discolour pigmented fish (11).CHILLED AND FROZEN RAW FISH surrounding gas. MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). a modified-atmosphere may contain any gas necessary for the desired effect. The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. the higher pH and reduction potential (Eh) of the fish muscle. gutted or filleted fish. High concentrations of carbon dioxide have the most pronounced microbial effect. and increase in-pack drip (12). the pathogens that may be able to grow before spoilage occurs. nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. phosphates. The microflora of meat is not the same as that of whole. without overt signs of spoilage. Gases used in MAP of fish most commonly include carbon dioxide and nitrogen. the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . an inert and odourless gas. and the promotion of a predominantly Gram-positive. slower-growing flora (11). the inhibition of common 'spoilage' bacteria (Pseudomonas. after additive application. Sulphur dioxide. but only in recent years has it seen a marked expansion in use and market share. VP is one of the oldest forms of altering the interior gaseous environment of a pack. Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. Modified-atmosphere preservation of fish was first reported in the 1930s. as they are able to grow at temperatures as low as 3. which are able to grow rapidly at low temperature.3 °C and produce toxins. In the United States. which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14).
and the use of a HACCP plan. The replacement gas should contain at least 40% carbon dioxide. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate.g. Subsequent storage must be at or below 3 °C. Only the highest-quality fish.3 °C at all points from packaging onwards. adequate product labelling in respect of storage temperature. The problem of water loss in MAP fish is common. adequate shelf life and cooking requirements. botulinum.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). this problem occurs to a lesser extent with meat. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. The growth of these bacteria is reduced by high carbon dioxide concentrations but. For fatty fish. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13).g. as already stated. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. at all times. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. Although all fish species need to be treated separately. These conditions include storage of the product at ≤ 3. 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. 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). 3. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. 5. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). from both a microbiological and a chemical standpoint. The addition of high levels of carbon dioxide to these fish. For other less fatty marine fish. In the United Kingdom. however. grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. should be used. owing to the dissolution of cellular structures. 2. 4. It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. TMAO) that can be reduced to produce 8 . trimethylamine and sulfides). use of high-quality raw fish. Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. greater advantages are seen. may result in unacceptable colour and textural change and hence little shelf life extension (11). however. but is recognised as a drawback of carbon dioxide application.
the skin turns from iridescent to dull and bleached with bacterial slime. TABLE 1. as traditional spoilage patterns of the external organs are typically very distinct. 18) and are probably the most widely used. to brown and slimy with an intense off-odour.III. texture.CHILLED AND FROZEN RAW FISH off-odours. so lower carbon dioxide concentrations may have better preservative effects. and this pattern is the basis of freshness grading schemes. C (C=reject) Significantly. The eyes turn from convex and clear to concave and opaque. A.4 1. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . Taste panel evaluation of cooked fish may be used to score or grade colour.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16.4. B. Various methods for whole fish freshness evaluation by a trained panel have been proposed. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. and metabolic activities of microorganisms present in the fish. with no smell. Degradation of whole fresh fish stored in ice generally follows a set pattern. 17. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. 1. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). the gills from pink and shiny. or for that matter evaluation of the taste of the fish. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. reactions caused by activities of the fish’s own enzymes. Over time. these are summarised in Table 1. smell and 9 . Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids).1 Spoilage Fresh fish spoilage and methods of evaluation Food spoilage can be considered as any change that renders the product unacceptable for human consumption (10). Loss of freshness and spoilage cannot be separated as processes.III Fish-quality scales Name of scale Torry Scale Quality Index Method European Grading Scheme Scoring system 10 to 0 (≤ 4 reject) 0 to 24 (arbitrary reject) Extra. and the flesh turns from bright and elastic to dull and soft (1). When working with fillets.
the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet.g. 23). 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. chemical. RT Freshmeter or Fishtester). all marine fish use/have TMAO (21). an osmoregulator. or simply a waste product present due to bioaccumulation (22). %). the Eh of fish muscle remains relatively high (20). rigor resolves. HxR and Hx) gives a quantity (K-value. After death. However. rigor mortis is the first noticeable change in the fish. which increases from 0% towards 100% with time. These processes are strongly influenced by the physical conditions of storage. After death. From being flaccid. post-rigor values are typically in the range of pH 5. spoiled and putrid. texture becomes soft and watery or hard and dry. By measuring the concentrations of the six components. the muscles relax again and the fish returns to a flaccid state. ammoniacal. possibly slightly metallic. fruity and/or bitter off-flavours. IMP. reduction in ATP and increase 10 . The process of ATP breakdown is used as an indicator of fish freshness (24). the pH drop of the muscle is correspondingly less. sulphurous and/or rancid smells. an anti-freeze. Because fish tend to have relatively little residual glycogen compared with mammals. a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. iii) Traces of sour. iv) Enhancement of the spoilage characteristic of phase iii. ADP. To a varying extent.8 to 6. this will generally correlate with the length and severity of rigor. the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). 19). 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. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical. biochemical and microbiological actions. Generally. is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. As the fish degrades. as little endogenous reduction of TMAO occurs (18). TMAO permits a high Eh to remain in the muscle tissue. or its remaining shelf life (1). During the initial storage. thus.FISH AND SEAFOOD taste of cooked fish. ii) Neutral taste.5 (18. development of sickly sweet. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. AMP. the Torrymeter. an intense odour compound. little flavour. the conductance generally increases. 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. cabbage-like. sea-weedy taste. bacterial reduction of TMAO to TMA. After some time.
as microbial spoilage is limited by the low storage temperature (1. 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). Evaluation of Total Volatile Base-Nitrogen (TVB-N. these measurements are usually used to quantify the amount of spoilage. also termed TVN). 31). For some fish species. or a specific fraction of the volatile bases. During spoilage. 27) produced by aerobic respiration and other forms of metal ion reduction (28). therefore. 7). for example the TMA fraction. so the measurement is effective only during the initial loss of freshness. as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. a correlation can be made between the spoilage of the fresh fish and the production of TVB-N. 26). however. not to describe freshness. Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. Chemical degradation continues after the initial post mortem phase. Comparing results for different fish species. however. Quality indices based upon the products of microbial metabolism do not explain changes in quality until microbial growth produces measurable changes in the fish. Lipid oxidation and other oxidative changes lead to oxidative rancidity. However. the importance of microbial action increases with time (5. is later significantly reduced by bacterial action. The Conway method (and variations of it) uses a strong inorganic base to volatilise the bases in the fish sample. 11 . Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. hypoxanthine. and are especially important in the spoilage of frozen fish. does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection. Oxidative rancidity is known to reduce the quality of fatty fish in particular (19. 18). colour changes. the residual acid is then titrated. 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. which may also be formed by bacteria (25).CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). 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. Neither does the change in pH during storage correlate well with the production of TVB-N. and a segregated weak acid to absorb them.
The sum of these changes may result in a different spoilage profile. For example. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks. Not all the bacteria growing on a food will lead to the production of objectionable characteristics.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. biochemical and microbiological reactions taking place within and on the fish. the slower the spoilage processes proceed (within limits).5. 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 factors are interactive and cannot be completely isolated.5.FISH AND SEAFOOD 1. The concept of specific spoilage organisms (SSOs) is not new.5 1.and temperate-water fish spoilage rates may be many. careful temperature control during storage is not only important in terms of quality loss. a minority are often associated with the majority of the spoilage. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. by altering the gas atmosphere (extrinsic). the results of altering one parameter may have far-reaching consequences on others. 34). For fresh fish iced immediately after capture and continually stored in ice. 1. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. 5). The reasons for the differences between tropical. the lower the temperature of storage. Also. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33.5. For fresh fish.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). 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.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. Fish spoil as a result of the chemical. 1.
It is thus an important spoilage organism of gadoid fish such as cod. catalase-positive.3. more especially. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. Pseudomonas spp. putrefaciens spoilage of fish is due to its biochemical action on muscle. obligate respiratory bacteria. therefore. the spoilage of a product may be strongly influenced by the conditions under which the product is held. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms.10% of the total flora of fresh fish from temperate marine waters. Members of the genus are able to produce pigments. although only since the late 1960s has it been realised that specific metabolites of Sh. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. 35). produce hydrogen sulphide (H2S) from cysteine. It is able to grow as fast as. i. 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. processed (e.3.1 Pseudomonas spp. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. 35. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. putrefaciens growth may be used as indicators of spoilage (30).2 Shewanella putrefaciens Sh. motile with polar flagella. These organisms are described as potential spoilers. (9) and they are considered SSO of iced freshwater fish (35). Sh. and may play some role in the spoilage of freshwater fish. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice.5. which produced the characteristic spoilage compounds of the fish (30. 13 .5. It is also present in fresh water. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp. or faster than the rest of the flora of ice-stored fresh marine fish (9). During the 1970s. aldehydes. the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed.e. frozen) fish products.g. 36). mediated spoilage is characterised by ‘fruity’.CHILLED AND FROZEN RAW FISH started in the 1940s. oxidase-positive. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s. 37). plus other characteristic compounds of the species responsible for spoilage. it is often isolated as about 1 . form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine. ‘oniony’ and ‘faecal’ odours from the production of ketones. As previously noted. 1. They are generally characterised as Gram-negative rods. on fish are diverse and in many cases species-specific. its ability to reduce TMAO to TMA. The importance of Sh.
additional psychrotrophic species such as Shewanalla baltica. at the time of rejection. Ph. putrefaciens was not present in such numbers (23. baltica has been identified as the main H2S producer in cod during cold storing (37). Ph. Research has shown that approximately 107 cfu/g of Ph. Shewanella gelidimarina. putrefaciens lies in its chemotaxis: the bacterium. does not show carbon-source chemotaxis. Sh.how to reduce a molecule that they are not able to take into their cells efficiently. Shewanella frigidimarina. 41). mesophilic strains of Sh. putrefaciens were identified as Shewanella algae. phosphoreum sufficient to cause spoilage was found. Shewanella waksmanii. they are heterogeneous. Shewanella oneidensis. and Shewanella aquimarina. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). with several new species being described. It is also able to produce hydrogen sulphide and a range of other off-odour compounds. whereas >108 cfu/g Sh. In terms of its taxonomy. Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39). This type of intoxication occurs when bacteria convert the histidine 14 . although.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. 41). The ferric reductase activity of Sh. and recent taxonomic description using modern molecular methods bifurcated this species. Shewanella olleyana. and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. Sh. Another distinct feature of the physiology of Sh. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). the bacteria have a significant problem to overcome . owing to its cell size and activity. Sh. In fact. 38). Shewanella denitrificans. phosphoreum has also been identified as responsible for histamine fish poisoning.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). Later. as the ferric iron is insoluble. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). unlike many other motile bacteria. putrefaciens were required. like all members of the Pseudomonadaceae. phosphoreum were required for 50% of taste panellists to reject a sample. but Sh.5. In 50% N2/50% CO2 MAP cod.3. putrefaciens growth has been collected (12. putrefaciens strains are characterised as microaerophiles or anaerobes. Shewanella affinis. This wide-ranging respiratory capability is thought to be unique (39). and Shewanella profunda were described. a population of Ph. was capable of being in a significant numerical minority on the MAP fish but still able to yield the majority of the TMA thought to be responsible for the rejection (23. Shewanella livingstonensis. The prevalence of the bacterium in so many environments. Initially. 1. it was proposed that this organism. it is strictly respiratory. putrefaciens has been studied.
thermosphacta is a well-characterised psychrophilic spoilage organism of meat. lightly preserved fish products are associated with botulism. 5. algal toxins. types B. bacteria and higher parasites all present some risk associated with fish consumption (1. Acetate production has been reported as a good indicator of spoilage by this organism. When inoculated into VP corned beef and sliced ham. Fewer still are capable of growing on fresh fish. a convenient but diverse species of bacterium with a number of different types. D.e. presents a potential risk.3. 5) reports that fresh fish consumption has never been shown to cause human botulism.1 Clostridium botulinum C. thermosphacta in the spoilage of some MAP fish. B. this is probably due to spoilage occurring before toxin elaboration.6. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters.3 °C (Group II) and these are non-proteolytic (i. E and F include bacteria with minimum growth temperatures of approximately 3. However. botulinum. Review of outbreak data suggests fresh fish to be safe.6 Pathogens: Growth and Survival Endogenous chemicals. B. Adult disease is caused by the production of the botulinum toxin. including antisera. B. botulinum recorded up to 1963. The heterogeneity of the types (A. human viruses. especially type E botulinum intoxication. Growing evidence suggests a role for B. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42).5. C1.3 days after having reached 108 cfu/g (42). Types A. E. Recent studies have investigated the dominance of B. 75% (with a 35% mortality rate) occurred in Japan. 1.CHILLED AND FROZEN RAW FISH present in fish into histamine. age. Some Ph. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). Huss (1. where a particular 15 . E. 1. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment. it is bacterial risks that increase after capture of the fish. 5).4 Brochothrix thermosphacta and lactic acid bacteria B. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. thermosphacta did not produce off-flavours until 2 . but with proper cooking. Of these. previous exposure and access to supportive treatment. 6). (and very rarely F and G) have been reported to cause human disease. Of 404 intoxication outbreaks of type E C. C2. these risks are substantially reduced (1. they do not produce significant product spoilage). F and G) is well documented (44). 1. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose.
and North America. The first report of the organism being a foodborne agent was with shirasu. including epidemics in India. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). 11). 1. V. accounted for the majority of cases (1. A unique clone of V. parahaemolyticus O3:K6 is responsible for many of the recent V. storage and preparation (11). 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. is small. Currently. again. vulnificus is associated with warm marine and estuarine waters. Since 1996. Proper chilling.g.6. there is no specific guideline that describes a minimum level of V. botulinum in the marine environment (isolated from around 90% of marine and environmental samples from northern European waters and its consequent presence in seafood (a surveyed incidence of as high as 65%) (2. parahaemolyticus cases have increased across the world. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. Proper temperature control (<3. 5). whether stored aerobically or in modified-atmospheres. and when in contact with contaminated water (no CDC reported cases from fish. Southeast Asia. France.3 °C) will eliminate all risk.2005). The bacterium causes primary septicaemia. and proper cooking (boiling for 1 minute. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. USA. Japan. the risk posed to the consumer of fresh fish. I-sushi. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . use of post harvest treatments and/or cooking of fresh seafood will reduce risks. were associated with finfish between 1978 and 1998 (46).g. Other lightly or semi-preserved (e. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). smoked. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. Despite the ubiquitous nature of type E C. In Japan. the majority are associated with products not cooked immediately before consumption. Generally. Russia. V. due to the cultural preference for raw fish dishes. 1978 . a Japanese boiled and semi-dried sardine dish. which was probably contaminated from an uncooked food source or an excreting foodhandler. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. salted or pickled) fish have also been associated with botulism. which is believed to be required for virulence. Only about 1% of marine isolates produce a thermostable haemolysin. parahaemolyticus is a marine organism. sushi) (5). the majority of outbreaks are caused by consumption of raw fish products (e. parahaemolyticus outbreaks. which can cause human gastroenteritis. This strain has been responsible for 50 to 80% of all V.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked.2 Vibrio parahaemolyticus and other vibrios V.
no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. Most cases have been sporadic rather than associated to large outbreaks. proper chill storage will eliminate any risk associated with this organism. V. It is known that Aeromonas spp. which is often fatal (>50%). but proper cooking should substantially reduce risk of infection. They are aquatic organisms with an epidemiology that is yet to be fully understood. Similar problems as those encountered for Aeromonas spp. the causative agent of cholera. 1. and fish and shellfish are probably the major reservoirs for the organism (50). though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. monocytogenes has been well documented as a foodborne human pathogen. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. 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. are associated with human diarrhoeal illnesses.20 °C. Some isolates are true psychrophiles. in its habitat. in assessing the risk of fishborne infection by P. is historically associated with faecally contaminated water. The major virulence factor appears to be the production of toxins with enterotoxic. cytotoxic. Since 2000.. coli on isolation media (50).4 Listeria monocytogenes L. cholerae O1.CHILLED AND FROZEN RAW FISH disorders). shigelloides is similar to Aeromonas spp. In the US. 17). There is no risk associated with the consumption of properly chilled and cooked fresh fish. and especially Aeromonas hydrophila. L. although lack of data concerning infectious doses and the possibility that foodborne virulence is linked to the immune state of the host mean that full risk assessment is impossible. others are psychrotrophic mesophiles (49). Chill storage may not eliminate growth.55 cases per 100. Although the psychrophiles are unable to grow at body temperature. but the bacterium is known to survive and grow in the shallow marine. shigelloides are encountered. 1. there is no risk from the consumption of properly handled and cooked fresh fish. and proper cooking will substantially reduce risk. infection. however.3 Aeromonas Aeromonas spp.26 to 0. listeriosis cases have been reportable to the CDC. monocytogenes incidence has been between 0. with minimum growth temperatures around 0 °C and optima of 15 . and especially estuarine environment. Despite this. their potential to produce disease-causing exotoxin in food has not been fully examined (48).6. P.6. Again. possibly due to under-reporting because of their likeness to E. sodium channel blocking and haemolytic activity (48).000 17 . 5.
6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. some Vibrio spp. Klebsiella pneumoniae and Hafnia alvei.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. Larvae of parasites such as Anisakis simplex are resistant to curing and marinating but can be easily destroyed by freezing at -17 to -20 °C for 2 hrs. The most frequently reported parasites of human importance in fish are round worms of the genera Anisakis and Pseudoterranova and tapeworms of the genus Diphyllobothrium. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death. but more especially by Morganella morganii. The organism poses a serious risk in chilled products not cooked before consumption. Fortunately. It is regularly isolated from seafoods (51). Infections are mostly associated with the consumption of raw or 18 . but cooking will significantly reduce the risk of infection. leads to the production of histamine. 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.pregnant women. at risk groups . especially pickled fish.98 minutes in cod and D60 of 4. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). it is not apparent to the consumer and it cannot be destroyed by cooking.6. Lightly preserved products. If fish are not properly refrigerated. Like other toxins. 1. full risk analysis is not possible (52). alcoholics. Being a psychrotroph. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. D60 of 1.6. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection.48 minutes in salmon have been reported (53).. Photobacterium spp. are exceedingly difficult to produce within the legal limits of histamine as the temperatures used during production can lead to rapid growth of histidine decarboxylating bacteria. but proper cooking will reduce any risk.FISH AND SEAFOOD persons. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. there is a possibility for growth of the organism on chill-stored fish. 1. Clostridium spp.. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . foetuses.2002 (55)) and is also common worldwide. neonates. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae. and Lactobacillus spp..
No.H. it may be desirable to test for Salmonella and V. 1997) Huss H. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF). coli counts where this method is preferred. long storage under refrigeration or one of the former prior to freezing. especially in warm-water areas and where fish are to be consumed raw. aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . 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. parahaemolyticus Staph. Gram L.IV) (57). Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease. For fish from inshore or inland waters of doubtful microbiological quality. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration.. Rome. 2003. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. 1. FAO. 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.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. volume.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. which in many cases will eliminate the risk.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. area or lot (CAC. In summary. parahaemolyticus (see Table 1. Ababouch L. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing.. EC.1997. 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. Faecal coliform counts may be used instead of E. TABLE1. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. 444.
Ashie I.R... The role of seafood in bacterial foodborne diseases.J. Fletcher R. Smith J. International Journal of Food Science and Technology.H. in FAO Fisheries Technical Paper No.FISH AND SEAFOOD 1. Huss H. 1989. Journal of Food Safety. 283-94..K.P. Kirby R. Codex Alimentarius Commission. Ahmed F. 12. 1996. 27. 3. Simpson B. 87121. 1997. Jeksrud W. 2.. 7. Rhodehamel E. Food and Agriculture Organisation of the United Nations.H. Reddy N..A. Kautter D. 334. 2. Food Control.8 1. Food and Agriculture Organisation. 8 (2).H. 5 (1). in FAO Fisheries Technical Paper No. 1996. Food and Agriculture Organisation of the United Nations. International Journal of Food Sciences and Techonology. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres. 12. 11. Gibbs P. Gram L. 10. 1992. Huss H. Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms. Nychas G. Assurance of seafood quality. Codex Alimentarius Commission. Davies A. 1994. 1994. 8. Armstrong D. Rosnes J. International Journal of Food Microbiology. Davey K. Rome. Huss H.. References Huss H.A.A. activities and safety. 24.. Sivertsvik M. Feldhusen F. Microbiological spoilage of fish and fish products. Ed. 4. 9. 2001.H. Rome. Incidence and growth of psychrotrophic Clostridium botulinum in foods.R. 348. 5-7. 33. Food and Agriculture Organisation. 14. Control of indigenous pathogenic bacterial in seafood. 2003.. 291-304. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth. Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef. 67-71.R.. Food Control. 20 .R.J. 1992. 243-60.. 6. Spoilage and shelf life extension of fresh fish and shell-fish.T. 2000. 87-118.N. Huss H. Incidence of foodborne pathogens in European fish. M. Gram L. and natural toxins. 37. International Journal of Food Microbiology.K. 1995.. Ed. 13. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). 1993.S. Microbes and Infection. 91-8. parasites. 121-37.E.J. 12.. Capell C. International Journal of Food Microbiology.. Davies A. Critical Reviews in Food Science and Nutrition. Italy. 5.. 19. Quality and quality changes in fresh fish.E. Food Control.H. 2002. 107-27. Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review.. Jehanno D. Dalgaard P. 36. 1651-60..
27. 27. 117-27. Westport.J. 18. Ed. 1987. 1995. Washington D... Conway E. An absorption apparatus for the micro-determination of certain volatile substances. International Journal of Food Microbiology. 19. 25. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish.. Sea Fish Industry Authority. in Advances in Seafood Biochemistry.. 23.. Edinburgh. Khayat A. 121-37. Food Technology. 305-17. 1415.. Flick G.M. 22. 354-8. 1982.E.edu/haccp/compendium/chapt08.C. National Advisory Committee on Microbiological Criteria for Foods.J. Microbial and biochemical spoilage of foods: an overview. Hendrie M.J. Ratnayake W.R. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere. Autolytic changes in fish post mortem. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout.J. 1995. 1959.. 22-36. Quality and Quality Changes. Adopted. Non-enzymatic oxidation of seafood lipids. International Journal of Food Microbiology.H.a review. Slade A. Gill T.htm Anon. 1996.. Letters in Applied Microbiology. Journal of Food Quality. 245-67. Biochemical Journal.. AVI Publishing. . 130-2.G. Huss H. 1985. in Chemistry and Biology of Marine Food Products. 33. Journal of Applied Bacteriology.. 37..CHILLED AND FROZEN RAW FISH 15. Technomic. March 20. 20. http://seafood. 1995. Huis in't Veld J.. Microbiological spoilage of fish and fish products.H.E. Dalgaard P. Yajima T. Part I . Surette M. 184.M. Hebard C. Journal of Agricultural and Food Chemistry. 29. 1992. in Fresh Fish. 41-50.. 28. 24. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products. Arai K. Martin R. The micro-determination of ammonia. Huss H.N. Martin R. Ackman R. Herbert R. Eds.E. 11. 34. Schwall D.. Eds. 33. Bacteria active in the spoilage of certain seafoods. 21 16. 1933. Gram L. Børresen T.. Chemical composition. Saito T. Qualitative and quantitative characterisation of spoilage bacteria from packed fish.A. in Fresh Fish. Flick G. Davies A. International Journal of Food Microbiology.M.E. 1971. Ed.M. Shewan J. Gibson D. 2007.H. 1-18.ucdavis. 19-22. Food and Agricultural Organisation of the United Nations. 1988. 42-54.J.. 21. Flick G. 1995. Reginstein J. Lancaster. Gill T.J.A.. Byrne A. Food and Agricultural Organisation of the United Nations. Rome. 36.E.E. 1992. 30. Rome.spoilage of fish. Quality and Quality Changes.J. Pedrosa-Menabrito A..H. Nature. 419-29. 26. Hebard C. 1996. Martin R. Changes in purine nucleotide of red lateral muscle of rainbow trout. Shelf life extension of fresh fish . Huss H. 17.S. 21. 1983. Leblanc P. 26. Lipid oxidation in seafood.J. National Advisory Committee on Microbiological Criteria for Foods. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage. 1.
43.L. Sumner J. 44. Archiv für Lebensmittelhygiene. 2002.F. 51-62.. Journal of Applied Bacteriology. 323-9. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish. Scanlan R. 36. 317. Ed. Nealson K. 37. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere. Lee J. Huss H. 33.M. Myers C...M. Journal of Bacteriology. Satomi M. 67.. 41. The inhibition by CO2 of the growth and metabolism of microorganisms. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens.J. Blackwell Scientific Publications. 22 . Annual Review of Microbiology. Iron and manganese in anaerobic respiration: environmental significance. 35.H. Dalgaard P. 39. Kell D. 6689-97.. 1989. Growth and activity of Shewanella putrefaciens isolated from spoiling fish. 328-9. 1992. 62-70. Myers J.. Role of proteolytic bacteria in fish spoilage.J. 4. 48. II. 311-43. 1984. Gorczyca E. 262-6. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. 9.. Venkateswaran K. Karnop G. physiology. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches. Environmental Biotechnology. Pseudomonas fluorescens.A. Gram L.europa. 3429-38. 109-36. 26. 1996.eu/LexUriServ/LexUriServ. Occurrence and importance of proteolytic bacteria as spoilage indicators. and an Achromobacter species. 334. The effect of storage temperature on shelf life. in Fresh Fish. 1989.. 71. Jørgensen B. 174. and regulation. 1985. 40. Italian Journal of Food Science.FISH AND SEAFOOD 31. Quality and Quality Changes.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J. Applied and Environmental Microbiology. 36.M..S. Magno-Orejana F. Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens. 1982. Brochothrix thermosphacta. FAO Fisheries Report. Drosinos E. 1973.H... Dixon N. Miller A. Fish spoilage bacteria – problems and solutions. 33. 72-80.B.. Saffarini D. International Journal of Food Microbiology. Food and Agricultural Organisation of the United Nations. 1994. 38.. 1995. Vogel B. Rome.. Nychas G. 18-21. Huss H.H. 34. 13.R. Oxford. Taussig M. Rome. 61-6. 2005. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia. Processes in Pathology and Microbiology. Brady P. Gram L. 32.R.. Applied Microbiology. Libbey L. 42. 1984. Cohen D. Dalgaard P..E.H.
Griffin P. Springer. Slutsker L. Loessner M. Kluwer Academic/Plenum Publishers.L. 52.H.C. Presence.N. 1-34. Altekruse S. Surveillance for Foodborne-Disease Outbreaks . 1991. 17-34. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets. 20. Food Technology. Ljungh A. 53. Whiting R. Mac Rae I. 55. 85-95. 1661-6. Ray B. Smith J. 23.. Miller A. The International Commission on Microbiological Specifications for Foods. 56. Lehane L. Morbidity and Mortality Weekly Report. Fish and Fish Products (Microorganisms in Fish Products).... E and F in salmon fillets stored under modified atmospheres at low and abused temperatures. Daniels N. Modern Food Microbiology. 49. 2003. 100-3. Hammond R.CHILLED AND FROZEN RAW FISH 45. Woodruff R. 181-8. Genigeorgis C. 1991. Sampling for Microbiological Analysis: Principles and Specific Application.... Wadström T. 12. Journal of Food Protection. Lynch M.K. Vibrio parahaemolyticus Infections in the United States. Painter J.A. Thompson S. 51 (4). 1973-1998. 181. 12.P.. The International Commission on Microbiological Specifications for Foods.. Use of risk assessment to reduce Listeriosis incidence. Microorganisms in Foods 2. Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B. 2000.. Applied and Environmental Microbiology. Braden C. 330-6.. International Journal of Food Microbiology.W. Olley J. 1994.A..M.M. detection and growth of Listeria monocytogenes in seafoods: a review. Myers M. Aeromonas and Plesiomonas as food.United States. Oxford. 181-96. USA.. 23 . 55 (SS10).M. Microbial Ecology of Food Commodities... 2005. 2005. Wilson S. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico. International Journal of Food Microbiology. The International Commission on Microbiological Specifications for Foods. 58. Journal of Infectious Diseases. Embarek B. 1987.J. in Microorganisms in Foods 6.. 48. International Journal of Food Microbiology.and waterborne pathogens. Bean N. 50. 51. Blackwell Scientific Publications. Panicker G. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures...C. 57. Majeed K.. MacKinnon L. International Journal of Food Microbiology.. 50. 2006.J.. 1-37. 2194200. 1986. 172-249.. 2000. Bej A. Histamine fish poisoning revisited. 1998-2002. 47. 1997.L. 54.. Bishop R. 46.P. Huss H. International Journal of Food Microbiology. 303-12. Jay J. New York. 1993. Lindroth S.H. 69. Garcia G.. Ed..K.K. Golden D. Embarek B.
D.. Villemure G. in Fish as Food. Smith G. Eds.G.9.P. Journal of Food Science. 1992. Hobbs G. 1. 271-85. Ed.. 1986... 1. 1984.. 1989. Simard R. Dalgaard P. 317 (Suppl. Gibson D. Olley J. Magno-Orejana F. Technomic Publishers. 1961. The Microbiology of Seawater Fish.A. in Torry Advisory Note No 93. Kitts D. Huss H. 1989. 51. Fisheries and Food. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. Cann D. McMeekin T.. 1985. 1997. Shinano H. Aberdeen.. Huss H. Gram L. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches.1 Further Reading Processing Anon. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. International Journal of Food Microbiology.9 1. Food and Agriculture Organisation. Journal of Food Protection.M. International Journal of Food Microbiology. Bulletin of the Faculty of Fisheries (Hokkaido University). Studies on the microbiological ecology of mackerel stored by the method of partial freezing. Torry Research Station. in FAO Fisheries Report No. International Journal of Food Microbiology. 35. Aberdeen.. Jones Y. 1992. Trolle G. 24 . Estimation of the bacteriological quality of fish by automated conductance measurements.H. 127-34. Akiba M. Huss H. International Journal of Food Microbiology. 62-70. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging.M.. Torry Research Station of the Ministry of Agriculture. 1.C... in Seafood Science and Technology.E. Gibson D. Food and Agriculture Organisation of the United Nations. Sumner J.D. Shahidi F.. Handling and processing of scad. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. Picard G.. Torry Research Station of the Ministry of Agriculture. Ravn Jørgensen B. 4.L.. 1988. 15. 6. 1984.1983.FISH AND SEAFOOD 1. 295-307. Fisheries and Food.H.. Taoukis P. Rome. Labuza T. Ross T. Lancaster. 73-89.2 Spoilage Shewan J. Microbiological quality and shelf life prediction of chilled fish.H.. 65-72. 55. Houston N.S.M. 13-32. Changes in microflora and chemical compounds in mackerel stored by partial freezing. Academic Press.9. Application of predictive microbiology to assure the quality and safety of fish and fish products.. Ogden I. 741-50. Ed.. Borgstrom G... 317-21. Florida. Cho Y.).
Amsterdam. 5. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. Gram L. Effect of culture age.9.K. Garcia M. International Journal of Food Microbiology.. Trolle G. Klausen N. Liston J.M. Blackie Academic & Professional. Otero A. 65-72. Eds.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products.J. 59. Huss H. 20023. Gram L.3 Pathogens Condon S. 322-6. http://eur-lex. International Journal of Food Microbiology. in Fish processing technology. Journal of Applied Bacteriology.europa.. Huss H.H. 1993. 1992. Elsevier Science. 1997.R.. Jacobsen M.eu/LexUriServ/LexUriServ. 1992.. 25 . Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C..CHILLED AND FROZEN RAW FISH Einarsson H. 1. Sala F. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. in Quality Assurance in the Fish Industry. Ed. 1. 1987.H.. Hall G.. 72.L. 479-88. Huss H.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. 4. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. London. 1987. Applied and Environmental Microbiology. 2197-203. Modified atmosphere packaging of fish and fish products. A rapid method for detection of histamine producing bacteria..9.H. 137-46.
smoked (hot) mackerel. or could be disguised by sauces.). Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2. 27 . clarity of eyes. emphasising the high quality of the seafood and the gourmet nature of the prepared products. colour of gills. smoked (cold) salmon. The same used to be said for the difference between fresh and frozen products. seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. colourings etc. This has the result that any microbes present in the product have the capability to grow (with subsequent spoilage) unless strict temperature controls are applied. and on-board freezing mean that products frozen at the point of capture are now perceived by the consumer as being of a better quality than a fresh product. This is no longer the case with many processed products now available in stores.g. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e. the exceptions being Sushimi (high quality raw fish). Few seafood products are sold ‘ready-to-eat’. The majority of seafood products produced require a cooking stage before consumption. 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.g. it does not necessarily apply to the toxins that may have been produced. and crustacea such as shrimp and crabs. due to the early cessation of spoilage. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr.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. Even though heating prior to consumption will eliminate pathogenic microbes. Sushimi tuna). although recent developments in improved supply chains. products of lesser quality. but still acceptable and meeting all safety criteria.2. smell etc.
prawns. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein. and are well known for their diet consisting of waste materials. 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. but these are now finding favour as products in their own right. They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. 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.FISH AND SEAFOOD Recent trends in the market for food and seafood products. Nevertheless. They are typically flown into export markets. Seafood is the term used descriptively to include all categories of product. and the consumer only has to heat (cook) the product. they are usually supplied filleted with head on. Microbiological hazards include Salmonella. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted. Shelf life on ice. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish. Listeria spp. molluscs. have shown the increasing importance of convenience foods. but should be equivalent to cold-water products. including freshwater fish. At the same time. Prepared natural fillets are fillets of fish without any additional coating. Doubts have been raised over the microbiological status and handling practices of reef-caught fish. Frozen fillets may have a surface layer of 28 . There is also a limited market for filleted material. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. 2. the processing industry has developed a wide range of value added products. and pathogenic Vibrio spp. they are usually of a better standard than species such as tilapia. The expected shelf life attainable for this format is usually up to 8 days. which can be farmed. This includes all categories of seafood including trout and other freshwater species. and these are increasingly being presented with an ever-expanding range of accompaniments. shellfish and molluscs. where the preparation has already been carried out... It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP). in particular. and this has produced a greater availability and diversity of products in the marketplace. or in MAP depends on catch conditions and treatment. as they tend to lose colour and sheen after defrosting. rather than frozen. etc. from both wild capture and aquaculture sources.
They are sold from wet fish counters.g. a protective glaze may be added to the frozen steak.g. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. 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. annatto with smoked haddock. sawdust with other woods adjusted for flavour profile and burn duration). 29 . Some products may be salted for flavour only (e. Hot-smoked fish are prepared from brined. Fish steaks can be cut from a natural fillet or loin (e. VP products) because of the high salt content acting as a barrier to spoilage. resulting in a product that is ready-to-eat without any further cooking. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes.g. haddock.2. This process in effect cooks the product and reduces or eliminates microbial contaminants. and the product then shaped and heat-processed to cook the product before being chilled. mackerel. Pâtés are prepared with raw fish. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. but may be increased (up to 1 month) by packaging methods like MAP or VP. Fresh fillets are sold from wet fish counters. be flavoured with herb dips or peppercorns. 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. The result is fish with a typical residual salt content of 1. tuna steaks) or band-sawn from a frozen block (e.g. or pre-packed in MAP or vacuum packaging (VP).2 days) longer than that of non-prepacked fish..5 . mackerel. cod). Again. in addition. salmon. which may. swordfish steaks). etc. which is approximately 30% (or 1 . and salmon. The shelf life for MAP fillets is typically up to 5 days. The expected shelf life of these products is up to 8 days (or longer for some chilled. or pre-packed frozen with added glaze. the ingredients premixed. Cold smoking is essentially a flavouring process. with a minimum residual salt content of 3.5% in the aqueous phase. natural fillets of fish such as herring. in VP or through mail order. and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. Added colours can be included with the brine solution to meet specific requirements for products e. An alternative process using a liquid smoke in the brine is claimed to result in a similar flavour with reduced water loss in product and improved yields.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze.5%.
haddock. Surimi is the insoluble protein fraction of fish after washing and processing. It is often used as a protein supplement. The majority are products made from various types of whitefish (cod. Natural prawns from cold or warm waters can include those peeled. This ‘catch-all’ category includes seafood salads. pasta or potato.g. 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. it can be used as a seafood analogue. rice. They are pasteurised products and treated as high risk raw materials. Breaded fillets/steaks and battered fillets are either natural or shaped fillets. coated in batter or breadcrumbs. cephalopods. 30 . or cut from a preformed frozen fish blocks into shapes.. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process. e. jellied eels. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. mixed whitefish. Other ingredients may be natural or reconstituted. and seafood mixes. addition of ingredients such as colours. fish nuggets/bites with dips and seafood-based snack products. Other. flavours. Scampi are tails of Dublin Bay prawn or langoustine. With further processing. taramasalata.FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate. ‘fingers’ before being coated in batter and/ or breadcrumbs. Band-saw dust and fish trimmings from fishblock preparation materials may be used. Fish cakes are products produced by mixing fish with potato and herbs. which may include molluscs. and may also include additional ingredients such as fresh herbs and spices. salmon) coated and sold as chilled products. Prawns. There is an increasing market for high value fresh or thawed fish (e.or a five-stage process.) Raw material is typically fish fillet.g. tuna. The protein source can be fish fillet or blocks. e. with an expected shelf life of around 7 days. rice or pasta. and heat-fixed. long-life seafood mixes using sorbate and benzoate as preservatives. binders. etc. The cakes are usually coated using a three. and in the shell. and formed into round cake shapes.g.
product handlers. 2). storage. and pathogenic flora associated with seafood. or visible damage from gaff marks (4). specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1).and turtlefriendly long lining to the less discriminatory netting. The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. Grading is based on size of fish (>25 kg). remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. sensory assessment of freshness etc. on the whole. as a result there is an increase in spoilage rate thereby reducing the product shelf life. i. A primary role in fish spoilage has been assigned to this organism. skin. After capture and during any subsequent processing. and ingredients added to the final product.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. Contamination will. is not used for sushi or other high risk (and high value) products. the microbial flora of seafood is associated with the gills. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. Recent public opinion has favoured the use of dolphin. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). including seasonal variation. searing or bar marking). and the potentially toxic by-products of histamine-producing bacteria (5.3 2. handling or preparation stage. and have emphasised the origin and sustainability of the fish as key marketing points. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. Only after death. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning.I. fat content. 6). and intestines of the fish. whilst gaping is also dependent on a range of other factors that effect the muscle composition. Dalgaard et al. see Table 2. Tuna that does not meet the highest of grades based on colour. and processing conditions such as dehydration or poor filleting practices.e.3. species (Bluefin>Bigeye>Yellowfin>Albacore).CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. The edible tissue of some species of fish are more susceptible to gaping than others. 31 . Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. In summary. For a summary of background information on the hazards associated with fisheries products and ingredients. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. which may become a focus of contamination deep within the muscle. will the saprophytic flora start to invade the surface flesh (1. it may be used for other products or markets depending on the product specification.
TABLE 2.FISH AND SEAFOOD 2. cereus B. Vibrio cholerae. moulds. yeasts Pathogenic microflora/toxins Salmonella. moulds Bacillus. cream. Moulds Enterobacteriaceae Salmonella. moulds Bacillus Bacillus. raw materials. pseudomonads. shelf life determination. enteric viruses 2. yeasts Lactobacilli. staphylococcal toxin. Fusarium. and milk Cheese Water/Ice Bacillus. moulds Bacillus.3.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. In addition. toxin.4 2. Enterobacteriaceae. 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. and so are the potential microbial contaminants associated with them. anaerobic spore-formers. Pseudomonas. Bacillus subtilis B.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. by the HACCP team or by a hazard and risk assessment programme. Escherichia coli. for chilled products. streptococci. histamine Cryptosporidium. Clostridium perfringens. Table 2. and associated spoilage and pathogenic microorganisms. Listeria. Staphylococcus. Aspergillus. 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. B. coryneforms. however. cereus Bacillus licheniformis.4.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. Shigella. Aeromonas. Clostridium botulinum. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter. coliforms. mushrooms. Bacillus cereus. intermediate processing. nuts and fruit Associated microflora Lactobacilli. 32 . Penicillium. Clostridium Lactobacilli. product assembly. protozoa and viruses B. be considered at an early stage during new product development. spore-formers. Staph.I provides an overview of common raw materials.
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. Effects on the microflora are minimal during tempering (7). The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. but it is assumed that any band sawing or dice process will introduce heat from blade friction.1 .4. Where the raw materials are frozen. Generic examples have been used to illustrate only the major points. 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. Examples of tempering techniques include the use of heated air. Rapid controlled cooling of heat-processed components will help to minimise spoilage. there are a number of distinct methods used. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. up to -5 °C so as to improve the cutting and handling properties during processing.4. All have the same intention. 2.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes. to evenly raise the temperature of the entire frozen product from -20 °C or lower. but it also poses a risk of cross-contaminating other products undergoing tempering. as a heat process is often required for functional or organoleptic reasons. Figures 2.2. Band-sawn materials or fish dice are normally promptly further processed.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.4.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. It is recommended that the principles of HACCP are applied to novel processes. this may be an incidental effect. line extensions and process reviews. For some components.3 Preparation of fish component of chilled and frozen recipe dishes 2. the core temperature. or the use of microwave technology.3. as the full complexity of the processes used for specific products cannot be covered in a general text. but will also need to ensure that the optimum shelf life can be safely obtained.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. and potential opportunities for growth of foodpoisoning bacteria. 2. and contamination from equipment to clean fish muscle. depending on the size of the fillet 33 .
1B . The processing of three examples types are described in Figure 2. On arrival at the processing plant.7 days. Raw material preparation stages for chilled or frozen coated fish products 34 . after landing. with the coating heat-fixed. 2. may not rise above 0 ºC during processing. are placed into plastic boxes and covered in ice to preserve their condition.3.1 (2. Fig.3.Fish Cakes). 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. by mincing and mixing with a binding agent (as described in section 2.Scampi. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process.1 Whole-fillet / scampi Scampi (Nephrops norvegicus).4.1. the scampi is washed in potable water to remove foreign bodies. 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. 2. quickfrozen.FISH AND SEAFOOD processed.1.2.4. chilled or sold chilled on defrost with an expected shelf life of 6 .1A .Shaped Fillets.1C .2). It is then size-graded. drained and then refrozen for storage until required for further processing. frozen in a nitrogen blast freezer and weighed into sacks for later processing. Frozen cores are usually batter-enrobed.1.
quickfrozen. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. more seriously. spoil more rapidly than the dry ingredients. or those where the trimmings. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. This is especially the case with block-sawn dust. Therefore.g.3.4. material will need to be transferred swiftly to a chiller. those that use frozen blocks of whole fillets. once the dry components have been rehydrated. that are to be reused for other foods.7 days. 35 . unless it is poorly handled. Fish cake cores may be batter-enrobed. additional spoilage organisms may be introduced. 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. Waste material produced from this type of processing is often reused in the third category (fish cakes). The mix is shaped and formed ready for packing or for enrobing.1. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. with an expected shelf life of 6 . It is for this reason that all by-products from primary processing.1. because of its nature. Where frozen blocks of fillets are used they will be “tempered” (temperature raised to around -7 °C) to facilitate efficient cutting with a band-saw to a specified thickness. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. chilled. The remaining dried ingredients are mixed with the fish. are handled and stored as any other food product. iii) Fresh/fillet/minced fish block. with the coating heat-fixed. ii) Fish-filleting waste . or promptly processed to avoid further quality loss.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.. pathogens such as Salmonella and Listeria.4. fish fingers).3. 2. or sold as a chilled product after thawing. but. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . and water is added to rehydrate the components.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. from potato flake.block-sawn dust. such as aerobic spore formers and Staphylococcus spp.2 Shaped products This type of product falls into two categories.off-cuts or belly flaps. The fish component of the cake will.
Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. The first stage.FISH AND SEAFOOD In all of the above examples.. any excess is removed with an air knife. and Enterobacteriaceae from flour. milk powder and spices. rusk or crumb.4. potentially. flour. a characteristic flora will develop from the batter components. with the coating heat-fixed.7 days. rapidly multiply if strict temperature control was not maintained. Natural fillets d. quickfrozen. and possibly Staphylococcus spp. e. raising agents. and to carry flavourings where required. Breaded or battered fish fillets are fully heat-processed before consumption. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. perfringens could. and if required. 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. which typically consists of flour. which can occasionally be isolated from protein-handling processes. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process. vibrios and other fishassociated spoilage organisms. Batter contains water. Over time. Spore-forming anaerobes such as C. such as Listeria and Vibrio spp. chilled. or sold as a chilled product after thawing. is the addition of a pre-dust. because of the intensive handling required during processing. lactobacilli from the milk powder and flour. which is product-dependent and not always required.g. Products are preformed and presented to the process (either three. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. Fish products. Tempered band-sawn materials b. and may include milk powder. from flour. The function of this is to improve the adhesion of subsequent coatings. fish cakes Portions or fillets may be batter-enrobed. absorb any excess moisture on the surface of the fish portion. Bacillus spp. i. some of this initial flora 36 . 2.2 illustrates the typical stages of enrobing or breading. A large variety of product formats and types can be batter-enrobed. egg. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. as well as ensuring that there is minimal contamination from human processors.4 Battering/enrobing process Figure 2. IQF/pre-formed fillets c. starch. with an expected shelf life of 6 . e. spices. Initially.g. a. If a crumb coating is required.or fivestage) frozen or chilled.e.
Fig. With inadequate temperature control of batter systems. The origin of contamination for the two organisms is often unclear. but it is believed that poor hygiene regarding product and food handlers.CHILLED AND FROZEN PREPARED FISH PRODUCTS associated with the ingredients will be diluted out. and both of these can produce heat-stable toxins that can survive a batter heat-fix process. cereus can proliferate under certain conditions. Flow diagram outline of a typical three.2. but because batter is heat-set. It is advisable that some control mechanism is employed to prevent toxinforming organisms from multiplying to dangerous levels. dry batter mixes are all potential sources.and five-stage enrobing/breading process The length of production run and temperature of the batter are key factors in deciding the balance of the dominant flora within the enrober. This can be achieved by frequent clean-downs and adequate hygiene. and.2. contaminants such as Enterobacteriacae can proliferate and develop during the process. time-limitation or production run temperature control. aureus and B. this is not of great significance to the microbiological standard of the finished product. cereus. Staph. replaced in part by process contaminants and bacterial flora from seafood. The FDA has issued clear guidelines on acceptable 37 . in the case of B.
38 . There are two main categories.2.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe.5. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7).1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal.1 ºC for no more than 3 hours is recommended (1).g. and must not show evidence of disease.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes. 2.4.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. pasta or potato. The vegetables must withstand processing. Fig. A temperature no greater than 10 ºC for 12 hours cumulatively or 21. Figure 2. The protein source can be fish fillet or block. canned fish may be used in their preparation owing to it having wider availability and low wastage. Flow diagram outline for recipe dish ingredients preparation 2. as this could introduce off-flavours and reduce shelf life. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products. rice.3.4. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. For example. to produce a specific product. but they are supplied without a source of carbohydrate e.
encourage more rapid decay and loss of structural integrity. For premium products. The opportunity for growth of L.lactobacilli. not just as accompaniments. the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. Washing with a biocide will at best achieve a 2 . coli O157. treated as a highrisk ingredient. spore-formers. Although the processes are claimed to be superior to using unmodified hypochlorite for washing. and may include sauce accompaniments. rice or pasta. chopping and mixing of material.e. and reduce spoilage organisms and potential pathogens. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. The majority are products made from various types of whitefish (cod. colour and water will occur.3 log reduction in bacterial numbers on the surface of vegetables. with cases of foodborne disease traced to Listeria monocytogenes. (7. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. but also as components in recipe meals. Once vegetables have been blanched. Blanching of vegetables can be an effective means of reducing or removing surface contamination. and may include molluscs or prawns that may or may not have undergone a minimal heat process. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. The presentation of vegetables has a significant effect on the product quality. In an attempt to improve washing and reduce chlorine taint. Raw material is typically fish fillet. coryneforms. they should be stored under temperature-controlled conditions. 39 . For example. will disperse the saprophytic flora of vegetables .4. or breadcrumbs. Salmonella typhi. with the accompanying release of additional nutrients by processing. Products must be washed thoroughly to remove pests and soil. However. for example. However. A number of examples of direct faecal contamination of vegetables exist in the literature (8). and protected from any post-processing contamination.5. any processing stage. Any assumptions regarding the application of good agricultural practice to produce should be made with caution. loss of quality.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.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. and E. all-year-round fresh vegetables are preferred. and mixed whitefish). haddock. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. if the process is not appropriately controlled. coliforms and pseudomonads throughout the mix. 8) will. even those with pH-modified dressings. For example. frozen block or canned fish. 2. pastry. control is more onerous. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components. i.
Sauces can be multi-component preparations or more simple preparations. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. a heat process in excess of 73 ºC will be required to rehydrate this component fully. cereus). An incident involving Staph.4. Appropriate handling facilities or equipment will be required to protect the product. cereus are well known and are still a common issue in the UK (9). such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. If the sauce contains modified starches for freeze/chill stability. Potato toppings made from flake or powder will normally have little associated spoilage flora. 11).1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. can germinate and rapidly multiply.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. 2. and then cooled slowly allowing germination and re-growth. B. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. and an increased risk of contamination from L. where composite meals are heat-treated. This is a particular issue especially where products are heat treated to eliminate vegetative cells. especially in those products that have a vegetable component or rice accompaniments. On hydration. a process equivalent to at least a 5D Listeria kill will be necessary. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp.6. aureus toxin is well documented (10. with the range always expanding as new tastes and cuisines are developed. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose.4.g. Alternatively.) to the product (12. cereus. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. spore-formers can proliferate if adequate control of temperature is not maintained. Sauces or marinades are added to fish to broaden appeal or to accompany components. Placing such products in a standard chill store to cool down could result in condensation. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. Typically. Products or dried ingredients contaminated with either spores of aerobic bacteria (e. 13).6 Preparation of non-fish components of chilled and frozen recipe dishes 2. but temperature-abused pasta can also spoil rapidly by mould growth. if subjected to temperatures between 20 and 40°C. For this reason all products. 40 . rely on the use of microbiologically sound ingredients. such as butter garnish with black pepper.
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. 13). and handling of processed sauce can be less rigorous than. 41 . Figures 2. Marinade-type products are becoming more popular. etc. 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. that of a dairy-based sauce. flour products. suspended in an acidified sauce with an oil component. 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.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig.herbs.2.4. The format uses flavour compounds .4 and 2. for instance. respectively.5 illustrate the recipe dish ingredients preparation and assembly and packaging. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. Without barriers. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. spices.
the method replaces air in the sealed packaging with a defined mixture of gases. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. which may restrict either microbial growth or enzyme activity. These include: 2.2.FISH AND SEAFOOD Fig. botulinum to grow.4. generally carbon dioxide and nitrogen.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions). unless the temperature is strictly controlled to below 5 °C throughout the supply chain.7.4. Flow diagram outline assembly and packaging of recipe dish 2. The oxygen-free conditions will however allow other anaerobic bacteria such as C. as it dissolves in the surface moisture to form a weak acid.7. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. and tuna loins are packed by placing the product in a plastic pouch and removing all the air.4.5.2 Modified-atmosphere packaging Originally developed for the red meat industry. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. 42 . 2.
Vibrionaceae will succeed (17 ). where the protein component is nearly always heat-processed. Spoilage will be most readily detected in proteinaceous components of ready meals. by a loss of texture. but they will also be readily adapted to the nutrient source and conditions. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15). Spoilage of this material is not expected to differ greatly from the processes described elsewhere. and finally flavour loss. is typically uncooked or at best lightly processed to retain eating characteristics. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp.5 Spoilage During processing. both organisms are capable of growth at chill temperatures. Shewanella putrefaciens and Ph. except for some crustacea. This is also true of chill storage and the distribution chain. Nonetheless. The case for growth and contamination of products with Aeromonas is stronger. the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. 43 ..10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. However. This is considered to be the one major difference between fish and meat products. Not only will these organisms have a numerical advantage over other proposed spoilage organisms. followed by moisture loss. and in influencing the final eating quality of the product. It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage. a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood. and Yersinia spp. The fish component of recipe dishes. At refrigeration temperatures (5 . phosphoreum. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18). (20).1 Chilled storage For chilled products or chilled components of frozen products.5. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’. but is not limited to. The literature has good information on spoilage of individual components of recipe meals. Aeromonas spp.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. Beyond this. 2. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). temperature control is important in preserving chilled integrity of ingredients. but little on in-pack spoilage. initially. in numerical terms. as this organism is a common contaminant of freshwater fish species (19). at slightly higher temperatures (10 ºC).
within dairy products are well known. It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage. For example.5.. where the variety of components and ingredients used in manufacture is varied and complex. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. This reinforces the need for prompt processing and temperature control of ingredients and finished products.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24). auto-oxidation of fats/oils).3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). Bacillus spp. Pseudomonas spp. 2. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. Still. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. botulinum. These have been proved to impart off-flavours. freezing also has a protective role in improving the safety of raw materials by. for example. and Aerobacter spp.. The conditions and freezing process most favourable for good-quality products will. producing characteristic off-flavours. the shelf life of products is limited by the natural spoilage process described earlier.FISH AND SEAFOOD 2. However. 44 . 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.. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. As a note of interest.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. Products that are developed with an extended shelf life beyond the 6 .5.g. 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. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). this should also take into account shelf life of product components used in the assembly of the final product. By retaining the intrinsic spoilage flora of seafood. however. Serratia spp. also be those that have the least effect on the viability of microorganisms (21). eliminating nematodes in products that do not have a heat treatment step.
water. farmed or free-living crustacea (30). agricultural run-off or sewage can contaminate in-shore species. 45 . Additional hazards from marine bio-toxins. These are C.. air. Hazards attributable to the non-seafood components are listed below and this information. 2.6. in some cases.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. Incidents in such diverse materials as cream. botulinum when held in favourable conditions for growth (28) i. and Campylobacter are recorded in filter feeding molluscs such as mussels. low oxygen or anaerobic conditions.e. along with previous discussion.104/ g should be treated with some concern (26). may be of benefit within any risk assessment for chilled and frozen prepared fish products. Therefore.1 Bacillus cereus This is a common contaminant of dust. 13. histamine intoxication.6. Cases quoting contamination of seafood with organisms such as S. and many raw or processed foods (26). have been linked with food poisoning and should also be considered. etc. soil. Counts of between 103 . This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. perfringens contaminating seafood will depend on the source of raw material. Vibrio spp. 2. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. permit rapid growth of B. and this issue may be of greater consequence for warmwater. A number of other Bacillus spp. Germination of spores requires favourable conditions. In addition. potato.6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. especially molluscan filter feeders. cereus. are dealt with elsewhere in this book. with pathogenic microorganisms or viruses. or a heat shock followed by holding at a temperature suitable for growth. spices and herbs and vegetable shoots have been reported (12. 27). perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). and some effects related to product type and time to toxin production have been noted.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. any risk of C. typhi.6.3 Clostridium perfringens A study examining C. botulinum type E and Vibrio parahaemolyticus (25). 2. Inappropriate holding times and temperatures will.
It is unclear. aureus enterotoxins in foods. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. 2. indigenous or other. which is not affected by any subsequent cooking process. 46 .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). 2. aureus (35). and will therefore cause “food poisoning” even in a product that is cooked before consumption. when introduced or inoculated into sterile components of chilled prepared recipe dishes. if any other potentially pathogenic organisms.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. at the present time. Chilling may help Shigella to survive. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32).5 Shigella spp. and inadequate cooking or holding within the temperature growth range (27). infected food handlers.6. the restrictions on growth rate seen with seafood will not apply.6. L. growth may occur in foods (26). L. but under the appropriate conditions.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. However. will colonise factory environments in a similar fashion except for possibly Vibrio spp. and this is believed to be due to a tolerance to brine used in the smoking process (32). Sources of contamination are many because of the widespread distribution of this organism in the environment. 2. but.6. These include inadequate refrigeration and preparing foods far in advance of planned service. These conditions will allow the production of the toxin. are considered primarily to be waterborne organisms but epidemiology indicates that contamination of vegetables and possibly in-shore caught seafood could act as vehicles for infection by Shigella (34). monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. monocytogenes as a contaminant in foods (31). An acceptance has now developed in the UK of the presence at low levels of L. Shigella spp. Rates of contamination with smoked seafood are much higher than those with unsmoked products. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33).
For instance. 2. unless the criteria is for a pathogenic organisms where only non detection is acceptable. 2. The following tables (Table 2. but these may be eliminated by a subsequent heat-treatment stage. and are a recognised means of providing data on manufacturing standards. pastry. etc. and their association with similar materials. (36). Microbial guidelines.6. Microbial levels at the end of manufacturing (m) will be set much lower than those at the end of shelf life (M) which in turn should still be safe to consume. but provide the industry with an indication of the microbial level at which action is required to ensure product safety. with respect to the number and types of ingredients used. such as batter mixes.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts. aureus enterotoxins and B. 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. It should be remembered that 47 .CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph. and not to final products. although due to long analysis times required. .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. 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. and as such the monitoring is for verification purposes rather than control. 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. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). as published by for example. although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40). cereus emetic toxin are often confused owing to similar symptoms. pasta. Microbiological specifications do not necessarily relate to product safety.II and 2.III based on IFST Guidelines (39)) provide example guidelines for seafood products. the fresh product is often consumed before the results are obtained. are not mandatory. Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems.
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.g. TABLE 2.FISH AND SEAFOOD the microbial criteria for other products (rice. coli GMP* <102 Maximum 104 TABLE 2. pasta.III IFST Guidelines for processed foods . monocytogenes B. coated fish.fish products. 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. L.II IFST Guidelines for part-cooked foods – e. fish fingers. Maximum= Levels at end of stated shelf life 48 . parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp. aureus V. batter etc) should also be consulted when establishing the specifications for raw materials and final product.
6.. Taylor S. 38 (11). Bartlett C. Beuchat L. 1997.. Gilbert R. 25 (1). Letters in Applied Microbiology. Tu A. 39 (10). International outbreak of staphylococcal food poisoning caused by contaminated lasagne. Communicable Disease Report.. Ohgiya S. 8. Ward D. Journal of Milk Food Technology. Journal of Food Science. 233-7. 70-2. 3. 204-16.M. Journal of Hygiene. 1997. 1962. in Fish Inspection Quality Control and HACCP: A Global Focus. Sakamoto T.. 1997. Kumeta H. Washington D. Huss H. Hoshino T. 1986. Bacterial histamine production as a function of temperature and time of incubation.J. Ishizaki K. Matsuyama H. Oxford. Aureli P. 166-73. 373-8. 24 (5). Hobbs G.W. 1976. Murrel H..M. 5. 11. International Commission on Microbiological Specifications for Foods. 10. 5 (3). 9. Eds. Ltd. 2.K. 47. Brown T.J. 1997. New York.. Taylor and Francis. Van Nostrand Reinhold. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. Bartram P. Communicable Disease Report. Letters in Applied Microbiology. Masuoka Y.. Yumoto I. Cambridge. Hackney C. Wienke A.. 96..R. 59 (2). Powers E.R. 167-93. Fish: microbiological spoilage and safety. Collette R. 1982. Microbiology of processed spices... The bacteriology of fresh and spoiling fish and some related chemical changes.. 663-87. Journal of Milk Food Technology. Outbreaks of foodborne illness in humans. 1991.8 1. 1976.R. 7.C.. 15. Shewan J. Christiansen T.H. 7 (24).. 1311-7.C. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. Woolaway M.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). Fish and Fisheries Products: Hazards and Controls Guidance. Lawyer R. 13. .. Pvt. London. References US Food and Drug Administration. 1. 67-73.L. Pathogenic microorganisms associated with fresh produce.L. 215. Importance of Photobacterium phosphoreum in relation to spoilage. Incidence and levels of Bacillus cereus in processed spices. 14. 2. Mejholm O.A.T. Latt T.. Animals and Spoilage.. England and Wales Quarterly Report. Food and Drug Administration. 1993. Proper processing and handling of raw tuna.R.. IBH Publishing Co.C. Food Science and Technology Today. Powers E. Kluwer Academic. 49 4. Journal of Food Protection. Recent Advances in Food Science. 1991. 2001...M. Behling A.E. Martin R.L. Dalgaard P..R. Slavin J. 668 -70. Microbiology of Marine Food Products. London. 12.G. Toxin Related Diseases: Poisons Originating from Plants.. 2005.
31.Yersinia..J. 1994.M. 351-72. 1987.A. Trolle G. Jemmi T. Clostridium perfringens in the environment. International Commission on Microbiological Specifications for Foods. 40 (5).. Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants. Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens. 28. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service. New York. Eds. 13 (1). Ed. Golden D. 24. 1986.. Walker S. Food Research International..C.J. 155-9. 4. 19..W.R.R. Blackie Academic and Professional. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures. 1974. Aeromonas and Verotoxigenic E. 50 17.FISH AND SEAFOOD 16. Report on Vacuum Packing and Associated Processes. Journal of Food Protection. 26.K. Effects of freezing and storage on microorganisms in frozen foods: a review. 1992. 11. in Advances in Fish Science and Technology. Blackie Academic Press. Is Aeromonas spp. 34-40. 2005. in Anaerobic Bacteria in Habitats Other than Man. 41-3. Gram L. Loessner M. U.. Halligan A.. Wood L. Granum P. Keusch A. Fresh and processed fish and shellfish . 472-506. Leistner L. International Journal of Food Microbiology. Leatherhead.C. 25. 1996.M. London. Glasgow. 30.J. 20. Speck M. 28 (4). Lund B.. perfringens? International Journal of Food Microbiology. 9. Leatherhead Food Publications. Barnes E. Springer. Journal of Food Safety. 333-6. 21..A Literature Survey. 51-158 Saxby M. Brooks J. 65. 23. Gaithersburg. Lund B. Food Taints and Off-flavours. 1996. Anon. 28. 1979. 27. PHLS Microbiology Digest. 59-72. Her Majesty’s Stationery Office. Applied Microbiology. Survey of the incidence of Aeromonas and Yersinia species in retail foods. 18.. 1977. SAB Symposium Series No. Gram L. Baird-Parker T. cereus and C. Microbiology in fishery science. Morgan D. 2000. Oxford.. 32. Huss H. 309-16. Mead G.J.J. Food Control. 655-60. Gould G. Jay M. 1992. Blackwell Scientific Publications. Huss H. Liston J. Curran D. Advisory Committee on the Microbiological Safety of Foods. Matches J.H.H. 22.. London. in The Microbiological Safety and Quality of Food. 274-86. Food Microbiology.. 29. Fishing News Books Ltd. Anaerobes in relation to foods of plant origin..E. coli (VTEC) . 1995. Andersson A. Modern Food Microbiology. Connel J.. Aspen Publishers. 1988. 25 (2). Food preservation by combined methods..L. Liston J. The Emerging pathogens . 1990. Ray B.V. a foodborne pathogen? Review of the clinical data. 4 (1).C. . 13. Eds. Ronner U. What problems does the food industry have with spore forming pathogens B. 1993. 1996.
34. 36. Kraft A.K.2. Principles and Practices for the Safe Processing of Foods. 1999.. http://www. London. Smith J.L. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. Development and Use of Microbiological Criteria for Foods. http://www. 39. Wolfe Publishing. 41. Phil Voysey.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. McCarthy S. U. 50 (9). CRC Press.K. Cambridge. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs. Varnam A.G. Shigella as a foodborne pathogen.pdf Shapton D.co.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. Foodborne Pathogens an Illustrated Text. British Retail Consortium. http://eur-lex. Journal of Food Protection. 2006.F.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33. Campden & Chorleywood Food Research Association. 1997.campden. 40. Shapton N. 372-6. Chilled Food Association. London. Psychrotrophic Bacteria in Foods: Disease and Spoilage.A. 38.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology. Journal of Food Protection. Gloucestershire. Chilled Food Association.uk/cday/posters2008/npd6. 1987. 1992. 37. 60 (4).H. Evans M. 1991. Institute of Food Science and Technology.eu/LexUriServ/ LexUriServ. 788-801.A.A.chilledfood. U.europa. 1998.. 51 . Microbiological Criteria. Woodhead Publishing Ltd. 35.
whelks. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). clams. where it is eventually swallowed and digested.1 3. squid. The phylum consists of five classes: Amphineura. For example. Box 3012 Bothell WA 98041-4421 United States of America 3. completely encloses the body.3. mussels. cockles.1 Definitions General features of the mollusc The phylum Mollusca. scallops. viruses. Gastropoda. Other species of shellfish such as crabs. while 53 . Jr. snails. and shrimp. lobster. abalones. In the process of filtering water. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells.7 litres of water per hour.6 litres. chemical contaminants and other impurities on the mucus of the gills. chitons. and Cephalopoda. mussels. S. which includes oysters. cockles. In these forms. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. 4). Scaphopoda. Pelecypods have little need for locomotion.S. O. The term shellfish in this context is limited to oysters. cockles and scallops. The class Pelecypoda includes all of the bivalves such as clams. Pacific Regional Laboratory Northwest U. P. composed of two valves hinged together dorsally. lying well within the mantle cavity. octopi. oysters. slugs. and 2. mussels. and scallops. Food and Drug Administration 22201 23rd Dr. Strong muscles. respectively. In most species.1. The class having species of economic and public health importance is the Pelecypoda. clams. oysters and clams can process about 3. they also trap bacteria. Plankton and algae form the bulk of their diet from seawater.E. MOLLUSCAN SHELLFISH Carlos Abeyta. the shell. can hold the shells tightly shut against enemies (1). Many microorganisms ingested by shellfish survive the digestive process (3. the adductors. Special tracts of cilia move this mucus toward the mouth. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. Pelecypoda.
3.1.North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp.FISH AND SEAFOOD commercially valuable. Whatever contaminants are in the water will eventually get into the shellfish.2 International commercial products Globally. are not as vulnerable to contamination through pollution of their beds as the bivalves. Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 . and usually at higher concentrations than found in their habitat. The habitat of oysters and mussels are sessile. dwelling on the bottom or attached to structures in the water column. but maintain contact with the water by means of a siphon tube (5). Clams are benthic and live burrowed in the mud.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 . Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific . many species of bivalves are edible (Table 3. The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3.South America Pacific – Japan Quahaug (also known as hard clam and hard shell clam) Coquina clam Hen clam Butter clam (also called the Washington clam) Mogal clam Little neck clam (also called rock Cockle Pacific littleneck) Atlantic – North American/Europe Atlantic – Europe/North America Japan Atlantic/PacificNorth America Donax spp.I).
MOLLUSCAN SHELLFISH Gulf clam Pismo clam Surf clam (also called bar clam) Freshwater clam Oysters Common oyster Portuguese oyster Blue point oyster (also called American oyster) Pacific oyster Europe Europe Atlantic – USA Ostrea edulis Crassostrea angulata Crassostrea virginica Gulf of Mexico Pacific – North America Atlantic – North America Titaria cordata Trivela stuttorum Spisula solidissima Japan Corbicula spp. Pacific – North America/ Australia/New Zealand Pacific – North America Crassostrea gigas Western oyster (also called Olympia oyster) Cockle Cockle Sydney rock oyster Rock oyster Dredged oyster Cockles Common cockle Ostrea lurida South America Japan Australia New Zealand New Zealand Ostrea chilensis Ostrea laperousei Crassostrea commercialis Crassostrea glomerta Ostrea lutaria North Atlantic – Europe/ North Africa Pacific – North America Cardium edule Cardium corbis Cardium aculeatum Cardium tuberculatum Spiny cockle Knotted cockle Mussels Blue mussel Atlantic/Mediterranean Atlantic/Mediterranean North Atlantic – Europe Pacific – New Zealand Pacific – North America Europe Atlantic/Mediterranean Mytilus edulis Common mussel Horse mussel Bearded horse mussel Mytilus californiaus Modiolus modiolus Modiolus barbatus 55 .
and other names. Different species are variously called quahog. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint.1. 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. extended above the surface to maintain water currents needed for respiration and feeding. or siphons. mostly marine bivalve molluscs. A mantle tissue encloses the body of the clam. Buried clams leave tubes. hard-shell or soft-shell clam. geoduck. cherrystone.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. laterally compressed.2. Most clams are only a few centimetres in their maximum dimensions.1 Clam Clam is the common name of a large group of often edible. but 56 . littleneck. The muscular foot is used to burrow in mud or sand.
Dried: shelled meats. Smoked: meats. then heat-processed. uncooked meats packed unprocessed. Live: in shells. 3.MOLLUSCAN SHELLFISH the giant clam of the Indian and Pacific oceans may be nearly 130 cm long and weigh 227 kg. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. or shelled meats. Soups: oyster stew. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts.2. sun-dried. and no siphon. These molluscs feed on minute plants and animals carried to them by the current. Oysters lack a foot or have a reduced foot. packed in weak brine. hermetically sealed. in cans with either brine or clam liquor. Live: in shell Fresh: in shell or shelled meats. uncooked. Frozen: shelled meats. washed. either whole or minced. smoked meats are also canned. skewered with bamboo sticks. Fresh: in shell. oyster soup. clam madrilène and clam liquor. trimmed and packed. Canned: meats removed from the shell by steaming. 57 . Frozen: in shell. Semi-preserved: meats cooked and packed in spiced vinegar. having rough and irregularly shaped shells that are closed by a single adductor muscle. after precooking and brining. all in cans. oyster bisque. Dried: Hoshigai (Japan) shelled meat.2 Oyster Oysters are marine bivalve molluscs. tidal waters or in brackish water in the mouths of rivers. Canned soups include clam chowder. Canned: clams are steamed open and the meats removed from the shells.1. or shelled meats. uncooked (shucked). the meats are smoked and packed in oil in cans or jars.
1. mussel paste. scallops can swim or leap by a series of rapid closings and openings of the valves. 3. flattened ear or wing at the hinge area. cooked meats packed in dry salt for transport. or packed with jelly. and the anterior end smaller and pointed. mussel in butter sauce.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. Semi-preserves: cooked meats with vinegar-acidified brine and spices. Unlike other sessile bivalve molluscs. They have filibranch gills. The shells are fluted and round with a broad. Fresh: shelled meats (often only the adductor muscle and the roe are eaten.1. or filaments. vinegar solution. are united. sauces.FISH AND SEAFOOD 3. scallops possess a single large adductor muscle. boiled. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves. The familiar marine mussel is asymmetrical.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. The adductor muscle is the only edible part of the animal.) Dried: peeled. Smoked: meats may be canned in edible oil. Salted: cooked meats bottled in brine. smouldered and afterwards dried (Japan). Bait: used live extensively for baiting fish hooks for line fishing. by interlocking tufts of hair-like cilia. Internally. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). the shell is often characterised with a lustrous nacreous or pearly lining inside their shells.2. The common scallop is about 5 cm long. gutted. in which the individual branches. also with other ingredients. Canned: cooked meats. Fresh: cooked meats.2. with the posterior ends of the shells (valves) broad and rounded. and held apart. 58 . brine. The posterior halves of the valves contain a large adductor muscle that holds the valves closed. In some species. Live: whole with shells. in mussel liquor.
clams and mussels. Salted: meats either lightly or heavily dry salted. The following pathogenic microorganisms causing illness in humans are characteristically associated with molluscan shellfish: Vibrio parahaemolyticus Vibrio cholerae O1 Vibrio fluvialis Vibrio hollisae Vibrio vulnificus Vibrio cholerae non-O1 Vibrio mimicus Vibrio furnissii 59 .MOLLUSCAN SHELLFISH Frozen: shelled meats. nutrient concentration and pollutants of the water column. Cockles range from about 1 to 15 cm in length and have two valves of equal size. and the environment. they accumulate pathogenic microorganisms from polluted waters. are vectors of disease transmission (8). Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms. Raw and partially cooked shellfish.2.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. mostly in oysters. also bottled in brine. Whatever contaminants are in the water will eventually get into the shellfish (7). Live: in shell. Canned: in brine. pH. 3. in sauce.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves. The microflora are dependent upon a number of factors such as season. Because molluscan shellfish are filter feeders. depending on length of journey. which affects the temperature.1. 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. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding. Canned: in own juice. Vinegar cured: bottled in malt vinegar after brining. 3. and butter. salinity. Fresh: meats removed from the shell by boiling.
. Flow diagrams of the major processing steps of molluscs are presented in Figures 3. but have never been cultured (14. 3. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. Alcaligenes spp. Pseudomonas spp.1 Shellstock 3. salinity. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. There are microorganisms. Bacillus spp.1. nutrients. spirochetes of the genus Cristispira and Saprospira. For example.1. mechanical harvesters. 13).1 Processing and its Effects on the Microflora Oysters. and cockles The processing of bivalves takes two forms. or by 60 . 15). 3.1 and 3.3. shellstock (live) which is shellfish in the shell.. and yeast cells of Rhodotorula rubra and Trichosporon spp.. observed microscopically. responding to the environment in which they are harvested.. 12. In summary. Flavobacterium spp. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora..3 3. mussels. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp..1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs.3. Vibrios. Water temperatures affect the concentrations of microflora in the water.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. whole or in part. Enterococci spp. Other microorganisms found in lesser numbers are the Achromobacter spp. and Aeromonads. Campylobacter coli Bacillus cereus Hepatitis non-A. Acinetobacter spp. dredges. The largest groups found are the Pseudomonads.. (9. Moraxella spp. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. and shucked shellfish that is shellfish. 10.3. 11. clams. Micrococcus spp. that have never been cultured on laboratory media.000 /g.1..2. from which one or both shells have been removed. They are termed viable non-culturable microorganisms. rakes. the initial flora composition and concentration of bivalves are highly variable.. At harvest. Temperature.
Shellstock may be wet-stored at the processing facility in tanks of clean seawater. or from other sources approved by the State Shellfish Control Agency (SSCA).3. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. Shellstock are placed in drycooler storage containers in the facility. If by boat. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock. Thirdly. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. The shellstock is transported dry. or immediately placed in refrigerated trucks. Generic flow diagram of live shellfish from harvester to point of sale 61 . Fig. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. usually in open-mesh containers and/or burlap sacks. in containers from which they can be retrieved easily. after harvesting.1). Water used for shellstock washing is from approved growing areas. or in bay-approved shellfish-growing waters prior to sale or processing. 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. Secondly. There are three primary reasons for wet storage: first. for transportation to the point of sale.1. by truck or boat.MOLLUSCAN SHELLFISH hand (Figure 3.
however. 21. The washing of sediment and detritus soon after harvesting will prevent further contamination. Over a period. bivalves tightly close their shells thus trapping water and microflora. The most important factor in controlling bacterial growth in shellstock is temperature. with prolonged storage the microbiological quality of the bivalve decreases. Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. with the exception of A. shellstock are not refrigerated on the harvest boat. and Citrobacter species) and Vibrios. An outbreak of infectious hepatitis in Sweden in 1956. handling. to avoid possible contamination of the shellstock.3. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. 16.1. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. 22).2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. when it reaches the processing facility. and temperature storage of shellstock to point of sale.1. Enterobacter. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. coli.FISH AND SEAFOOD 3. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. Harvest conditions are another factor that influences the growth of bacteria in shellstock. without adequate refrigeration. Generally. 25). Furthermore. are prevented. there is evidence that in the absence of adequate refrigeration. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. and thus contaminating the shucked meat at the point of sale. vulnificus. for a considerable length of time. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. Generally.2 ºC (45 ºF) or below (19). Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. 24. and the sanitary practices applied during harvesting and shipping. 26). hydrophila (18). Klebsiella. At harvest. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. If long delays occur before reaching the processing facility and/or refrigeration. the bacterial counts in the meats will increase. Water used for shellstock washing should be of good sanitary quality. At this temperature the multiplication of faecal coliforms (E. 17). Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. some pathogenic organisms will survive and multiply in shellfish (20. including V. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). 23. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). shellstock are placed in refrigerated trucks for transport or placed in wet storage. synthetic seawater for the purpose of de-sanding and salt uptake.
Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. After washing of meats. for the effects of microflora during the processing of shellstock. In the blower-washer. washing. Containers are placed in packing boxes for shipping at refrigerated temperatures. are applied to the shucking operation of shellfish. Shuckers routinely use either cotton or rubber gloves during the shucking operation. thus contributing to high bacterial counts in the final product (28). clothing becomes very soiled. whereas cotton become wet and 63 . Meats are collected in either perforated or non-perforated buckets for delivery to the washing station.1. Due to the nature of the shucking operation. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). lobsters.2. The process of shucking involves forcing a knife between the valves and cutting the adductor muscle of one valve freeing the shellfish from one valve. the tank is opened through a drain gate and chute. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank. the finished product may become contaminated.3. perforated tray in which shucked shellfish are spray washed and/or drained) or in a blower-washer (a tank-like device for immersion washing of shucked shellfish). or subjected to splash from foot traffic. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported.2.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. 3.1 Processing description The processing of shucked shellfish is limited to washing of shellstock. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. and packing shellfish into containers (Figure 3. through which the washed meats are discharged for rinsing and then packed in containers. Studies have shown the use of rubber gloves reduces the contamination level. Thus.1.2 Effects of microflora during processing The same considerations as previously discussed. 3.3.3. In addition. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process.2).1. If shellfish are not reasonably clean at the time of processing. Under these conditions if the shuckers enter the packing room.2 Shucked shellfish 3. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. shellstock is subjected to contamination when stored on the floor in standing water. removal of shellfish (shucking). and fish. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. washing and culling shellfish prior to storage is essential. To remove the shellfish attached to the remaining valve.
Shucked meats are thoroughly drained. 64 . Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. Shucking containers are rinsed clean with running water. 3. which may lead to contamination of product. aprons. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. within one hour.2 ºC or less. 2. Separate rooms or lockers for storage of clothing. 4. There is a tendency to store such articles on the shucking benches or in packing rooms.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). 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. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. Fig 3. and sanitised before each filling.2. and gloves help prevent the contamination of shellfish.
Pseudomonas. As described previously. is stored in the bivalve as a source of energy. The initial flora found in shellfish is the major factor of spoilage in bivalves. whereas bacterium such as Pseudomonas. 34. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. Temperature abuse will result in high bacterial counts. particularly oysters stored at 7 ºC. rough. Much of the work has been done with oysters and clams. Glycogen is hydrolysed by the initial flora found in shellfish. in oysters. anywhere during the process of shellfish from harvesting to the point of sale.47 to 6. streptococci. abundant gas production. spoilage will occur. are an excellent medium for growth of bacteria. showed that organisms such as Achromobacter. blowers and other utensils that have cracked. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow. For example. where there is a breakdown of temperature control or prolonged storage of shellfish. All equipment coming in contact with the shellfish such as colanders. skimmers. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. Shucked shellfish are an excellent medium for the growth of bacteria.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. these include lactobacilli. Micrococcus. 65 . Alcaligenes. Earlier studies.2 ºC or less.8% (37). In summary. 31. and yeasts. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. particularly shucked meats.2 ºC. and proteolysis (10). clean gloves. a carbohydrate. Lactobacilli seem to be the predominant microflora in shellfish. Proteus. at the turn of the century. This process changes the bacterial profile to microorganisms favouring low pH. The environment has an effect on the bacterial population found in shellfish. glycogen levels range from 0. 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. and packing the shellfish in a timely manner. Glycogen. Achromobacter and Flavobacterium levels decrease during storage. 32). Shellfish. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. Flavobacterium. shucking pails. Controls that should be applied include using clean packing equipment. 3. thus producing acid and gas (38). These organisms may play a significant role in the spoilage processes. 35. 33. 36). Shucked shellfish are held and transported at temperatures of 7. The spoilage process in shellfish may be divided into three stages: increase of acidity.
Sporadic cases are frequent along the coast of the United States. 41). there is a recent report of low levels of V. headache and fever.5 cases/100. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. This bacterium is present in all the major processing steps of a shellfish plant. approximately 23 cfu/g. and to the point of sale. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. however. Products involved were from the same processor.48 hours with abdominal pain. during processing.000 population/year (46).FISH AND SEAFOOD 3. Diseases caused by Vibrios are enteric in nature. It is a common marine bacterium. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. from harvesting to finished product (i. and estimates based from a hospital study show less than 0. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. prevention of cross-contamination. parahaemolyticus possibly producing illnesses (40). Major outbreaks are associated with the warmer months. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). 3. diarrhoea. 3. 66 .1 Vibrio parahaemolyticus V. of V. V. These organisms can be present in the shellfish from the point of harvesting. where outbreaks occur regularly.e. coastal areas). Fortunately. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39).1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. and harvesting of shellfish in the cooler months. They can be introduced to the shellfish from the environment or added during processing.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. parahaemolyticus. 44).1. although non-hemolytic strains have been associated with illness (43. parahaemolyticus causes gastroenteritis lasting 24 . nausea. and very common in Japan.5. Between May and September 1997. primarily in brackish or saltwater. lot and retail outlet. parahaemolyticus from shellfish is low. only a small percentage of strains in the marine environment are potentially pathogenic (45). ranging from epidemic cholera to sporadic cases of diarrhoea. The incidence of V.5. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. shellstock and packing of shellfish in jars) (29). One of 250 cases reported involved shucked products with low levels. V. widespread in both polluted and unpolluted waters.
The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. fever. Oysters are the most hazardous food associated with V. nausea. 49). cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. with a fatality rate of 50% for individuals with septicaemia. They are sensitive to cold. chills. are particularly susceptible to primary septicaemia. however. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. vulnificus is by rapid refrigeration during warm weather months. V. Healthy individuals are susceptible to gastroenteritis. Onset of symptoms occurs in 48 hours and can be severe. in the range of 0. and preventing cross contamination once the product is cooked. The bacterium is an estuarine species that causes wound infections. The organism attaches to the intestine and release of a toxin is suspected. 3. V. cholerae non-01 is ubiquitous in the estuarine environment.48 hours of onset. diabetes.0. Control of V. 55. or immunosuppression. All isolates are considered pathogenic. 3. like V. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. 54. It is one of the most severe foodborne infectious diseases. 46. Hazards from this bacterium can be controlled by thorough cooking of shellfish. Inadequate refrigeration will accelerate the growth of V. 23 of 24 major USA-West Coast estuaries were positive for V. 46. It causes diarrhoea. parahaemolyticus. lasting 6 . 53. In one study. The occurrence of V.5.MOLLUSCAN SHELLFISH Control of V.5. Again.3 Vibrio cholerae non-01 Illness caused by V. 56).2 Vibrio vulnificus The symptoms of V. these individuals are advised not to eat raw shellfish. and rapidly become inactive at low storage temperatures.II (8. 57. vulnificus are fever. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. parahaemolyticus growth in shellfish meats is temperature dependent.4 . those who have liver disease. V. gastroenteritis. High-risk individuals.1. Control of oyster-associated 67 . cholerae non-01 (51). or primary septicaemia.7 days. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). Death has been reported to occur within 36 hours from onset. Primarily. 48). leukaemia. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50).1.000 population/year (8. cirrhosis.8 case/100. the organism has been found in the Gulf of Mexico. abdominal cramps. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. vulnificus. and vomiting. cholerae non-01 has been associated with septicaemia. parahaemolyticus the estimated annual incidence rate is very low. which have been implicated in shellfish-related disease outbreaks (31). cholerae. and nausea within 24 . 58).
or cross-contaminated shellfish (57. and shock. parahaemolyticus and V.2 Campylobacter jejuni C. the pathogenic severity is lower than V. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. imported product from a country experiencing an epidemic. Onset of disease is 6 hours to 5 days. processors should know the product source. nausea. 3. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. undercooked. jejuni requires reduced levels of oxygen (3 . There is evidence of strains of V. mimicus. fever. The presence of these strains appears to be in low numbers and non-virulent (59).5. cholerae 01 is transmitted by faecal contamination of food or water. diarrhoea. Illnesses are associated with the warmer months. V. 61. nausea. and the transmission of the disease by consumption of raw. these organisms cause gastroenteritis lasting 24 . They are common marine bacteria. however.10 days. The frequency of infection of these organisms is no different from other Vibrios. acidity and heat. V. Control of infection from these organisms is no different from the other Vibrios. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). It is considered a fragile and sensitive organism to environmental stresses such as oxygen.5%) and 2 . and southern Asia including India and Bangladesh. vomiting.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. is the same as previously discussed for the non-01 strains.10% carbon dioxide for optimal growth conditions. It causes diarrhoea. low pH. and illness lasts 7 .FISH AND SEAFOOD V.5 Other Vibrios spp. The control of V. drying. 63). vulnificus. e. Gulf coast environment. 68 . Illness can include abdominal cramps. 3. salinity. Poor sanitation and contaminated water supplies will spread the disease. associated with shellfish-borne illness outbreaks.S.5. dehydration. In general.1. V. fluvialis.48 hours with abdominal pain. In developing countries. (8. In addition. particularly in oysters. widespread in both polluted and unpolluted waters. 62.5. cholerae 01 in molluscan shellfish. respectively.1. cholerae 01. preventing crosscontamination of cooked seafood. cholerae 01 establishing in the U. and nausea. cholerae non-01 illnesses is by cooking seafood thoroughly. 60). hollisae.g. Death may occur after severe fluid and electrolyte loss. headache and fever. 3. There are several other Vibrio spp. abdominal pain. including V. Onset of illness is from 3 to 5 days after ingestion of contaminated food. and V.
83.3. V. 73). 81. Campylobacter appear to survive well in oysters during storage. 3. 78) and P. shigelloides (79. 66).500 cells/gram of Campylobacter may cause illness (64. parahaemolyticus.5. Campylobacter spp. 82.MOLLUSCAN SHELLFISH TABLE 3. of Cases 4 362 0 1 0 a Includes V. from Pacific shellstock oysters purchased from a retail grocery store (74). C. infection occurred among participants of a firemen's banquet. Although. C.II Molluscan shellfish associated outbreaks caused by Vibrio spp. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. 80) are ubiquitous in the marine environment. however. however. The seasonality of 69 . jejuni does not multiply in shellstock at refrigerated temperatures. control of C. To reduce the risk of shellfish-borne campylobacteriosis. cholerae 01 and non-01. vulnificus.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. fluviallis. V. and V. ranging from 0. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). This finding is significant in that C. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters.4 to 114 cells. V. have been isolated from shellfish beds throughout the world (67. 65. 70. 71). 85). V. of Incidents 4 279 0 1 0 No. and their immediate proper refrigeration. jejuni were recovered. 76. hydrophila group (75. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. 68. 72. mimicus. They have been implicated in limited shellfishassociated gastroenteritis cases (8. although their role as a pathogen is highly controversial and circumstantial. hollisae Infective dose varies with host susceptibility and virulence of the strain. 84. Recently. Human feeding studies suggest that about 400 .1 Aeromonas hydrophila group and Plesiomonas shigelloides A. they survive well at refrigerated temperatures.3 Other organisms of concern 3. whereas at 10 ºC Campylobacter survive only for 5 days (71). 77. In one case. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. jejuni begins in the harvest of shellstock from approved waters. 69.5.
FDA/Administrative Guidelines 3 Source (91). FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish.0001 negative for the presence3 Under review4 Source (89). dried milk/whey drink powders. being isolated more often during the warmer month.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. negative for heat-labile toxin (LT) or heat-stable enterotoxin (ST) 1 Negative for the presence1 Negative for the presence of toxin producing 01 or non-01 organisms 1 <10. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. 3. Control of these organisms in shellfish products is limited. coli Enterotoxigenic E. coli Salmonella V.000 to 3. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. 2 Source (90). vulnificus 1 Acceptable Levels in Molluscan Shellfish <500.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10.000 cfu/g1 <230/100 g2 <1000 per gram. prepared sandwiches.600. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. Note: Although molluscan shellfish are not included in the selected high risk food. and where indicated are intended to be used for official control purposes. jejuni V. soy products. They are easily destroyed with mild cooking temperatures. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. 70 . Similar results were seen by other investigators (86. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. 87) of the outgrowth of A. non-dairy frozen desserts.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. cholerae V. hydrophila group stored at refrigeration temperatures. Caution should be exercised by individuals with haematological malignancies (88). 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. vulnificus in molluscan shellfish.000/gram (29). aureus C. Test Standard Plate Counts E. FDA/ Administrative Guidelines. In one study. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. parahaemolyticus Staph.
References The Phylum Mollusca. in Microbiology of Marine Food Products. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. Gill S.A. Colwell R. Eds. Jr. Bacterial flora of the market oyster.C. 19-34. Thompson C. Cook D. Bernard F. 7. 1981. Walker W. 755-76. 8. Tubiash H. Journal of Food Protection. 447-52. 1926. 1936.E. 6. Fate of enteric bacteria in estuarine sediments and oyster faeces. 30. 15..J. Cook D. 1991.M.R.. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. U. 36. The American Oyster.E. Saunders Company. Rowse A. Lee J.S.. W..A. 13.7 1.S. 104-9. American Journal of Public Health. Vanderzant C. 26. 1964. Tall B.. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. 72-8. Rippey S.F.L. Nauman R. Ed.C. 64. 1984.W. 419-25..K. 10. Proceedings of the National Shellfisheries Association. Philadelphia. Pfeifer D.A. Hood M. water and sediment from Galveston Bay. 30. 1983. Fish and Wildlife Service.MOLLUSCAN SHELLFISH 3. 16. American Journal of Hygiene.K. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). 46. Microbiology of bivalve molluscan shellfish. Effects of harvesting waters and storage conditions on yeast population in shellfish. 33643. Ward D. Eds. 11. 42. 1970. in Multilingual Dictionary of Fish and Fish Products. Fleet G. Wiley-Blackwell.. 1982. 13. Government Printing Office. 1995. 58. Smith F.N.. Liston J. 82-7. 1963. Washington D. Biology of the oyster in relation to sanitation.R. Hackney C. Galtsoff P. 8. 3.W. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). A typhoid fever epidemic caused by carrier bootlegging oysters. 29. Ray S. 633-40.D. New York. 7. 5. 1968. 71-6. 1994. 6. Jr. Viability and release Salmonella charity and Escherichia coli from oyster faeces. 544-8. 17. Clinical Microbiology Review. Microbiological characteristics of Dungeness crab (Cancer magister). USA. 9.H.S. Applied Microbiology. 33-6. Applied Microbiology... in General Zoology.R. Infectious diseases associated with molluscan shellfish consumption. Applied and Environmental Microbiology. Villee C. 1975. 4. 71 . Old H. American Journal of Public Health. Van Nostrand Reinhold. Journal of Milk and Food Technology. 105-8. Organisation for Economic Co-operation and Development. Eliot C. Veliger. 12.R. Applied and Environmental Microbiology.B. 245-7. in Fishery Bulletin 1964 No. Schmidt C. 1960. Galtsoff P. 44.R. 14.R. Journal of the Mississippi Academy of Sciences.S. Lovelace T. 1940. Colwell R. 1973. 2.. 231-51...
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38. 53. Stott R..M. 2891-3.F. Washington Report October 14. 73 . Obara Y. Hlady W. Kato T. 204-5. Stroh E. 38. Jacobson M. Miyamoto Y.M. Kaysner C.. Abbott S. Takizawa K. Powers C. Journal of Clinical Microbiology. New York. 1976. West Coast States and British Columbia.B. Infectionskrankheiten und Hygien. Hoge C. 2.. 128-31..L. Loomis A. Abeyta C. Shellfish Specialist.P. 60. Academic Press. 1989.T.. Berry F.. 1989.. 35.M. U.D.W. 2820-2. Vanderzant C. 1961.. Peeler R. 36. Journal of Infectious Disease. 1937. 1979. Yokohama Medical Bulletin. 41. Janda J. Kaysner C. Kelly C. 46. 43..C.G. Food and Drug Administration. Public Health Reports. 273-80. Jr.M. Ishibashi M. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States west coast. Fieger E. 27.D. Leitch J. Bacteriological control of oysters during processing and marketing.S. Bacteriological examination of oysters stored at low temperatures. Urease-positive.. Grodner R. 69. Journal of Bacteriology. 1926. Journal of Clinical Microbiology. 1349-51. 353-7. Geiger J. 39.. 1969. Bacteriological findings in Baltimore oysters. Marine Fisheries Review. 97. Journal of Bacteriology. Sample T. 25. 34.N. Bothell. Studies on pathogenic halophilic bacteria.A. Arcisz W. 1954. 37. 1914..S. Zentralblatt für Bakteriologie. 13-7...A. Novak A. 1916. 41. 1997. Joseph S. Journal of Infectious Disease.. 1. Watsky D.MOLLUSCAN SHELLFISH 32. Vibrio parahaemolyticus illnesses. DePaola A.. Journal of Food Science. 160. 133-47.. 33. Epidemiology and spectrum of Vibrio infections in a Chesapeake Bay community. Tanikawa E. 107-8. Ward W.. 47. Bulletin of the Johns Hopkins Hospital. The bacterial flora of market oysters. 313-22. In vitro haemolytic characteristics of Vibrio parahaemolyticus: Its close correlation with human pathogenicity. kanagawa-negative Vibrio parahaemolyticus from patients and the environment in the Pacific Northwest.. 1147-9. 1989. 45. 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. Vibrio infections associated with raw oyster consumption in Florida. 27. Sidwell V. Serological and hemolytic characteristics of Vibrio parahaemolyticus from marine sources... Thompson C. 1987. 1958. 100. U.... Takeda Y. Volume 1. Ed. The bacterial content of market oysters. Geographic and monthly variation in composition of oysters. Yamai S.W. Crassostrea virginica. Journal of Food Protection. Microbiology of shellfish decomposition. 985-93. 44. Borgstrom G. Morris J.. 40. (2). 1997. in Fish as Food. Israel E. 42.M. 41. Kelly M. Parasitenkunde.E.A. Joseph M.. Wekell M.. Jr..A.. Libonati J.L. Akiyama S. Applied Environmental Microbiology.G. Takikawa I..
T. Abeyta C. 8.. Twedt R..M.. Roderick G.T. 312. 1980.M.. 91-103..A.G.. Adams M.C. Feldman R. 1988... Zywno S.I. Allegra D.. Morris J. Hebert S. 45.. 2050-3. 55. 94. Caraway C.R. sediment and waters of the U.G. Applied and Environmental Microbiology.R...B.. Lockman H. 1985.. Spite G. Vibrios in the Louisiana Gulf Coast environment. Craig J.A. Jr. 1.A. Kaper J. 1984. Wilson R. Kaper J. Colwell R. Conte J. Wathen H. DePaola A.J. Wachsmuth I. Wazenski T. 56. Review of Infectious Disease. Brenner F.. Joseph S. Maden J. Blake P. Applied and Environmental Microbiology..G. Francis D.A.W..A.T. Cholera . 1981. Jr.J. Characterisation of Vibrios cholerae isolated from oysters..M. Non-O group 1 Vibrio cholerae gastroenteritis in the United States: Clinical. 656-8. New England Journal of Medicine....R. Barrett T. 10. Clinical features and an epidemiological study of Vibrio vulnificus infections.A. 275-83. 299-312. Feeley J....M. Wekell M.E. Siebeling. New York.. Safrin S.. Morris J. 46. 57. 1012-7. Duran W. Snyder J.... Jr. Twedt R.T. Applied and Environmental Microbiology. Illness caused by Vibrio damsela and Vibrio hollisae.W. 1979. 1984.. Incidence of Vibrio cholerae from estuaries of the United States West coast.G. Hickman F. 1983. 558-61. serology. and enterotoxin production of Vibrio cholerae in Chesapeake Bay. John Wiley & Sons Inc.A..N. Roberts N.F. Blake P. 1981...R. McPhearson R. 74 . epidemiologic.. Bradford H. Jacobs R.. An epidemiological study of V. Presnell M.B. 305-9.. 51.T..F.. 60.G. 41..W.P.F. 305. 59. 1982.. 1985. Hunt J.E. 53. 37. Gulf Coast. Non-01 Vibrio cholerae in shellfish. Ecology...C. R.G. 1982...O. Davis B.. Motes M. 5972..O..D. 253. cholerae in Louisiana.A possible endemic focus in the United States. Kaysner C.J. Vibrio vulnificus: Man and the sea. Becker R.FISH AND SEAFOOD 48. Leitch J.K. Bradford H. 1987. Blake P. DePaola A.M. Pons V.O. Tacket C. Jr. Journal of the American Medical Association.S. 58.. Hug A. Hollis D. 50... and laboratory characteristics of sporadic cases. 1344-8. Wilson T. McFarland L.G. Black R.... 343-50. Cholera and other vibrioses in the United States. Lancet. McCay C.. Ecological relationship between Vibrio cholerae and planktonic crustacean copepods.W.M. Morris J. Jr. 49. 149. Becker S...E. Applied and Environmental Microbiology... Jr.M. Stott R. Rahman R.D. 1475-8. Peeler J.G. Plaard R. Microbial Ecology.L. Johnston J. Jr.M. McFarland L. Journal of Infectious Disease.B..... 1294-7. 53. Annals of Internal Medicine. 52. Ed. 802-6. Non-01 Vibrio cholerae bacteraemia: Case report and review. Blake P. Riddle C... 1983.. Puhr N. Morris J. Miller H. Colwell R. Tacket C.A.E. Journal of Food Protection.V. Small P. West P.. Hug M. Colwell R. New England Journal of Medicine. Weaver R. Lee J.M. in Vibrios in the Environment. 54. 61.A.
.A.. Kaysner C.P. 323-5. Journal of Infectious Disease.M.. Feldman R.J. 256.. London.. 71.G. (jejuni. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster. Blake P. infections in man. Robinson D. coli and lari) from shellfish and the marine environment. Wekell M... International Journal of Food Microbiology... Public Health Laboratory Service. Helicobacters and Related Organisms. Moore J. Ireland. British Medical Journal. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation. Feldman R. New York. Eds. C. 1987.L. Austin S. 13. 1993.. Abeyta C.. Jr. Newell D. Clements M. Campylobacter gastroenteritis associated with raw clams.W. 72. in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections. 1996. in Campylobacters.M. Jr. Brenner D. Ketley J.... Pearson A.K. in Campylobacters.S. 68. Wilson I.M. 472-9.J. from patients with diarrhoea. Alam A. Jones D.R.A..R. 63. Proudford R.M. New York. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N.G. Rowe B. Blaser M. 621-4.. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness. 64.I. Eds. 1980. 4. 80.. 607-9..R. Wekell M.G.D. Abeyta C.. Infective dose of Campylobacter jejuni in milk. Morris G.M. Journal of Clinical Microbiology. 6-9 September 1983. Griffin M.E. 73. Brussels. Wilson T.. Isolation of vibrio-like group. A method for isolation of Campylobacter spp. 169-71. Levine M. Blaser M.. Levine M.G. Plenum Press. Clements M. Plenum Press.A.E... 99.. 1983..J. Journal of Food Protection. Huq M. Dalley E.M.R.K. 66..R. Hughes T. Huges T. Davis J. 75 . Kaysner C. EF-6. 51-64. Disease from infection with Vibrio mimicus.A.. Wilson I. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp. Ketley J.K.. Fitzpatrick M. Shandera W.. Journal of the Medical Society of New Jersey. Eds.X. Murphy P. 4. 157. Jr. 65.MOLLUSCAN SHELLFISH 62. a newly recognised Vibrio species. Black R.M.. Abeyta C.. 1983.A. Microbial Ecology in Health and Disease.A.E.J..D.G. Skirrows M. Experimental Campylobacter jejuni infection in humans.F. 69... Wareing D.S. Black R.G. Davis B. Murphy P.B. S41. 4. Journal of Rapid Methods and Automation in Microbiology.. 1995.H.. Matsuda M. 70..A. 1980. Newell D.. Arumugaswamy R.P. 1584.L. Wareing D.. 67.E.G..M. 101-4.A. Kaysner. 1981. Helicobacters and Related Organisms. Studies of Campylobacter jejuni infection in volunteers. Johnston J. Stott R. 282. Moore J. 1996. 1988. Wilson T.A. Deeter F.M. 56. 1991. Hunt J. 257.. 11.. Annals of Internal Medicine.
Plesiomonas enteric infections in the United States. Seidler R. Bark D. Report (1/13/84). Prevalence and distribution of Aeromonas hydrophila in the United States.J. Applied and Environmental Microbiology.L. Aeromonas hydrophila: ecology and toxigenicity of isolates from an estuary.A. 1985. 79. Oyster borne disease outbreaks in St. Kaysner C.H.. 52.C. 81. Holmerg S. Hood M.A.. 27. Esch G.A. Trost P. 979-85.. 78. Singelton F..M. The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish. Effects of processing and storing oyster meats on concentrations of indicator bacteria.T. Applied and Environmental Microbiology. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. 1981. Herrington T. Stelma G. 1997. Kelly M..D. 76 . Kain K. Sullivan J..M.. GA. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness. Hirsch R. Colwell R. 39. 55. Journal of Food Protection. 105.D... Annals of Internal Medicine.M.M. 80..D... 1989.. Robins-Browne R.W.R. 359-77. Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species.. Beaman J.L.J. Peterson D.. Partridge K. 449-57. III. Tenge B. 82.. Kaper J. 85. 50. Hunt J.W.. Journal of Food Protection.. Petersburg. Infection and Immunity. 76.M. Atlanta.... Van Damme L. 731-8. Journal of Food Protection. Abeyta C. Miller M. Tall B. 1990. Blake P.B.. Journal of Applied Bacteriology.FISH AND SEAFOOD 74. Levine M. 86..M. 1978.. 1988. 343-9. Clinical features. Abeyta C. Fla. 1984. 1987.L. Koburger J. epidemiology.. 47..J.A. 84. Tzipori S. Burke V. Vandepitte J.A... 36.P. Incidence of motile aeromonads from United States west coast shellfish growing estuaries. Journal of Food Protection.J. Robinson J. 47.. 998-1001.W. Journal of Rapid Methods and Automation in Microbiology. 7-12.C. Baker R.. Wekell M. Applied and Environmental Microbiology. Herrington D.. 1980... vibrios and Aeromonas hydrophila. Journal of Clinical Microbiology.M. Gracey M. 849-55. 77.A. 48.. Farmer J. 475-9. Jr. 1146-9. 1986. Kaysner C. 83.R. Wachsmuth I. Plesiomonas shigelloides: An opportunistic food and waterborne pathogen. In vitro and in vivo pathogenicity of Plesiomonas shigelloides... Wekell M. Frequent isolation of Edwardsiella tarda and Plesiomonas shigelloides from healthy Zairese freshwater fish: A possible source of sporadic diarrhoea in the tropics. 53. Ruple A. and treatment of Plesiomonas shigelloides diarrhoea. 75. 5. 1984. Fliermans C. Hickman-Brenner F. Kaysner C. 1989.F.. Jr. Lockman H.K. Jr.A. 223-47. Department of Health and Human Services/Public Health Service/Food and Drug Administration... 598-601. Wekell M. 690-4. 52. Stott R. Hazan T. Abeyta C. 87..A. Cook D. Journal of Food Protection.N. Cooper M.
Clams. 1997. 560.C. Inc.R.. U.600 (CPG 7119. Quality Control.S. Food and Drug Administration.E.. 90. in National Shellfish Sanitation Program Manual of Operations. mussels. Chapter 5: Foods. Ed. Ed. U.1987.S.8 Further Reading Ahmed F.C.. Food and Drug Administration. Food and Drug Administration.H. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. Pittsurgh. U. and HACCP: A Global Focus. Food and Drug Administration. Program 7303. The Microbiological Safety and Quality of Food.L. Smith J. Food and Drug Administration. Lancaster. Food and Drug Administration Compliance Policy Guides...E.S. Food and Drug Administration Compliance Program Guidance Manual. Food and Drug Administration. 89. Slavin J. U.A. Aspen Publishers Inc.M. Food and Drug Administration. Ed. Seafood Safety.Pathogen monitoring of selected high risk foods (FY 88/89).W. Washington D. in U.S. Food and Drug Administration. Collette R.C. Burdge D. 1991. fresh or frozenadulteration by bacteriological contamination Sec.W. U.S. Sherlock C..S. Lund B.S.. Baird-Parker T. National Academy Press. Government Printing Office . in U. 1995.S. Gould G.12). Washington D. U. Technomic Publishing Co. 63-8.MOLLUSCAN SHELLFISH 88. Part I Sanitation of Shellfish Growing Areas.. Washington D. 1987. Martin R. Fish Inspection. oysters. 7. 77 .. 3. 2000. Gaithersburg.C. 1995.030. 91.
. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. Shrimps.4.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda.. peeled product. with shells and heads on or off are usually packed. and frozen in 1 and 2. jointed limbs and a chitinous exoskeleton. Crayfish. packed and frozen in 0. (deep-water shrimp). crayfish and shrimp (1). Pandalus borealis (deep-water shrimp).0 kg polythene bags.g..5 and 1. crabs. lobsters. Such products are usually only partially cooked before freezing. the front pair ends in claws. M. 79 . and are usually aquatic.5 kilo pack (boxes or bags). prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). and Trachypenaeus spp. lobsters and crabs have ten legs. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. Species eaten from tropical waters include Penaeus spp. e. They possess a segmented body. Pandalus montagui and Crangon crangon (inshore shrimp). Peeled ready-to-cook shrimp are raw. with additional water to form a glaze.5 and 1. and available in 0. Peeled and cooked shrimp are ready-to-eat after controlled thawing. Parapeneopsis spp.0 kg polythene bags. CRUSTACEAN SHELLFISH Linda Nicolaides.Ph.
2 Initial Microflora As with finfish.g. for crabs and lobsters. packed into polythene bags. their feeding and living habits.. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp. Cooked and chilled. storage above the recommended storage temperature. which means that the haemolymph can be a reservoir of bacteria. Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP). only live carcasses are processed. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors.FISH AND SEAFOOD Crabs. the geography of the area they are captured from or are grown in. 4. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. shrimp die immediately after capture. e. boiled to kill the crustaceans and cook their meat prior to consumption. However. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. during the peeling of cooked shrimp. Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function. In comparison. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed. Pasteurised crabmeat is edible meat removed from the exoskeleton by hand. particularly after it has been cooked. The structure of the circulatory system of crabs is not closed. For example. traditionally. stored and transported as frozen product. which include the environment from which they have been captured or harvested. the potential exists for crosscontamination from both the processor and from the environment. Following capture or harvest. particularly for members of the genus Vibrio (2).g. As with finfish. so that spoilage may occur during landing and transportation to the processing plant. Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance. with the increasing demand for ready-to-eat/cook crustacean products. the season. hence. Shewanella putrefaciens and 80 . Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria. or frozen crab. tropical waters. and the temperature and quality of the waters in which they exist. it is important that shrimp are handled and iced correctly in order to keep them in as fresh and uncontaminated a state as possible. e. lobster tails and cooked crabmeat presented decoratively in their shells. spoilage bacteria are of little importance as. If these products are subjected to temperature abuse. lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption.
lobsters and crayfish are harvested from cages or pots containing bait.. or are found in brackish water around the shores. or cooked. bacterial contamination and spoilage are inevitable. When the shrimp arrive at the processing plant they are washed and then sorted into size. After capture and transport to the processing plant. chilled sea water. In contrast. uncooked prawns and shrimp is presented in Figure 4. peeled and cooked. Vibrio spp. graded according to size.. As with finfish. the genera associated with tropical species include Enterobacteriaceae. They are then taken to market alive in either holding tanks. A large proportion of shrimps are produced by aquaculture. however. Hence. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots. Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. or transported directly to processing plants. corynebacteria. with “head on” or “head off” whereby the edible tail is removed from the head. the rate of deterioration of the crustacean shellfish by these organisms is reduced.1). The claws are usually bound together to prevent the animals fighting and causing damage to each other. In comparison shrimps die quickly after capture. especially if they are stored in ice or iced water.3.1 Prawns and shrimps An example of the process used for preparing frozen. Shrimps can be processed whole. or with the shell and central vein removed. and Achromobacter spp. Pseudomonas spp.for example. 4. up to 16 days for tropical species compared with 8 . Shrimp will then be glazed and frozen in a raw state. 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. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice .10 days for temperate water species (3). These animals can survive out of water if the surrounding environment is kept cool and damp. from the time of capture and processing. they are washed.1. Alcaligenes. thus. 4. gills and thorax. chilled 81 . Micrococcus spp. whether GHP are in place. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. glazed and frozen (see Figure 4.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group.3 Processing and its Effects on the Microflora Crabs. Shrimp might be further processed with the shell on.. so they are either sold on the local market or are processed in local processing plants. The rate of spoilage will be a function of time and temperature.
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. and packed to meet the customer’s specification. 4. After cooking.frozen uncooked prawns 82 . Fig. should be followed to ensure that the product is not re-contaminated with microorganisms present in the processing environment or from the workers. Good Manufacturing Practice (GMP). spices and cooking batter. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour.FISH AND SEAFOOD frozen. Flow diagram .1. Breaded products introduce an added risk of spore forming bacteria. including GHP.
indicative of protein degradation and the growth of the principal spoilage bacterium Sh. with the growing demand for ready-to-eat products from crab and lobster meat. they include Sh. wet and ‘soapy’ or ‘jammy’ to the touch. the joint between the tail and carapace is open and the limbs hang loosely. whilst boiled crustaceans have a pleasant slightly sweet smell. An offensive odour is also produced. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. and a strong ammoniacal odour is produced.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. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step.CRUSTACEAN SHELLFISH 4. immediately prior to boiling. If hygienic procedures are not followed. bacteria of public health significance. or in hermatically sealed cans that have undergone an appropriate heat treatment. 4.4. putrefaciens. there is no associated spoilage problem. putrefaciens and Pseudomonas spp. The shell will be soft. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. The shells have a crisp. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. If boiled. contaminating microflora may include enteric bacteria. 83 .2 Cooked crabs and lobsters As indicated earlier. In comparison. the meat is either repacked and sold in the shells. 4. Heat is produced if the shrimps decompose in a confined space. since crabs and lobsters are traditionally consumed immediately after cooking.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. If the tail drops.4. With spoiled crabs and lobsters. 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. bright and dry appearance. slightly iodine smell. for crabs. there is no indication of life. dull and sticky.3. and have a sweetish. viruses or parasites. or packaged and processed. spoiling prawns and shrimps have shells that are soft. Once removed and cleaned. However.
The above results prompted the FDA to develop a compliance programme for the pathogen. shrimps and cockles sold (17).2 Listeria spp. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11).5% of frozen raw shrimps from different countries (13). even if GMP is followed. cooked and smoked fish. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. shrimp and shellfish (11). Hence. 16). in 24. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. 4. monocytogenes was absent from 40 samples of cooked prawns. 6. The presence of L. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. later surveys in 1990 recovered the pathogen from retail raw. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. An import alert was put in place in June 1987. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12).1 Pathogens: Growth and Survival Salmonella spp. and usually for both. 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. Salmonella cannot be completely removed from raw shrimp during processing.5. fish fingers. 7). 5. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. in domestic/imported shrimps (15). Following this. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). monocytogenes in cooked ready-to-eat foods. Further surveys identified Listeria spp. a common pond fertiliser used in aquaculture.5 4. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) .FISH AND SEAFOOD 4. However.5.cooked prior to consumption. Although foodborne illness had not been associated with the product. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . as well as for Salmonella. although efficient cooking will quickly eliminate it (6). nearly 30 years elapsed before the potential hazard of L. animal manure. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. Moreover.
although contamination by types A and B may occur from the environment. Vibrio cholerae.5. aureus because of the ubiquity of this organism in the processing environment.5 Vibrio spp. the pathogen can grow at temperatures as low as 3 °C. combined with the degree of handling of these types of product. 25). so is able to survive heat treatments that eliminate vegetative cells. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. Researchers have demonstrated that inoculated shrimp supported toxin production by C. whilst boils and cuts are also common sources of the pathogen. pasteurisation and cooking. Crabs are covered in a chitin exoskeleton. aureus on their hands. is widely distributed in aquatic environments. Crustaceans have been linked with food intoxications attributed to Staph. 24.5. The pathogen has resistant spores. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. Once the competing microflora has been removed by a heat treatment. However. e. which can become colonised by V. botulinum type E is commonly associated with fish and fishery products. The former. An epidemic that began in South America in 1991. 4. monocytogenes has been attributed to crustaceans. V. cholerae non-O1. Control should be achieved by promoting GHP and staff training programmes. 19). 22) as well as from undercooked crustacea when harvested from contaminated waters (23.5. therefore. cholerae.g.CRUSTACEAN SHELLFISH old. especially in products that have been vacuum packaged. the causative agent of cholera. about 50 . 4. V. it is important that the cooking stage is controlled to eliminate the pathogen from the product and that procedures are in place to prevent any cross-contamination of the cooked product. cholerae is divided into two groups. was associated with seafood consumption. To date. as part of the natural bacterial flora. pregnant women and the immuno-compromised. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. 4. cholerae O1 and V. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21.4 Clostridium botulinum C. particularly crabmeat (18. 85 . aureus is not a member of the intrinsic flora associated with crustacean shellfish.70% of healthy individuals carry toxin-producing strains of Staph. botulinum type E at 10 °C but not at 4 °C (20).3 Staphylococcus aureus Staph. no incidence of foodborne illness caused by L. The main source of contamination is the hands of processing personnel.
8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. although A.g. occurs in crustaceans from warm environments.8% of samples contained C. non-B Hepatitis viruses (31). Of these. Contamination by this organism can be controlled by rapid chilling of crustaceans after harvesting and avoidance of time/temperature abuse during distribution and processing chains.5. however. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. in a survey carried out in the United States between 1977 and 86 . 31. Hepatitis A virus presents the most serious hazard. vulnificus is an invasive and lethal pathogen associated with wound infections. However.7 Aeromonas hydrophila and Aeromonas sobria A. hydrophila is able to grow well at refrigeration temperatures (0 .9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. but has the potential to cause fatal foodborne illness (26). jejuni has not. the potential hazard exists as C. However.5. hydrophila has been isolated from prawns (29). or freezing at a temperature of -23 °C for 7 days (30. parahaemolyticus. V.6 Campylobacter jejuni and Campylobacter coli C. hydrophila and A. Astroviruses and non-A.30 MPN/g) (27).5. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. Caliciviruses. Quantitative levels were below limits of detection in all cases (<0. 4. a marine halophile. 32). Norovirus. been implicated in a case of foodborne infection associated with crustaceans. whilst 5. 4. Parasitic infections are controlled by either cooking infected material prior to consumption. to date. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. 4. e.5. V. A. coli. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). particularly trematodes (6). Japan. It is more commonly implicated in countries where crustacean shellfish are eaten raw.2 °C) so that GHP coupled with controlled low temperature storage is essential to prevent the pathogen from reaching levels that may present a hazard. 4. South America.
No foodborne incidents have been attributed to this source to date.11 Chemical pollutants and biotoxins Researchers have demonstrated that crabs feeding in waters contaminated with algae do have potentially high levels of algal toxin domoic acid (DA) in their viscera. 4.5.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33). consistent with that in the stomach.CRUSTACEAN SHELLFISH 1981. Provisions laid down concerning the use of additives would also apply to these types of products. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34).5. which results in a persistent and apparently permanent loss of short-term memory (36). The risk of such hazards is far higher with fin fish and molluscan shellfish. 38. are included in the Commission Regulation (EC) No. 4. 39).1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. Norwalk viruses can also withstand heating to 60 °C. although it is one of the European Union (EU) legislated allergens. A summary of microbiological criteria for crustacean shellfish is listed below. 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. which could be transferred to the meat during unskilled removal from the shell (35). or chlorine concentrations similar to those found in drinking water. The toxin causes amnesic shellfish poisoning (ASP) when consumed. 2073/2005.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. 4. only 0. produced or imported into the European Union (EU). They can survive low pH levels.6. 87 . Selected parts from this regulation is presented below. from farm to fork (37. 4.
FISH AND SEAFOOD
184.108.40.206 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 220.127.116.11.1 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. 18.104.22.168.2 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. 22.214.171.124 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.5. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. where the salt replaces moisture in the fish flesh. 93 . The processes take anywhere from a few hours to a few days or even months. their combinations and/or variations. fish is immersed for a few hours in a relatively low concentration brine. alternating fish layers with a layer of salt. and the extracted moisture is allowed to drain away. Fish can be salted in three different ways or their combination: Dry salting. Usually. and a preliminary operation to some smoking. SMOKED AND DRIED FISH Dr. both to preserve and to create a variety of new products. drying. and before significant spoilage takes place. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. drying and smoking processes. and marinating processes. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. split and opened out flat. and is sometimes known as ‘pickling’.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. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. dependent on the country or the type of product. The fish is headed. CURED. rather than for preservation itself. and salting is done by stacking the fish in layers. Wet salting or brining. Salting is both a method of preserving fish. It includes the salting. Fish curing is practised worldwide.
as well as other flavourings (sugar. the preservative effect is due to the low pH used in the process. Food products can be dried in three different ways: Air or contact drying. Fish in contact with refrigerated plates freeze. are added to the marinade. This is a mixture of a dry and a wet salting process. it can also be named ‘pickling’ or ‘blood pickling’. depending on the type of product. spices). Smoking is an ancient process for preserving and flavouring food. The fish is mixed with dry salt but in a watertight container. When the process is running in a closed system. The product is dried by contact with heated surfaces or by radiation. It has found limited application with fish. the ice sublimes and the vapour is removed from the fish by vacuum pumps. usually vinegar. Traditionally. without other preservation methods applied after boiling. is used as a preservative instead of salt. this was done naturally by exposure of the fish to the air. such as ‘gravad’ fish. This is the common method of drying fish. thus reducing the water activity of the product. Freeze drying. evaporated water is removed by vacuum pump. It is not commonly used for fish products. or boiling it with dry salt and water. under normal pressure. Salt. Salted-boiled fish are the products obtained on either boiling fish in a brine. The products are usually consumed raw. as concentrated brines are formed. Heat is transferred to fish from heated air or a heated surface. prior to or during the boiling process. and the fish can be immersed in such brines for long periods. Thus. 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 . As the water is drawn out of the flesh by the salt a brine is formed. Usually. After the freezedrying process. this ‘brine’ can be collected and placed again over the fish several times. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. Salt-boiling. thus making the product unsuitable for microbial growth. and the air movement removes the vapour. Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. Included in this group are some lightly preserved products where no acid is added to the marinade. Marinades are acidified brines. where an organic acid. Marinating or pickling. the product rehydrates quickly but does not retain reabsorbed moisture on cooking. Vacuum drying. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms.FISH AND SEAFOOD Dry / wet salting. Longer shelf life is achieved in the products when salt is added. thus delaying spoilage for a few days.
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. However. It is commonly used in place of traditional smoke. the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. process temperatures are over 60 °C. and filtered to remove tars and particulate matter. It can be smoked either traditionally. SMOKED AND DRIED FISH of smoke. The smoke itself may contain spores of moulds. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. the salt used in the processes may contain halophilic bacteria. which impart the flavour of smoke to fish. Usually.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. After the salting step. Smoking can be either traditional. In this process. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke. The secondary objective is to impart sensory characteristics such as smoke colour and smoke flavour to the product. 5. by exposure to the wood or sawdust smoke. smoking is now used mainly to give the fish a characteristic flavour. In this process. and the preservative effect is only slight. It can be injected into the product. temperatures below 30 °C are used. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. or added to the brine with other flavourings. sporeforming bacteria or osmophilic moulds. or by means of smoke-flavouring processes with liquid smoke. Smoked-flavoured fish. or liquid smoke. Usually. Clear smoke. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour. fish can be smoked by means other than the direct action of smoke. or by means of smoke flavouring processes. With the exception of some Asian countries. Cold-smoking. by exposure to the wood or sawdust smoke. leaving only elevated levels of carbon monoxide. such as injection or dipping in a solution of liquid smoke. the fish is subjected to heat for a period that does not coagulate protein.CURED. The process was designed so that the smoke is filtered to remove all smoke particles. 95 . Is an illegal method (in the European Union (EU)) employed to treat tuna to ensure a bright red colour is maintained during storage and transport. Hot-smoking. including pathogens.
2 g of salt per litre of brine). Salt in the water phase links the two conditions of moisture and saltiness. a 0º brine saturation means pure water. Thus. it is important to understand the concept of salt concentration.4% salt (Table 5.0 158. etc.I).4 52.0 . For smoking salmon. it is impossible to establish standardised processing stages in the curing of fish. When a milder brine. TABLE 5.FISH AND SEAFOOD 5. The dissolution of salt in water is dependent on the temperature. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.8 211. This should be distinguished from the salt content.0 g of salt per litre of brine). or salt in the water phase.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) 26.3 Processing and its Effects on the Microflora Owing to the wide variety of products.2 237. Different brine strengths are required according to the curing processes and/or the type of product. processes and producers’ private recipes. only the most common processes can be described here. is employed. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. and salt content. smoking or marinating processes. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. When working with salting.8 79.6 g of salt or more per litre of brine) may cause salt burn in this way. the fish is usually salted with a 70 .8 211. all using salting steps. The strength of the solution is described as degrees of saturation (brineometer degrees). it is important to state the temperature. Salt concentration means the percentage of salt in the water contained in the fish. sometimes the salt crystallises onto the surface of the fish.4 184.2 105. it causes swelling because the fish absorbs water from the salt solution (1).6 264.6 132. about 50º (132. % Salt in the water phase = % salt content % salt content + % moisture x 100 Another important point to remember when wet salting is used is the concentration or strength of a brine. a 90º or above brine (237. Therefore.80º brine (184. At 18 °C a 100º brine is produced when the solution contains 26. When a 100º brine is used.
but the concentration of copper and iron must be kept under 0. however.1 Dry salting / Kench curing Usually. 5. lean white fish is used for this purpose. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. are recommended to help make the product whiter. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. and the salt has penetrated throughout the fish (3). This is because fatty fish turn rancid quickly. having a short section at the end of the tail.1 Salting Salting of fish for preservation purposes has been used since pre-history. 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.0. The salt used should be of food-grade quality. it tends to lead to a slight yellowing and softening of the product. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away.1. Usually it is followed by other preservation techniques such as drying and/or smoking. can be dried further to produce ‘klipfish’ or ‘bacalão’. for which there is a high demand in Mediterranean countries.15 . when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). respectively.3. and salt can speed this oxidation process. The calcium and magnesium impurities. if very pure sodium chloride is used for curing purposes. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. outgrowth of halophilic bacteria may occur.3. which include calcium and magnesium. Salting can be done in a number of different ways. low in calcium and magnesium. 97 . when the air humidity is high. producing characteristic pink patches.1 ppm and 10 ppm. However. On the other hand. caught in North Atlantic waters.35% and 0. and for a heavy salting a ratio of 1:3 salt:fish is used (1). ‘Green cured’ split cod with about 60% of the backbone removed.15% calcium and magnesium. Also.1. addition of levels of 0. When pure salt is used. See Figure 5. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. This condition is known as ‘putty fish’ (4).CURED. SMOKED AND DRIED FISH 5. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. the fish is restacked with fresh salt after about a week. The most common fish used in the northern hemisphere is cod (Gadus morhua). respectively (1). and a ‘putty’ fish product is also obtained (1). For a light salting. ‘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. mainly in tropical conditions. the fish is mixed with one part of salt per eight parts fish. and such delays. and essentially free from iron and copper (3). are likely to cause hydroscopicity of the product and.
Fig. when a light brine is used. Where fish are salted in their whole form. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. it appears that water activities much less than 0. Only small fish can be salted whole. such as 98 . eviscerated and/or split). the process is mainly for flavouring purposes prior to the use of other preservation processes. although toxin production would be unlikely (4). The greatest barriers to salt penetration are the skin and scales.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat. Food-poisoning organisms.90 are rarely achieved. Processing of dried salted cod 5. such as Staphylococcus aureus could survive and grow at such water activities in the product.3.5.2 Wet salting or brining As referred to above.1.1. For large fish it is advisable to split it before salting. so that the risk of spoilage is high.
98 5. headed and gutted.10 1.41 6.91 2.88 5.56 6. a saturated brine is used.78) (1).03 2.11 3.44 2.25 5.76 4.46 4.35 4.11 4. There are several variations of this process.0 3. The fish are kept immersed in their own brine near saturation point for several days.85 3. limiting the amount of rancidity that can develop.96 1.3.41 4. See Table 5.5 3.76 99 . sardines.3 Dry / wet salting In this process.49 4.0 4.26 2. after being drained and pressed at 465 kg/m2 for 15 hours.23 8. thus.5 4.98 6.76 4.60 2.73 1.26 7.54 5.76 4.05 7.74 9.65 2.26 4. and can be collected and replaced again over the fish.78 6.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) 2.25 5.33 5.95 3. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.30 2.CURED.45 3. they can be stored at 28 °C.64 4.17 5.61 3.98 6. the product obtained is called gaspé-cured (5). The main objective of the brining method of salting fish is to limit air penetration during processing. and a red burgundy colour will develop in the product.51 3.47 6.78 2.26 1.41 5. the brine is then replaced over the fish.26 3.09 8.03 4.II.51 5. can be cured in a saturated brine solution for 6 days and.0 2.23 2.09 8.85 3.41 3. Histamine formation is of importance when using scombroid and some clupeidean species.76 4.00 3. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance. Fresh sardines. Anchovies are the main fish used for these purposes.1. the liquid that comes out of the fish forms a brine. TABLE 5.86 2. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .06 4.04 5.11 9. with pressure being applied to the top of the container to help the brine to drain away.19 3. SMOKED AND DRIED FISH smoking.23 2. For example.90 (1). This is why it is the preferred method for fatty fish.80 5.5 3. Histamineforming bacteria do not generally grow at water activities below 0. with a shelf life of about 4 weeks (water activity (aw) about 0.99 2. but as it is placed in containers. but mackerel.70 3.44 2.16 7.22 6.84 1.17 4. the fish is dry-salted. For preservation purposes.66 5.76 4. herrings and other fatty fish can be used.5 10. Here.70 6.61 3.66 5. the fish enzymes play an important role by maturing or ripening the product.3 days.23 3. if split fish is dry-salted. 5.
the occasional implication of pindang in cases of foodborne illness. However. giving a cooked appearance to the product. but it has further smoking. Usually. Ceviche. only the nonspore-forming organisms are eliminated in the process and. usually requiring refrigeration below 10 °C for longer-term storage. Traditional jellied eels are a product where pieces of fish are cooked in boiling water. The first process can give a longer shelf life. salt. or can be boiled by being placed in a basket. There is epidemiological evidence that these products have been the cause of some 100 . usually flavoured with onions and hot chillies. and even death.FISH AND SEAFOOD 5. it can be repacked in milder brine (1 . drying and salt-boiling steps until a hard translucent block is obtained (4).3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. and packed (3). whereas the latter method preserves the fish for only a few days. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. until the flesh is soft. is made of different varieties of fish. Another variation uses sour cream instead of the second brine step.3.3. Once cooled. Afterwards.6 days without refrigeration (1). In a second step. Other preservation techniques can be used for extending the shelf life of these products. and spices) is added to the brine. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. All these products are ‘semi-preserves’. They will have a salt content >6% (w/w) in the water phase. a Peruvian product. Another salt-boiled Indonesian product. As referred to above. has a shelf life of several years. containing 1 kg of salt for every 25 kg of fish.2 months. and 6% salt. and lasts for 3 . 5. The lime juice plays the role of the vinegar. up to 1 . they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. After this. which is dipped into a boiling brine for 10 . because of the salt content of the product. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is.30 minutes.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. A much firmer product is obtained when the fish is salted prior to acidification. underlines the need for careful control (4). placed in a marinade of lime juice. flavouring (sugar. Herring can also be cured for several days in an 80 to 90º brine with 2. then packed in barrels with a 70º brine. or a pH <5 and they are stabilised for a limited period. herring or other fatty species are used. ‘ikan kaju’. outgrowth of bacterial spores after processing may occur (4).5% 120grain distilled vinegar.7%) for as long as 3 weeks.2% acetic acid and 2 . even if the product is hermetically sealed.4% of salt) for distribution and sale (1). they are packed for consumption within 2 weeks at chill temperature (6). The fish can be soaked in a high concentration of vinegar (about 4 .
There are several products named marinades (matjes. South America and the Caribbean. They are an important dried product. In some coastal areas of Portugal. mixed with salt. etc. but sometimes it is dried without prior salting. such as squid. air flows naturally. Stockfish products have a golden brown colour and are very hard. Stockfish is extensively used in Portugal. gravad fish. depending on the thickness. The most common method used for fish products is the air-drying process.CURED. gutted. split and dried gadoid species) are traditionally made in cold climates. still made without prior salting on a large scale.3. jellyfish. sugar and spices. Foodborne pathogens. such as Listeria monocytogenes and Clostridium botulinum. are likely to survive in these mild-processed products. some fish species such as moray eel and varieties of dog fish. Provided the product is kept in dry and fairly cool conditions. after frying. In Scandinavian countries. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. It is important to assure the quality of the raw material. At pH levels below 4.5. a very fine delicacy for Oriental food lovers. 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).) where no acid is added to the process. are sundried and. the fish is dried naturally in a relatively cold environment. 5. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. used as an appetiser. by using either solar heaters or mechanical dryers. 101 . marinated products will keep for several months at a temperature of 4 °C. the temperature and the amount of pressure applied (5). it will last for many years (5). the fish is dried after salting or brining. and a fine product is obtained. Usually. Stockfish (dressed. Italy and Spain (5).4 Air drying This is probably the oldest method of fish preservation. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). Some bacteria and enzymes are still active in the presence of acid and salt. Gravad fish is a Scandinavian product made of salmon fillets. and the stacks are placed in the refrigerator. under a light pressure. bacterial toxins and parasites (7). It takes about 2 days to marinate completely. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. Dried octopus is also a very common dish in the interior of the country. Fresh dill is placed between the layers of fish. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). Matjes are made of fat young herring. Shark pectoral fins.0) (7). sea cucumbers and seaweeds. which are placed in a mixture of salt. are commonly dried in Asian countries. Other products. and the flesh will break down. In the first case. are generally air-dried at sea. Dried shrimps are widely used in Asia. 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.
The fish must be stored in reasonably dry conditions to avoid moisture absorption. aureus) or spores (e. there is an increasing demand from people that have emigrated to industrialised nations for traditional products. the protein must be set or denatured with low-temperature drying before applying higher temperatures. and it is exported to Portugal. some of which are listed in Table 5. more commonly nowadays. the diffusion of the bulk water to the surface of the fish is blocked. botulinum growth might occur in the bulk of the product. typically in stews and soups.75 (1). Spain. dried salted cod (bacalão) is now mainly produced in Norway. Italy and South America. However.g. skin up. France. and increasing amounts of such products are making their way into the market place. botulinum). spoilage and possibly C. After dry salting.83 (9)). by warm air circulating in a drying chamber.the lower the humidity. tarpaulin. the better the moisture transfer. making a water barrier and consequently. nor subsequent cooking operations will affect the presence of toxins (e. the quality is more difficult to control. Whilst the low aw of the final product will inhibit microbial growth. A highly demanded dried fish product in Mediterranean countries is dried salted cod. The amount of moisture transfer from the fish to the air is dependent on the relative humidity . the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or. and has the primary objective of creating a product with specific taste and textural characteristics. Thus. To prevent this. and the water must be changed several times before cooking. the product can be held for months at room temperature. but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). neither it. 102 . that may have contaminated the product during the drying process. Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources. In conclusion. It is important to balance the drying rates and quality/safety issues.FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. 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. if the rate of water removal is too high in comparison with the diffusion rate. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain. toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0. They must be soaked in water. When no more water can be removed from the fish.g. Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s.III. Staph. Whilst the production of dried fish products is tightly controlled in industrialised countries. Dried salted products must have a water activity below 0. C. Iceland and Canada. the fish must be dried slowly enough to avoid case-hardening. which leads to mould growth (aflatoxins. Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. the skin will form a hard case (case-hardening). The product is dried to a final water content of about 25 to 38% (3). In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. or wooden drying rack to dry in the sun.
3. such as Escherichia coli.CURED. botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. aureus (toxin) C. herring. making full description impossible here. Hardwoods are considered better because of the milder flavour they impart to the food. and the use of smoke generated by superheated steam or liquid smoke. swordfish. mussels and many others from tropical countries. It can be used for cold or hot smoking. white mangrove and abura has an inhibitory effect on some microorganisms. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns.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. SMOKED AND DRIED FISH TABLE 5. sprats. using electrostatic filtration of the smoke. Blends of various woods can be formulated to produce sawdust with specific flavours. is mainly a process for flavouring fish rather than a preservation technique. cod. The smoking procedures vary widely. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. However. eel. Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . softwood colours the product quickly. 5. The quality of the smoke is related to the type of wood used. trout. There are many fish species that are used for smoking purposes . mackerel.5 Smoking Smoking today.salmon. Top-quality fish must always be used. cod roes. Staph. aureus and Saccharomyces cerevisiae (10). Maintaining the temperatures of pyrolysis between 425 and 200 °C. oysters. It has been reported that the smoke from mahogany. tuna. haddock. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10). Aroma and flavour are a blend of smoke components (3). In the mechanical kiln. clams. as referred to above. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish.
as it retains more moisture. smoked cod fillets and finnan haddock are usually cooked before consumption. which is important for the appearance of these products (4). it will take up salt twice as fast as fresh fish (10). or quick-frozen and stored at -18 °C (2). Brining at 10 15 °C is recommended (10). are boiled before smoking (12). after being smoked need to be chilled immediately to 4 °C before being packed. Also. however. This difference is crucial for safety issues because the flesh is not cooked. If frozen fish is used. filleted. owing to the elution of soluble proteins. such as smoked salmon-trout. giving the product the desired appearance and flavour (4).and hot-smoking is the temperature of the process. Thus. wooden boxes or master cartons (10). temperatures must be kept under 30 °C throughout the process.18 hours at 4 °C. tuna and swordfish. the highly desirable cold-smoked salmon. thus allowing the deposit of smoke on the fish surface.1 Cold-smoking The major difference between cold. The salting period is dependent on the size of the fish and its presentation (whole. the excess salt is removed with water. Some herring products. they can be vacuum packaged. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. 70 – 80 º brine is commonly used. all cold-smoked products should be cooked before consumption. Thus. If wet salting is chosen. but a ‘safety’ (i. Sugar. Wet salting or brining is preferable to dry-salting when smoking fillets because. Cod roes. skin off or on.5. it will reduce the salty flavour. 5.FISH AND SEAFOOD water flow system with chlorinated water (25 . pasteurisation) temperature is not achieved in any stage of the process. spices. the following step is salting. The fish can be dry-salted using a weight of salt 5 . an attractive glossy pellicle on the fish fillet surface is achieved after draining. covering the fish surface completely (10). or packed in retortable foil pouches. In cold-smoking. skin on or off). When frozen/thawed fish is used. Depending on the type of product.10% of the weight of the fish. However. gutted or filleted. but this concentration will again depend on the type of product and the length of the brining stage. usually dry-salted and cold-smoked. permitted colourings or smoke flavours can also be added to the product in the brining step. The maintenance of temperatures below 30 °C avoids protein coagulation. Sugar can be added together with the salt. thawing must be done slowly for 12 .e. and the cold smoke is denser. Drying should be minimal at the start.or hot-smoked. low temperatures will slow the uptake of salt. it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11). The length of the smoking process for such products is much greater than for the hotsmoked fish. 104 .3. All the smoked products. Whether the product is prepared headed. but it will increase towards the end of the process (10). canned or bottled. whether cold.50 ppm) should always be done just before process initiation (3). as well as many other products prepared in the same way. are not cooked prior to consumption. shrink packaged.
This temperature should be maintained during storage and distribution (3). a partial cooking period at 50 °C.CURED. and their sensitivity to salt (13). refrigerated storage and vacuum packaging still allow its growth (12). To avoid growth of non-proteolytic C. in order to produce toxin. if present. On the other hand. Neither the smoking temperature nor the salt content is enough to kill the organism. gravad salmon (also called gravad lax or gravlax). monocytogenes. which underlines the danger of storage at abuse temperatures of >10 °C (13). secondly. condensation will occur inside the pack. and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). such as Yersinia enterocolitica. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). Length of the smoking process at each stage will depend on the species and the type of product. Afterwards.2. can be isolated from seafood. During smoking. Vacuum packing is recommended only for coldsmoked salmon. there is no point in the process that can fully assure the absence of L. The exception is vacuum packed fish. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). See Figure 5. These products should be vacuum packed at room temperature. the products are cooled. but the proteolytic types A and B.5. as. The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. Pathogenic vibrios. if a 105 . A typical temperature set is as follows: firstly.3. if they are taken from a chill room at 3 °C and packed. botulinum type E present an even longer time than before to grow and produce toxin (14). further cooled to 3 °C within 12 hours and then packed. Hot-smoked products require a longer cooling stage. thus allowing any psychrotrophic C. are of concern when smoking fish from tropical waters (13). If the refrigeration chain is maintained. botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. a final cooking period at 80 °C (12). Although other human pathogens that can grow at refrigeration temperature. can grow with salt levels of 10% in the water phase. typically mesophilic. thirdly. Furthermore. Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. but they should be cooled to a temperature of 10 °C within 3 hours. 5. and hotsmoked salmon (kippered) (3). Smoked salmon fillets are usually vacuum packed whole or after slicing. Smoked products can be also packed under controlled-atmosphere packaging. risk from the presence of Bacillus cereus and Clostridium perfringens can also be excluded as potential hazards because of the relatively high temperatures they require to grow to.2 Hot-smoking Hot-smoking must use temperatures between 70 and 80 °C at any stage of the process to ensure protein coagulation throughout the product. SMOKED AND DRIED FISH During cold-smoking. a drying period at 30 °C. stored at 4 °C and packed.
Fig. and moulds will grow on the product (12). 5.FISH AND SEAFOOD product is packed too warm without being cooled to room temperature. Process of cold-smoked sliced salmon 106 .2. it will have a reduced shelf life.
Mackerel. 17. Aspergillus ostianus. 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). They will keep well frozen. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. 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. Boiled or steamed clams. Aspergillus tamarii) and several Penicillium spp. apart from the improved appearance. When refrigerated at 3 °C. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. then hot smoking and finally sun drying it. (Penicillum cyclopium and Penicillium puterellii) (19). Katsuobushi. including ‘umami’ (16. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. 22. Aspergillus oryzae. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). Eels.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. are usually 80º brined for 5 . but seems.3.CURED. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). fatty fish will remain in good condition for 6 days. synthetic smoke flavours. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. including Eurotium repens and Eurotium rubrum. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). a very expensive and fashionable smoked product in the Netherlands. rewetted and remoulded to give the desired shape and consistency. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. break down of fats and the moderation of smoke flavour. 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). moulding it into long thick sections. white fish about 8 days. similar to masmeen. 23). The classical liquid smoke is 107 .5. The product is removed from the drying cycle.6 hours and smoked for 3 . In terms of safety. Natural smoke extracts. The production method is similar to masmeen. 18). cooking in brine. and substances unconnected with smoke but with a smoky flavour or smell. chilled or freeze-thawed. and many closely related Aspergilli (Aspergillus ochraceus. resulting in a characteristic flavour (20. They are generally not cooked prior to consumption. 5. 21. are used. to have no further advantage over other packing methods for other fish (12). Vacuum packaging retards the onset of rancidity when fatty fish is used. Growth of these moulds on the tuna results in the extraction of moisture.3½ hours using the typical temperature set for hot-smoking referred to above (12). but mechanical drying is used instead of sun drying. are usually hot-smoked after being immersed in 80º brine or dry-salted (6).
established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). both of which are considered potential carcinogens. or smoke extracts in organic solvents.4 Spoilage Brining.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. It has been shown that liquid smoke can reduce the formation of PAHs (3). According to some recent research. 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. Liquid smoke was considered to provide advantages of safety. applying it as a dip after brining. consistency of flavour and colour.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. and less than 0.8% and still provide protection for 7 days (25). Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. 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. The European Scientific Commitee for Food. the salt level could effectively be reduced from 4. spraying is preferable because it is easiest to control flavour and acceptable with this method (3). so as to impart an unnatural red colouration to the flesh. When liquid smoke was used. 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.06 μg/kg of benzoanthracene in the final foodstuff (28).3. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). 5. In experiments comparing traditionally smoked fish with liquid smoke. or applying it as an atomised spray within a modern automatic smoke kiln. 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).03 μg/kg of benzopyrene. 5. drying. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish.5. smoking. injecting it into the fish flesh. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. Fresh fish spoilage may be measured by 108 . There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines.6 to 2. nutritional neutrality and antioxidant activity (27).
but. the consequent reduction in the pH will help to inhibit spoilage organisms. and high concentrations are usually required to inhibit their growth fully. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. the fish flora are transferred to the brine mix. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). Upon salt packing. however. Often. Usually. these halophilic organisms may grow and cause spoilage. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure). a very dry product is required to produce 'shelf-stability'.. When heavy brining or salting is important for product shelf life. Micrococcus spp. When acid marinating or smoking. and of smoke to add chemical preservatives. a product may become undesirable for consumption as a result of changes in its sensory characteristics. the production process will modify the microbial flora present on the product. therefore. marination or drying).. Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2.g. as the fish are added. otherwise.000 cfu/ml of brine may build up. a process similar in microbiological effect to that of saturated brining. Most marinated and smoked fish products are first brined or salt-pickled. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). spoilage is due to endogenous microorganisms and enzymes of the fish.CURED. the bulk of the microflora comprises Bacillus spp. for some. contact with the bacteria 109 . chemical oxidation. Stronger and longer brining has an effect like that of pickling. or extended refrigerated storage. which are found in the salt. limits the types of microorganism capable of growing. Even saturated brines allow some halophilic organisms to grow. Spoilage needs to be carefully defined for each product. which may also be produced to describe the quality attributes and their loss over time for processed fish. Within the limits of safety. Brining. also. The use of drying and/or salt to lower the water activity. For some products. and coryneform bacteria. Once these changes are defined. both of these will affect the spoilage patterns. contaminant bacteria from other materials used to produce the product. or their products require refrigerated storage. The microflora attached to the fish is relatively unaffected by light brines used before smoking. for others. Once placed into brines. SMOKED AND DRIED FISH grading schemes or quality indices. the shelf life may be estimated and investigation made into the causes of the loss of sensory quality. In salts. pickling and dry-salting all introduce bacteria to the fish. these bacteria tend to continue to dominate the flora in the liquid. Some brines are re-used over a period of days and a load of 10. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt.3%). and. the fish are cut and splayed to allow greater surface contact with the salt. the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. rock and manufactured salts did not usually contain halophilic bacteria (29).
as the water activity of saturated brine solutions is low enough to inhibit most bacteria. Studies conducted on dried and salted fish from Indonesia (31.. The condition describes the growth of the halophilic moulds (Wallemia sebi. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage. Aspergillus wentii. as little if any growth will occur below 5 °C on salted fish (29). ‘dun’ is not characterised by the production of offensive odours.g. sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). lactic acid bacteria (LAB) and moulds. 34) have shown that P. 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). were also isolated. At the pre-drying stage. This condition is often associated with case-hardening. repens and E. sometimes covering the whole of the fish with a whitish. it was found in about 50% of samples examined. E. Therefore.FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. Several Penicillium spp. ‘pink’ and ‘dun’. or fat breakdown) caused by enzymes rather than by microbial growth. (syn. gaspé). Eurotium amstelodami. spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. and still produce a reasonable dried product. niger. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. Sporendonema sebi)). flavus. W. (syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. which. The metabolism of the organisms will produce off-odours. it is possible to wash off the infected parts. rancidity due to lipolysis. like the ‘pink’ organisms. including A. 33). halophilica was isolated 110 . though only the latter two species were considered likely to have grown on the fish (32). The bacteria form coloured colonies on the surface of salted fish prior to drying. A. B. together with a range of Aspergilli. Only halophilic microbes may grow. although it may occur in other salted fish. powdery growth. or after drying if the fish is stored in excessive humidity. and Halobacterium spp. yeasts. Basipetospora halophila. yeasts and fungi. and Aspergillus penicilloides. The microbiology of heavily salted products is dominated by Gram-positive. Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration. A number of spoilage organisms can grow and produce conditions such as ‘sliming’. ‘Pink’ describes a condition associated with surface growth of the pigmented halophilic bacteria Halococcus spp. may also be found in the salt used in production. The bacteria are osmosensitive and will lyse in water. Aspergillus sydowii. pisce was the main fungus isolated. if not too contaminated. rubrum are also common isolates from dried fish. spore-forming bacteria. most notably Penicillium citrinum and Penicillium thomii. protein breakdown. Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. the best way of controlling the organisms is to refrigerate the fish. Unlike ‘pink’. including hydrogen sulphide (26). halotolerant or halophilic micrococci. 32.
Aspergillus clavatus and Aspergillus niger. LAB. so the bacteria spoiling these products will be psychrotrophic or psychrophilic. 36). as already stated. The preservative effect of the smoke will inhibit some spoilage microorganisms. will have a different residual post-smoking flora from 'hot-smoked' fish. For more information on spoilage see Table 5. where a high internal temperature is reached.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations.5% sodium chloride in the water phase will inhibit some bacteria.IV. but. The salt in the water phase must be high enough to inhibit the growth of C. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage. however. 111 . Vacuum packing limits chemical oxidation. It has been reported that the type of salting (dry. Pseudomonadaceae. Aspergillus spp. flavus. Table 5. but this will not prevent the growth of spoilage bacteria. around the water activity of saturated sodium chloride. not the growth of spoilage organisms. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. Fish processed by cold-smoking. and microbial spoilage is more likely. Marine vibrios were the dominant flora in smoked salmon that was drysalted.. In Vietnam.CURED. Cold-smoked fish will contain microbial flora representative of that of the raw fish. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life. able to grow on dried salted fish. botulinum type E. Smoked fish products tend to have a light brine treatment. with a lower aw limit for growth of 0. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants.747. 33. which does not create the salty conditions required for spoilage by those halophilic microorganisms. The brining to 3 . has been isolated from dried fish (35). The heat during hot-smoking will kill vegetative microorganisms but not all spores. including A. Refrigeration is necessary. it was. Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. SMOKED AND DRIED FISH infrequently at low levels.
LAB. halophila Halophilic spoilage organisms Halophilic spoilage organisms.not used/unchanged. Dun.FISH AND SEAFOOD TABLE 5. 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. boiled. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. sebi and B.IV Spoilage of salted. 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. and Halobacterium spp. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted. + to +++ strength of preservative effect 4 Post-process contaminants 112 .variable. . Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms.
types A and B are not affected by 3 . Before.5% to 5%. 9 and 8 of the outbreaks. botulinum Staph. The concentration required to prevent growth at room temperature can vary from as low as 3.8 1.5 5.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). respectively.97 0. botulinum type E Staph. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. Examination of 165 outbreaks of botulism caused by fish products showed that smoked.5 5. botulinum.8 0. although the combination of the two factors. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). and C.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades. It has been shown that. popular demand for healthy foods has led to a change in the tastes preferred by the public and. sodium 113 . Since neither the smoking nor the drying parts of this curing process are particularly severe.75 0. the products were heavily salted.5-10 4.5-8. The presence of salt in cured products has a great effect on the growth of the bacteria.5 0-11 3. pressure on the traditional methods of curing. could not grow and produce toxin.5 4.9 0. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. SMOKED AND DRIED FISH TABLE 5. aureus Yeasts Moulds Lactic acid bacteria pH 5.97 0. but if the fish products are stored at temperatures <10 °C.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. the importance of cold-chain integrity must be emphasised. when present in the raw material.5-10. smoked and dried.86 0. the non-proteolytic type E strains can grow in up to 5% sodium chloride. the fermented products being the most dangerous group (7).0-9.5. under optimal conditions.CURED.61 Organisms Most Pseudomonadaceae C.5-8. botulinum type E. salted and pickled products accounted for 10. botulinum type E and production of toxin. However. In the United States. with this change. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. and as botulism is a serious hazard in foods. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13).
but.4.94 (7). they will grow rapidly (7). its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. coli all occur on fish products as a result of contamination from animal/human reservoirs. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). 5. As with other organisms. the risk of infection can be eliminated by proper cooking.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. Thus. has been reported (42). 5. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 .5% salt in the water phase. Sliced smoked fish products.5.or cold-smoked fish should have at least 3.5. are particularly susceptible to contamination and. It is a halotolerant bacterium able to grow at water activities as low as 0. related to the consumption of a raw smoked fish meal. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin.30% of the population positive for enterotoxin-producing strains (41).5. Infection by Shigella sonnei has increased in the UK. if it contains sodium nitrite ≥100 ppm. if the person handling the product is a carrier of Staph. The FDA established that hot. In foodstuffs. as they require more handling. if storage temperatures are over 10 °C.0 . Once the product is contaminated. However. after processing. An outbreak of Shigella in an Italian cruise ship. Shigella and E. the organism can survive most of the light curing processes. when the bacterial load of the product is lower. As they are poor competitors. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. they do not grow well in the raw material. if recontamination occurs and storage conditions are favourable. throat and skin. the organism may contaminate the product. and the minimum pH for growth is 4. the human nose. The organism is mesophilic with a minimum growth temperature of 7 °C. aureus. 5. Usually. but.3 Enterobacteriaceae The pathogens Salmonella. but requires higher temperatures for toxin production (>10 °C).0% salt in the water phase (30).4 Listeria monocytogenes The psychrotrophic L. more importantly.86 and 10% sodium chloride. it should contain not less than 3. gills and guts as well as in several lightly preserved fish 114 . growth and toxin formation may occur.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. aureus growth.5. these parameters change as a result of the interference of food components and other limiting factors that can act in an additive or synergistic way to prevent Staph.
in the EU Fisheries R&D-FLAIR program.45 °C. cold-smoked fish. monocytogenes pathogenic for only specific segments at risk. Some LAB.0. when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). As they are heat-sensitive. from freshwater fish. The use of LAB in fish preservation is a topic that has been studied in projects (UP. Of microbiological concern in the northern hemisphere are two parasites . Until 1997.514.9. marinades (ceviche) (43) and gravad fish (44). both being detected in cold-smoked salmon. which considered L. favours growth of psychrotrophic LAB. Its limiting conditions for growth are 0 . which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). such as ‘lightly preserved’ fish products.92 .2.95 (45). Unlike some European countries. widely used as starter cultures in food processes. or nematodes. has also been detected in cod (7). have been shown to have an anti-listerial activity in many food products.5 Parasites Despite the presence of parasites in fish being very common. Pseudoterranova dicipiens. SMOKED AND DRIED FISH products such as cold-smoked fish. 5. from seawater fish.6 and water activities below 0. do not survive 1 min at 60 °C. a number of fish products.12%. They have complicated life cycles. in combination with curing agents and refrigerated storage. The Health Protection Agency (HPA) in the UK considered that it was unacceptable for readyto-eat foods (including all the products that are consumed without prior cooking) to contain any serogroup of L. which means that the process must conform to a level of ‘no detectable L. the USA established ‘zero tolerance’ for the organism. the only concern to public health is when eating raw or uncooked fish products. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. and other local traditional products (7). As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. researchers are looking for other ways of preventing the pathogen’s occurrence or growth.5. most of them are of little concern to public health. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod.8 . salt levels of 8 . are considered unsafe. a round worm. Two outbreaks in the United States have been reported to be linked to salmon (10). a short period of freezing either of the raw material or of 115 . monocytogenes’ in 25 g of the finished product. and in CT95-1207FAIR). the CT 3162-FAIR program. pH 4. Thus. and the tape worm Diphyllobothrium spp.CURED. monocytogenes in levels at or above 102 per gram (46). gravad fish. there were no reported outbreaks related to cured fish.the round worm Anisakis spp. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. The round worms. which. lightly salted caviar. which include intermediate hosts. This includes matjes herring.
<100 20 . cholerae Vibrio parahaemolyticus L. texture. monocytogenes Staph. for example. aureus C.VI Guidelines for the microbiological quality of ready-to-eat smoked fish. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E. salami. not detected in 25 g Campylobacter spp. perfringens B.FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). coli O157 & other VTEC Vibrio.<103 N/A 100 .<104 100 .6 Published Microbiological Criteria TABLE 5.<100 200 .<100 10 . (not L. ‡ On occasions some strains may be pathogenic. 5.<107 ≥107 N/A <100 <20 100 .<104 20 . These foods fall into category 5. Acceptability is based on appearance. † Guidelines for aerobic colony counts may not apply to certain fermented foods. E. soft cheese.<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. monocytogenes) (cfu/g unless stated) <106 106 . cereus and Bacillus subtilis# <20 <20** <20 <10 20 . the organism should be identified # N/A denotes not applicable 116 . and unpasteurised yoghurt. and the levels or absence of indicator organisms or pathogens. coli (total) Listeria spp.100 >100 N/A Pathogens Salmonella spp.<100 20 .<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . smell. taramasalata and cooked shellfish sampled at the point of sale (adapted from (11)) Criterion Satisfactory Borderline limit of acceptability (cfu/g unless stated) Unsatisfactory Unacceptable/ potential hazard* (cfu/g unless stated) (cfu/g unless stated) Aerobic plate count† (30 °C.<100 >104 >100 N/A N/A <20 20 . If the Bacillus counts exceed 104/g.<104 104 .
References Regenstein J.. Birnie K. New York. Eds.A. Leatherhead Publishing. New York. 4. Food spoilage fungi. 1997. Horner W. 2007. Torry Advisory Note No. in The Seafood Industry. Assurance of Seafood Quality.. Leatherhead. Ed. 1994. Bannerman A. Martin R.7 1. Horne J. Torry Advisory Note No. Fernandes R. 2. Catching. Van Nostrand Reinhold. Torry Advisory Note No.M. in Fish Processing Technology. McLay R. Chapman & Hall.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. 117 . Doré I. Urner Barry Publications Inc. Preservation of fish by curing (drying. 82 (revised). 9. 1984. USA. Hall G. Moody M. Huss H. 8. Wareing P. 6.. or presence of toxin 0. Wareing P. 3273. botulinum * Limit of tolerance 1x 103 ETEC/g. Van Nostrand Reinhold. parahaemolyticus Vibrio vulnificus C. cured and dried fish. 37(revised). aureus * Vibrio cholerae V..W. Hot Smoking of Fish. London. in Micro-Facts: The Working Companion for Food Microbiologists.. coli (ETEC) L.E. salting and smoking). * Staph. Flick G. Regenstein C.M. 1982. SMOKED AND DRIED FISH TABLE 5. monocytogenes Salmonella spp.CURED.F.F. FAO Fisheries Technical Paper. Introduction to Fish Technology. 1990. Marinades. 1979.. 334. Eds. 381-406. 56. 7. 1993. 1991. except in the viscera of dungeness crab. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. Smoked. Flick G. 3. The Smoked and Cured Seafood Guide. 120-38.H.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. Fernandes R. 254-82 5. Handling and Processing Eels. 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.
88.E. Doi M.. 958-60. 1995. Biotechnology and Biochemistry. 6 (4).M. 1992. Bioscience. Patel T.. 16. Yamauchi H. 843. Technomic Publishers. Jeppesen V. Doe P. 553-72.A. 2002. 2008. 22..R. Martin A. Doi M. 508. 1981.. Kinoshita Y... Hayashi K. 1980. 108 (3). 335-40. 1994. 12.. Food Reviews International.. Kanayama T. 1982. 14. Ninomiya Y. Embarek P.. 56 (6). Microbiological control for fish smoking operations. Agricultural and Biological Chemistry. Bannerman A. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors. Bioscience. Biotechnology and Biochemistry. 6 (6).. Horne J.M. Control of biological hazards in cold smoked salmon production. Matsui M.E. 2324-5. 22 (revised).. 118 . Matsui M. 1989. 1984.. The discovery of umami.C. 23. 51-81. 1977. Sakakibara H. Hosokawa M. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor. Huss H. Packing Fish in a Modified Atmosphere. Dillon R.B. 24.. Shuto Y. Druschitz A. Ogiwara Y. Perusing Asian soup options.W. Paranjpye R. Lindemann B. 15. 2007. 53 (4). Ed. 5 (revised). Teeny F. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour. 25. Chapman & Hall.. Shiemori Y. 935-41.C. Bannerman A. Botulism and Fishery Products. 1995. 17. 1998. Torry Advisory Note No. Halpern B. Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species. London. Kinoshita Y. 18.. Torry Advisory Note No. 840-6. 1997.. 1051-5. Torry Advisory Note No. Cann D..H. 13. 20. Food Chemistry. Yajima I. 18. Lancaster. 1990. 61 (8). in Fisheries Processing.. Shuto Y.FISH AND SEAFOOD 10. Cann D. Yokayama S. Eklund M. Fish Drying and Smoking: Production and Quality. Aoshima H. 45 (10). 19. 59 (12). Torry Advisory Note No. Bioscience.. and Biochemistry. Martin A. Shuto Y. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour. Peterson M..F. Ninomiya M. What’s in a name? Are MSG and umami the same? Chemical Senses.P.M. 17 (9). O-methylation of 2. Doi M.. 21.. Biotechnological Applications.. Food Control. Biotechnology.. Food Product Design..K. Recommendations for the Preparation of Smoked Salmon. 11. Doi M. Flavour constituents of dried bonito (Katsuobushi). Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish. Pelroy G. 27 (9).. 1386-7. Processing Cod Roes. Mitou M. Journal of Food Protection...
Boca Raton. Luxembourg. Food and Drug Administration.D. Journal of the Fisheries Research Board of Canada. Importance of autolysis and microbiological activity on quality of cold-smoked salmon.. Sirvas S. Simini B. Ed. Oxford. Hansen L. Cox J.I. 1996. Hocking A.. 22.A. Food Technology. Pitt J.V. Ed. 35. Wheeler K. Academic Press. 1996. in Fish and Fishery Products Hazards and Controls Guide. 37. Huss H. The control of botulism in hot-smoked trout and mackerel. 38 (3). 74. Shewan J. Golden D.F. 218-25. An outbreak of Shigella. Journal of Food Technology. 1941.1988. Tour highlights production and uses of smoke-based flavours.A. 39. 41. 34. 29 (2). CRC. 1994. Journal of Applied Bacteriology. 181-8. Diyaolu S. in Fish as Food. 3 (4). Borgstrøm G.. The microbiology of sea-water fish. Hocking A. Adebajo L.CURED.M.. 119 . Journal of Tropical Medicine and Hygiene.F. CEC.T. US Food and Drug Administration. Scientific Committee for Food... 1986. 1985. Anggawati A.K. Antimicrobial properties of smoke. Røntved S.I. 1991. Washington DC. 74 (2). 348. London. V.S. Fungi and Food Spoilage. 107-11. Maga J..A. National Academic Press. 36. Fuchs R. Taylor L. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. 88-90. 29.. 49 (1). Institute of Medicine. 1994. Lancet.H. 311-7..Y. 43. Fungal flora of South Vietnamese fish and rice. 27686. Smoke flavourings... Hess E. Townsend J. 2005. 1997.. Food and Drug Administration. 1995. Pitt J.M. Jay J. 742. Pitt J. Committee on Evaluation of the Safety of Fishery Products. Hocking A. Sprouse R.. 27.. 5. Springer Science. Nahrung. Mycotaxon.D. 38. Loessner M. Ed. Wheeler K. 28. Food Microbiology. Gill T. Studies on salt fish. Rockville. 33. Letters in Applied Microbiology. in Reports of the Scientific Committee for Food (34th Series). 1979. Ed. New York. Lucas F. 74. Press Inc. 1961. Blackie.J. New species of fungi from Indonesian dried fish. 1991.O. 32.B. 14. ceviche. Incidence of Listeria monocytogenes in an acidified product.E. 123-9. 42.C. Commission of the European Communities. 12.. Pszczola D.D. 1971. Modern Food Microbiology. 1993..I. Fungi associated with Indonesian dried fish.A. 164-9. Cann D. 70-2. Seafood Safety. Studies on Sporendonema epizoum from ‘dun’ salt fish.D. 30.M. Ahmed F.E. Food Research International. 197-208. Hocking A. SMOKED AND DRIED FISH 26.. 1995. 40.R. 351-7. Smoked and smoke-flavoured fishery products. in Smoke in Food Processing. 31. Frank M.. 98100.. Interactions among xerophilic fungi associated with dried salted fish.
. 26 (4)... in Fish and Fishery Products Hazards and Controls Guide. 188 (3). Fletcher R. Hurst A. 1980. 55 (4). Food and Drug Administration.N.A. Moral A.. Association of Food and Drug Officials.C. 5-7. Pathogenic microorganisms of importance in seafood.. 47. Donovan T. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus.L. 863-6.cfsan. US FDA Centre for Food Safety & Applied Nutrition.A.. Wooten R. de Louvois J. Elsevier Science Publishers B. 1991.. 197-209. Round Worms in Fish. 2001. 5 (1). Eklund M. Incidence and growth of psychrotrophic Clostridium botulinum in food.S. 1980. 45. Tham W.8 Further Reading Gibson D.V. Communicable Disease and Public Health. Danielsson-Tham M.H. Pelaez C. 3. Journal of the Association of Food and Drug Officials. 120 . Loncarevic S.J. 35 (11) 2904-7. http://www.FISH AND SEAFOOD 44. Little C. Pelroy G. Journal of Food Protection.. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions. Bolton F. Huang Y. 46. 833-41.A. Roberts D. US FDA Centre for Food Safety & Applied Nutrition. Journal of Clinical Microbiology.. Cured. Loncarevic S.html Gilbert R.. Amsterdam. Nye K. 1982. Harrison M. Hobbs G.. Davies A. and smoked fish establishments good manufacturing practices. Food Control. Beltran A.. Peterson M. 48... Journal of Food Protection.D. Washington D.. 1992.. Susuki E. in smoked and "gravad" fish.. 80.R.. Jacobsen M. 5.. Hughes A.M.. Advances in Food Research. An outbreak of listeriosis suspected to have been caused by rainbow trout.. Prevalence of Listeria monocytogenes and other Listeria spp. Eds.fda. Acta Veterinaria Scandinavia.. 2000. Eklow A. 135-85. Clostridium botulinum and its importance in fishery products. 232-6. 1989. 58 (8).. 37 (1). Appendix 5: FDA and EPA Guidance Levels.C... Ed.E. 1976. Tham W. Zeitschrift für Lebensmittel-Untersuchung und –Forschung.J. in Quality Assurance in the Fish Industry. 1994. 22.. 507-10. 163-7. Huss H..W. Torry Advisory Note No. 13-8.gov/~comm/haccp4x5. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon. Paranjpye R.W. Ribeiro C. Mentzing L-O. Cann D.. 1997. Persson I.M. Garren D. Richards J. Unnerstad H. 1996.. 45 (9). 49-59. Danielsson-Tham M-L. Canadian Journal of Microbiology. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale. 1995. Liston J.. Gibbs P. Ericsson H. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects. salted.
Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout.. 217-36. Applied Microbiology. Letters in Applied Microbiology.. Pelroy G.T. 1994. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. Lashbrook L. Peterson M. 25. SMOKED AND DRIED FISH Sutherland J. 1995. solute. Beuchat L. Ben Embarek P. Caugant D. Elsevier.P. 4389-95. detection and growth of Listeria monocytogenes in seafoods: a review.. and salt concentration on growth and survival of Vibrio parahaemolyticus. van Spreckens K. Keusch A. Contamination pattern of Listeria monocytogenes and other Listeria spp. 27 (6). Beuchat L. Dallas H. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. 309-16. Slade A. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. 844-6.R.J. 11. Jemmi T. Kramer D.. Nicolaides L.S.178-80. Roberts T. Bayliss A. Ratnam S. 1994. 502-8.. 1992.. Journal of Food Protection. Tran T. Journal of Food Safety. Histamine production by psychrophilic flora. 1995.CURED. Davies A. Eds. and temperature on the growth of Vibrio parahaemolyticus.R. Hitchins A. 394-6. International Journal of Food Microbiology. International Journal of Food Microbiology.W.N.. 1987.. Prevalence of Listeria in smoked fish.A.. Rørvik L. Hudson J. Incidence of Listeria in hot and cold-smoked fish. 58 (5).. Competition of food bacteria with Listeria monocytogenes during enrichment culture. Combined effects of water activity. 19-27. Applied Microbiology.. Food Technology. 1974. Interacting effects of pH. Taylor S. 94-8. Dillon R.. 1991.. temperature. New York. Presence.K. 309-18. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. 23 (1). Yndestad M. 1994.. in a salmon slaughterhouse and smoked salmon processing plant.. Food Microbiology. 11. 55 (11). 1075-80. 42.E. 21 (3). 14. Fuchs R. 293301.. Paranjpye R. 1995. Poysky F.A. Liston J. 1994. Letters in Applied Microbiology. 121 . pH and sodium chloride. 866-70. Marine toxins of microbial origin. Journal of Food Protection. Letters in Applied Microbiology.A. in Seafood Quality Determination.. 61 (12).. 1734. Eklund M. 25 (5). International Journal of Food Microbiology. 19. 1973. 1992. Patel T. 1988.R.C. Poindexter C.
they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. North American and other markets and are. typically in the range 15 . Those to which only salt is added tend to be mainly autolytic.6.I. As with most traditional products that are still produced principally on a cottage industry or domestic scale.1. FERMENTED FISH Prof. Carbohydrates are added to some fish sauces during their preparation.1 Definitions As applied to fish products. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6. there are numerous variants of some common themes and a host of local names used to describe them. For example. Essentially. in economic terms. they are exported to European. 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. koji. The relative importance of these two activities depends on the formulation of the product. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. though some are produced elsewhere.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. Fermented fish products are largely confined to east and south-east Asia. The fish sauces are produced on the largest scale.25% and are used mainly as a condiment. producing the fish sauces and pastes. 123 . the most important of all fermented fish products. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. added during preparation. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. lactic fermentation can play a significant role. whereas in those where a carbohydrate source such as rice or sugar is also added.
fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. In Europe. 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.FISH AND SEAFOOD TABLE 6. made from fish livers in Greece. salty taste. made from whole fish in France. 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. may be direct descendants.1. These have long since fallen from use. 124 . and the more widely known Worcestershire sauce a very distant relative. although garos.III. where the bacterial production of lactic acid is a major feature.II. jeotkals (shrimp) nam-pla (pa) Budu Patis nam-pla nuoc-mam padec Belachan (shrimps) bagoong kapi mam-ca man-tom (shrimps) Laos Malaysia Philippines Thailand Vietnam Even within the limitations of fish and salt as the sole ingredients. Fermented fish products are of very minor importance outside Asia. a variety of products can be produced within a single country. to products analogous to other lactic fermented foods such as salami.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. This is well exemplified by some of the fish/salt products of Thailand described in Table 6. and pissala. 6. A selection of Asian products is presented in Table 6.
pla-som. burong-ayungi. shrimps.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. mam-chao. e. e. tai-suchi phaak.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 . crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 .g.8 months Main dish TABLE 6. som-fak/ som-fug 125 Malaysia Philippines Thailand . funa-suchi. pa-chao. burong-dalag.FERMENTED FISH TABLE 6. pla-chao.g. som-pa-keng pekasam. 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.3 months Main dish Tai-pla Bowels 1:3 6 .12 months 10% raw sugar added. 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 . mam-seeing sikhae som-kay-pa-eun. pa-khem. som-pa. burong-bangus pla-ra. ayu-sushi. mam-pa-kor. cencalok burong-isda.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx.
and their microflora tends to reflect their local environment more than that of marine species. and the initial microflora is likely to be considerably larger in these cases. A modern account of their production. or buried salmon. made from herring. fermented trout. in a survey of fermented fish products in north-east Thailand. in Sweden. Here.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform.the flooded rice fields. Table 6. the most popular varieties of which are rakørret. and counts ranging from below 104 /g to in excess of 107 have been reported. is a traditional. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. freshwater fish are the usual raw material. states that sugar is often included with the salt.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.FISH AND SEAFOOD In Northern Europe. however. some fermented fish products remain popular.IV Fish species used in lactic fermented fish products of the Philippines Product names Burong ayungin Burong bangus Burong dala Burong gurami Burong hito Burong kanduli Burong tilapya Balao balao Burong hipon Local Ayungin Bangus Dalag Gurami Hito Kanduli Tilapya Tagunton Suwahe English Silver perch Milkfish Mudfish Goramy Catfish Sea catfish Tilapia Shrimp Shrimp Scientific name Therapon plumbeus Chanos chanos Ophicephalus striatus Osphronemus goramy Clarias batrachus Arias manillensis Tilapia nilotica Macrobrachium spp. TABLE 6. paddy ponds beside rice fields used for collecting fish when the field has dried up. 126 . The diversity of fish used in fermented products (see. for example. which has been increasing in popularity over the last 50 years. it was found that several different sources were used for the fish . and old descriptions of their production often refer to the addition of whey. malt or flour (1). 6. These are often described as fish/salt products. relatively mild-tasting product. rakefisk or surfisk. especially in fish/salt products. There are also more heavily fermented products. Penaeus indicua Many of the fish used in fermentation are uneviscerated. Norwegian gravlaks. and a large local freshwater reservoir (2). For example. which employ lower levels of salt than Asian products. in Norway and surstrømming. 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.
Endomycopsis and Saccharomyces . however. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. sufficient to saturate the water phase present and produce an aw of 0. and studies 127 . Other halophilic species have been identified from fish/salt products. the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. When this is a simple carbohydrate source such as sugar. (3. or boiled or roasted rice. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. mean that microorganisms have been generally held to play little or no role in the process that follows. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. they can contribute to the initial microflora.74 or below. angkak (Philippines and Thailand) and look-pang (Thailand) are used. principally species of the genera Rhizopus. More recently. after one month. Similar results were seen with patis. In some cases. in many cases. Generally. were found to be the predominant isolates. which included nam-pla and patis. Mucor. the Philippine fish sauce.1 Processing and its Effects on the Microflora Fish sauces and pastes Detailed flow charts for several fish/salt fermented products are presented as Figures 6.3 6. Bacillus spp. The amount of salt used is often only vaguely defined but is generally sufficient to reduce the aw below levels at which most bacteria associated with the raw material will grow and. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). It also demonstrated their significant proteolytic activity during the first month of fermentation. traditional starter culture preparations such as koji (Japan). where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. 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 – 6. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). either directly or as the partially saccharified rice they produce. and the anaerobic conditions that prevail in the bulk of the material. 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.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). however. These preparations contain a mixed microflora of moulds and yeasts.4. This. 4). In another microbiological study of four fish sauces. 6. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7).FERMENTED FISH Where other ingredients are used.3.
6.2.1. Patis (Condiment) 128 . Belacan (Condiment) Fig.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 . 6.12). Fig.
6. thought to be largely the result of autolytic breakdown of the fish tissues. In fish sauce production.FERMENTED FISH Fig. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. however. To produce semi-solid fish pastes as opposed to fish sauces. the 129 . eviscerated fish are sometimes used to slow the autolytic process. 14).3. Ika-Shiokara (side dish) Fig. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13.4. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are.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. for example. filtered and bottled. 6.8. snack) 130 . In contrast to free-swimming fish. 6. Fig. Figure 6.5.3. Hoi-Malangpu-Dong (side dish.5). Some examples of products and their formulation are presented in Table 6.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. 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. which is eventually separated from the residual fish tissues. particularly glutamate. volatile fatty acids and nucleotides.6 – 6.V and Figures 6. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur.
side dish) 131 . N. north. south of Thailand.3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. NE. central. C NE. the prefix 'pla' is replaced by 'kung'.12 months Shelf life Use 1 .V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C.FERMENTED FISH TABLE 6.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0.6. N S 5 . NE 10 . S.12 months Main dish 2 Pla-jao C. C NE. Fig.03 Preparation time 6 .10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 . 3:1:0.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0.4 are also produced using shrimps.7 days 5 . NE Formulation Fish (whole or pieces): salt: roasted rice.2-4 Fish (pieces): salt:khaomark (fermented rice). north-east. In these cases. Kung-Chom (staple food.12 days 3 . N.5-1.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. N.6. C.
(from Arroyo et al. their growth and metabolism are nonetheless slowed. then this will take longer to develop than if lower salt levels are used. This is readily explained by the susceptibility of LAB to elevated salt concentrations.10%. but there is considerable variation in this. Phillipines Journal of Food Science and Technology. 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. giving a reduced rate of acid generation.3. 2.salt and lactic acid. of the two factors contributing to shelf life .1 The use of higher salt levels results in a longer production phase but a better keeping quality product. Red burong isda 132 . the salt is of far greater importance in determining the product’s long-term stability. 106) Fig. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. It also indicates that.7.2. If acidity is a desired sensory property of the product. typically in the range 2 . The LAB isolated from these products are those with more marked salt tolerance but.. these products are salted at lower levels than the fish sauces and pastes. while they are able to grow at high salt levels.VI).6.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content. Generally.
4.8 .3 .8 .6.9 .5 .5.3 5.2 4.9. 106) Fig.5 3.0 Shelf life 3 weeks 6 .5.5 .FERMENTED FISH (from Arroyo et al.8.6.9 4. 133 .17.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16). 2.3 years 1 .6. White balao-balao TABLE 6..8 pH 4.5.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.1 .0 .12 months 1 .7 .0 . Philippines Journal of Food Science and Technology.2 126.96.36.199 4.0 .6 3.5.4 .5.1 4.9 4.8 2.
In som-fak 9% of LAB isolated were able to ferment rice whereas 19% could ferment garlic and omission of garlic from the formulation resulted in lower acid production during fermentation (20. the rice may be added in a partially saccharified form such as kao-mark (Thailand). such as salt content. In several products.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. also lower the unit cost of the product and this is probably a significant factor in many cases. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. For example. a common ingredient in these products.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. inulin.FISH AND SEAFOOD 6.0 . Addition of carbohydrate will. TABLE 6.5) despite other factors. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. Where they are used. When added as boiled or roasted rice alone. Garlic.1 .4.9) compared with the white product (pH 4. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. Rice is the most common carbohydrate source used. This results in a more active lactic fermentation and a lower pH.2. also serves as a valuable source of the more readily fermented carbohydrate. Although the ability to ferment starch is not widespread in the LAB. alternatively. 19). being similar (18.3. 21). nare sushi pla-chao pla-paeng-daeng pa-chao Country Philippines Japan Thailand Thailand Laos Saccharified rice kao-mark Kha-namak Starter culture ang-kak koji ang-kak - 134 . they increase the amount of soluble sugars produced and thus the range of LAB that can grow.VII). although the final population of LAB may be similar. 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).3. of course.
the level of salt employed is on the high side for fish/salt/carbohydrate products.9%. allowing other spoilage organisms to grow.7 to below 4.17. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). which ranged from 7. 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). There is little published information on the factors that limit the shelf life of the low-salt products.5. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. presumably correlating with the salt content.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). In a successful fermentation. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. 6. The growth of yeasts and moulds could. and there is a marked regional variation in the preferred type. 135 .2. the pH drops from 6. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products.5 in the first 2 . The mixture is allowed to ferment for 2 . will not reliably exclude air. reporting the product’s pH to range from 4. Excessive growth and acid production by the LAB could render products too sour for acceptability. in turn. Results of a product survey were consistent with this. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB.3 days. around 12%. Clearly. Traditional packing techniques used with lactic fermented fish products. 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. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). the Korean fermented fish product. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. is something of an exception since millet is used along with garlic as the carbohydrate source. but they are likely to behave similarly to lactic fermented products produced from other commodities.5:9:3. such as wrapping in banana leaves.8 . markedly extending the shelf life (25). lead to an increase in pH. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage.7 up to 6. Generally.2 at the end of fermentation.FERMENTED FISH Sikhae. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. thereafter declining to about 4.3 weeks at around 20 °C. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases.
parahaemolyticus has been reported growing down to a pH of 4. and consumed without cooking. although these limits apply when all other factors are optimal (26). In Japan. After capture. Because of the severity of the illness it causes. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. fish products are the most common source of botulism. As in Norway. During fermentation. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. viable numbers will probably decline quite rapidly. the high salt concentrations preclude the growth of all food-poisoning bacteria and those initially present would not survive well the prolonged exposure to high salt during production. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). which accounted for 88% of outbreaks in the period 1952 .87. 6. outbreaks of 136 . The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and.8 and at salt levels as high 10%. There is little in the way of documented information associating fermented fish products with foodborne illness. 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. however. V. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. but we are unaware of any reports of this happening. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. In the fish sauces and pastes.FISH AND SEAFOOD Products where a substantial population of moulds and yeasts is deliberately added in the form of saccharified rice or a starter culture would spoil as a result of their continuing activity breaking down the rice and producing unacceptable levels of alcohol in the product. typically 30 °C. are Vibrio parahaemolyticus and Clostridium botulinum. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. Several measures have been suggested for reducing the risk of botulism. under conditions inimical to growth and high ambient storage temperatures. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. The bacterial pathogens most commonly associated with the living environment of fish.
. whale meat. Institute of Nutrition. walrus nose. One study with the trematode Haplorchis taichui.M. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. and warm weather can result in rapid growth of C. Kasemsarn B. Where these products are consumed without final cooking. Microbiology and chemistry of fermented fish. 1966. which is generally consumed raw (29). References Riddervold A. Oslo. Greenland and Alaska.FERMENTED FISH botulism have become less frequent in recent years. 137 .. particularly in south-east Asia and the western Pacific regions (30). Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. Puwastien P. Studies to determine the effect of traditional processing techniques on the infectivity of metacercariae was identified as a priority research area by a WHO study group (30). 3. Yhoung-Aree J. Lutefisk. Salting levels are often insufficiently low to prevent growth of C. Mahidol University. 31. and these are often used in the production of fermented fish products. Saisithi P.. Dollar A. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada.6 1. though 27% survival was still recorded (33). Thailand. affecting an estimated 40 million people. 2.O. Raw materials subject to this process include seal flipper. 105-10. Rakefisk and Herring in the Norwegian Tradition. Novus Press. although the absence of any source of fermentable carbohydrate means that the process is putrefactive. 1990. Fermented Fish Products Study (Thailand) Phase 1.. while increasing the acetic acid content of the formulation had little or no effect (32). botulinum and the production of toxin. Liston J. giving an alkaline fermentation. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. Dhanamitta S. Many of these products are described as fermented. and salmon heads and eggs. 1989. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. Foodborne trematode infections are emerging as a major public health problem. Journal of Food Science. botulinum. Baseline Study (1987-1988). probably reflecting greater awareness of the critical factors that determine the product’s safety (27). they could act as an important vehicle of transmission. beaver tail and paw. Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. which will persist into the product. salt content was found to have the greatest impact on the viability of larvae. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31). With this particular organism.. 6.
353-63.A. Im W. Halophiles in Thai fish sauce (Nam pla). Thawai C. 1957. Velankar N..R. 37. 185-92. Itoh T.K. Dhamcharee B. 7. Olympia M. 63. 61-73. in General and Applied Aspects of Halophilic Microorganisms. Thongthai C. Tropical Science. Lee S. Nov. Yasir M. 934-9. 106-8. Microflora of four fermented fish sauces. 1986... Aslam Z..T. Kaneko Y. 1975. 80.. Applied Microbiology. 16. Hiraga K.. isolated from jeotgal.. 6338. Process Biochemistry. Bioscience.R. 43. Biotechnology and Biochemistry. 138 .. 25. 70. 16A. Salinococcus jeotgali spp.. Rattagool P. Some chemical changes in fish silage.. Tanasupawat S. Pakdeeto A.FISH AND SEAFOOD 4. Suntinanalert P.K. Saono S. Journal of Food Technology.. International Journal of Systematic and Evolutionary Microbiology. 1979.. 1395-401. SK33 isolated from fish sauce fermentation. Phil.. Dong-Ho K. Ono H... 14. Constantinides S. Yongsawatdigul J.. Takano M. Orejana F. Crisan E..H. from fermented shrimp paste in Thailand. 124-30. 1976.. Journal of Food Science and Technology. Lim J.. Fermented fish products of south east Asia. 29. 13. Tsunemi M. 141-2.. Jae-Hun K. 643-7. Rodriguez-Valera F. Canberra. 12. a traditional Korean fermented seafood.. Kyong-Haeng L. International Journal of Systematic and Evolutionary Microbiology. Dymsza H. SR5-3 isolated from fish sauce: purification and characterisation.. Sinsuwan S. 1991.. Protein hydrolysate from fish. Takada K. 381-8. 1995. Chung Y. Fukuda H. Government of Australia.R.M. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries. 3649. Tanasupawat S. Plenum Press.. Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme. microbiological and chemical properties of salted and fermented squid.H. Hull R. 11.T.. Journal of Fermentation and Bioengineering. 11.. Journal of Scientific and Industrial Research (India). 2000.. 6. A halophilic serine proteinase from Halobacillus spp. 17. Kudo T. 5. 9. Moonmangmee S.V. 2007. 15.... Ed. Effects of gamma radiation on sensory qualities. Reddi P. 8. Production and characterisation of NaCl-activated proteinases from Virgibacillus spp. Oda K. 57. 1972. 57. 17-30. 10. Namwong S. Nov. 2008.D. Sands A. 3..W.J. Cooke R. Catheptic activity of fish muscle. Lentibacillus kapialis spp. 2006. Journal of Food Science. Yook H. Rodtong S. Byun M.. Journal of Food Protection.. Backhoff P..S. New York. Adams M. Ahn H. Liston J.M... 1985. 2007.. Agents of proteolysis and its inhibition in patis (fish sauce).
1990. 22. 1983..W. Tanaka M.FERMENTED FISH 18. in Microorganisms in Foods.L. 307-14. Food Chemistry. 30. Paludan-Müller C.... Yamprayoon J. Huss H. 138-44.. 426-35. 21. Moody L.. 104-6. 1999. Ostergaard A. 46. 19. 23. 3-10. Tropical Science. New York. 1987. Caldo G. Control of foodborne trematode infections.A.. 1983. Valyasevi R. Fung C. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper. Characterisation of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation. Caldo G. Blackie. 105-12.D. Erikson U. Hauschild A.. Rustad T. 29. 28. 38. Ed. Two outbreaks of botulism associated with fermented salmon roe – British Columbia. Dodds K.K.A.. Epidemiology of human foodborne botulism. 1995. 92. 24...W. Canada Communicable Disease Report. in Clostridium botulinum: Ecology and Control in Foods. 2002... 2002.H. in Clostridium botulinum: Ecology and Control in Foods. 26. Yeast-flora in red burong-isda a fermented fish food from the Philippines. Volume 5: Microbiological Specifications of Food Pathogens.H. burong-isda.. 27. Dawar M. World Health Organisation Technical Report Series. 1-4. 1992. Journal of Food Science and Technology. Kozaki M. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak. Sakai H.L. International Commission on Microbiological Specifications for Foods.. Wainwright R. Gram L. Saisithi P. 25. a Thai low-salt fermented fish product. Journal of Agricultural Science (Tokyo). Marcel Dekker. Eds.H. Wedell-Neergaard C.. Vibrio parahaemolyticus. 28 (6). Huss H... 28.B.. Huss H. Journal of Applied Microbiology.H. New York. Gram L. Journal of Agricultural Science (Tokyo). Paludan-Müller C. International Commission on Microbiological Specifications for Foods.. 1998. London.. 305-22. Visessanguan W. a Thai low-salt fish product. Gram L. Martin J. 3. ASEAN Food Journal. in the Philippines. Sakai H. International Journal of Food Microbiology. Hazards from northern native foods.. Fermentation and spoilage of som fak. 20. Eds.H..B. 2007. Benjakul S. Marcel Dekker. 1815. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation.D.W. Riebroy S. The fermented fish food. 139 . 69-104. Traditional fermented fish products with special reference to Thai products. 219-29...M. 1996. Isaac-Renton J. Owens J. Kozaki M. Ben Embarek P. WHO Study Group. Hauschild A. Hauschild A. 103. 27. Patrick D. Avurhi J..H. 28.. 1992. 274-86. Dodds K.
23. Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom.. Srivatanakul P. Bloemsma B. Kikukawa H. 32... Nout M. International Journal of Cancer. Nithikathkul C. Uchida K. International Journal of Food Science & Technology..R. 15998. Horst K. Ruamsuk A.R. Fermentation and Food Safety. Survival of Haplorchis taichui metacercariae in lab-pla.. 33. 661-70.FISH AND SEAFOOD 31. Jedpiyawongse A. northeast Thailand. 117. Aspen Publishers Inc.7 Further Reading Adams M.H.. 140 . Survival of Anisakis larvae in marinated herring fillets... Todoroki T. Wongsawad C... Chuboon S. 2001. Kittiwatanachot P.. Adams M. Sriplung H. Roepstorff A. Honjo S. Progress in Industrial Microbiology.. 1986.. Hanai S. Huss H. Gaithersburg. 854-60. 110-1.. 29. 2005. 6.J.. Miwa M. Southeast Asian Journal of Tropical Medicine and Public Health. 1995. 36. 2005.R. Deerasamee S. Sripa B.. Fermented flesh foods. Thai traditional food preparation...
most notably the azaspiracids and the cyclic imines as well as some additional polyethers. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. Ph.. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins. Since the original chapter in this series was published (6). produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. a number of new toxins and toxin families have been identified. Nevertheless. several scientific reports have described their efficacy and potential.C. While some of these toxins only occur in seafood from a restricted geographical region.D. Wright. An emerging technology is the use of immunosensors (1 . 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. The manual on harmful marine algae and their products. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. C. 5). including the well known marine toxins 141 . Increasingly sophisticated instrumentation (e. Although most of these antibody-based test kits have yet to receive international regulatory approval.. physiology and detection of marine and freshwater toxins has also appeared (8). and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue.7.M. and a second updated edition dealing with the pharmacology. Scientific and Cultural Organisation) has been updated (7). FISH AND SHELLFISH TOXINS Jeffrey L. It remains to be seen if this approach gains momentum in the future. the export of exotic seafood that may contain such toxins can lead to unexpected public health issues.3) and some practical applications for detecting shellfish toxins have been reported (4.g. known and unknown. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins.
a great variety of shellfish from all latitudes. dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). Saxitoxin Group instead of the Paralytic Shellfish Poisons). 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. and a review on the risk assessment of marine toxins has also been produced (14). and another. 7.1).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.2. Not all the toxins found in shellfish extracts are microalgal products. The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). neoSTX and Gonyautoxins (GTX) 1-4.g.FISH AND SEAFOOD has been produced (9). 7. PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). all of which contain a tetrahydropurine skeleton (Figure 7. can become contaminated with these toxins. Around twenty PSP toxins are known. and a tropical species Pyrodinium bahamense (16). Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. with intoxications occurring in many countries even today (15). Many of these species are extremely common and are found throughout the world in a variety of cold. Gymnodinium catenatum. even crustaceans such as lobsters. and sub-tropical waters and habitats (15). In addition. Consequently. some are the result of biotransformation processes by the filter-feeding host. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11). The Ncarbamoyl sulfates are readily hydrolysed to STX. 142 .1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. All PSPs degrade rapidly in alkaline conditions. 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. temperate.
this is never sufficient to render the seafood safe to consume.30 minutes of consumption. However. 143 . 7. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. Neurophysiological studies have shown that PSP toxins all possess the same mode of action though some derivatives such as STX and neoSTX are considerably more potent than the C-toxins (15). tongue. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7.2.2 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. and humans seem to be particularly susceptible. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. and fingers within 5 . Human symptoms begin with tingling and numbness of the lips. 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. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis.1.
15. After some development and various trials. including Japan. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19).3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. 7. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. and while simple and relatively inexpensive to operate. 23. specifically 144 . One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. a criticism has been that such methods are time consuming. Alternative HPLC methods have subsequently reduced this to a single HPLC run. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation).8 µg/g) of shellfish or seafood tissue. 24) (Figure 7. and over ten analogs are known (22. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. China.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). However it remains as the monitoring method of choice in many countries. 7. Since its discovery. Taiwan. the detection limit (4 µg/g) of the method is close to the safe level and does not provide any information on the profile of PSP toxins present in the extract. other derivatives have been reported. and islands of the South Pacific. Most intoxication events have occurred in South East Asia.FISH AND SEAFOOD 7.3.2). Thailand. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12.2. and has also been evaluated by a number of EU laboratories. 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. Two approaches have been taken here. The mouse bioassay has been in place for decades and the method has been standardised. While the chemical detection methods are most promising for the future. TTX most commonly occurs in many species of puffer fish. as well as other parts. 17).1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. a pre-column method has received Association of Analytical Communities (AOAC) approval (18).
2.2 mg of purified toxin can be lethal. 27). raising speculation that TTX is produced by a bacterial source. molluscs. preventing depolarisation and propagation of action potentials in nerve cells. the blue-ringed octopus.3. 7. 25. 7. All of the observed toxicity is secondary to the action potential blockade. and once again the mouse bioassay has been used to detect the presence of the toxin. gonads and roe (24). Typically this has been accomplished using HPLC followed by post 145 . Structure of tetrodotoxin 7. hypotension. The onset of symptoms usually occurs rapidly.2 Toxicology Like the PSP toxins. 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. and even newts. Severe poisoning causes paralysis and respiratory failure. Since then. and in mild cases results in distal muscle weakness. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. where puffer fish of the genus Fugu are a delicacy.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions.3. and blocks diffusion of sodium through the sodium channel. respiratory failure and coma. nausea and vomiting. An oral dose of 1 . In Japan. horseshoe crabs. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. and more recently by an actinomycete Norcardiopsis dassonvillei (26). A review of TTX detection methods has been published (12. and ingestion of larger amounts leads to cardiovascular effects including bradycardia.FISH AND SHELLFISH TOXINS in the liver. Fig. 26). this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. it is served in speciality restaurants with qualified chefs to properly prepare the fish. 23.
3. Structure of domoic acid and three naturally occurring photoisomers 7. but 146 . These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. but it is known that such congeners are less toxic than DA itself (30). Currently 12 isomers of DA have been reported (see Figure 7. but all of them are considerably less toxic than DA itself (29). Fig.3 for examples). 7. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. Immunoassay methods have also been investigated. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide.4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA).FISH AND SEAFOOD column oxidation to the fluorescent derivatives. Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28).1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). need further development (24). and while promising. 7.4. a polar glutamate agonist. However this method has been criticised for being complex. 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).
but any impairment of the blood brain barrier results in additional risk. and clams.FISH AND SHELLFISH TOXINS only three strains (P. 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.4. cormorants and pelicans (34). Pseudonitzschia australis. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). Levels of DA in the other strains are generally <1 pg/cell. including persistent short term memory deficits (36 . 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. and is mainly distributed in the blood. Recently. Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. southern and western portions of Ireland. 33). 147 . another diatom species Nitzschia navisvaringica. seizures. Victims display a variety of symptoms including headache.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. 7. scallops. which has been well documented in studies along the west coast of the US. and sea animals such as sea lions and even whales (35). Another factor is the movement of the toxin through the food chain. it has been reported that cooking or steaming contaminated mussels merely has the effect of spreading the toxin to other tissues of the shellfish as a result of disruption of the digestive gland. and review of the toxicologic pathology of DA has been published (36).10% of the administered dose. To a great extent. The original 1987 event was followed some years later by a more widespread episode along the western coast of the US and today toxin-producing strains of Pseudonitzschia have been found in the Gulf coast of the US and along the western shores of the US and Canada. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. and this has led to ASP toxicity in birds.. 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 in locations around New Zealand (32. For example. multiseries. as does impaired renal function which increases serum concentration and residence time of the toxin. Okinawa and more northerly parts of Japan. has been reported as a medium-level producer of DA (33). The oral absorption of DA is around 5 . nausea. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. Penetration of the blood brain barrier is poor in healthy subjects.39). the Mediterranean. the Philippines. disorientation and coma. found in the Pacific around Vietnam.
The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA).4. and this is typically the method of choice. and dinophysistoxin-2 (DTX-2) (Figure 7. 48. An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. shellfish tissue can be efficiently extracted with aqueous methanol (1:1).4). In the context of this chapter. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. The benthic Prorocentrum spp. 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. However. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. 7. are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. 41). the heterotrophic dinoflagellates Dinophysis spp. Alternatively. Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. Since then. as can LC/MS.FISH AND SEAFOOD 7. dinophysistoxin-1 (DTX-1). australis. although in the latter study the probes for Pseudonitzschia pungens. DTX-2 was discovered in contaminated Irish shellfish (46).30 ng/g) with a rapid clean-up step using a strong anion exchange resin.. 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). but just over 15 years ago a third toxin.. produce a variety of other 148 . 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. To further complicate matters. and consequently were often overlooked as the real source of the toxins. Dinophysis spp. and an immunosensor method has also been reported (4). the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. and these methods have been reviewed (40. 49). Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). in Monterey Bay.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. Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). California. The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. though this method is most commonly used to analyse plankton and seawater (42). Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp.
Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp.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. although occasionally fatty acyl esters linked through the 7-OH group (referred to as DTX-3 compounds) are also found and are likely a result of bioconversion in shellfish tissue. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. In this case Dinophysis was suspected as the culprit organism. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. Dinophysis caudata. are also recognised as a threat to human health. Dinophysis acuta. and include Dinophysis acuminata. 149 . are recognised as DSP toxin producers. which include the pectenotoxins and the yessotoxins. DTX-5. 7. This was probably not the first DSP episode.5. Fig. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). Dinophysis fortii.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. While these esters are not toxic in vitro. 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.. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. Structure of okadaic acid and the two related DSP toxins 7. and Dinophysis norvegica among others (47. Lately these compounds. Today over ten Dinophysis spp. Prorocentrum maculosum. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. 49). such as Prorocentrum lima. and Prorocentrum belizeanum. In comparison. DSP toxins produced by benthic Prorocentrum spp. The parent acids are the only DSP congeners to be found in shellfish tissue.4.
New Zealand and Chile.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. A regulatory level of 80 . Due to the lipophilic nature of the toxins. although a comparative study of in vitro phosphatase inhibition with mouse bioassay data has shown that OA is about twice as potent as DTX-2 (51).5. and abdominal pain.5. and a detection limit of 100 µg/kg has been reported (12). All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. DSP episodes have been reported worldwide and are included in many monitoring programs (47). 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. 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). even with the minimal of clean up steps (48).3 days. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. DSP episodes have been reported from Japan. and while this provides critical seafood safety information. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. which can lead to an underestimation of the total DSP content in a sample.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . In recent years. and DTX-2. it was agreed that the established mouse bioassay (52) with an observation time of 24 hours is currently the preferred method for the detection of the acute toxicity of acetone soluble DSP toxins. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. 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. nausea. it does not provide information on the DSP toxin profile.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. Human symptoms include diarrhoea. DSP-Check is a commercially 150 . Since the late 1970s. vomiting. 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. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). various European countries. 7. 7. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity.
Within this group. Fig. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness. However. with good agreement with the mouse bioassay and LC–based methods (54.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. for many years these compounds were included in the DSP toxin complex. 7. Some doubt remains as to the significance of these compounds as marine or shellfish toxins with respect to human health since the PTXs for example are co-extracted with the DSP toxins. and another rapid antibodybased test kit for DSP toxins has also been reported (53). As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49).6. 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.5 and 7. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . over 20 PTXs are known (56) and more than 30 YTXs have been reported (57.6 Lipophilic Shellfish Toxins (LST) Lipophilic extraction of toxic shellfish for DSP toxins often results in the simultaneous extraction of other lipophilic polyethers such as the pectenotoxins (PTXs) and yessotoxins (YTXs). fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays. Some representative examples of both groups of toxins are shown in Figures 7. In yet another approach. 58).5. Indeed. 55). but with the increased availability of pure toxins permitting detailed toxicology studies it is now recognised that both these groups of compounds possess a different toxic profile from the DSP toxins. 7. with a claimed detection limit of 20 µg/kg.
which may co-occur in blooms with other organisms. Japan.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). Structure of the two parent yessotoxins. 57.6. eastern Canada. the toxin content and profile may vary among different strains. While 152 . PTX-1 and PTX-2 are frequently the most common derivatives to occur and PTX-2 is the most potent following intraperitoneal (IP) injection (230 µg/kg body weight).FISH AND SEAFOOD Fig. As is typical for many algal species.6. 58).2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). many parts of Europe and the Mediterranean. 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.6. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. YTX and homo-YTX 7. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. 7. 7. New Zealand and South America (49.
but less so following oral dosing (61). For many years it was believed that 153 . Although there is no evidence that yessotoxins are responsible for human illness. 7. it may be compromised by other biological materials in the matrix.6. The structures and chemical data including the molecular weight of each member have been collated (56). 58).FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. some of which are found as glycosides and sulfates. 7. though its cell effects may be linked to modulation of calcium channels (62). but a fluorescent derivative has been prepared for others. Many YTX toxins contain a conjugated diene system. and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative.7). Estimates of acute toxicity of the parent compound following IP injection vary considerably. chemical analysis is best carried out by LC/MS and several approaches have been described (57.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. However. a regulatory level of 1 mg/kg shellfish tissue has been set. Some of the PTXs are potent cytotoxins in vitro (60). 64). The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds. which aids detection by LC/UV. the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present. the PTXs show considerably less toxicity when administered orally. Their toxicology and mechanism of action remain uncertain. There are no clear cases of human illness resulting from ingestion of PTXs (59). Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. but various congeners are acutely toxic in the mouse bioassay following IP injection. The YTX group has not been reported to cause human illness.3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue. Because of the number and complexity of the YTX group. The crude extract may be used directly in the mouse bioassay. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7. which is further compounded by differences in the experimental design. While LC/UV detection is possible for PTXs. 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.
and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf. the same toxins have been reported to be produced by three genera of Raphidophytes. oysters.FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida. and cockles (65. verruculosa off the mid Atlantic coast of the U. 7. PbTx-1 and PbTx-2. Structure of the two parent brevetoxins. brevetoxins are found in mussels. Many of these species are associated with massive fish kills.7. 66).1 Origins and distribution The brevetoxins are produced by the dinoflagellate Karenia brevis (formerly known as Gymondinium breve and Pytochodiscus brevis) a dinoflagellate found in the Gulf of Mexico. 66). In the Gulf of Mexico. 154 . namely Chatonella marina. but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65.7. Interestingly. 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”.S. They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. was chemically characterised as PbTx-2 (67). which are found in Japanese waters (66). These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). whelks. Fig. Fibrocapsa japonica. and off the coast of New Zealand (65). and one of the brevetoxin metabolites BTX-B2 7. and Heretosigma akashiwo.
8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. While the mouse bioassay continues to be used. In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. 7. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . and diarrhoea. the competitive ELISA method correlated most closely with the mouse bioassay results. 69). 7. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. which can result in a cascade of associated events in a cell (66). Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. The mouse bioassay is used as the primary detection method. this family of toxins has proven to be a challenge both to identify and monitor. The mean time to onset of symptoms is around 3 hours.7. with hot spots in the coral reef areas of the Caribbean and French Polynesia. vertigo.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). and in severe cases seizure. 7. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). shellfish homogenates. Ingestion of brevetoxins results in a variety of symptoms including parathesia. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. malaise. nausea.7. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73).FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). 66. The human health implications following exposure to NSPs have been reviewed (69). and mammalian body fluids without pretreatment or dilution (72). In this application. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). 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. with a mean duration of symptoms of around 1 day.
8). Fig.1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins. complex. a group of closely related polyethers that are considered the source of ciguatoxins.FISH AND SEAFOOD transferred through the food chain to the larger carnivores. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins. and only a few structures have been identified. green and brown seaweeds and also occur in sediments and coral debris. 7. CTXs are large. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7.8. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red. These include species belonging to the genera Ostreopsis and Coolia. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. The diversity of benthic phytoplankton that exist on reefs further complicates the picture. Distribution through the food chain begins with 156 . most likely a consequence of biotransformation of the parent toxins as they pass up the food chain.8. lipid soluble polyethers. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. but more are suspected to exist. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin. A detailed comprehensive review of ciguatoxins has recently been published (74).
groupers. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. sharks. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. Local island populations attempt to avoid poisoning by catching only smaller fish. only trace amounts of CTXs can cause illness following consumption of contaminated fish. which can return following the consumption of alcohol and certain foods such as oily fish (74. there are neither standards nor an official testing program. myalgia. weeks or even months (74 . and there are suggestions that such incidents are increasing and spreading worldwide (74). Other symptoms include a metallic taste. 7.8. and temperature sensation reversal. 75). A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . and in fact CTX poisoning is often only identified after consumption of contaminated fish. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. where cold objects feel hot and vice versa.1 µg of ciguatoxin can result in human illness (74. which in turn are consumed by carnivores such as snappers. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. The recommended method to extract fish tissue has been described in detail (74) and the initial warm acetone extract is partitioned against hexane and diethyl ether where the CTXs are concentrated. barracudas.76). all of which can be linked to the effect of CTX on excitable membranes. and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. although not all CTXs are equipotent. Patients have reported symptoms lasting weeks and even months. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. and although the gastronomical problems may dissipate within a day or so. Like the brevetoxins. These include paresthesias (numbness and tingling of the extremities).FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish.3 Detection methods As a consequence of their potency. 78). The onset of symptoms can occur rapidly within 24 hours. Another symptom that can develop over time is extreme fatigue.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. 75. Even when suspect samples are available. and the sensation of loose teeth (77). and it is estimated that ingestion of as little as 0. arthralgia. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. the neurological symptoms can persist for days. 7.8. which can be confused with chronic fatigue syndrome. and moray eels. Globally. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. pruritus.
Acetone extracts of fish tissue are subjected to solvent partitioning. but this only has application in a clinical setting (81). A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine. at C-3 and C-23). In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74).FISH AND SEAFOOD Lethal Dose (MLD) 0. following consumption of contaminated mussels Mytelius edulis (82). another in vitro sodium channel-based assay. Immunoassays may be an alternative detection method (79).9).80%). Usually the major AZA derivatives found in shellfish are AZA1 (50 . although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). has been developed. and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. which are based on an LC-MS/MS approach.9). This is then subjected to LCMS/MS analysis. 158 . and following synthesis of the molecule some time later. Chemical detection methods are severely compromised by the lack of standards. Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. minor revisions of a few stereocenters were made. high-throughput screen (74). Consequently. which features characteristic fragment ions corresponding to sequential losses of three molecules of water.25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required.20%) present in lesser amounts (Figure 7. with AZA2 (10 . and the corrected structure was published in 2004 (86) (Figure 7. Recently.9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. followed by clean-up on silica and aminopropyl cartridges. and only a few structures (AZA 1 . and another dealing with the chemistry and detection methods has been published (85).30%) and AZA3 (5 . 84). 7. almost 20 additional derivatives have been reported (83 . only a few laboratories are able to undertake such analyses. Recent reviews of this new group of toxins have appeared (83.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. Since the original discovery and identification of the toxin.g. The majority of these have been proposed based on LC/MS analytical data.85). The original structure proposed revealed an entirely new class of shellfish toxin. specific for CTX activity.
and to date this has been the only source from which these toxins have been found at toxic levels. but current data indicates that unlike the DSP compounds. The bivalves affected include mussels (Mytilus edulis. and to date no long-term effects have been recorded (83.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. Scandinavia.3 days. 7. Donax spp. diarrhoea and stomach cramps.). cassipes (83). Ostrea edulis). it has been impossible to establish that P. Portugal.FISH AND SHELLFISH TOXINS Fig. and most recently in a crustacean (Cancer pagurus). clams (Tapes philipinarum. these molluscs were never considered to be the biogenetic source. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. AZA toxins do 159 . which can persist for 2 . AZA occurrence has been reported by several European countries including France.9.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea.9. cockles (Cerastoderma edule). There has been considerable debate over the source of azaspiracids. Following the original episode. Ensis siliqua. and off the coast of Morocco (83. 90). In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. vomiting.9. 89). Mytilus galloprovincialis). it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). 7. which are ingested by P. scallops (Pecten maximus). These symptoms are not unlike those experienced following the consumption of DSP toxins. However. and some time after the original episode in Ireland. 88. as in other cases of shellfish toxins. 84. oysters (Crassostrea gigas. cassipes produces AZAs in culture. While this result is not necessarily in debate. Structure of the three principal azaspiracids found in shellfish 7.
Once again. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . Importantly these pathological outcomes were reported as being different from those caused by DSP. and ASP. abnormalities in the thymus and spleen. and erosion and bleeding in the stomach. and closely related ptierotoxins (Figures 7. though the possibility exists that they may inhibit other phosphatases. Based on IP minimum lethal dose data. and necrosis of epithelial cells on the microvilli (91). AZAs also display cytotoxicity towards various mammalian cell lines (83. 7. the spirolides. PSP. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. the lack of sufficient material and other logistical problems have prevented complete studies of any chronic effects arising from long term exposure to low levels of the AZA toxins. parenchymal cell pyknosis in the pancreas. but all contain a spirocyclic or cyclic imine function within their structure. 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. However. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg). The mouse bioassay has been used to monitor shellfish extracts.9. which are particularly amenable to detection by positive ion electrospray mass spectrometry.10 – 7. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. 7. 85). Using the same approach. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. 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. 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). the pinnatoxins.12).10 Cyclic imines These toxins are represented by several distinct structural types.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. 91. 92).
10. 7.. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . 161 . two additional congeners. At the time of the initial observation of gymnodimine. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin. 95). Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94. Pacific oyster.1.1 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. surf clam and New Zealand abalone. In only two cases (gymnodimines and spirolides) has a planktonic source been identified.520 Da (94. scallops. 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. Some time later. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. but they are included here for completeness. cockles. they are sometimes referred to as emerging toxins of uncertain human health concern. Subsequent taxonomic studies revised this again to a new dinoflagellate K. 7.10. selliformis (97). Structure of the cyclic imine gymnodimine 7.FISH AND SHELLFISH TOXINS health.10). Fig. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. 95).10. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). 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.
and so spectroscopic detection by UV methods is not favoured. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. Based on this diminished oral toxicity. Oral administration of the toxin to mice by gavage resulted in about an 8.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds. it has been stated that regulation against gymnodimine is not required (98).FISH AND SEAFOOD 7. A later study using purified toxin established an LD50 dose (Lethal Dose. The time lapse between ingestion and death is between 5 and 15 minutes. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay.10. Since their original discovery which resulted in the characterisation of spirolides A-F (99.10.1. Most cyclic imines lack an extended chromophore. 7. the first step in the process is typically an initial clean-up step using a short C18 cartridge and elution with water containing increasing amounts of methanol.1. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode.100). and this is the detection method of choice. There is a rapid onset of symptoms including respiratory distress. finishing with 100% methanol.to10-fold reduction of toxicity (LD50 600 . 7. Reduction of the imine group eradicated the toxicity. However. together with a lack of evidence of human intoxication. almost a dozen derivatives in total have been identified to date (94. Whether isolated from dinoflagellate cells directly or from shellfish tissue. paralysis of the hind quarters and eventually complete immobility. and lack of responsiveness to stimuli. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection.3 Detection methods The characteristic “fast-acting. They were originally discovered in extracts of mussels (M.11). edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. 50%) of 96 µg/kg. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 .10. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. 95). 95) (Figure 7. abdominal breathing.900 µg/kg). In particular. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food.
While oral toxicity is considerably less than by IP injection.707 µg/kg.2. by gavage (87 . They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . All data reported here are for desmethyl spirolide C. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98).10.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia.11. the western coast of Scotland. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). 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. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 .8 µg/kg). Interestingly. 7.11. Structure of four spirolide toxins A-D 7. southern Ireland. fasted mice).166 µg/kg. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada.2. Fig. the gulf of Maine. fasted mice). 95).10. 7.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). or by oral ingestion (381 . in fjords in Denmark and Norway. those spirolides possessing the vicinal dimethyl grouping (see Figure 7.2 Toxicology The spirolides are the most studied group of the cyclic imines. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins.
10. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7.12. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C.FISH AND SEAFOOD (100).3 Detection methods As discussed for gymnodimine. While the mechanism of action remains to be clearly defined. A number of methods following this procedure have been described (94. consistent with modulation of acetyl cholinergic receptors (103). 7. the mouse bioassay can be used to detect these fast acting toxins.10. which includes two diastereomers. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. To date.12). Fig. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . and in fact these compounds share a number of other structural features.B contain a spiro imine moiety similar to that found in the spirolides. four members of the group have been identified – two of them being the C-34 diastereomers. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. Three members of this group have been reported.3 Pinnatoxins and pteriatoxins Pinnatoxins A . 7. 102). However.2. 7. 95. 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.
3 Detection methods No formal detection methods for these compounds have been reported. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures.10. Despite this progress.100 µg/kg). but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. while pinnatoxin D was essentially inactive. Only the acute toxicity in mice following IP injection has been reported.FISH AND SHELLFISH TOXINS 7.2 Toxicology There is little information describing the toxicology of these compounds (98). 7. 7. However.3.10.most likely a dinoflagellate.3. there is no compelling evidence to suggest that they have resulted in human toxicity (98).1 Origins and distribution Shellfish of the genus Pinna. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). and more advanced means of detection. While this is recognised as a real possibility for a wide range of toxins. arise from an algal source . like the gymnodimines and spirolides. Coupled with this. The LD99 (Lethal Dose.10. 7. There is strong circumstantial evidence to suspect that these toxins.11 Conclusion There has been considerable progress in the detection of known marine toxins. 99%) values for pinnatoxins A-C are in the range (22 . but this remains to be conclusively established (106). This is generally a result of increased monitoring and continually improving detection technologies and methods. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. have long been suspected of causing human illness after ingestion. which in many cases are now found to be more widespread than previously believed. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins.180 µg/kg). With increasing knowledge of the chemistry of toxins. 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. The LD99 dose for the pteriatoxins is reported to be in the range (8 .3. new toxin groups such as the azaspiracids and cyclic imines have also been reported. which occupy tropical waters around Okinawa and the south east coast of China. to date action or 165 .
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Naar J. Gandini C.. Botana L. Boca Raton. Squicciarini D. Lenzi A. Ed.. 179-85. Ciguatera fish poisoning: treatment prevention and management.. 68.. Toxicon. Bourdelais A. Poon-King T. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. Literature review of Florida red tide: implications for human health. Backer L.P. Butera R. 75. Baden D.S.S.W. Locatelli C. Toxicon.. Bourdelais A.. 707-23.S.A.H. in FAO Food and Nutrition Paper 80. 99-115.. 3. Neurologic Shellfish Poisoning (NSP).. Clark R.C. 110. Botana L. Jacocks H. Fernandez M. Dickey R. 131.. 2008. 69.E. 2004. Wang Z. Johnson D.M. Watkins S. Granholm A. Reich A. Pierce R... Food and Agriculture Organisation.A.. Food and Agriculture Organisation..FISH AND SHELLFISH TOXINS 65. West Indian Medical Journal. Bottein M-Y.. Kirkpatrick B. Freidman M. Schrank K. 71. Manzo L.. Chest... in Seafood and Freshwater Toxins. The molecular and integrative basis to brevetoxin toxicity.. 1598-603. Harmful Algae. Dechraoui M-Y.D. Aerosolised red tide toxins (Brevetoxins) and asthma. Zaias J.. 6. Fleming L. New fish-killing alga in coastal Delaware produces neurotoxins. Kirkpatrick B. 66. Food and Agriculture Organisation of the United Nations.. Wanner A.. 2008. toxicology.. Ed. 76. and detection.A. CRC Press. Muscle Nerve.E.S.G. 50. Weisman R.. Kubanek J. shellfish and mammalian body fluid.J.. Ramsdell J. Boca Raton.C.G. 73.M. 2008... Ed. 825-34. 2007.B..... Fleming L.. Rome. Lancaster J.E. 50. Abbott J. 479-516.. Baden D. 2002. Granade R.E. 72.. Poli M. 117-71.. Baden D. Benson J. Naar J. 2004. Ramsdell J. Baden D.G... Ed. Tomas C... Whitney P. Poon-King C. 2002.N. Henry M. 23. 220-6.. Cheng Y. Naar J. are potent ichthyotoxic neurotoxins that accumulate in fish. Flewelling L.R..M.G. Bossart G. Ciguatera toxins: Chemistry.. Kubanek J. Tomas C.. Naar J. 74. 519-50. 2008.. 171 . 77... Marine Biotoxins. Landsberg J. Steidinger K.. Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2.. Pierce R. Pierce R. Backer L. Prockop L. 465-70. Flewelling L. Marine Drugs. 551-8..E. Chen A. Wolny J. Abraham W. CRC Press. Bienfang P. Reich A.. 2007. Botana L..G. Mild ciguatera poisoning: case reports with neurophysiological evaluations.. Bean J. 2004.J. Abraham W. 70. Gannon D...P. Cheng Y. Buonocore M.. Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater.. 67. Environmental Health Perspectives. 187-94. Environmental Health Perspectives. in Seafood and Freshwater Toxins. Baden D. Ayyar R. Brevetoxins... like ciguatoxins. 2000. in Seafood and Freshwater Toxins..M... Fleming L.. Boca Raton. 53. 2007. Detection of brevetoxins in the twenty-first century.A. 456-79...M.. Zaias J...P.. 110. Dickey R.D. Backer L. CRC Press.
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.
procedures. Establish a system to monitor control of the CCP. hygiene managers. It is recommended by the Codex Alimentarius that the practical application of the HACCP principles be approached by breaking the seven principles down into a 12-stage logic sequence. ingredient and packaging buyers. 2. Establish procedures for verification to confirm that the HACCP system is working effectively. Determine the CCPs.A point. 6.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. 4. 5.The application of methods. Each stage is discussed below in detail.Obtaining evidence that the elements of the HACCP plan are effective. procedure. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. 8. Establish the critical limit(s). from primary production to final consumption.3. tests and other evaluations. Verification . 8. quality assurance or control. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. Conduct a hazard analysis.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . food chemistry or engineering.1 is a flow diagram illustrating this 12-stage logic sequence. food microbiology. Establish documentation concerning all procedures and records appropriate to these principles and their application. The team should also include 177 . 7. operation or stage in the food chain including raw materials. 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. Validation . in addition to monitoring to determine compliance with the HACCP plan.1 Assemble HACCP team HACCP requires management commitment of resources to the process. Figure 8. 3.
Fig. it may be obtained from external sources. If expert advice is not available on-site. biological. 8.FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations. 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.g.1. Logic sequence for application of HACCP 178 . chemical and/or physical).
If the processing does not include a kill step. such as enteric viruses in shellfish.).3. The description should include information such as the product name. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation.3. However. freezing. storage and distribution conditions and instructions for use.3. pH. aquaculture drug residues and potentially pathogenic Vibrio spp. such as children or the elderly. 8. processing conditions (e. is a cooking process required?).g. packaging.3. marine toxins associated with particular fish and shellfish species.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora.g. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice.2 Describe the product It is important to have a complete understanding of the product. 8. 8. It is also important to identify the consumer target groups.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. which should be described in detail.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. In addition. or in aquaculture operations. fermentation. Vulnerable groups of the population. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments. etc. including practices onboard fishing vessels. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . shelf life. physical and chemical structure (including water activity (aw). there may be species-specific hazards related to certain fish and shellfish. contaminated fish and shellfish should not be used.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. Expertise on such inherent hazards.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.3. 8. etc.3. heat treatment.1. These factors are likely to have a significant effect on some potential hazards.). naturally present on fish from warm waters (> 15 °C). challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. composition. may need to be considered specifically.
and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards. The team should then conduct a hazard analysis to identify which hazards are of such a nature that their elimination or reduction to an acceptable level is essential to the production of safe food. time/temperature data. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. survival or multiplication of pathogenic microorganisms. ingredient specifications. it should also include details of CCP steps.3. and identify any measures to control identified hazards The HACCP team should list all hazards that may reasonably be expected to occur at each step in the production process. and therefore a detailed description of the procedure applied must be available. conduct a hazard analysis. or circumstances of any reclaim or rework activity.3. 8. If HACCP studies are applied to proposed new process lines or products. once determined. 8.6 List all potential hazards associated with each step. The operation should be observed at all stages. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised. However. because the hazard analysis and decisions regarding CCPs are based on these data. For example.3. 180 ● ● .5 On-site confirmation of the flow diagram The HACCP team should confirm that the flow diagram matches the process that is actually being carried out. it is essential that the details of each step are fully appreciated and recorded. It is essential that the diagram is accurate.FISH AND SEAFOOD product. It may include details of the factory and equipment layout. cleaning and hygiene procedures and storage conditions. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. features of equipment design. 8. the qualitative and/or quantitative evaluation of the presence of hazards. Ideally.4.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters.
1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products. Note: it is important at this stage that no attempt is made to identify CCPs. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. Therefore it is not possible. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual. and there are an equally varied range of potential hazards associated with them. Hazards introduced during processing of fish products depend very much on the characteristics of the process. Pathogens may be part of the resident microflora of the living animal (e.6. Salmonella spp. botulinum.g. For example. and viruses).). There may also be inherent hazards from parasites such as the roundworm Anisakis simplex and from marine toxins such as ciguatera in reef fish (derived from microalgae) and scombrotoxin (biogenic amines) development in fish containing high levels of histidine. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage. the number of consumers potentially exposed and their vulnerability. 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. 181 . For example. since this may interfere with the analysis. One control measure may be relevant to several process steps. the hurdle effect.3. The incidence of psychrotrophic. any food safety objectives or manufacturer’s food safety requirements. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards. or may originate from polluted water or from post-capture contamination (e.g. or desirable. where a hazard is repeated.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. and the potential growth of Listeria monocytogenes on smoked fish. The HACCP team should then determine what control measures exist that can be applied for each hazard. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. 8. Vibrio spp.
and the reader is referred to the appropriate product chapter. More than one critical limit may be defined for a single step. structured approach to decision making.8.3. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. biological or sensory parameters. it is not possible to generalise on likely CCPs. Where possible.7. and its use may not always be appropriate. However. it is necessary to specify both time and temperature for a thermal process. However. 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. given the enormous variety of fish products and processes in use.FISH AND SEAFOOD 8. 8. chemical. For example. 8. If a significant hazard has been identified at a step where control is required for safety. then the process must be modified to include a control measure. critical limits should be specified and validated for each CCP.3.2) to provide a logical.3.8 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. application of the decision tree should be flexible. 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 .1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits.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.3. Criteria used to set critical limits must be measurable and may include physical.1 Fish and seafood Again.
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.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
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.
1992. Oxford. International Journal of Food Microbiology.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H. Blackwell Scientific Publications. 3 (1).H. 1992.. 188 .L. Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety. Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. 33-44. 15 (1/2). Geneva. Development and use of the HACCP concept in fish processing. Mayes T. 1988. Bryan F. 14-9. WHO. Food Control. 1992. World Health Organisation. Microorganisms in Foods. International Commission on Microbiological Specifications for Foods. Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality.
Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . under which food is produced to optimise public health and to prevent. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. however. 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. including microbiological criteria. from structural conditions in the factory or process facility. The new legislation was designed to establish conditions. A number of standards are provided in food legislation. and one that plays an important role in most countries’ food legislation. EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr. to sale or supply to the final consumer. final product specifications. transport and delivery vehicles requirements. to personnel requirements. the existence of microbiological standards alone cannot protect consumer health. eliminate or acceptably control pathogen contamination of food.9. with the intention of introducing consistency and clarity throughout the food production chain from primary production. have been replaced by several linked measures on food safety rules and associated animal health controls. and conditions of raw materials. Hygiene is a general concept that covers a wide subject area.1 Introduction Hygiene is an important aspect of ensuring food safety. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials. 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.
as last amended by Regulation (EC) No 806/2003. handling and offering for sale or supply to the consumer. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. p. Subsequently. 9. N. p.15). 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. p. 41). transportation. A number of more detailed implementing and transitional measures have been adopted at EC level. 24. processing. 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. 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.2 Legislative Structure From 1 January 2006. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. and sets out what those enforcing the provisions have to do.1991. distribution. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No.1). storing. J. 190 • • . Regulation 854/2004 relates to the organisation of official controls on products of animal origin.9.1992. as last amended by Regulation (EC) No 806/2003.B.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. European Communities L 268. 24. existing hygiene Directives including those below were repealed: • Council Directive 91/492/EEC of 15 July 1991 laying down the health conditions for the production and the placing on the market of live bivalve molluscs (Off. 7.9. packaging.7.1991. 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. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. manufacturing.
Additionally the requirements of EC Regulation 853/2004 must be complied with.I. 2073/2005. domestic preparation or handling. national legislation in the form of a Statutory Instrument (S. 852/2004.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. of stock farming. 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. etc. 2006 No.g. revocation of existing implementing legislation. There are some exemptions. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. and of hunting and fishing. right through to the implementation of procedures based on HACCP principles. 19.14.1993.7. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. Although the regulations have the force of law. 1 .) in England.I. 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. is required to give effect to the EU regulations. The Food Hygiene (England) Regulations 2006 (S. the legislation is directly applicable law. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. and binding in its entirety on all member states from the date of entry into force. food storage that is for private or domestic consumption. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. Wales and Northern Ireland. penalties and powers of entry.11) The EU hygiene regulations apply to all stages of food production including primary production.3 Regulation (EC) No. 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. for example. As regulations. which will be covered later in this chapter. Primary products are defined as products of primary production including products of the soil. and equivalent legislation in Scotland. with primary production. 9. 191 . This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. Wales and Northern Ireland). setting offences. p. e.
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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: 188.8.131.52 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.
184.108.40.206 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.
220.127.116.11 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.
18.104.22.168 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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22.214.171.124 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.
126.96.36.199 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.
188.8.131.52 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.
184.108.40.206 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. 220.127.116.11 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.
18.104.22.168 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 22.214.171.124 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). 126.96.36.199 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:
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. Conditioning means the storage of live bivalve molluscs coming from class A production areas. or in natural sites. live tunicates and live marine gastropods. heading. shucking. and chopping. to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing. in particular as a result of feeding on plankton containing toxins. Fresh fishery products means unprocessed fishery products. reptiles and frogs) whether wild or farmed and including all edible forms. 197 . and all mammals. 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. slicing. after preparatory work such as bleeding.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. such as gutting. to remove mud. heading. slicing. purification centres or dispatch centres in tanks or any other installation containing clean seawater. whether whole or prepared. where necessary. shelling. live echinoderms. gutting and removal of fins and. parts and products of such animals. filleting. mincing or processing. followed by wrapping or packaging. if necessary: chilling or freezing: filleting. 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. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness. where appropriate. including products packaged under vacuum or in a modifiedatmosphere that have not undergone any treatment to ensure preservation other than chilling. slime or sand. skinning.
Extracts of the requirements of Annex II. please refer to Regulation (EC) 853/2004.1.FISH AND SEAFOOD 9. (ii) However. In this case. 198 . Section I of Regulation (EC) 853/2004. Section I of Regulation (EC) 853/2004 are given as follows: 9. 9.4. for other codes. EÜ.2. (iii) Food business operators must have in place systems and procedures to identify food business operators from whom they have received and to whom they have delivered products of animal origin. the code would be UK. 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.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. (iii) Additionally. EC. and the characters easily decipherable and be clearly displayed.1. the mark must show the approval number of the establishment.4. the new mark must show the approval number of the establishment where this occurred. a new mark must be applied to the product. EB or WE.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. EK. the food business operator may apply the same identification mark to both types of food. EF. the mark must be oval shaped and include the abbreviation CE. EY. (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. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. (iv) If the identification mark is applied in an establishment within the Community. when a product's packaging and/or wrapping is removed or is further processed in another establishment. For the UK. EG. EO.2 Form of the identification mark (i) The mark must be legible and indelible.4.2.2. EK. ES.4.2 Fishery product requirements 9.
9. it applies to those animals when not placed on the market live. or be printed on a label affixed to the product. a) requirements for establishments. Part A and Chapter V apply to retail. echinoderms.4. the mark may be applied to the outer surface of the container or packaging. Chapter III. granulate and powdered products of animal origin carried in bulk. the approval number of the establishment and the abbreviation for the community where appropriate. they would need to be obtained following Section VII on live bivalve molluscs. Section VIII of Regulation 853/2004. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . wrapping or packaging in another establishment. processing. reference should be made to the actual Regulation. Parts A. This section does not apply to bivalve molluscs. specifically relating to fishery products are given below.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. its wrapping or the packaging. (ii) For products of animal origin that are placed in transport containers or large packages. (iii) In the case of liquid. i. including vessels.LEGISLATION 9. Extracts of the requirements of Annex III. and are intended for further handling.1. as amended. for full requirements. The requirements of this section accompany those in Regulation (EC) 852/2004.2. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. and fishery products carried in bulk. (v) When the mark is applied directly to products of animal origin. the wrapping or the packaging. tunicates and marine gastropods when placed on the market live. it is sufficient to apply the mark to the exterior of that package only.4.3 Method of marking (i) The mark is to be applied directly to the product.e. Therefore. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code.2. C and D. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. the colourings used must be EC authorised colours. With the exception of Chapters I and II. Chapter IV. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer.
clean seawater may be used for the handling and washing of fishery products. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter. There is a derogation from point a. Chapter VII of Regulation 852/2004. removing fins. gutting. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc.1 Chapter I .2. heading. Structural and equipment requirements refer to the following points 1. (a) primary production covers the farming. The definition of fishery products covers primary production as well as its associated operations.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. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 .2.e.. 2. if a vessel has water coming in that is used with fishery products. this supplements the requirements of Annex II. A. from the place of production to the first establishment of destination.FISH AND SEAFOOD c) for the water supply.) 9.4. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material. i. bleeding. refrigeration and wrapping. it must be placed somewhere that will not contaminate the water supply. or to handle or process them after harvesting. fishing and collection of live fishery products with a view to their being placed on the market. 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.
to ensure that melt water does not stay in contact with the products. where necessary.LEGISLATION (i) Such vessels need to have holds. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. any waste processing unit needs to have a separate hold to store the waste. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. Storage areas for the finished products also need to be sufficiently large and cleanable. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. and not more than 0 °C after 16 hours. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. A temperature recording device must be present and placed where the temperature in the hold is the highest. Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C. (ii) Holds must be separated from the engine compartments and from the crew quarters by partitioning that prevent any contamination of the stored fishery products. (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. 4. 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. 3. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. and allow the monitoring and. recording of temperatures. where necessary. Also. 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. (iii) In vessels equipped for chilling fishery products in cooled clean seawater. 201 .
other than those kept alive. 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. e) In cases where whole fish intended for canning is frozen in brine. 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 product must be kept at no more than -9 °C. if this is not possible. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption. must be chilled or. chilled and wrapped. (ii) In the case of food business operators responsible for auction and wholesale markets etc. c) Fishery products.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. where fishery products are displayed for sale. B.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. Chapter II .2. provided no damage occurs to the flesh. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. landed as soon as possible. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. and products washed thoroughly.2. The brine must not be a source of contamination. Handlers may use spiked instruments to move large fish or fish that might injure them. 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.4.Requirements during and after landing 9.
They may also continue to be transported in cooled water after landing and be transported from aquaculture establishments until they reach the first establishment on land that carries out an activity other than transport or sorting. handling fishery products A. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. They should be wrapped. 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.3 Chapter III . and where necessary. (ii) Heading and gutting must be carried out hygienically. B. they must be stored under ice in appropriate facilities. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. part I. points 1 and 2. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices.2. including vessels. Chapter I. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice.4. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises.Requirements for establishments. 854/2004. Fillets and slices must then be removed from work tables as soon as possible after preparation. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . (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. prepared or processed immediately after arrival at an establishment on land. C. packaged and chilled as quickly as possible after preparation.LEGISLATION consumption. dispatched.2. Additionally. 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. 9. C. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed.
(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. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. except when supplied to the final consumer.4 Chapter IV . 9. D. (iii) sprat.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.4.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. they must be cooled rapidly. (iv) (wild) Atlantic and Pacific salmon.2. (b) fishery products from the following species. If no other method of preservation is used. (ii) Compliance with the above is not necessary if epidemiological data indicates that the fishing grounds of origin do not present a health hazard in relation to the presence of parasites and this has been authorised by the competent authority. and (c) marinated and/or salted fishery products. or a stabilising treatment. (iii) A document from the manufacturer. if the processing is insufficient to destroy nematode larvae.Requirements for certain processed fishery products A. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . (ii) mackerel. Once produced. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. cooling must continue until a temperature 204 . stating the type of process they have undergone.2. 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. raw materials must have no guts. must accompany fishery products referred to above when placed on the market.
Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. pressing. B. B. and the raw material is processed within 36 hours after loading. there is an exception: food business operator may refrain from chilling the fishery products when whole fishery products are used directly in the preparation of fish oil for human consumption. raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. processing. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. However. Shelling/shucking is to be done hygienically.2. centrifugation. the following conditions need to be met: A. refining and purification steps before being placed on the market for the final consumer.LEGISLATION approaching that of melting ice is reached. Organoleptic properties of fishery products An organoleptic examination of fishery products must be carried out to ensure that fishery products comply with any freshness criteria (such criteria have yet to be set). provided that the freshness criteria are met and the total volatile basic nitrogen (TVB-N) value of the unprocessed fishery products does not exceed the limits set out in point 1 of Chapter I of Section II of Annex II to Regulation (EC) No.4.2. 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. and after this. C. 2073/2005). 205 . (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. 9. 2074/2005).5 Chapter V . 2073/2005.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. 2074/2005.
6 Chapter VI .2. prepared and processed fishery products belonging to the family Gempylidae. 9. 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 .Storage of fishery products (i) Fresh fishery products. 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.FISH AND SEAFOOD D.2. Fishery products contaminated with parasites may not be placed on the market for human consumption. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. 9.2. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. it should be kept at no more than -9 °C. and when fishery products are wrapped on board fishing vessels. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. The scientific name must accompany the common name on the label. E. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market.4.2. thawed unprocessed fishery products. In the case of whole frozen fish in brine intended for the manufacture of canned food. Molidae. Fresh. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae.7 Chapter VII .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.4. Diodontidae and Canthigasteridae. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing.
(c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. melt water must not remain in contact with the products. 9. species belonging to the Gadidae family.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability.2.2. Helicolenus dactylopterus. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp.2.4. 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. 9.8 Chapter VIII . The temperature of frozen fishery products must not fluctuate above -15°C.4. (d) 60 mg of nitrogen/100 g of whole fishery products used directly for the preparation of fish oil for human consumption as referred to in the second sub-paragraph of Part B(1) of Chapter IV of Section VIII of Annex III to 207 . 2074/2005. Sebastichthys capensis. (ii) If fishery products are kept under ice.3 Other information: Rules relating to visual inspections for the detection of parasites in fishery products are stated in Annex II (obligations on the competent authorities) of Regulation (EC) No. 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. provided that the journey is short and the competent authority allows this. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated.).Transport of fishery products The above points for Chapter VII apply in relation to transport. species belonging to the Merlucciidae family.
for full requirements. this Section also applies to live echinoderms. Section VII of Regulation 853/2004. The sample must consist of about 100 g of flesh.4. they supplement the requirements of Annex II to that Regulation.1 Chapter I . reference should be made to the actual regulation. and point 3 of Chapter VII applies to retail. VI. taken from at least three different points and mixed together by grinding.4.FISH AND SEAFOOD Regulation (EC) No 853/2004. as amended. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. 9. specifically relating to bivalve molluscs are given below. (b) In the case of other operations. The requirements of this Section accompany those laid down in Regulation (EC) No 852/2004: (a) In the case of operations that take place before live bivalve molluscs arrive at a dispatch or purification centre. 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. (b) and (c) of Part B(1) of that Chapter. VIII and IX. 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. please refer directly to the regulation.3 The regulation details specific hygiene requirements for bivalve molluscs. Chapter IX applies to pectinidae harvested outside those areas. however. they supplement the requirements of Annex I to that Regulation. 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. where the raw material complies with points (a). tunicates and marine gastropods. The copy of 208 . Extracts of the requirements of Annex III. Chapters V. shellfish species etc. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation.3. Apart from the provisions on purification.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.
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. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. (ii) Provided the requirements of chapter V are met. 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.Class A means they can be placed directly on the market . collection from: . 9. Transport needs to allow sufficient drainage for best survival conditions. B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with. Requirements for 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.4.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority. B.2 Chapter II . 209 . Requirements for production areas (i) The competent authority classes (classed A.3. C.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market .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. poles or other fixed means are to identify the boundaries of the sites.Hygiene requirements for the production and harvesting of live bivalve molluscs A. processing or relaying.
Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. (iv) yessotoxins.3. 160 micrograms of okadaic acid equivalents per kilogram.4.3.4. Before dispatch. 9.5 Chapter V . requirements if there are many batches of live bivalve molluscs.4. B. for: (i) paralytic shellfish poison (PSP).4 Chapter IV . 210 . the purification system to allow them to resume /maintain filter-feeding activity. Requirements for purification centres There are requirements for live bivalve molluscs to be washed. the levels of marine biotoxins should not exceed the following limits. 800 micrograms per kilogram.3 Chapter III . dinophysistoxins and pectenotoxins together. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. (ii) amnesic shellfish poison (ASP). 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.Hygiene requirements for purification and dispatch centres A. and there are requirements for tanks and water storage containers. containers and a label to identify purification. Requirements for dispatch centres Handling should not result in contamination to any degree.3. 1 milligram of yessotoxin equivalent per kilogram.FISH AND SEAFOOD 9.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc.
4. including the identification mark. 160 micrograms of azaspiracid equivalents per kilogram. must be waterproof. dispatch centre or processing establishment.Identification marking and labelling In addition to general identification marking (contained in 9. the species of bivalve mollusc and date of packaging (at least the date and month) are required. 211 .2. the label.188.8.131.52.7 Chapter VII .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. 9. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction.3. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.3.9 Chapter IX .Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9. 9. Requirements of Chapters III and IV are to be complied with. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. Registration document requirements also apply. by way of derogation from directive 2000/13/EC. Finally. they cannot be re-immersed in or sprayed with water.3. Also. 9.3.6 Chapter VI .LEGISLATION (v) azaspiracids. (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.1).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.8 Chapter VIII .
Therefore the health standards for fishery products as previously stated in Annex III. 9. Chapter V of Regulation 853/2004 apply. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures.5 Regulation (EC) No. 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. This refers to official controls or production and placing on the market of fishery products. echinoderms.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. or - - 212 . 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. (ii) fishery products not complying with the requirement concerning biotoxins. Fishery products in particular must comply with the requirements of Annex III of the regulation. In this regulation. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. they derive from: (i) poisonous fish. chemical. Section VIII. or (iii) bivalve molluscs. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004. 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.FISH AND SEAFOOD 9.5.
Live bivalve molluscs from Class C areas must not exceed 46. In the remaining 10 % of samples. Sampling plans and frequency are required.Official controls concerning live bivalve molluscs from classified production areas A. section VII. as previously stated. coli per 100 g of flesh and intravalvular liquid.5.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. that microbiological criteria and chemical contaminants amongst other requirements are met. coli per 100 g of flesh and intravalvular liquid. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. live echinoderms.2. - - - - B. live tunicates and live marine gastropods. 213 . coli per 100 g of flesh and intravalvular liquid. Classification of production and relaying areas: This section reinforces that specified in Annex III. Regulation (EC) 854/2004. A summary is given as follows: 9.1 Chapter II . three dilutions Most Probable Number (MPN) test specified in ISO 16649-3.5. in 90% of the samples. Live bivalve molluscs from Class B areas must not exceed. chapter V is reinforced. Annex II 9. three dilutions MPN test specified in ISO 16649-3. The reference method for this analysis is the five-tube. The reference method for this analysis is the five-tube.000 E. Also. Monitoring of classified relaying and production areas This is to ensure there is no malpractice.2 This states official controls for the production and placing on the market of live bivalve molluscs.600 E. 000 E. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. 4.
5.2. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying. which has applied since 1 January 2006. the Member States may permit the import of bivalve molluscs and fishery products from the countries listed respectively in Annex I and Annex II to regulation (EC) 2076/2005.2 Chapter III . E. Procedures for re-opening a closed production area are stated D. establishes microbiological criteria for a range of foods. 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. 9. the competent authority is to close the production area. F.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. 214 . 9. 9. opening or closing a production area. boundaries or class of a production area or its closure.5.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions.FISH AND SEAFOOD C. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. 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.6 Regulation (EC) No. Decisions after monitoring There are specific requirements whereby if sampling results indicate that health standards for molluscs are exceeded or there is a risk to human health. 2073/2005.
volume. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product. based on the absence. 9. with chapter 2. The criteria are outlined in Table 9.I. a batch of foodstuffs or a process.LEGISLATION The aim of this legislation is to complement food hygiene requirements. 9. presence or number of microorganisms.II. The relevant criteria are outlined in Table 9. and/or on the quantity of their toxins or metabolites.6. per unit(s) of mass. 215 . and the legislation applies to all businesses involved in food production and handling.1 Food safety criteria Chapter 1 of the regulation focuses on food safety criteria that cover foods such as ready to eat foods and fishery products and live bivalve molluscs. area or batch.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process. ensuring that foods being placed on the market do not pose a risk to human health. Two kinds of criteria have been established: food safety criteria.4 referring to fishery products.6.
monocytogenes.TABLE 9.1 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.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life . other than those intended for infants and for special medical purposes(4) (8) L. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1. monocytogenes 1. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1.3 Ready-to-eat foods unable to support the growth of 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. monocytogenes L. monocytogenes.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.
tunicates and gastropods 1(16) 0 230 MPN/100 g of flesh and intravalvular liquid 100 mg/kg 200 mg/kg HPLC (19) ISO TS 16649-3 Salmonella 1.27 Fishery products which have undergone enzyme maturation treatment in brine.25 Live bivalve molluscs and live echinoderms.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1. coli (15) 1. manufactured from fish species associated with a high amount of histidine (17) Histamine . tunicates and gastropods 9 (18) 2 E.Food category Microorganisms Sampling plan(1) n 5 0 Absence in 25 g EN/ISO 6579 c m M Products placed on the market during their shelf life Products placed on the market during their shelf life Products placed on the market during their shelf life HPLC (19) Products placed on the market during their shelf life 400 g mg/kg Limit(2) Analytical reference method(3) Stage where the criterion applies 1.17 Live bivalve molluscs and live echinoderms.
Engraulidae.92. to the satisfaction of the competent authority. spirits and similar products. — fresh. Other categories of products can also belong to this category. products with pH ≤ 5.25. — — bottled or packed waters. Single samples may be taken at retail level. uncut and unprocessed vegetables and fruits. shall not apply. Particularly fish species of the families: Scombridae. Assay of biogenic amines involved in fish decomposition. excluding sprouted seeds. m = M. 2. beer.1 . that the product will not exceed the limit 100 cfu/g throughout the shelf life. including cocoa and chocolate products. c = number of sample units giving values between m and M For point 1. J. The most recent edition of the standard shall be used. 5 This criterion shall apply if the manufacturer is able to demonstrate. 1996. live bivalve molluscs.. monocytogenes. soft drinks. — bread.4 or aw ≤ 0. according to which the whole batch is to be deemed unsafe. References: 1. AOAC Internat.. This criterion shall apply to products before they have left the immediate control of the producing food business operator. 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. Valle M. biscuits and similar products. Bouquelet S.. Products with pH ≤ 4. 43-9. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter.. Dervin C.94.1. Malle P. Clupeidae. Malle P. subject to scientific justification.. Duflos G. when recontamination is not possible after this treatment (for example. 79. Coryfenidae. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. Scombresosidae. A pooled sample comprising a minimum of 10 individual animals. cider. sugar. products heat treated in their final package). Bouquelet S. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. wine. coli is used here as an indicator of faecal contamination. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . 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.0 and aw ≤ 0. to the satisfaction of the competent authority. Pomatomidae. E.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. products with a shelf-life of less than five days shall be automatically considered to belong to this category. when he is not able to demonstrate. honey and confectionery.
if all the values observed indicate the absence of the bacterium. monocytogenes before the food has left the immediate control of the producing food business operator when he is not able to demonstrate that the product will not exceed the limit of 100 cfu/g throughout the shelf life: — satisfactory. excluding live bivalve molluscs and live echinoderms. Salmonella in different food categories: — satisfactory. 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. if all the values observed indicate the absence of the bacterium. 1999. 82. — unsatisfactory. — unsatisfactory. — unsatisfactory. no values observed exceed the limit of M. if any of the values are > the limit. if all the values observed are ≤ the limit. if the following requirements are fulfilled: 1. AOAC Internat. where the limit refers to a pooled sample. Histamine in fishery products from fish species associated with a high amount of histidine: — satisfactory. L. tunicates and gastropods in relation to testing E. — unsatisfactory. Interpretation of the test results relating to Table 9. 1097-101. L. a maximum of c/n values observed are between m and M 3. monocytogenes in other ready-to-eat foods: — 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. L. — unsatisfactory. coli. J. the mean value observed is ≤ m 2. 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.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus. monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory.I The limits given refer to each sample unit tested. if the presence of the bacterium is detected in any of the sample units. if the presence of the bacterium is detected in any of the sample units. if all the values observed indicate the absence of the bacterium. The test results demonstrate the microbiological quality of the batch tested(1). (1) 219 .
TABLE 9.4.1 Shelled and E.II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2. coli 5 shucked products of cooked crustaceans and molluscan shellfish 2 100 cfu/g 1000 cfu/g EN/ISO 6888-1 or 2 FISH AND SEAFOOD Coagulasepositive staphylococci 5 Improvements in production hygiene 220 .
and the rest of the values observed are ≤ m. c = number of sample units giving values between m and M The most recent edition of the standard shall be used. and the rest of the values observed are ≤ m. if all the values observed are ≤ m. The defendant may be required to produce well-founded scientific proof to support his claims.I.7 Food Hygiene (England) Regulations 2006.I. with chill holding tolerance periods.LEGISLATION 1 n = number of units comprising the sample. the defendant will need to prove that the food was on service or display. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. which overrides the 8 °C requirement. — acceptable. if a maximum of c/n values are between m and M. — unsatisfactory. Schedule 4 of the Food Hygiene (England) Regulations 2006. if one or more of the values observed are > M or more than c/n values are between m and M. if a maximum of c/n values are between m and M.1 In the U. 2006 No.K) Temperature control requirements 9. 14 (Hygiene requirements specific to the U. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. 14 details temperature control requirements for foods in general. — unsatisfactory. 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. had not been 221 . The regulations prescribe a chilled food holding temperature of 8 °C or less. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. 2 Interpretation of the test results relating to Table 9. For example. S.7.II The limits given refer to each sample unit tested. 2006 No. if one or more of the values observed are > M or more than c/n values are between m and M. The test results demonstrate the microbiological quality of the process tested. if all the values observed are ≤ m. S.K. E. The regulations provide for defences in relation to upward variations of the 8 °C temperature. 9. tolerance periods for chill-holding of foods and hot-holding variations. — acceptable.
3). The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment.8 Guidance In the U. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. 2006 No. 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. Alternatively. 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. 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. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. or. 9. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety.S.K. 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.food. was kept at above 8 °C or the recommended temperature for an unavoidable reason.I. 2006 No.gov.I. 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. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S. such as that below. and was kept at above 8 °C or the recommended temperature. 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. for a limited period consistent with food safety. For other food. S.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing.
L139. 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. 22) as amended by Regulation (EC) Nos. 25.2004. Regulation(EC) No. 2075/2005 and Regulation (EC) No. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41.10. L283.12.LEGISLATION 9. 852/2004 is now set out in a Corrigendum (OJ No. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No.2004. Regulation (EC) No. L139. 853/2004 is now set out in a Corrigendum (OJ No. 30. 1. 30.4. Regulation (EC) No. 25. 1662/2006. 1441/2007. as read with the corrigenda at OJ No.4. Regulation (EEC) 2136/89. 854/2004 is now set out in a Corrigendum (OJ No. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No.6. 223 .6. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito. laying down common marketing standards for preserved sardines. 1663/2006. 2074/2005. namely.2004. L338.9 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. 2074/2005 and Regulation (EC) No. 1). 2073/2005 on microbiological criteria for foodstuffs (OJ No. L226. 83) as amended by Regulation Nos 882/2004. 62) as amended by Regulation (EC) No. these are not be discussed here. 1688/2005. 3) as amended by Regulation (EC) No. 25. 1019/2008 and as read with Regulation 2073/2005.10 Further Reading Regulation (EC) No. 2076/2005. L139.2006. as amended. 9. 30. 2076/2005. 2076/2005. L226.2004. Commission Regulation (EC) No. 2074/2005. 206). The revised text of Regulation (EC) No. Regulation (EC) No. L226. 22. The revised text of Regulation (EC) No. 55). 2076/2005.6. Regulation (EC) No. Regulation (EC) No.2004.4.2005. The revised text of Regulation (EC) No. 14. 2074/2005.2004.
(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. 21. 6) 224 . 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17. 22.2005. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.2000. 22) Commission regulation (EC) No 2065/2001 laying down detailed rules for the application of Council Regulation (EC) No 104/2000 as regards informing consumers about fishery and aquaculture products as amended by Commission Regulation (EC) No 1792/2006 of 23 October 2006 (OJ L 278. derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004.12.10.12. 22. 23.2001.1.FISH AND SEAFOOD Commission Regulation (EC) No 2074/2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC)No 882/2004 of the European Parliament and of the Council.2005. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004.
5 pH Under otherwise optimal conditions.1.5 minutes.3 . and D-values at 45 and 51 °C have been reported as 29. and 2. PATHOGEN PROFILES 10.1 10.10. a 10 to 1. Morphology Gram-negative rods. 0.0 .3 minutes.000-fold increase in number is observed after one week in food systems (1).10 (3). 10. At 5 °C. At pH 4. no growth is observed (1).1 Aeromonas spp.5. 10.1.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.1.1. respectively (1).0 x 1. under optimal incubation temperatures.1. 225 . 10.5 µm 10. The minimum pH value for growth is thought to be pH 184.108.40.206 Temperature Aeromonas grows at temperatures of 2 . The optimum growth temperature is 28 °C (2).45 °C (1). Aeromonas has the potential to grow over a pH range of ca 4 .2 Oxygen requirements Facultative anaerobe.3. Some death will occur at this pH if the temperature is 4 °C.
2 Clostridium botulinum Seven different types of C. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C.5 .0 µm.1. to a lesser extent. Group I will not grow at temperatures of 10 ºC or less. 10.2. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. Types A. botulinum are known.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 . at 85 ºC for 5 minutes.2% sodium chloride (NaCl) (1).1 Morphology Gram-positive. botulinum is prevented. but Group II strains are psychrotrophic.3 Temperature All strains of C.30 minutes. 6).22.2. All type A strains are proteolytic and type E strains are usually non-proteolytic. 10. All C. botulinum are not particularly heat resistant. F are the types that are responsible for most cases of human botulism (5.7 .2. A to G. B. being capable of slow growth and toxin production at low temperatures . but the interior of the food may do so. Thus an aerobically packed product may not support the growth of the organism on the surface.even as low as 3 ºC (7).2 Oxygen requirements Although C. which may be inactivated by heating at 80 ºC for 20 . forming at least seven different toxins.2. and groups I and II are those responsible for the botulism cases.4 x 1. or at 90 ºC for a few seconds.2. E and. 0. many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth.4 Heat resistance The vegetative cells of C. botulinum is an obligate anaerobe.FISH AND SEAFOOD 10. and is sensitive to > 4. spore-forming rods. 226 . but the spores of this organism are more so. 10. There are four main groupings of the organism. 10. 10.5% NaCl (4). botulinum grow reasonably well in the temperature range of 20 45 ºC. types B and F can be either. botulinum types produce heat labile toxins.
In commercial situations. 11)).97) (10. These values have been established under carefully controlled laboratory conditions.7 Characteristics of C. the minimum aw for growth may be lower.3.3. parcooked meals with pH value > 5.94 for Group I and 0.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e. Note also that if humectants other than salt are used.0 µm.3 10. 10. particularly by cooking. vacuum packed.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4.0.3 .0 . These concentrations correspond to limiting aw of 0. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains.like other clostridia . 10. However. botulinum spores The most heat resistant spores of Group I C. therefore. 227 .10. safety margins must be introduced to allow for process variability. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. pH and temperature.2. It will not.21 minutes) (9). 9). where oxygen levels are reduced.6 and pH 5. grow on the surface of foods unless they are vacuum or gas-packed.is an obligate anaerobe.PATHOGEN PROFILES 10. 10.0 and aw > 0.1.1 Clostridium perfringens Morphology Gram-positive spore-forming rods. perfringens .2.2. 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. botulinum depends on solute. respectively (8. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms.g.97 for Group II (5).9 x 2.2 Oxygen requirements C. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. they may survive mild heat treatments (70 . 0. 10.
although C. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance.6 Aw/Sodium chloride C. lower concentrations may be effective in combination with other factors.3.3 Temperature The most significant characteristic of C. 0 . perfringens is not tolerant of low water activities. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8.it is destroyed by heating at 60 ºC for 10 minutes (14.93 and 0.3. perfringens appears to be 0. the enterotoxin is not heat resistant . 10. (It is unusual for spores to be formed in foods after the growth of this organism (14)). perfringens spores The spores of C. 18). death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. the limits for aw are affected by temperature. While some growth can occur at 50 ºC. however. The lowest aw recorded to support the growth of C. and their heat resistance is also affected by the heating substrate. Salt concentrations of 6 . It grows best at pH values between 6 and 7 (the same pH as most meats). 13).3. Under otherwise ideal conditions. 10. pH. At cold temperatures. etc. perfringens in vacuum packed beef (18).50 ºC. perfringens strains. Spores.5 minutes (17).5 pH C. The D-value for C. type of solute. 13). 15. perfringens has the potential ability to grow within the temperature range 15 . will survive greater extremes of pH (and aw) (12. depending on strain and other conditions.45 ºC. perfringens can vary quite considerably in their heat resistance.7 Characteristics of C.8% inhibit growth of most C. Some studies indicate that the presence of 3% NaCl delays growth of C. perfringens is not a tolerant organism with respect to pH. 10.3. 10. Its optimum temperature for growth is 43 .3.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. vegetative cells die rapidly (12).97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. perfringens. In addition. As in the case of other factors limiting the growth/survival of this organism.3.10 ºC. Recorded heat 228 . perfringens in roast beef at 60 ºC is 14. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures. 16).FISH AND SEAFOOD 10.
4.5 x 0. and temperature. Its optimum growth temperature. 10.4 ºC is probably a more likely minimum for growth in foods (19).4. 1. The upper temperature limit for the growth of L.albeit slowly .4 .PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17.2 Oxygen requirements Aerobe or microaerophilic.4. however. 10.4.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. 60 and 65 ºC were reported as 10. D-values for L. -0.especially at refrigeration temperatures.4 10. monocytogenes in crawfish tail meat at 55.2 has been suggested as the lowest working limit (22).6 .98 and 0. 10. Under ideal conditions. pH < 5. Morphology Gram-positive short rods. the organism is able to grow at pH values well below pH 5 (pH 4. it is not a sporeformer.8 minutes for heat sensitive spores (12). growth at low temperatures can be very slow. 229 . It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.4.2. using hydrochloric acid as an acidulant). However. so can be destroyed by pasteurisation. 10. monocytogenes is reported to be 45 ºC (20). the lowest limit for growth is likely to be considerably higher .2. 10.3 is the lowest value where growth has been recorded. however.3 .0.1 Listeria spp. monocytogenes is not a particularly heat resistant organism. In foods.at refrigeration temperatures down to.4 Heat resistance L. or even below 0 ºC.5 . and where acetic acid is used as an acidulant. 0. respectively (21). requiring days or weeks to reach maximum numbers.19 minutes.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.23.0 µm. being potentially capable of growing .64.0 minutes for heat resistant spores to 1. is between 30 and 37 ºC.
1 Salmonella spp. It is likely to survive. 230 .83 (20).97.4. but has been shown to be able to grow at an aw level of 0.12% NaCl or more). peritrichous flagella.0 x 1 . 10.0.5.0 µm. 10.5 . at salt levels found in foods (10 .5 . the optimum being 30 ºC (23).6 10. or even grow.5. 25).3.8 .6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24. 10. 25).4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).3 Temperature Plesiomonas can grow in the temperature range of 8 .5 10.90. 10. monocytogenes is quite tolerant of high NaCl/low water activities.5.6 Aw/Sodium chloride L.5 pH Plesiomonas has a pH range for growth of pH 4.FISH AND SEAFOOD 10.1 Plesiomonas Morphology Gram-negative. 0. 10.1.2 Oxygen requirements Facultative anaerobe. 10.5. It grows best at aw of ≥ 0. Morphology Gram-negative short rods.5 (24.0 .3 µm.5.5.45 ºC. 0. The bacterium may survive for long periods at aw values as low as 0.8. 10.7 x 1.6.
5 ºC.PATHOGEN PROFILES 10. However. Growth is slow at temperatures below about 10 ºC. However. Salmonella senftenberg 775W is about 10 .4% NaCl.0. Salmonellae are quite resistant to drying. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. 10. temperature and pH.48 ºC. D60 ºC values normally range from about 1 to 10 minutes. the optimum being 35 .7.8 . It is not.6.6. under otherwise ideal conditions. some strains are able to grow at temperatures as low as 4 °C (26).5. strains vary in their ability to withstand heating. The growth of Salmonella is generally inhibited by the presence of 3 . depending on serotype. Furthermore.4 Heat resistance Salmonella is not a spore-forming organism. therefore.3 Temperature Salmonellae can grow in the temperature range of 7 .6. 10. and with an appropriate acid. depending on the type of acid and temperature.20 times more heat resistant than the average strain of Salmonella at high aw (27).6 Aw/Sodium chloride Where all other conditions are favourable. and appropriate heat treatments must be determined experimentally for low-aw foods.6. Salmonella has the potential to grow at aw levels as low as 0.37 ºC. 231 . or possibly 0.93 (as reported in dried meat and dehydrated soup). a heat resistant organism. although salt tolerance increases with increasing temperature (28). with a z-value of 4 . The optimal pH for Salmonella growth is between 6. Some death will occur at pH values of less than about 4.945. 10.6. 10.5.9.5 pH Salmonella has a pH range for growth of pH 3. substrate.5 .2 Oxygen requirements Facultative anaerobe. Salmonellae are quite resistant to freezing. pasteurisation and equivalent treatments will destroy the organism under normal circumstances.
aureus viability. 10. aw. 10. aureus than citric acid).7 10.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH. It can survive well at low temperatures.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. 31). the D60 ºC value is 2 .0.7. Staph.2 to 9.7. Freezing and thawing have little effect on Staph. Most strains of staphylococci can grow within the pH range 4. aureus can grow within the temperature range 7 – 48. aureus is more limited under anaerobic than aerobic conditions. 10.9 µm diameter.20 minutes. with an optimum of 35 .3 Temperature Under otherwise ideal conditions.7. 232 .7 . but may cause some cell damage (30). In meat macerate.FISH AND SEAFOOD 10. however. stressed cells can also be less tolerant of heating.3 (optimum 7. fat content. The following relate to limits for growth only.2 Oxygen requirements Facultative anaerobe.1 Staphylococcus aureus Morphology Gram-positive cocci. depending on aw.7.5 ºC.7. Under most circumstances. etc. As is the case with other bacteria.5). aw and temperature (sensitivity to acid increases with temperature). the organism is heat sensitive and will be destroyed by pasteurisation. under otherwise ideal conditions (29. The limits for toxin production are also narrower than for growth.7. NB: The growth of Staph.0 . 0. 10.).37 ºC (29).
10% NaCl. It can grow over the aw range 0.6 Aw/Sodium chloride Staph.1 Vibrio cholerae Morphology Gram-negative curved rods.8.87 and > 0. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant.5 .99 little or no toxin production in anaerobically packed foods. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31. aureus can grow in 7 .8.86 is the generally recognised minimum in foods (32). with a temperature range for growth of 10 .5) (29. 10.8 10. 31) between 0. pH.7. Staphylococci are more resistant to salt present in foods than other organisms. 0.>0. In general. However. 10. and solute.PATHOGEN PROFILES 10.0.7 Limits permitting toxin production 10 .2 Oxygen requirements Facultative anaerobe.0 . 34). an aw of 0.4 .6 µm 10.8 x 1.99 aerobically under otherwise optimal conditions.43 ºC (35).83 . atmosphere.7. An effect of increasing salt concentration is to raise the minimum pH of growth.2 .9.0 (optimum 7.8.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. but certain strains can grow in 20% NaCl. strain. In general. * dependent on temperature. 233 .2. 10.7. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities. Staph.3 Temperature The optimum temperature for growth has been reported as 30 ºC.
cholerae can grow over the aw range 0.65 minutes (35). there is evidence that V.1 .5 ºC).97 .8.9.6 Aw/Sodium chloride V. it has a pH range for growth of pH 5 . 10.6 (35. but only moderately sensitive to freezing (40). 10. parahaemolyticus is easily destroyed by drying. cholerae in crab meat homogenate at 60 ºC is 2. 234 . but not acid conditions. with an optimum of 0. 10.998. cholerae is sensitive to heat at temperatures of >45 ºC. cholerae may persist on frozen fish for >180 days (36).1 Vibrio parahaemolyticus Morphology Gram-negative curved rods.9.9. 10.8. 10. 10. It can grow in the presence of 0.5 pH This organism is tolerant of high pH values.9 10. 37.2 Oxygen requirements Facultative anaerobe.0% NaCl (35). The D-value for V. declining in number during storage. respectively. with an optimum of 37 ºC (39). with optimum at 7.0.984 (35). and it is sensitive to refrigeration temperatures (0 .6.8. are 5 and 43 ºC. 38). under otherwise favourable conditions. V.9.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.FISH AND SEAFOOD Also.4.4 Heat resistance V.
4% (39. 41).11.8 . 235 .4 Heat resistance V. 10.10.5 pH The pH range for growth is 4.98 (35.2 Oxygen requirements Facultative anaerobe.922. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43). parahaemolyticus are not heat resistant.29 minutes. vulnificus is heat sensitive. with an optimum pH of 7. depending on solute.6 Aw/Sodium chloride V. 10.10. 41). 10. The optimum concentration of NaCl equates to aw of approximately 0. The organism can survive for long periods at freezer temperatures (42).10 Vibrio vulnificus 10. parahaemolyticus will grow at aw levels as low as 0.02 .4 Heat resistance V.10. The D-values at 55 ºC for clam and crab homogenate are 0.5% NaCl. V.6 .1 Morphology Gram-negative curved rods.9. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0. with a temperature range for growth of 8 . respectively (39).3 Temperature The optimum temperature for growth has been reported at 37 ºC. respectively.9.5 minutes.9. 10.6 (41). and are readily destroyed by cooking.43 ºC (42). 10.0. 10.8.10. but the optimum concentration is 2 . and 2.PATHOGEN PROFILES 10. parahaemolyticus is unable to grow in the absence of salt.
Dodds K. Faris A. Johnson E. Clostridium botulinum. Clostridium botulinum and Clostridium perfringens. Volume 1: Diseases Caused by Bacteria. 1993. Lund B. New York.A. Peck M. Bier J. Murrell K. Blackie. with an optimum pH of 7.10. Ed.L. 357-67. New York.M. 121-76.R.M.O. 10. in Microorganisms in Foods. Betts G..L. References International Commission on Microbiological Specifications for Foods. 2005. Cliver D. London. 1994. Notermans S. 1993. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’. 2003. in Food Microbiology: Fundamentals and Frontiers. Eds. Dodds K. Food Control. International Commission on Microbiological Specifications for Foods.. Aeromonas. Stelma G. New York. 1989.W.11 1. Austin J. Clostridium botulinum.W.H.. 57-63.. Murrell K. Aeromonas hydrophila. Smith J.L.. Cliver D. Marcel Dekker.E.D.P. Novak J. Gorman J..H. 1989. 329-49. New York. Gaze J. Abeyta C. 1996. 2001. in Clostridium botulinum: Ecology and Control in Foods.. Marcel Dekker.. 10. Beuchat L.N. Diseases Caused by Bacteria. ASM Press. in Foodborne Bacterial Pathogens..8 (42).. 1994. Aeromonas species.R.5.. Hauschild A.O. Washington D.D. Clostridium botulinum. in Foodborne Pathogens. in International Handbook of Foodborne Pathogens. 279-303.H. 383-408.. New York.C.. Eds. 6. Marcel Dekker.P. 3.H.5 pH The pH range for growth is 5 .W. Miliotis M.D.N..5 . Microbiology and Molecular Biology. 10. Eds. in Clostridium botulinum: Ecology and Control in Foods.. Aeromonas hydrophila group.. Kovacek K. Hauschild A.FISH AND SEAFOOD 10. Eds. New York.W.L. Kim J. in Foodborne Bacterial Pathogens. 1-19. 236 . Foegeding P. Eds. Stelma G. Great Britain. Marcel Dekker Inc.5% (42).K. Dodds K.W.10.P. with an optimum of 2. 6 (1).10. Doyle M. 7. 97-131.. Marcel Dekker. Bhunia A. Juneja V. 5-19.6 Aw/Sodium chloride V. Palumbo S.R.. Ed. Principles of control.. Doyle M. Hui Y. 11. Volume 5: Microbiological Specifications of Food Pathogens. 2.. Caister Academic Press. Hui Y. Potential hazards associated with REPFEDS.M. Marcel Dekker. Ed.. New York. Eds. Gorham J. 1-27. in Foodborne Disease Handbook. Volume 1. in Foodborne Disease Handbook. Doyle M. vulnificus grows over the range 0. 1995. 8. Marcel Dekker. Montville T. 5.H.. 9.. Fratamico P. 111-89. Hauschild A.0% NaCl.D. Eds.J. 4.H.
G. Listeria monocytogenes. 129-38.H. Montville T. 22.. 1994. Clostridium perfringens.H.O. 208-13. in Foodborne Diseases. in Foodborne Disease Handbook.K. Beuchat L.D. Eds. London. Johnson E.. London.. Washington D.. 26.C. International Commission on Microbiological Specifications for Foods.. 20. 2001. Labbe R. Ed. in Microorganisms in Foods. New York. Plesiomonas shigelloides. New York.G. Eds. Juneja V. 237 . Marcel Dekker. 311-25.R. Clostridium perfringens.H. in Foodborne Bacterial Pathogens.T. Elsevier. in Microorganisms in Foods. Eds. Washington D. 1990. Varnam A. 1989. London. Evans M. Montville T. 24.A. London. Banks J.A. Emery D. Kim C. Clostridium perfringens. Ed. Halvorson D. International Commission on Microbiological Specifications for Foods. New York. Cliver D.. in Food Microbiology: Fundamentals and Frontiers. 2007. 1996.J.R. Marth E. Clostridium perfringens food poisoning. ASM Press. 1990. Koburger J.M. 201-8. London. 1991.. Diseases Caused by Bacteria. Plesiomonas shigelloides. Clostridium perfringens. 112-25. International Commission on Microbiological Specifications for Foods. Volume 1.H. 16. 1989. Blackie. 229-40.. in Foodborne Bacterial Pathogens. Wrigley D. 15. International Commission on Microbiological Specifications for Foods.A.PATHOGEN PROFILES 12.G. Doyle M... Academic Press. 18. Listeria monocytogenes. 141-82. Murrell K. in Food Microbiology . 68 (2). 735-42. Cliver D. New York. 336 (8721). Marcel Dekker. 351-82.. Evans M.P. 2005. Lund B. Listeriosis and Food Safety.. Matthews K.P. Labbe R. Hui Y. Rinke H. 2006. 133-67. Walker S.P.An Introduction. 157-62.J.P. 23. 1990. Marcel Dekker.J.. Riemann H. Ed. Ed. Listeria.. Wolfe Publishing Ltd. Gorham J. McClane B. Varnam A. Ed. Marcel Dekker. 191-243. 1996. Archer P.O. Volume 5: Microbiological Specifications of Food Pathogens. Cliver D.. International Commission on Microbiological Specifications for Foods. Ed. Volume 5: Microbiological Specifications of Food Pathogens.A.V. in Foodborne Pathogens: An Illustrated Text. 1989. ASM Press. Volume 5: Microbiological Specifications of Food Pathogens. 137-64. 25. Growth of Listeria monocytogenes at refrigeration temperatures. 1996.J. 982-6. Ryser E. 33. London. 13. Blackie. Lancet. Journal of Applied Bacteriology.O..R. in Microorganisms in Foods. Nagaraja K. 19.M.. Eds. Clostridium perfringens gastroenteritis. Plesiomonas. in Foodborne Infection and Intoxication. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs.A. 21. Blackie. Doyle M.. International Commission on Microbiological Specifications for Foods. 14. Avian Disease. Eds. 17. Doyle M. Foodborne disease due to Bacillus and Clostridium species.C.
. 28. 1989. 36. 38. 2005. International Commission on Microbiological Specifications for Foods. Crowcroft N. Ed. Bergdoll M. International Commission on Microbiological Specifications for Foods. in Foodborne Disease. 985-98. London. Cliver D. 642. 1983. Riemann H. Volume 5. Caister Academic Press. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems.P. Ed. in Microorganisms in Foods. Eds. Tohyama Y.M. Smith J. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures.. in Foodborne Bacterial Pathogens. London. Blackie. Doyle M. Woodhead Publishing Ltd. International Commission on Microbiological Specifications for Foods. 327-52. 327-445. Bhunia A.A. Marcel Dekker. Twedt R. Staphylococcus aureus. Jay J. Microbiological Specifications of Food Pathogens. 543-68.. Cleland D. Stress Response and Resistance. Volume 5. 1930. Academic Press. 13 (11).K. 2005. International Commission on Microbiological Specifications for Foods. Ed. 2005. Ed. R15764. Doyle M. Springer Science. Applied and Environmental Microbiology. Jay J. Ed. Griffiths M. 8. Staphylococcal Food Poisoning. 238 .M. 1989. Blackie. Cambridge. Lee-Wong A. Wymondham. CDR Review.R. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. Salmonella. in Microorganisms in Foods. Staphylococcal gastroenteritis. Oliver J. 1996. Blackie. 45 (3). Golden D. New York. New York.S. 29. in Understanding Pathogen Behaviour Virulence. 33... Vibrio parahaemolyticus. Dairy. Vibrio cholerae. Cliver D. 32. Staphylococcal intoxications. Eds.A. London. Microbiological Specifications of Food Pathogens. Food and Environmental Sanitation.. Loessner M. London. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning. 85-106. 545-66. 426-35.S.J. 523. 273-84. Academic Press. Volume 5: Microbiological Specifications of Food Pathogens. in Foodborne Pathogens: Microbiology and Molecular Biology. D’Aoust J-Y. Warner R. Golden D. 30. 331-57. International Commission on Microbiological Specifications for Foods.L.O.M. 37. Japanese Journal of Experimental Medicine. Ed.P. 39.C. 35. Stewart G.J. Vibrio parahaemolyticus.62.M. 4 (13).P. 217-64. International Commission on Microbiological Specifications for Foods. New York.A.. 1996. 1996.O.. Reed G. Salmonellae. 40.H. Eds. 1993. 414-25.C. in Foodborne Infections and Intoxications. Wilkinson B. London.S. Bergdoll M.. in Foodborne Pathogens. in Modern Food Microbiology. 1990. in Microorganisms in Foods. 1994. Marcel Dekker. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). Gustafson J. Cholera: current epidemiology. 34. 31.FISH AND SEAFOOD 27. Ed. Fratamico P. 2005.D. Loessner M..
in Microorganisms in Foods..M.A. Microbiological Specifications of Food Pathogens. 239 . 42. London.W. Springer Science. Cook D.J.. in Modern Food Microbiology..M.D. Loessner M. Ed.J.A. International Commission on Microbiological Specifications for Foods. Loessner M. 436-9. 657-78.. Vibrio vulnificus. Golden D. 985-9. and Campylobacter species. Ruple A. Yersinia. 43.. Foodborne gastroenteritis caused by Vibrio. 1996.PATHOGEN PROFILES 41. Journal of Food Protection. Jay J. Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters. 2005. Golden D. 55 (12). New York. Blackie. Jay J. 1992. Volume 5. International Commission on Microbiological Specifications for Foods.
FISH AND SEAFOOD 240 .
gsi.org/ .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.gsi.defra.org.uk Web site: firstname.lastname@example.org or v.org.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: email@example.com/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: firstname.lastname@example.org 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 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: firstname.lastname@example.org Web site: www.uk Web site: www.
suite. Andersens Boulevard 37.dardni.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: email@example.com/index/fisheriesfarming-and-food.org Web site: www.is Web site: www.no Web site: www.C.htm Irish Fish Processors and Exporters Association 25 Kincora Avenue Clontarf Dublin 3 Eire Ireland Tel: + 353 1 833 7882/ 02770249 Fax: + 353 1 833 7882 Email: tormg.gov.FISH AND SEAFOOD Department of Agriculture and Rural Development.dk Web site: www.dk Web site: www.fao.dk Danmarks Fiskehandlere H.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.fiskeriforskning.is/ 242 .eapo.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@firstname.lastname@example.org/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .ie/templates/homepage.uk Web site: www. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.com Web site: www.matis.com/ Danish Institute for Fisheries Technology and Aquaculture (DIFTA) illemoesvej The North Sea Centre DK-9850 Hirtshals Denmark Tel: + 45 98 944300 Fax: + 45 98 942226 Email: difta@difta. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: email@example.com. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www. 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/fishery/ European Association of Fish Producers’ Organisations H.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: post@fiskeriforskning.
uk Web site: www.org Web site: www.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.nz/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.org.gov.nz Web site: www.com Web site: www.csiro.cmar.org Other Sources of Information British Standards Institution 389 Chiswick High Road London W4 4AL United Kingdom Tel: + 44 (0) 208 9969001 Fax: + 44 (0) 208 9967001 Email: email@example.com/ 243 .uk Web site: www.bsi-global.gov.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 Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: email@example.com New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: firstname.lastname@example.org 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/en/index.co.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: firstname.lastname@example.org.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 Web site: www.org.seafood.dh.org.ac.hpa.
who.int/foodsafety World Health Organization (WHO) Headquarters CH .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: firstname.lastname@example.org/index.nhs.int Web site: www. jsp Food Safety 10 .scot.uk Web site: www.de 244 .uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: email@example.com Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: firstname.lastname@example.org Web site: www. 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.euro.who.nhs.sik.org Web site: www.hps.uk Web site: www.int Web site: http://www.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/ SIK .scot.uk/scieh.int.co.se/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: firstname.lastname@example.org Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: email@example.com.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.net/web/indexen.euro.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.
fpa-food.CONTACTS US FDA .gov/ World Health Organization (WHO): Food safety programmes and projects http://www.org/ Royal Institute of Public Health (UK) http://www.sofht.ift.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.co.foodscience.uk/ Society of Food Hygiene and Technology (UK) http://www.org/ Institute of Food Technologists http://www.gov/ Institute of Food Technologists 525 W.foodprotection.org.uk/ 245 .org/ The Association of Food.gov/foodsafety/ Food Safety Authority of Ireland http://www.csiro.riph.ie/ Food Science Australia http://www.eu/pol/food/index_en.fda.htm Centre for Disease Control and Prevention (CDC) (US) http://www.org Web site: http://www.int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa.who.fsai.org/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.cfsan.FoodSafety.cdc. Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: firstname.lastname@example.org/ International Association for Food Protection http://www.ift. Beverage and Consumer Products Companies http://www.
FISH AND SEAFOOD 246 .
as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. as autochthonous bacterial flora of fish 3 Aeromonas spp. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. 69-70 Aeromonas hydrophila. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. in crustacean shellfish 86-7 ATP.INDEX Achromobacter spp. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. 111 Astroviruses. 102 Bacillus spp. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. preserved by dry/wet salting 99 Anisakis spp. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. 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 . in crustacean shellfish 87 Alteromonas spp. growth and survival in crustacean shellfish 86 Aeromonas sobria. causing spoilage of dried fish 110. planktonic source of spirolides in shellfish 163 Allergens. source of PSP toxins 142 Alexandrium ostenfeldii. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. 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. to test microbiological criteria 19 Aeromonadaceae.
in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage. importance in lactic acid fermentation in fish products 130. 137. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. 136. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. of recipe fish meals 43 Cholera. for PSP toxins 144 Chemical indicators. of fish – effects on microflora 5-6 Carbohydrate. 226 Breading process 36-8 Brevetoxins. 98-9 Brochothrix thermosphacta. 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. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. 113. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. 68-9 Capture and handling. of fresh fish spoilage 11 Chemical pollutants. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. 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. 131 Carbon dioxide. in crustacean shellfish 86-7 Campylobacter coli. as autochthonous bacterial flora of fish 3 Clostridium botulinum. fish sauce 123 Basipetospora halophila. preserved by marinating 100 Chemical detection methods. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. associated with consumption of fish/fish products 16. 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 .
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. to determine fresh fish spoilage 11 Eels. processing 334 Cockles. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . as autochthonous bacterial flora of fish 3 Crabs. to determine fresh fish spoilage 11 Corynebacteria. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. source of azaspiracids 159 source of DSP toxins 148-9. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. 152 Diphyllobothrium. growth and survival in crabmeat 84 growth and survival in cured. as contaminants in scampi 36. for detection of DA 148 ELISA kit (DSP-Check). of fish – different processes 94 Dun. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. 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. 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.INDEX pathogen profile 226-7 Clostridium perfringens. hot-smoking 107 ELISA assay. 149-50. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). to detect NSP toxins 155 ELISA kit (Biosense). 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. smoked and dried fish 114 Enzymic spoilage. adverse condition on poorly preserved fish 110 EC grading scheme. 106 Contacts 241-5 Conway method. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. of prepared fish products 44 Escherichia coli.
non-B) viruses. as source of ciguatoxins 156 Garlic. 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. for evaluation of fish quality 9 Eurotium. fish sauce 123 HACCP. to measure fish skin resistance 10 Flavobacterium. preserved by marinating 101 Gravlaks. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. specific EU hygiene rules 196-224 Fresh fish. causing pink condition on fish surface 110 in fish sauce 127 Handling. as source of inulin in lactic acid fermentation in fish products 134 Garos. member of cyclic imine group 161 Gymnodinium catenatum. of fish – effects on microflora 5-6 Hepatitis (non-A. mould causing spoilage of dried fish 110 Evaluation. 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 fish at sea 5 Fatty fish. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. source of PSP toxins 142 Gyoshoyu. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. of fresh fish spoilage 911 Evisceration. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . of prepared fish products 44 Gambiertoxins. 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. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. concerning crustacean shellfish 87-8 European Grading Scheme. 130-5 Fish-spoilage bacteria 12 Fishtester. processing 35 Fish component. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. fermented fish product 126 Green cured split cod 97. 102 Gymnodimines.FISH AND SEAFOOD EU legislation.
outbreak caused by gravid 115 Lobster. 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. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. for fresh and frozen fish (Table) 20 Micrococcus spp. preserved by marinating 101 Microbiological criteria. definition 1 Legislation. preserved by salt-boiling 100 Karenia brevis. enabled by carbohydrate source in fish products 130. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 . as source of yessotoxins 152 Lipid content. hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. 135 I-zushi. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. food hygiene 189-224 Lingulodinium polyedrum. as autochthonous bacterial flora of fish 3 Lean fish. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. as source of botulism 136 International legislation. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. preserved by salt-boiling 100 Ionising radiation. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. in fish fillet 1 Lipid oxidation. concerning crustacean shellfis 88 Jellied eels. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149. smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. to reduce rate of fish spoilage 12 Ikan kaju. for microbial inactivation in fish products 127. 100-1 Masmeen 107 Matjes.
contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P. as TTX producer 145 Norovirus. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay.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. 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. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. associated with consumption of fermented fish products 137 Oxidative rancidity. fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. in crustacean shellfish 867 Norwalk virus. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. caused by lipid oxidation 11 Oysters. of crustacean shellfish – effects of processing 81-3 of cured. reduction in smoking process 108 Norcardiopsis dassonvillei. shigelloides. 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.
225-39 Pathogens. in fish fillet 1 Proteus. spoilage organism in oysters 65 Protoceratium reticulatum.INDEX Parasites. adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164. 108 Polypaecilum pisce. 145-6 definitions 28-30 non-seafood ingredients 32 Processing. as component for seafood ready meals 40 Pathogen profiles. contact details 241-2 Prorocentrum spp. planktonic source of azaspiracids 159 Pseudomonadaceae. detection methods 165 origins and distribution 165 toxicology 165 Pissala. causing spoilage of dried fish 110-1 Photobacterium phosphoreum. 197 Plesiomonas. 69-70 Polycyclic aromatic hydrocarbons. as source of yessotoxins 152 Protoperidinium cassipes. as source of DSP toxins 148-9 Protein content. 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. 163. 81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 . 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. 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. 100-1 Pindang. effects on microflora of fish 5-9 Professional bodies. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation. preserved by salt-boiling 100 Pink. as source of toxins 159. as component for seafood ready meals 40 Prawns. fermented fish product 124 Plankton. 152-3 Penicillium spp. mould causing spoilage of dried fish 110 Potato. for detection of PSP toxins 144. smoked and dried fish 115 Pasta. smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. in molluscan shellfish 60 Pseudomonas spp. as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. reduction in smoking process 103. pathogen profile 230 in molluscan shellfish 5960.
processing 34. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. description and uses 58-9 Scampi. antibody-based kit to detect STX 144 RT Freshmeter. as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products. as source of tetrotoxin 144-5 Putty fish.) 41 ingredients preparation. assembly and packaging. detection methods 165 member of cyclic imine group 164 origins and distribution 165 toxicology 165 Puffer fish.FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries. processing 35 Shellstock. chilled and frozen 1-25 Recipe dish. processing and effects on microflora 60-1 Shewanella. preserved in saturated brine 99 Sauce. 36 Seafood. fermented fish product 126 as source of botulism 136 Rakørret. for microbial inactivation in fish products 127.) 41 Recipe fish products 38-40 Rhodotorula rubra. 83 254 . as carbohydrate source in lactic acid fermentation in fish products 134 as component for seafood ready meals 40 RIDASCREEN. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. flow diagram (Fig. importance in lactic acid fermentation in fish products 130 importance in salting. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 growth and survival in cured. detected in cod 115 Pteriatoxins. source of PSP toxins 142 Quality Index Method. to measure fish skin resistance 10 Salmon. resulting from poorly salted cod 110 Pyrodinium. hazard in salting of fish 97 Salt concentration. flow diagram (Fig. vacuum packing 105 Salmonella spp. fermented fish product 126 Raphidophytes. smoking and marinating of fish 96 Salt-boiling 94. 135 Rakefisk. growth and survival in chilled and frozen raw fish 19 Pseudoterranova dicipiens. producers of brevetoxins 154 Raw fish. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova. for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops. for evaluation of fish quality 9 Radiation. 100 Salting 97-9 definitions 93-4 Sardines. in molluscan shellfish 60 Rice.
as measure of fish degradation 10 Sliming. 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. concerning crustacean shellfish 89 Vacuum packaging. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru.INDEX Shigella spp. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 . fish and shellfish 141-74 Trade associations. 13. for evaluation of fish quality 9 Torrymeter. 14 Torry Scale. as contaminant in scampi 36 as contaminant in enrobing process 37 as hazard in dry salting of fish 89 growth and survival in chilled and frozen prepared fish products 46-7 growth and survival in crustacean shellfish 84 growth and survival in cured. of fish after capture 5 Surstrømming. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. as indicator of fresh fish spoilage 11 Toxins. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. fermented fish product 126 Taste panel evaluation. role in fish degradation 10. in molluscan shellfish 60 Tuna. 101 Storage. of crustacean shellfish 80. fish sauce 123 Shrimps. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. grading of 31 hot-smoking 107 US legislation. processing and effects on microflora 63-4 Skin resistance. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. contact details 241-2 Trematode infections foodborne 137 Trichosporon.
as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses. growth and survival in chilled and frozen raw fish 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 fermented fish 136 growth and survival in molluscan shellfish 66-67 pathogen profile 234-5 Vibrio vulnificus. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. 152-3 256 . 68 Vibrio cholerae non-01. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01. growth and survival in chilled and frozen raw fish 16-7 growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 67 pathogen profile 235-6 Vibrionaceae. of shellstock 61. 62 Yersinia spp. 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. processing for seafood recipe meal centres 39 Vibrio spp. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus.
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