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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
thank you for all the support. for his scientific input and Victoria Emerton.Goldwyn. I thank the contributing authors for their collaboration. The books are intended as a source of information for microbiologists and food scientists working in the food industry and responsible for food safety. and growth characteristics for key pathogens associated with the product. published by Leatherhead Food International and RSC Publishing. Fish and Seafood. I dedicate this book to my husband . and Meat Products. Technical Team Leader for her valuable editorial contribution. Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged. I also acknowledge and thank Jackie Apps. potential hazards associated with the consumption of a range of products. and Ann Pernet for the work put into typesetting and indexing. both in the UK and elsewhere.FOREWORD The Microbiology Handbook series includes Dairy Products. and a detailed list of contacts for various food authorities. which has been the chief factor in ensuring the completion of this book. Rhea Fernandes Leatherhead Food International v . All handbooks also contain a review of the related legislation in Europe and UK. Alison Turner. My gratitude is extended to Dr Peter Wareing. guides to HACCP. The books in the series are designed as easy-to-use guides to the microorganisms found in foods. Training Manager. Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product.
6 Pathogens: Growth and Survival 2.6 Published Microbiological Criteria 3.3 Processing and its Effects on the Microflora 4.8 References MOLLUSCAN SHELLFISH 3.7 Published Microbiological Criteria 2.1 Definitions 4. 4.4 Spoilage 1.8 Further Reading CRUSTACEAN SHELLFISH 4.6 Published Microbiological Criteria 4.2 Initial Microflora 3.3 Initial Microflora 2.5 Factors Affecting Fresh Fish Spoilage 1.CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.4 Processing and its Effects on the Microflora 2.7 References 3.7 References 4.2 Initial Microflora 4.3 Processing and its Effects on the Microflora 3.3 Processing and its Effects on the Microflora 1.7 Published Microbiological Criteria 1.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.6 Pathogens: Growth and Survival 1.1 Definitions 1. vii .4 Spoilage 4.1 Definitions 3.4 Spoilage 3.5 Pathogens: Growth and Survival 3. 3.5 Spoilage 2.8 References 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. CHILLED AND FROZEN RAW FISH 1.2 Initial Microflora 1.5 Pathogens: Growth and Survival 4.2 Definitions 2.
4 Implementation and Review of the HACCP Plan 8.3 Processing and its Effects on the Microflora 5.4 Amnesic Shellfish Poisoning (ASP) Toxins 7.1 Introduction 189 9.7 Food Hygiene (England) Regulations 2006.5 Pathogens: Growth and Survival 6. CURED. 14 (Hygiene requirements specific to the UK) 221 9.2 Paralytic Shellfish Poisoning (PSP) Toxins 7. 7.3 Stages of a HACCP Study 8.4 Regulation (EC) No. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.1 Definitions 5. 852/2004 on the General Hygiene of Foodstuffs 191 9.1 Definitions 6.3 Processing and its Effects on the Microflora 6.2 Definitions 8.5. 2006 No. 9.2 Legislative Structure 190 9. 8.11 Conclusion 7.1 Introduction 8. 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 9.7 Further Reading FISH AND SHELLFISH TOXINS 7.3 Regulation (EC) No.10 Further Reading 223 viii .4 Spoilage 6.8 Ciguatoxins 7.6 Published Microbiological Criteria 5. SMOKED AND DRIED FISH 5.6 References 6.8 Guidance 222 9.5 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 6.8 Further Reading FERMENTED FISH 6.5 Pathogens: Growth and Survival 5.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.2 Initial Microflora 5.9 Other Relevant Legislation 223 9.3 Tetrodotoxin 7.6 Lipophilic Shellfish Toxins (LST) 7. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9. S.7 References 5.10 Cyclic Imines 7.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.4 Spoilage 5. EU FOOD HYGIENE LEGISLATION 189 9.5 Regulation (EC) No.I.9 Azaspiracids 7.1 Introduction 7.2 Initial Microflora 6.6 Regulation (EC) No.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.
7 Staphylococcus aureus 10.4 Listeria spp.5 Plesiomonas 10. 10. 10.10 PATHOGEN PROFILES 10.9 Vibrio parahaemolyticus 10.3 Clostridium perfringens 10.2 Clostridium botulinum 10.6 Salmonella spp.10 Vibrio vulnificus 10.1 Aeromonas spp.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .8 Vibrio cholerae 10. 10.
Poultry. Cross N. the consumption of fish and seafood has increased. Clark S. fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins.A. Wiley. Chandan R. Wageningen Academic Publishers. Milk. Further Reading General Fish/Seafood Microbiology Cross N.M. 2006. crustacean shellfish. cured. The second edition of this handbook is a review of the entire book for currency of information. rivers and lakes whose water can range from pristine to contaminated. Eds. 2007. Saebo A. Oehlenschlager J.INTRODUCTION Fish and seafood are a main source of animal protein in the diet. The Microbiology Handbook. Nollet L. Safety and Processing of Wild and Farmed Fish.. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs. Jacobsen C. Luten J..H. Seafood Research from Fish to Dish: Quality. Seafood and Vegetables. Wageningen.. Bekaert K.. Meat.. smoked and dried fish. thus. in Handbook of Food Products Manufacturing: Health. and fish and shellfish toxins. xi . for fish. chilled and frozen prepared fish.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish. Hui Y. season. molluscan shellfish.. fermented fish..B. Geographical region. 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. Seafood processing: basic sanitation practices. 94767. and. Catches can be gathered from seas. Often contamination is from human and animal sources.. Hoboken. Because of their health advantages over red meats.
Gram L. New York. in Seafood from Producer to Consumer: Integrated Approach to Quality. Proceedings of the International Seafood Conference. 1997. 329-65.Y. 1997. Predictive microbiological modelling and seafood quality.Y. Proceedings of the International Seafood Conference.L. Spoilage and shelf-life extension of fresh fish and shellfish. November 1995.L.. Ohlsson T. Biogenic amines in fish and shellfish. Cambridge. Johnson J.. Blackwell Publishing. Woodhead Publishing Ltd.M. 2002. Oxford. Taylor S.H.K. 245-66 Bremner H.International Commission on Microbiological Specifications for Foods. 60 (4). Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products.B. and Clostridium botulinum in smoked fish and shellfish. 36 (1-2). in Minimal Processing Technologies in the Food Industry. Noordwijkerhout. Hui Y. Blackie. 167-77. 91-8.N. 2004. Bruinsma B. Salmonella spp.H.B. in Fish Processing Technology..D. November 1995. Stanfield P.E. Ed. Huss H. Rawles D. Marcel Dekker.. Ed. 431-43. 1997.A. Elsevier.S.. Seafood: processing. 543-61.. 121-37.. 8 (2). 2003. Control of indigenous pathogenic bacteria in seafood. Fish and fish products (Microorganisms in fish products). Gorham J.H. Modified-atmosphere packaging of fish and fish products. Dalgaard P.S. 1997.. Academic Press. Tong P. Luten J. 174-249.H. Stanfield P. Ed. Gudmundsson M. Kluwer Academic / Plenum Publishers. in Seafood from Producer to Consumer: Integrated Approach to Quality. Huss H.. 87121. 2005. 318-23. Seafood processing: basic sanitation practices. Heinitz M. Elsevier. Smith J. 2002. Safety and Quality Issues in Fish Processing. in Food Processing: Principles and Applications. Huss H.. Gram L. Microbiological spoilage of fish and fish products. Eds. Noordwijkerhout. in Microorganisms in Foods 6: Microbial Ecology of Food Commodities. Microbiology of fish and fish products. 200-23. Ventresca P.. Luten J. Eds.A..A.P. The incidence of Listeria spp. Amsterdam. Oxford. Martin R. Cambridge.M. 1996.L.. Minimal processing in practice: seafood. Eds. 1996. The fish industry. 1998. 459-72. International Journal of Food Microbiology. Simpson B. McCarthy S. 413-30. Hafsteinsson H. Hall G. Ashie I. Nip W-K. New York. 61 (3). Vo1ume 39. 1996. Amsterdam.J. Journal of Food Protection. 1997.. Blackwell Publishing. basic sanitation practices. Smith J.H. Dalgaard P. xii . in Advances in Food and Nutrition Research. Woodhead Publishing Ltd. London. International Commission on Microbiological Specifications for Foods. Bengtsson N. in Kosher Food Production. Blech Z. Journal of Food Protection. Ed. Ed. Food Control.R. 2004. Flick G. London. 33 (1)..R. in Food Plant Sanitation... Blech Z. Ed. CRC Critical Reviews in Food Science and Nutrition. Hui Y. 372-6.. Davies A.
Evaluation of the bacteriological quality of seafood. Gram L. Bacterial pathogens transmitted by seafood. Microbiological quality of fishery products. Poultry and Fish Products. August 1991. 305-17. Copenhagen. 26 (3). in Fisheries Processing: Biotechnological Applications. Elsevier. in Quality Attributes and Their Measurement in Meat. 1994. Garrett E. Seafood toxins of microbiological origin.N. London. Liston J. and specifications for seafood products. Liston J. Liston J. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. 1994. 2257. poultry and fish. 1994. Hobbs G. 54-61. Amsterdam. Jakobsen M. International Journal of Food Microbiology. Bonnell A. 1994. guidelines. Jakobsen M. International Journal of Food Microbiology. 121-8. 11 (2). Copenhagen. Elsevier.H. Dalgaard P. Modelling of microbial activity and prediction of shelf-life for packed fish.S. 26 (3).R. Liston J. Huss H.. 5662. Qualitative and quantitative characterization of spoilage bacteria from packed fish. Microbial growth and its control in meat. Pearson A. 1-17. Volume 5: Microbiological Specifications of Food Pathogens. Dalgaard P.International Commission on Microbiological Specifications for Foods.. Application of predictive microbiology to assure the quality and safety of fish and fish products. Ed.R. 1989. London.D.. 1992. in Food and Beverage Technology International: USA. The microbiology of seafood products.. Microbial hazards of seafood consumption. 269-82.. 1990. Elsevier. Chapman & Hall. 16 (1). Eds. 1992. International Commission on Microbiological Specifications for Foods. Sterling Publications Ltd. London. 1996.. Ross T. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. 265-79. Twigg B. Bacterial spoilage of seafood. Copenhagen. Fish spoilage and safety. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Amsterdam. Eds. Berlin. 5 (3).. 44 (12). Food Science and Technology Today. Ed. 118-20. Bernoth E-M.A. Dutson T. August 1991. Bonnell A.H. 166-73.. xiii . Institut fur Veterinarmedizin. in Public Health Aspects of Seafood-borne Zoonotic Diseases. 46 (1). Jakobsen M.. Glasgow. Food Technology. 1991.A.M. in Microorganisms in Foods. Olley J. August 1991. Ward D. Gilbert R. Liston J. Ward D.M. Chapman and Hall. 459-78. Activities Report of the R&D Associates. Martin A. 93-105. Ed. 1995.M. Blackie. 25-39. in Quality Assurance in Seafood Processing: A Practical Guide. Blackie.D. Huss H. Ed. Eds. Eds. 319-33. Microbiological indicators and seafood processing.H.J. McMeekin T. Amsterdam. 1994. Sofos J. Evaluation of the bacteriological quality of seafood. Huss H. Ed. 1992. Fish: microbiological spoilage and safety. Gibson D.R. A new look at microbiological standards. International Journal of Food Microbiology. 359-403. Gram L. 1995. 90-106. PHLS Microbiology Digest. 1992. London. 1990.
C.. 46 (1). Seafood-borne bacterial pathogens of marine origin. Robinson R. 1986. and specifications and inspection of seafood products. 1985.R. 199-226. Norris J. Food Technology. Simmonds C. 1988. 42 (3). 103.a review.. Ed. guidelines. 42 (3). xiv . London. Hackney C. Ed. London.S. 71-117. CSIRO Food Research Quarterly.. Ed. Journal of Food Quality.spoilage of fish. Eyles M.. Microbiological standards. Part I . Applied Science Publishers. Dicharry A.K. in Microbiology of Frozen Foods.J. 11 (2). Lamprecht E.M. Wiley. 169-208. Microbiological hazards associated with fishery products. 117-27. Microbiology of fish. Food Technology. The bacteriology of fish handling and processing. Volume 1. Elsevier Applied Science Publishers.K. Hodgkiss W. in Developments in Food Microbiology. 1982. in Essays in Agricultural and Food Microbiology. Hobbs G. 90-3. Microbiology of frozen fish and related products. Hobbs G. 1988.R. Garrett E. 8-16. 104-9. Regenstein J. 1988. Davies R.Pedrosa-Menabrito A. Shelf-life extension of fresh fish . Chichester. 1987.
lean fish are those fish that store most of their fat in the liver. migratory patterns. giving it its characteristically flaky structure. while fatty fish have fat cells distributed along their bodies. Fish fillets are a poor source of carbohydrates. In addition. “fish” will be used for fresh and seawater finfish. The lipid fraction is the component showing the greatest variation. crustaceans. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. and other forms of aquatic animal as part of their ‘fish’ definition. but typically present a low fat percent when compared to other meats. The connective tissue content of fish is also lower than that found in meat. fins and a streamline body. offering less than 0.1. In addition.20%. typically 3 and 15% of total weight.21%. Protein content in fish fillet varies typically from 16 . This super class of vertebrates includes all the bony and cartilaginous finfish. CHILLED AND FROZEN RAW FISH Associate Prof. Fish are an important part of a healthy diet since they contain high quality protein. Fish flesh is dominated by the abundance of white muscle in relatively short segments.2 . In this chapter. Muscle composition and structure of fish also differ from those found in other meat. Fish can be divided into fatty and lean fish. it shows a typical season pattern especially in migratory species such as herring or mackerel. typically fluctuates between 0. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. Fish fillet composition can vary significantly within the same species due to feed intake. and excludes molluscs and Crustacea. and spawning season. respectively (1). and is mostly interspersed between the muscle fibres. However. The lipid content.5% (1). ‘fish’ also refers to the flesh of such animals used as food. which can be up to 67%. most fish contain omega 3-fatty acids and other essential nutrients.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. 1 .
and maintained at temperatures just below the freezing point.FISH AND SEAFOOD Chilled fish is fish that has been cooled to. marbling. but no further control is exercised during storage. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases. firmness. transportation and sale. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. Evisceration is the removal of the viscera from a fish. Controlled-atmosphere packaging (CAP) refers to packaging in an atmosphere where the composition of the gases is continuously controlled during storage. during storage. DMA is dimethyl amine. at -2 to -4 °C. Quality refers to palatability and organoleptic characteristics such as tenderness. colour. transportation and sale. and flavour based on the maturity. renders that fish objectionable to consumers. and maintained at or below -2 °C (normally below -12 °C) during storage. frozen or preserved. juiciness. The proportion of each component is fixed when the mixture is introduced. and texture of the fish. 2 . through their development on fish. usually carbon dioxide (CO2). but not below 3 °C during storage. This technique is primarily used for bulk storage. nitrogen (N2) and oxygen (O2). Superchilled fish is fish that has been cooled to. hence. Frozen fish is fish that has been cooled to. and maintained at or below 7 °C. spoilage microflora describes an association of microorganisms that. Fresh fish is raw fish that has not been processed. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. Spoilage describes changes that render fish objectionable to consumers. Organoleptic refers to qualities such as appearance. colour. odour and texture. transportation and sale.
2 Initial Microflora The subsurface flesh of live. Flavobacterium. since they heavily depend on the type of sample analysed and the protocol used for isolation. some species (most Vibrios) require sodium chloride for growth. as with other vertebrates. the natural fish microflora tends to reflect the microbial communities of the surrounding waters. Clostridium. Shewanella and Pseudomonas. Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp. More accurate. Gram-positive organisms such as Bacillus. potential human pathogens may be present in both scenarios. Moraxella. microorganisms colonise the skin. In general. Human pathogenic bacteria can be part of the initial microflora of fish. gills and the gastrointestinal tract of fish. posing a concern for seafoodborne illnesses (2). Micrococcus. For example.) are also common aquatic bacteria. Lactobacillus and coryneforms can also be found in varying proportions (1). processing or storage (Table 1. and typical of the fish flora. 3 . In fact. healthy fish is considered sterile and should not present any bacteria or other microorganisms.I).II) (3). The number and diversity of microbes associated with fish depend on the geographical location. These pathogens can be divided into two groups: organisms naturally present on fish (Table 1. It is difficult to estimate how many microorganisms are typically associated with fish. On the contrary. and those that although not autochthonous to the aquatic environment. Gastrointestinal tracts and gills typically yield high bacteria numbers. Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters. 1. The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. molecular-based methods have not yet been used to address this issue. 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.CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed. However. are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest. It must be noted that mesophilic bacteria can rapidly overgrow psychrotropic organisms. although these are influenced by water quality and feed. several culture media containing sodium chloride and more than one incubation temperature should be used. whenever possible. It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish.
1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.109 cells/g) * Some sources considered L.19 µg/kg bodyweight from < 102 . 4 . Toxin levels 0.102 101 .35 °C 8 . However.106 cells/g.103 High (107 .II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil.>106 Salmonella spp. 105 . Escherichia coli Yersinia enterocolitica 101 .37 °C High for most species.0. numbers of pathogenic bacteria in raw fish tend to be low. vulnificus) 5 . sewage. 4).14 . stream water. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 .26 °C 10 . In addition. and mud Ubiquitous. monocytogenes as part of the natural aquatic flora It is apparent from the above that there is potentially a very diverse range of organisms present on fish. birds.37 °C Unknown Unknown TABLE 1.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp.1 . Plesiomonas shigelloides 3 . during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria. Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp.106 cells/g (exception: V. estuarine environments.FISH AND SEAFOOD TABLE 1. and risk associated with the consumption of seafood is low (2.
and especially when fish is stored on board for longer periods. After capture. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria. icing or freezing. in which the health of each animal can be assessed prior to slaughter. birds and soil. and the killing process is designed to minimise stress. with very little control over the condition of the fish at the time of death or the duration of the killing process. with fish such as tuna and cod being more commonly eviscerated than sardines. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. Increasingly.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. hook and line.a practice that may have both advantages and disadvantages. human hands and clothing.3 1. Fish may be eviscerated prior to storage at sea . The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. This contrasts greatly with the meat industry. in recent decades. and conditions of storage. finfish will almost invariably come into contact with nets. aquaculture practices have been expanding worldwide. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms.3. In addition. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating. has an effect on the amount of stress and physical damage that the fish will suffer during capture. however. However. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating. The length of time that set nets have been in the water or the time trawlers’ nets are towed. offering better control of fish health prior to. In eviscerated fish. in which the bacterial load 5 . ropes. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans.1 Processing and its Effects on the Microflora Capture. If evisceration is carried out at sea. mackerel or herring. decks. the cuttings provide areas of exposed flesh that are open to microbial attack. or traps. handling and processing Wild finfish are usually caught by net. and their re-use will increase the crosscontamination of other fish with such microorganisms. 1. cortisone levels increase during prolonged stress and can alter the fillet quality. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside. The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea. Physical damage such as loss of scales. with the production of off-odours. During capture and storage. boxes and/or baskets. chilled brine or refrigerated seawater at -2 °C. potential contaminants added during handling and processing. digestion and off-flavours in uneviscerated fish. and spoilage. and during harvest.
oxygen etc. if there is some oxygen demand from the product. What is more. in time. and to limit the loss of quality (5).2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. often in the absence of adequate refrigeration. for example. MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. however. These containers are also used for displaying the fish during auction at the quayside. Anaerobic atmospheres have less effect on fresh fish shelf life.). and specific spoilage organisms may use other terminal electron acceptors naturally present in the fish (trimethylamine-N-oxide (TMAO). Physical barriers not only protect from physical damage.3. and of the quality of the water used for ice and to wash fish. but if applied properly should extend the life of the product. Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. fish have a higher post mortem pH. a reduction in the oxygen tension is achieved which. These changes can be deleterious. The atmospheric conditions surrounding a product may be passively or actively altered. and minimising handling will all reduce microbial cross-contamination. In MAP. 1. allowing more growth of spoilage organisms. not part of the normal flora of the meat animals (8). will result in fully anoxic conditions. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. By vacuum packing foods. they are usually process contaminants. nitrogen.FISH AND SEAFOOD can be substantial. careful and sanitary handling during processing is required to reduce the risk of contamination with potential human pathogens. Monitoring of the seawater for algal growth in order to limit the risk of algal toxin ingestion. but isolate the food in an environment different from the bulk atmosphere. cleaning of the work environment. use of effective detergents and disinfectants. from water activity (aw) through to physical damage. Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. 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. ferric ion (Fe3+)). However. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9). 7). Packaging changes the intrinsic and extrinsic parameters affecting a product. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. partial fermentation of sugars occur leading to lower pH. As with all foods. Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. In the absence of oxygen. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. by actively altering the composition of the 6 .
nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains. which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). phosphates. The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). Flavobacterium. without overt signs of spoilage.CHILLED AND FROZEN RAW FISH surrounding gas. sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. which are able to grow rapidly at low temperature. after additive application. the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . and the promotion of a predominantly Gram-positive. the inhibition of common 'spoilage' bacteria (Pseudomonas. and the problems of muscle structure damage by the modified-atmosphere (11). MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). The products. may not be considered ‘fresh’ fish. High concentrations of carbon dioxide have the most pronounced microbial effect. botulinum. In the United States. an inert and odourless gas. Many additives have been tried in conjunction with changed atmosphere. the pathogens that may be able to grow before spoilage occurs. discolour pigmented fish (11). Replacement of oxygen with nitrogen. Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). Modified-atmosphere preservation of fish was first reported in the 1930s. a modified-atmosphere may contain any gas necessary for the desired effect. gutted or filleted fish. but can dissolve into fish liquids and deform packages. Sulphur dioxide. including salt. The microflora of meat is not the same as that of whole. VP is one of the oldest forms of altering the interior gaseous environment of a pack. slower-growing flora (11). although less information on their effectiveness is available.3 °C and produce toxins. and increase in-pack drip (12). as they are able to grow at temperatures as low as 3. but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. 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. but only in recent years has it seen a marked expansion in use and market share. Micrococcus and Moraxella). Gases used in MAP of fish most commonly include carbon dioxide and nitrogen.
and the use of a HACCP plan. In the United Kingdom. botulinum. Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). The addition of high levels of carbon dioxide to these fish. 4. this problem occurs to a lesser extent with meat.g. but is recognised as a drawback of carbon dioxide application. TMAO) that can be reduced to produce 8 . oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. The growth of these bacteria is reduced by high carbon dioxide concentrations but. at all times. Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. The replacement gas should contain at least 40% carbon dioxide. grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. owing to the dissolution of cellular structures. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. adequate shelf life and cooking requirements. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13). adequate product labelling in respect of storage temperature. 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. 3.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). Storage studies have shown that the extension in shelf life of MAP cod fillets was proportional to carbon dioxide concentration up to around 50% (V/V). however.3 °C at all points from packaging onwards. however. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. For fatty fish. as already stated. These conditions include storage of the product at ≤ 3. trimethylamine and sulfides). 5. Although all fish species need to be treated separately. from both a microbiological and a chemical standpoint. may result in unacceptable colour and textural change and hence little shelf life extension (11).g. Subsequent storage must be at or below 3 °C. The problem of water loss in MAP fish is common. 2. It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. use of high-quality raw fish. should be used. Only the highest-quality fish. greater advantages are seen. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). For other less fatty marine fish. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 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). texture. reactions caused by activities of the fish’s own enzymes. Taste panel evaluation of cooked fish may be used to score or grade colour. Various methods for whole fish freshness evaluation by a trained panel have been proposed. smell and 9 . and metabolic activities of microorganisms present in the fish. and this pattern is the basis of freshness grading schemes. these are summarised in Table 1. C (C=reject) Significantly.III. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. The eyes turn from convex and clear to concave and opaque.CHILLED AND FROZEN RAW FISH off-odours. with no smell. evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis.1 Spoilage Fresh fish spoilage and methods of evaluation Food spoilage can be considered as any change that renders the product unacceptable for human consumption (10).4 1. and the flesh turns from bright and elastic to dull and soft (1). Degradation of whole fresh fish stored in ice generally follows a set pattern. A. the skin turns from iridescent to dull and bleached with bacterial slime. 17. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. or for that matter evaluation of the taste of the fish. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 .4.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids). to brown and slimy with an intense off-odour. Loss of freshness and spoilage cannot be separated as processes. so lower carbon dioxide concentrations may have better preservative effects. TABLE 1. as traditional spoilage patterns of the external organs are typically very distinct. 1. Over time. the gills from pink and shiny.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. When working with fillets. 18) and are probably the most widely used. B.
the pH drop of the muscle is correspondingly less. ADP. the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. HxR and Hx) gives a quantity (K-value. bacterial reduction of TMAO to TMA. a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP. an osmoregulator. To a varying extent.g. ammoniacal. possibly slightly metallic. 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). sea-weedy taste. iv) Enhancement of the spoilage characteristic of phase iii. sulphurous and/or rancid smells. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical. an intense odour compound. the muscles relax again and the fish returns to a flaccid state. rigor mortis is the first noticeable change in the fish. 19). The process of ATP breakdown is used as an indicator of fish freshness (24). AMP. IMP. the conductance generally increases. 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. rigor resolves. Because fish tend to have relatively little residual glycogen compared with mammals. little flavour. After death. or its remaining shelf life (1). %). which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product.5 (18. biochemical and microbiological actions. thus. or simply a waste product present due to bioaccumulation (22). By measuring the concentrations of the six components. 23). These processes are strongly influenced by the physical conditions of storage. reduction in ATP and increase 10 . the Torrymeter. From being flaccid. the Eh of fish muscle remains relatively high (20). As the fish degrades. development of sickly sweet. RT Freshmeter or Fishtester). Generally.8 to 6. which increases from 0% towards 100% with time. chemical. fruity and/or bitter off-flavours. ii) Neutral taste. iii) Traces of sour. an anti-freeze. After death. spoiled and putrid. is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20. However. post-rigor values are typically in the range of pH 5. texture becomes soft and watery or hard and dry.FISH AND SEAFOOD taste of cooked fish. as little endogenous reduction of TMAO occurs (18). 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. all marine fish use/have TMAO (21). During the initial storage. After some time. TMAO permits a high Eh to remain in the muscle tissue. cabbage-like. 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.
This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. 7). 31). 27) produced by aerobic respiration and other forms of metal ion reduction (28). as microbial spoilage is limited by the low storage temperature (1. Lipid oxidation and other oxidative changes lead to oxidative rancidity. Chemical degradation continues after the initial post mortem phase. 18). as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. 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. therefore. and a segregated weak acid to absorb them. 11 . During spoilage. so the measurement is effective only during the initial loss of freshness. hypoxanthine. does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection. these measurements are usually used to quantify the amount of spoilage. 26). The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. for example the TMA fraction. the residual acid is then titrated. Neither does the change in pH during storage correlate well with the production of TVB-N. the importance of microbial action increases with time (5. The Conway method (and variations of it) uses a strong inorganic base to volatilise the bases in the fish sample. also termed TVN). Evaluation of Total Volatile Base-Nitrogen (TVB-N. Quality indices based upon the products of microbial metabolism do not explain changes in quality until microbial growth produces measurable changes in the fish. Oxidative rancidity is known to reduce the quality of fatty fish in particular (19. Comparing results for different fish species. However.CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. 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. colour changes. is later significantly reduced by bacterial action. using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. which may also be formed by bacteria (25). Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. and are especially important in the spoilage of frozen fish. the majority of volatile bases are produced from the soluble non-protein nitrogen of the fish (free amino acids and other low-molecular-weight nitrogenous compounds). or a specific fraction of the volatile bases. however. For some fish species. however. a correlation can be made between the spoilage of the fresh fish and the production of TVB-N. not to describe freshness.
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. 34). but also crucial for the assurance of consumer safety. biochemical and microbiological reactions taking place within and on the fish. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33. The factors are interactive and cannot be completely isolated. the slower the spoilage processes proceed (within limits).5. For example. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. a minority are often associated with the majority of the spoilage. the results of altering one parameter may have far-reaching consequences on others. including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. Fish spoil as a result of the chemical. For fresh fish iced immediately after capture and continually stored in ice. Not all the bacteria growing on a food will lead to the production of objectionable characteristics. The sum of these changes may result in a different spoilage profile. by altering the gas atmosphere (extrinsic).5. careful temperature control during storage is not only important in terms of quality loss.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 (1.5. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. Also. 1. The reasons for the differences between tropical.5 1. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 .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 AND SEAFOOD 1.2 Fish-spoilage bacteria The parameters affecting the multiplication of microorganisms in foods have been categorised into two general groups: intrinsic (inherent qualities of the food) and extrinsic (qualities of the food environment) (35). The concept of specific spoilage organisms (SSOs) is not new. 1. the lower the temperature of storage. 5). The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks. For fresh fish.and temperate-water fish spoilage rates may be many.
oxidase-positive. ‘oniony’ and ‘faecal’ odours from the production of ketones. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms.g. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp.10% of the total flora of fresh fish from temperate marine waters. on fish are diverse and in many cases species-specific. plus other characteristic compounds of the species responsible for spoilage. aldehydes. its ability to reduce TMAO to TMA. the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. Members of the genus are able to produce pigments. The importance of Sh. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s.3. Pseudomonas spp. putrefaciens spoilage of fish is due to its biochemical action on muscle. Sh. It is thus an important spoilage organism of gadoid fish such as cod. mediated spoilage is characterised by ‘fruity’. catalase-positive. 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. As previously noted.2 Shewanella putrefaciens Sh. produce hydrogen sulphide (H2S) from cysteine. it is often isolated as about 1 . putrefaciens growth may be used as indicators of spoilage (30). frozen) fish products. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. although only since the late 1960s has it been realised that specific metabolites of Sh. During the 1970s. processed (e. They are generally characterised as Gram-negative rods. 35. the spoilage of a product may be strongly influenced by the conditions under which the product is held. It is also present in fresh water. and may play some role in the spoilage of freshwater fish. motile with polar flagella. 1. It is able to grow as fast as. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. which produced the characteristic spoilage compounds of the fish (30.5. 13 .5. i. 37). (9) and they are considered SSO of iced freshwater fish (35). 35).3. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine.CHILLED AND FROZEN RAW FISH started in the 1940s.1 Pseudomonas spp. more especially. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. 36). or faster than the rest of the flora of ice-stored fresh marine fish (9). The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp.e. therefore. These organisms are described as potential spoilers. obligate respiratory bacteria.
but Sh. Sh. putrefaciens were identified as Shewanella algae. The ferric reductase activity of Sh. and recent taxonomic description using modern molecular methods bifurcated this species. This wide-ranging respiratory capability is thought to be unique (39). with several new species being described. Shewanella denitrificans. putrefaciens were required. putrefaciens strains are characterised as microaerophiles or anaerobes. like all members of the Pseudomonadaceae. Research has shown that approximately 107 cfu/g of Ph. putrefaciens lies in its chemotaxis: the bacterium. Shewanella frigidimarina.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). phosphoreum has also been identified as responsible for histamine fish poisoning. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). they are heterogeneous. Later. putrefaciens growth has been collected (12. In 50% N2/50% CO2 MAP cod.3. a population of Ph. additional psychrotrophic species such as Shewanalla baltica. In fact. 41). Sh. does not show carbon-source chemotaxis. putrefaciens was not present in such numbers (23. Sh.5.how to reduce a molecule that they are not able to take into their cells efficiently. Shewanella waksmanii. unlike many other motile bacteria. the bacteria have a significant problem to overcome . baltica has been identified as the main H2S producer in cod during cold storing (37). Initially. It is also able to produce hydrogen sulphide and a range of other off-odour compounds. In terms of its taxonomy. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). it is strictly respiratory. Another distinct feature of the physiology of Sh. whereas >108 cfu/g Sh. phosphoreum were required for 50% of taste panellists to reject a sample. owing to its cell size and activity. putrefaciens has been studied. Shewanella gelidimarina. at the time of rejection. and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. as the ferric iron is insoluble. 41). mesophilic strains of Sh. Shewanella olleyana. although. 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. phosphoreum sufficient to cause spoilage was found. 38). and Shewanella profunda were described. Shewanella affinis. Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). Shewanella livingstonensis. Ph. The prevalence of the bacterium in so many environments. Shewanella oneidensis. it was proposed that this organism. Ph. and Shewanella aquimarina. 1. This type of intoxication occurs when bacteria convert the histidine 14 . but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39).
presents a potential risk. (and very rarely F and G) have been reported to cause human disease. Review of outbreak data suggests fresh fish to be safe. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment.3. they do not produce significant product spoilage).6 Pathogens: Growth and Survival Endogenous chemicals. Of 404 intoxication outbreaks of type E C. including antisera. 5) reports that fresh fish consumption has never been shown to cause human botulism. a convenient but diverse species of bacterium with a number of different types. lightly preserved fish products are associated with botulism. E and F include bacteria with minimum growth temperatures of approximately 3. B. D.CHILLED AND FROZEN RAW FISH present in fish into histamine. these risks are substantially reduced (1. Some Ph. algal toxins. this is probably due to spoilage occurring before toxin elaboration. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. Adult disease is caused by the production of the botulinum toxin. 1. However. botulinum recorded up to 1963. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43). thermosphacta in the spoilage of some MAP fish. especially type E botulinum intoxication. age.1 Clostridium botulinum C. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. Recent studies have investigated the dominance of B.6. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. it is bacterial risks that increase after capture of the fish. 5). B. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). previous exposure and access to supportive treatment. but with proper cooking.3 days after having reached 108 cfu/g (42). Growing evidence suggests a role for B. where a particular 15 . 75% (with a 35% mortality rate) occurred in Japan. Fewer still are capable of growing on fresh fish. Acetate production has been reported as a good indicator of spoilage by this organism. Huss (1.4 Brochothrix thermosphacta and lactic acid bacteria B.5. C1. Of these. E. B. bacteria and higher parasites all present some risk associated with fish consumption (1. 1. The heterogeneity of the types (A. 6). botulinum. human viruses.3 °C (Group II) and these are non-proteolytic (i. When inoculated into VP corned beef and sliced ham. thermosphacta did not produce off-flavours until 2 . Types A. types B. F and G) is well documented (44).e. 1. 5. C2. E.
there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. including epidemics in India. and proper cooking (boiling for 1 minute. Southeast Asia. parahaemolyticus outbreaks. Proper temperature control (<3. Other lightly or semi-preserved (e. 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. Only about 1% of marine isolates produce a thermostable haemolysin. again. 5). In Japan. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. due to the cultural preference for raw fish dishes. 1. the majority of outbreaks are caused by consumption of raw fish products (e. salted or pickled) fish have also been associated with botulism. parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. 11).6. 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. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk. and when in contact with contaminated water (no CDC reported cases from fish. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45). Proper chilling.3 °C) will eliminate all risk. 1978 . is small. but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. storage and preparation (11). parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). were associated with finfish between 1978 and 1998 (46). The bacterium causes primary septicaemia.2005).2 Vibrio parahaemolyticus and other vibrios V. parahaemolyticus cases have increased across the world. smoked. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. which is believed to be required for virulence. Currently. there is no specific guideline that describes a minimum level of V. Russia. a Japanese boiled and semi-dried sardine dish. sushi) (5). and North America.g.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. V. parahaemolyticus O3:K6 is responsible for many of the recent V. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . France. The first report of the organism being a foodborne agent was with shirasu. whether stored aerobically or in modified-atmospheres. This strain has been responsible for 50 to 80% of all V. the risk posed to the consumer of fresh fish. which was probably contaminated from an uncooked food source or an excreting foodhandler. accounted for the majority of cases (1. Since 1996. I-sushi. USA. vulnificus is associated with warm marine and estuarine waters. the majority are associated with products not cooked immediately before consumption. which can cause human gastroenteritis. Generally. A unique clone of V. Japan. Despite the ubiquitous nature of type E C. V. parahaemolyticus is a marine organism.g.
1. which is often fatal (>50%).6.55 cases per 100. and proper cooking will substantially reduce risk. monocytogenes has been well documented as a foodborne human pathogen. L.. Some isolates are true psychrophiles. there is no risk from the consumption of properly handled and cooked fresh fish. listeriosis cases have been reportable to the CDC. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. with minimum growth temperatures around 0 °C and optima of 15 .CHILLED AND FROZEN RAW FISH disorders). are present on fish and can grow to significant numbers during storage (1. however. Chill storage may not eliminate growth. and especially Aeromonas hydrophila. infection.3 Aeromonas Aeromonas spp. in its habitat. is historically associated with faecally contaminated water. possibly due to under-reporting because of their likeness to E.6. monocytogenes incidence has been between 0. Similar problems as those encountered for Aeromonas spp. and especially estuarine environment. The major virulence factor appears to be the production of toxins with enterotoxic. It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. Since 2000. Again. They are aquatic organisms with an epidemiology that is yet to be fully understood. It is known that Aeromonas spp. 1. Although the psychrophiles are unable to grow at body temperature. 17). in assessing the risk of fishborne infection by P. Most cases have been sporadic rather than associated to large outbreaks. their potential to produce disease-causing exotoxin in food has not been fully examined (48). but the bacterium is known to survive and grow in the shallow marine. There is no risk associated with the consumption of properly chilled and cooked fresh fish. sodium channel blocking and haemolytic activity (48). shigelloides is similar to Aeromonas spp.4 Listeria monocytogenes L. P. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. Despite this. the causative agent of cholera. but proper cooking should substantially reduce risk of infection. although lack of data concerning infectious doses and the possibility that foodborne virulence is linked to the immune state of the host mean that full risk assessment is impossible.20 °C. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis. cytotoxic. are associated with human diarrhoeal illnesses. shigelloides are encountered. V.000 17 . coli on isolation media (50). cholerae O1.26 to 0. and fish and shellfish are probably the major reservoirs for the organism (50). 5. In the US. proper chill storage will eliminate any risk associated with this organism. others are psychrotrophic mesophiles (49).
Being a psychrotroph.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. Like other toxins. Klebsiella pneumoniae and Hafnia alvei. Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . Photobacterium spp. Larvae of parasites such as Anisakis simplex are resistant to curing and marinating but can be easily destroyed by freezing at -17 to -20 °C for 2 hrs. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. Clostridium spp. neonates.. full risk analysis is not possible (52). but proper cooking will reduce any risk. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death.6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans. some Vibrio spp. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses.. and Lactobacillus spp. Infections are mostly associated with the consumption of raw or 18 .. within high histidine-containing fish (especially members of the Scombridae and Scomberesocidae families) is responsible for an intoxication termed scombroid poisoning (54). Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae.FISH AND SEAFOOD persons.48 minutes in salmon have been reported (53). there is a possibility for growth of the organism on chill-stored fish. Lightly preserved products. It is regularly isolated from seafoods (51). If fish are not properly refrigerated. 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.98 minutes in cod and D60 of 4. it is not apparent to the consumer and it cannot be destroyed by cooking.6. alcoholics. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). especially pickled fish.pregnant women. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection.6. Fortunately. leads to the production of histamine. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied. 1. at risk groups . The organism poses a serious risk in chilled products not cooked before consumption. but cooking will significantly reduce the risk of infection. 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 more especially by Morganella morganii.2002 (55)) and is also common worldwide. 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. 1. D60 of 1. foetuses.
Faecal coliform counts may be used instead of E.H. TABLE1. No. aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . 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. it may be desirable to test for Salmonella and V.IV) (57). Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. 2003. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). including parasites and/or quantity of their toxins / metabolites per unit of mass. coli counts where this method is preferred. 1. area or lot (CAC. parahaemolyticus (see Table 1.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. volume. Rome. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. 444. EC. 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. Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers. In summary.7 Published Microbiological Criteria Microbiological criteria for food defines the acceptability of a product or a food lot based on the absence or presence or number of microorganisms.1997. especially in warm-water areas and where fish are to be consumed raw.. For fish from inshore or inland waters of doubtful microbiological quality. 1997) Huss H. which in many cases will eliminate the risk. parahaemolyticus Staph.. long storage under refrigeration or one of the former prior to freezing. Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease. Gram L. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration.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. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it. FAO. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF). Ababouch L.
H. 1992. Food Control. Microbiological spoilage of fish and fish products.. International Journal of Food Sciences and Techonology. 67-71. 2000. Food Control. Jehanno D. Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review. 2002.A. 10. 5 (1). Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef. Microbes and Infection. Ed.R. 334.K. in FAO Fisheries Technical Paper No. 2. 8.J. Dalgaard P. Huss H.E. Rhodehamel E. 5. 1996.R. Incidence of foodborne pathogens in European fish. 6.. 1994. 19.. 291-304. 1993. Incidence and growth of psychrotrophic Clostridium botulinum in foods. 87121. Spoilage and shelf life extension of fresh fish and shell-fish. activities and safety. Kirby R. Jeksrud W. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003). Armstrong D. 8 (2). 1996. Food and Agriculture Organisation of the United Nations. 4. 24. 2003. Italy... 27. 20 .H.S. 14. 9. 12.A. Davey K. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth.J. 11.H. 2001. 7.. Codex Alimentarius Commission. Huss H. 283-94. Fletcher R. 243-60.A.E.. 121-37. Simpson B. 12. Smith J. 37. in FAO Fisheries Technical Paper No. 1992. Journal of Food Safety. M. Food and Agriculture Organisation. Food and Agriculture Organisation of the United Nations. Food and Agriculture Organisation. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres.T.N.J. International Journal of Food Science and Technology.. International Journal of Food Microbiology.. Rome. Huss H. Food Control. References Huss H. Ed. Reddy N... Capell C. Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms.R. Rosnes J. Gibbs P. and natural toxins.H. Codex Alimentarius Commission.8 1. Rome. Sivertsvik M. Assurance of seafood quality.H.. Critical Reviews in Food Science and Nutrition. Control of indigenous pathogenic bacterial in seafood. Feldhusen F. 3. 1651-60. Ashie I..FISH AND SEAFOOD 1. 1997. International Journal of Food Microbiology. Nychas G. 1995. Davies A. Gram L. 36. 107-27. 12. Quality and quality changes in fresh fish.R. 348. Ahmed F. 87-118.P. parasites. 13. Kautter D. The role of seafood in bacterial foodborne diseases. Davies A.. Huss H. 2. 91-8. 1994.. Gram L..K. 5-7. 1989. International Journal of Food Microbiology. 33.
International Journal of Food Microbiology. March 20. Food Technology. Gill T.. 21. 24. Bacteria active in the spoilage of certain seafoods. Surette M. Reginstein J. Børresen T. 29. in Fresh Fish.E. The micro-determination of ammonia. Hendrie M.G. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere. Letters in Applied Microbiology. Herbert R.N. 1. Conway E. . Hebard C. Khayat A. International Journal of Food Microbiology. Dalgaard P.M. Huis in't Veld J. Pedrosa-Menabrito A. Byrne A. 245-67. 1995.. 1959.H. 21 16.E.J. Schwall D. 27. in Fresh Fish.E. Hebard C. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout. Rome.J. 19-22. Huss H. Martin R. Nature. 1415. 1995.A.S. Yajima T. 25.A. 1933. 1995.. Huss H. 28.J. 130-2. Technomic. Ed. Autolytic changes in fish post mortem.. in Advances in Seafood Biochemistry. Leblanc P. Sea Fish Industry Authority. 1992. Gram L.M. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage. 1992.J. 17. 1996. 37. Flick G. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products. 41-50. Biochemical Journal. Slade A. Journal of Applied Bacteriology. Food and Agricultural Organisation of the United Nations.. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish.R. Lipid oxidation in seafood. Washington D. 21. Flick G.J.. Edinburgh. 1-18.. in Chemistry and Biology of Marine Food Products. Eds.htm Anon. Ed. 42-54. 1971. Ackman R. International Journal of Food Microbiology. 1985. 184. Chemical composition.E. Microbiological spoilage of fish and fish products. An absorption apparatus for the micro-determination of certain volatile substances.C. Martin R. Adopted.M.. 1987. Journal of Agricultural and Food Chemistry. Gibson D. 23. 1983. National Advisory Committee on Microbiological Criteria for Foods. Gill T. Part I .. 305-17. Non-enzymatic oxidation of seafood lipids. Saito T. 22. 354-8. Westport. 2007. Lancaster. 11. 33..H.a review. 117-27. Food and Agricultural Organisation of the United Nations. 1988..CHILLED AND FROZEN RAW FISH 15.H.H. Shelf life extension of fresh fish .J. Changes in purine nucleotide of red lateral muscle of rainbow trout. Davies A. 33.. Martin R.J. 26.M. Arai K. Eds. 1982. 1996. 20.E. 36. Flick G. Ratnayake W. AVI Publishing. Huss H. Quality and Quality Changes..E.J. 22-36. 26. National Advisory Committee on Microbiological Criteria for Foods.edu/haccp/compendium/chapt08. Journal of Food Quality. Qualitative and quantitative characterisation of spoilage bacteria from packed fish. 27.ucdavis. Shewan J.. 18. 121-37. Quality and Quality Changes. Rome... 30. http://seafood..spoilage of fish. Microbial and biochemical spoilage of foods: an overview. 419-29. 34. 19.. 1995.
Huss H. Pseudomonas fluorescens. FAO Fisheries Report. 1996. Libbey L. and regulation. Iron and manganese in anaerobic respiration: environmental significance. Gram L.A. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia. 26. Satomi M... Sumner J... Taussig M.S. 42. Occurrence and importance of proteolytic bacteria as spoilage indicators. Archiv für Lebensmittelhygiene. The inhibition by CO2 of the growth and metabolism of microorganisms..M. Karnop G. 1984. Myers C. Dalgaard P. Oxford. 22 . International Journal of Food Microbiology. Growth and activity of Shewanella putrefaciens isolated from spoiling fish.. Fish spoilage bacteria – problems and solutions. 1985. Miller A. 317. 1989.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J.eu/LexUriServ/LexUriServ.R. Drosinos E. Brady P... 37.H.. Scanlan R. 41.M. Applied Microbiology. Quality and Quality Changes. 34.. Environmental Biotechnology. 18-21. 36. Food and Agricultural Organisation of the United Nations. Gram L.R. 35.. 323-9. 1994. 48.. and an Achromobacter species. in Fresh Fish. Nealson K. 1995. Dixon N. Magno-Orejana F. Nychas G.europa.J. 1982. 262-6. The effect of storage temperature on shelf life. 6689-97. 1992. Blackwell Scientific Publications. Annual Review of Microbiology. 44. 2005.H. 51-62. 61-6.. Lee J. Saffarini D. 3429-38. Dalgaard P. 174. 109-36. II. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere. 13.H. Jørgensen B. Processes in Pathology and Microbiology. Myers J. Applied and Environmental Microbiology. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. Cohen D. 36.. 67. 71. 39.H. Gorczyca E. Rome. 1973. physiology.F. Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens. 33.J. Italian Journal of Food Science. Journal of Bacteriology. Vogel B. 4.FISH AND SEAFOOD 31. 32. 40.M.B. Kell D.. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish. 9. 72-80. Venkateswaran K. Rome. 311-43. 328-9. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens. 38.E. Huss H. Brochothrix thermosphacta. Journal of Applied Bacteriology. 43. 62-70. 1984. 33. Ed.. 2002. 1989. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches. Role of proteolytic bacteria in fish spoilage. 334.L.
.. in Microorganisms in Foods 6..P. Microbial Ecology of Food Commodities.. 2005. Microorganisms in Foods 2.. 20. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico. Embarek B.J. 51 (4).. Bean N. Food Technology. Embarek B.A. Lehane L... Majeed K. 1991. Genigeorgis C. 2005. Ed. Garcia G. Mac Rae I.CHILLED AND FROZEN RAW FISH 45. International Journal of Food Microbiology. Lindroth S. 46. 57. 1997. 2003. 1998-2002. 53. Huss H. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures. Presence. Bishop R.L. 55. Daniels N. Sampling for Microbiological Analysis: Principles and Specific Application. 48. Fish and Fish Products (Microorganisms in Fish Products). 55 (SS10). 12.C. Ljungh A. 23 . Morbidity and Mortality Weekly Report. 69. Smith J. Ray B. 1993..L. International Journal of Food Microbiology.. Bej A. USA.. The International Commission on Microbiological Specifications for Foods. The International Commission on Microbiological Specifications for Foods. 100-3. Use of risk assessment to reduce Listeriosis incidence. 1973-1998. 181. Jay J. 17-34. Braden C. International Journal of Food Microbiology. 51. 2000..N. Vibrio parahaemolyticus Infections in the United States.. 1986. 181-96. Lynch M. Panicker G. 172-249. 52. Slutsker L. 330-6.. 85-95. Oxford. 12.. Wilson S.. 181-8. Painter J. Loessner M.K. Kluwer Academic/Plenum Publishers.M. 2000. 23. New York. Blackwell Scientific Publications. Hammond R. MacKinnon L.. 1994. Woodruff R. detection and growth of Listeria monocytogenes in seafoods: a review. Modern Food Microbiology. 50.A.K.. 1-37. 303-12. E and F in salmon fillets stored under modified atmospheres at low and abused temperatures.. 2194200. Histamine fish poisoning revisited.H. 1991. Journal of Infectious Diseases. 49.M. 2006.and waterborne pathogens.H. 47. International Journal of Food Microbiology. Journal of Food Protection. Olley J. Surveillance for Foodborne-Disease Outbreaks . Thompson S. Springer.M. 1-34. 50. 1661-6. 58.P. Myers M.J..K. Golden D.. Applied and Environmental Microbiology.. International Journal of Food Microbiology..C. 56. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets. Miller A. Altekruse S. Wadström T. Griffin P... 54.United States.W. The International Commission on Microbiological Specifications for Foods. Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B. Aeromonas and Plesiomonas as food. 1987. Whiting R.
International Journal of Food Microbiology.. Aberdeen. 6. 1984. Food and Agriculture Organisation of the United Nations. 1.. Kitts D. McMeekin T. Magno-Orejana F.H.FISH AND SEAFOOD 1. Lancaster. 55. 1984. 51.1983. Fisheries and Food.. 1992. Hobbs G. Academic Press. in FAO Fisheries Report No. Journal of Food Protection. 13-32. 1988. 4.1 Further Reading Processing Anon. Eds. Ed.C..D... 1992. 1989. 1986. Jones Y. Rome.S. Dalgaard P. Taoukis P.9. Cann D. 24 . Gram L. Torry Research Station. Torry Research Station of the Ministry of Agriculture. Studies on the microbiological ecology of mackerel stored by the method of partial freezing.M. Torry Research Station of the Ministry of Agriculture. 15. Ravn Jørgensen B.. 35. 295-307. Food and Agriculture Organisation. 1989..P. Ross T. 317 (Suppl. 65-72. Picard G.D. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. Shahidi F. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. International Journal of Food Microbiology...2 Spoilage Shewan J. Smith G.).E.9 1. 62-70. Huss H.. in Seafood Science and Technology. Borgstrom G. Huss H.. Gibson D. Cho Y. Sumner J.M. Huss H. Bulletin of the Faculty of Fisheries (Hokkaido University).A. The Microbiology of Seawater Fish. International Journal of Food Microbiology. Labuza T. Journal of Food Science. in Torry Advisory Note No 93.. 271-85..L. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. Akiba M. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches. Technomic Publishers. Shinano H.. 741-50. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. Ed. Estimation of the bacteriological quality of fish by automated conductance measurements.G.9. Houston N. Simard R. International Journal of Food Microbiology. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. Ogden I. Trolle G.. Florida.M. 1. Villemure G. 1997.. Olley J. Microbiological quality and shelf life prediction of chilled fish. Gibson D. 317-21.H..H.. Aberdeen. 73-89.. Changes in microflora and chemical compounds in mackerel stored by partial freezing. 127-34. 1. 1961. in Fish as Food. Application of predictive microbiology to assure the quality and safety of fish and fish products. 1985. Fisheries and Food. Handling and processing of scad.
. http://eur-lex. International Journal of Food Microbiology.M. Jacobsen M. 20023.J.eu/LexUriServ/LexUriServ. London.R. 1. Liston J.. 4.europa.. Amsterdam. Elsevier Science. 2197-203.H. Hall G. 65-72. 1. 137-46. Eds. Trolle G.3 Pathogens Condon S. Gram L.K. Journal of Applied Bacteriology..9. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish.9.4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. 1992. in Quality Assurance in the Fish Industry. Huss H. 5. Klausen N. A rapid method for detection of histamine producing bacteria. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. Huss H. Sala F. 25 . Gram L. in Fish processing technology. Applied and Environmental Microbiology.. 322-6. Effect of culture age. Huss H. 1987. 1987. 59. Otero A.L. International Journal of Food Microbiology. 72. 1997.. Garcia M.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A..H. Modified atmosphere packaging of fish and fish products. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. Ed. 479-88.. 1993. 1992. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. Blackie Academic & Professional.H.CHILLED AND FROZEN RAW FISH Einarsson H.
Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2.1 Introduction It was often thought that whilst the highest quality seafood products are processed and sold as whole fish and/ or fresh fillets (e. or could be disguised by sauces. 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. products of lesser quality. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e. although recent developments in improved supply chains. colour of gills. smell etc. Even though heating prior to consumption will eliminate pathogenic microbes. smoked (cold) salmon. Few seafood products are sold ‘ready-to-eat’. The majority of seafood products produced require a cooking stage before consumption. Sushimi tuna). due to the early cessation of spoilage. 27 . 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.g.2. the exceptions being Sushimi (high quality raw fish). This is no longer the case with many processed products now available in stores. seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. 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. it does not necessarily apply to the toxins that may have been produced. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr. smoked (hot) mackerel. colourings etc. but still acceptable and meeting all safety criteria.). emphasising the high quality of the seafood and the gourmet nature of the prepared products. clarity of eyes. The same used to be said for the difference between fresh and frozen products. and crustacea such as shrimp and crabs.
. they are usually of a better standard than species such as tilapia. molluscs. They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. from both wild capture and aquaculture sources. They are typically flown into export markets. shellfish and molluscs. often using the simplicity of a microwave oven. The expected shelf life attainable for this format is usually up to 8 days. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein.. and these are increasingly being presented with an ever-expanding range of accompaniments. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP). and this has produced a greater availability and diversity of products in the marketplace. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted. and are well known for their diet consisting of waste materials. Shelf life on ice. rather than frozen.FISH AND SEAFOOD Recent trends in the market for food and seafood products. Cold-water fish microbiological specifications are inappropriate for assessing freshness of warm-water fish. There is also a limited market for filleted material. Listeria spp. Nevertheless. Prepared natural fillets are fillets of fish without any additional coating. which can be farmed. as they tend to lose colour and sheen after defrosting. At the same time. Doubts have been raised over the microbiological status and handling practices of reef-caught fish. Frozen fillets may have a surface layer of 28 . prawns. and it is worth noting the additional hazard to fish handlers from Vibrio vulnificus from injuries received during handling and filleting of spiny tropical fish. and the consumer only has to heat (cook) the product. but these are now finding favour as products in their own right. they are usually supplied filleted with head on. Microbiological hazards include Salmonella. including freshwater fish. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. Seafood is the term used descriptively to include all categories of product. the processing industry has developed a wide range of value added products. in particular. This includes all categories of seafood including trout and other freshwater species. have shown the increasing importance of convenience foods. but should be equivalent to cold-water products. 2. where the preparation has already been carried out.2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. and pathogenic Vibrio spp. or in MAP depends on catch conditions and treatment. etc.
and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. Added colours can be included with the brine solution to meet specific requirements for products e. or pre-packed frozen with added glaze.. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. swordfish steaks). 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. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes.g. Cold smoking is essentially a flavouring process. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product.2. which is approximately 30% (or 1 . Some products may be salted for flavour only (e.5% in the aqueous phase. cod). or pre-packed in MAP or vacuum packaging (VP). be flavoured with herb dips or peppercorns. etc. An alternative process using a liquid smoke in the brine is claimed to result in a similar flavour with reduced water loss in product and improved yields.5%. 29 . sawdust with other woods adjusted for flavour profile and burn duration). mackerel. with a minimum residual salt content of 3. The result is fish with a typical residual salt content of 1. 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. resulting in a product that is ready-to-eat without any further cooking.g. This process in effect cooks the product and reduces or eliminates microbial contaminants. but may be increased (up to 1 month) by packaging methods like MAP or VP. Fish steaks can be cut from a natural fillet or loin (e.2 days) longer than that of non-prepacked fish. which may.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. haddock. the ingredients premixed. Again. Hot-smoked fish are prepared from brined. salmon.5 . A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes. in VP or through mail order. The shelf life for MAP fillets is typically up to 5 days. natural fillets of fish such as herring. mackerel. and salmon. The expected shelf life of these products is up to 8 days (or longer for some chilled.g. annatto with smoked haddock. VP products) because of the high salt content acting as a barrier to spoilage. Pâtés are prepared with raw fish.g. tuna steaks) or band-sawn from a frozen block (e. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. They are sold from wet fish counters. Fresh fillets are sold from wet fish counters. and the product then shaped and heat-processed to cook the product before being chilled. in addition. a protective glaze may be added to the frozen steak.
with an expected shelf life of around 7 days. rice. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process. Natural prawns from cold or warm waters can include those peeled. it can be used as a seafood analogue. fish nuggets/bites with dips and seafood-based snack products. tuna. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. long-life seafood mixes using sorbate and benzoate as preservatives. rice or pasta. They are pasteurised products and treated as high risk raw materials. and seafood mixes. 30 . salmon) coated and sold as chilled products. With further processing. and formed into round cake shapes.or a five-stage process. ‘fingers’ before being coated in batter and/ or breadcrumbs. pasta or potato. There is an increasing market for high value fresh or thawed fish (e. and in the shell.. This ‘catch-all’ category includes seafood salads. flavours. mixed whitefish. e. e. The majority are products made from various types of whitefish (cod.g. and may also include additional ingredients such as fresh herbs and spices.FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate. 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. Breaded fillets/steaks and battered fillets are either natural or shaped fillets. haddock.g. Other. and heat-fixed. which may include molluscs. It is often used as a protein supplement.g.) Raw material is typically fish fillet. taramasalata. coated in batter or breadcrumbs. The protein source can be fish fillet or blocks. Fish cakes are products produced by mixing fish with potato and herbs. Surimi is the insoluble protein fraction of fish after washing and processing. Scampi are tails of Dublin Bay prawn or langoustine. binders. etc. jellied eels. or cut from a preformed frozen fish blocks into shapes. Prawns. cephalopods. Band-saw dust and fish trimmings from fishblock preparation materials may be used. addition of ingredients such as colours. Other ingredients may be natural or reconstituted. The cakes are usually coated using a three.
specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). 6). the microbial flora of seafood is associated with the gills. Contamination will. For a summary of background information on the hazards associated with fisheries products and ingredients. and the potentially toxic by-products of histamine-producing bacteria (5.3. After capture and during any subsequent processing. and ingredients added to the final product.e. 2). Only after death. as a result there is an increase in spoilage rate thereby reducing the product shelf life. 31 . skin. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning. will the saprophytic flora start to invade the surface flesh (1. A primary role in fish spoilage has been assigned to this organism. The edible tissue of some species of fish are more susceptible to gaping than others. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. which may become a focus of contamination deep within the muscle. or visible damage from gaff marks (4). Tuna that does not meet the highest of grades based on colour. Grading is based on size of fish (>25 kg). and pathogenic flora associated with seafood. Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. Recent public opinion has favoured the use of dolphin. whilst gaping is also dependent on a range of other factors that effect the muscle composition. The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants.I. fat content. product handlers. and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. Dalgaard et al. on the whole. handling or preparation stage.and turtlefriendly long lining to the less discriminatory netting. sensory assessment of freshness etc. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). In summary. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. searing or bar marking). and intestines of the fish. it may be used for other products or markets depending on the product specification.3 2. species (Bluefin>Bigeye>Yellowfin>Albacore). storage. is not used for sushi or other high risk (and high value) products. remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. and processing conditions such as dehydration or poor filleting practices.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. including seasonal variation. see Table 2. and have emphasised the origin and sustainability of the fish as key marketing points.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. i.
by the HACCP team or by a hazard and risk assessment programme. pseudomonads. nuts and fruit Associated microflora Lactobacilli. toxin. moulds Bacillus. Clostridium Lactobacilli. product assembly. mushrooms.I provides an overview of common raw materials.4 2. Pseudomonas. and milk Cheese Water/Ice Bacillus. moulds Bacillus. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter. be considered at an early stage during new product development. Clostridium perfringens. coliforms. Penicillium. cereus Bacillus licheniformis. cereus Staphylococcus aureus. Vibrio cholerae. histamine Cryptosporidium.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients.FISH AND SEAFOOD 2. Enterobacteriaceae. Clostridium botulinum. staphylococcal toxin. for chilled products. Aspergillus. TABLE 2. In addition.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. raw materials. and associated spoilage and pathogenic microorganisms. yeasts Pathogenic microflora/toxins Salmonella. Table 2. B. protozoa and viruses B. cream. Bacillus cereus. shelf life determination. Bacillus subtilis B. streptococci. Staphylococcus. Aeromonas. Shigella. Listeria.4. however. moulds. Escherichia coli. anaerobic spore-formers. moulds Bacillus Bacillus. cereus B. spore-formers. coryneforms. intermediate processing. enteric viruses 2. 32 .3. Fusarium. Moulds Enterobacteriaceae Salmonella. and so are the potential microbial contaminants associated with them. 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. 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. and.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. Staph. yeasts Lactobacilli.
1 . defined holding time at a chill temperature (0 – 3 °C). but will also need to ensure that the optimum shelf life can be safely obtained. Rapid controlled cooling of heat-processed components will help to minimise spoilage. this may be an incidental effect.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. line extensions and process reviews. 2. It is recommended that the principles of HACCP are applied to novel processes. to evenly raise the temperature of the entire frozen product from -20 °C or lower. Examples of tempering techniques include the use of heated air.2 Raw material tempering Whenever frozen seafood is used as the raw material it is normally tempered before processing. there are a number of distinct methods used. Generic examples have been used to illustrate only the major points. up to -5 °C so as to improve the cutting and handling properties during processing. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. 2.3 Preparation of fish component of chilled and frozen recipe dishes 2. Effects on the microflora are minimal during tempering (7). or the use of microwave technology. the core temperature. but it also poses a risk of cross-contaminating other products undergoing tempering. Band-sawn materials or fish dice are normally promptly further processed.3. depending on the size of the fillet 33 .4. For some components. and potential opportunities for growth of foodpoisoning bacteria. but it is assumed that any band sawing or dice process will introduce heat from blade friction.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. Block-sawn dust that is required for use in fish cakes is swiftly transferred to a chiller to avoid water loss and spoilage from process contaminants.4. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface.5 illustrate basic principles in the preparation of chilled and frozen recipe dishes.2. as a heat process is often required for functional or organoleptic reasons. Figures 2. Where the raw materials are frozen. as the full complexity of the processes used for specific products cannot be covered in a general text. and contamination from equipment to clean fish muscle. All have the same intention. It is critical that the time and product temperature during tempering be controlled to avoid elevated temperatures (>5 °C) on the product surface allowing microbial growth or the onset of spoilage in raw material.4.
Scampi. chilled or sold chilled on defrost with an expected shelf life of 6 . It is then size-graded. frozen in a nitrogen blast freezer and weighed into sacks for later processing. are placed into plastic boxes and covered in ice to preserve their condition.FISH AND SEAFOOD processed. Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen.1C .2. Fig.4. may not rise above 0 ºC during processing. The “blown” meat is graded.Shaped Fillets. quickfrozen.3. On arrival at the processing plant.1.1 Whole-fillet / scampi Scampi (Nephrops norvegicus). Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product. 2.1.7 days. Raw material preparation stages for chilled or frozen coated fish products 34 .2). 2.1B . Frozen cores are usually batter-enrobed. The processing of three examples types are described in Figure 2. the scampi is washed in potable water to remove foreign bodies.1 (2. after landing.4. by mincing and mixing with a binding agent (as described in section 2. drained and then refrozen for storage until required for further processing. with the coating heat-fixed.3.Fish Cakes). 2. This material is transferred to a processing facility with the ice replaced as necessary to ensure that the product is maintained at the temperature of melting ice.1A .1.
2 Shaped products This type of product falls into two categories.4. Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. ii) Fish-filleting waste . fish fingers). Fish cake cores may be batter-enrobed. such as aerobic spore formers and Staphylococcus spp. but. The mix is shaped and formed ready for packing or for enrobing. Where frozen blocks of fillets are used they will be “tempered” (temperature raised to around -7 °C) to facilitate efficient cutting with a band-saw to a specified thickness. that are to be reused for other foods. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. Waste material produced from this type of processing is often reused in the third category (fish cakes).CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. pathogens such as Salmonella and Listeria. Therefore. The fish component of the cake will. The remaining dried ingredients are mixed with the fish. additional spoilage organisms may be introduced. those that use frozen blocks of whole fillets. with the coating heat-fixed. and water is added to rehydrate the components. unless it is poorly handled.off-cuts or belly flaps. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. quickfrozen.7 days.1. from potato flake.1. or sold as a chilled product after thawing. or promptly processed to avoid further quality loss. or those where the trimmings. It is for this reason that all by-products from primary processing. are handled and stored as any other food product.. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process .block-sawn dust. because of its nature. iii) Fresh/fillet/minced fish block. as the physical structure of the material has been destroyed.4. 2. This is especially the case with block-sawn dust. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. material will need to be transferred swiftly to a chiller. with an expected shelf life of 6 . chilled. once the dry components have been rehydrated.3.3.g. spoil more rapidly than the dry ingredients. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or. 35 . more seriously.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.
e.4 Battering/enrobing process Figure 2.. some of this initial flora 36 . with the coating heat-fixed. If a crumb coating is required.7 days. Tempered band-sawn materials b. raising agents.4. and possibly Staphylococcus spp. flour. starch. Spore-forming anaerobes such as C. milk powder and spices. egg. and Enterobacteriaceae from flour.g. Breaded or battered fish fillets are fully heat-processed before consumption. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process. is the addition of a pre-dust. perfringens could.2 illustrates the typical stages of enrobing or breading. from flour. rapidly multiply if strict temperature control was not maintained. such as Listeria and Vibrio spp. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. Fish products.or fivestage) frozen or chilled. as well as ensuring that there is minimal contamination from human processors. The function of this is to improve the adhesion of subsequent coatings. vibrios and other fishassociated spoilage organisms. Natural fillets d. a characteristic flora will develop from the batter components. e. any excess is removed with an air knife. Initially.FISH AND SEAFOOD In all of the above examples. or sold as a chilled product after thawing. spices. Over time. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. i. and may include milk powder. because of the intensive handling required during processing. The first stage.g. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. and to carry flavourings where required. and if required. fish cakes Portions or fillets may be batter-enrobed. e. quickfrozen. which typically consists of flour. potentially. rusk or crumb. which can occasionally be isolated from protein-handling processes. Products are preformed and presented to the process (either three. which is product-dependent and not always required. with an expected shelf life of 6 . The batter in the enrober is continuously recirculated and fresh ingredients (normally a dry pre-mixed recipe) are constantly added to a pre-mix vessel with potable water. A large variety of product formats and types can be batter-enrobed. 2. a. absorb any excess moisture on the surface of the fish portion. lactobacilli from the milk powder and flour. IQF/pre-formed fillets c. Bacillus spp. chilled. Batter contains water.
replaced in part by process contaminants and bacterial flora from seafood. but it is believed that poor hygiene regarding product and food handlers. dry batter mixes are all potential sources.2. With inadequate temperature control of batter systems. cereus.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. contaminants such as Enterobacteriacae can proliferate and develop during the process. aureus and B. It is advisable that some control mechanism is employed to prevent toxinforming organisms from multiplying to dangerous levels. time-limitation or production run temperature control. and both of these can produce heat-stable toxins that can survive a batter heat-fix process. This can be achieved by frequent clean-downs and adequate hygiene. this is not of great significance to the microbiological standard of the finished product. Fig. Staph. The FDA has issued clear guidelines on acceptable 37 . and.2. in the case of B. but because batter is heat-set.CHILLED AND FROZEN PREPARED FISH PRODUCTS associated with the ingredients will be diluted out. The origin of contamination for the two organisms is often unclear. cereus can proliferate under certain conditions. Flow diagram outline of a typical three.
There are two main categories. and must not show evidence of disease. 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. Fig. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. as this could introduce off-flavours and reduce shelf life.g.4.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes.4. Flow diagram outline for recipe dish ingredients preparation 2.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. Figure 2. 38 .3. A temperature no greater than 10 ºC for 12 hours cumulatively or 21.5. pasta or potato. but they are supplied without a source of carbohydrate e.2. to produce a specific product. The protein source can be fish fillet or block. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products.1 ºC for no more than 3 hours is recommended (1).5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe. canned fish may be used in their preparation owing to it having wider availability and low wastage. rice. For example. The vegetables must withstand processing. 2.
However. for example. and may include molluscs or prawns that may or may not have undergone a minimal heat process. and protected from any post-processing contamination. Products must be washed thoroughly to remove pests and soil. all-year-round fresh vegetables are preferred. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. Any assumptions regarding the application of good agricultural practice to produce should be made with caution. any processing stage.lactobacilli. will disperse the saprophytic flora of vegetables . colour and water will occur. A number of examples of direct faecal contamination of vegetables exist in the literature (8). rice or pasta. haddock. or breadcrumbs. For premium products. Although the processes are claimed to be superior to using unmodified hypochlorite for washing.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. they should be stored under temperature-controlled conditions. not just as accompaniments. control is more onerous. encourage more rapid decay and loss of structural integrity. coli O157. frozen block or canned fish. even those with pH-modified dressings. but also as components in recipe meals. if the process is not appropriately controlled. Raw material is typically fish fillet. coliforms and pseudomonads throughout the mix. and reduce spoilage organisms and potential pathogens. spore-formers. loss of quality. However.3 log reduction in bacterial numbers on the surface of vegetables.4. i. Blanching of vegetables can be an effective means of reducing or removing surface contamination. For example. pastry. 2. The opportunity for growth of L. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. with the accompanying release of additional nutrients by processing. For example. and E. Once vegetables have been blanched. The presentation of vegetables has a significant effect on the product quality.e. In an attempt to improve washing and reduce chlorine taint.5. Salmonella typhi. coryneforms. the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. and mixed whitefish). 39 . Washing with a biocide will at best achieve a 2 . 8) will. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components. and may include sauce accompaniments. (7. The majority are products made from various types of whitefish (cod. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. chopping and mixing of material. treated as a highrisk ingredient. with cases of foodborne disease traced to Listeria monocytogenes.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings.
13). Typically.) to the product (12. cereus. Alternatively. An incident involving Staph. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. On hydration. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. 40 . rely on the use of microbiologically sound ingredients. especially in those products that have a vegetable component or rice accompaniments. and an increased risk of contamination from L. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. 11). 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. and then cooled slowly allowing germination and re-growth. Sauces or marinades are added to fish to broaden appeal or to accompany components. cereus). the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. For this reason all products. 2. B. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B. if subjected to temperatures between 20 and 40°C.6. Products or dried ingredients contaminated with either spores of aerobic bacteria (e. a heat process in excess of 73 ºC will be required to rehydrate this component fully. If the sauce contains modified starches for freeze/chill stability. Potato toppings made from flake or powder will normally have little associated spoilage flora.4.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. a process equivalent to at least a 5D Listeria kill will be necessary. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC.g. cereus are well known and are still a common issue in the UK (9). such as butter garnish with black pepper. Sauces can be multi-component preparations or more simple preparations. can germinate and rapidly multiply. Appropriate handling facilities or equipment will be required to protect the product.4. where composite meals are heat-treated. Placing such products in a standard chill store to cool down could result in condensation.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. spore-formers can proliferate if adequate control of temperature is not maintained. but temperature-abused pasta can also spoil rapidly by mould growth.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. This is a particular issue especially where products are heat treated to eliminate vegetative cells. aureus toxin is well documented (10.
41 .2.4 and 2. It is expected that this product format would have a restricted shelf life to limit potential for growth of psychrotrophic bacilli or any survival of enteric pathogens (12.5 illustrate the recipe dish ingredients preparation and assembly and packaging. the sauce component will spoil rapidly and the possibility of vegetative pathogens from post-process contamination or raw components growing to significant numbers cannot be ruled out. respectively. that of a dairy-based sauce. for instance. Materials prepared with regard to good hygienic practice need only be assembled in a tray or pouch before chilling or freezing in readiness for distribution. flour products.herbs.4. suspended in an acidified sauce with an oil component. and handling of processed sauce can be less rigorous than. etc. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. Marinade-type products are becoming more popular. 13). Figures 2. The format uses flavour compounds . Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. Without barriers. spices.
4.2. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. These include: 2.2 Modified-atmosphere packaging Originally developed for the red meat industry. The oxygen-free conditions will however allow other anaerobic bacteria such as C.FISH AND SEAFOOD Fig. and tuna loins are packed by placing the product in a plastic pouch and removing all the air. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. as it dissolves in the surface moisture to form a weak acid. unless the temperature is strictly controlled to below 5 °C throughout the supply chain.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. Flow diagram outline assembly and packaging of recipe dish 2. which may restrict either microbial growth or enzyme activity. botulinum to grow. generally carbon dioxide and nitrogen. 42 .5.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions). 184.108.40.206.4. the method replaces air in the sealed packaging with a defined mixture of gases.
5 Spoilage During processing. except for some crustacea. initially. Beyond this. Aeromonas spp.. 2. It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage. but little on in-pack spoilage. (20). Not only will these organisms have a numerical advantage over other proposed spoilage organisms. The literature has good information on spoilage of individual components of recipe meals. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18). a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood. but they will also be readily adapted to the nutrient source and conditions. Spoilage will be most readily detected in proteinaceous components of ready meals. Nonetheless. the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. is typically uncooked or at best lightly processed to retain eating characteristics. and in influencing the final eating quality of the product. The case for growth and contamination of products with Aeromonas is stronger. where the protein component is nearly always heat-processed. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. 43 . temperature control is important in preserving chilled integrity of ingredients. This is considered to be the one major difference between fish and meat products. and finally flavour loss.5. and Yersinia spp. at slightly higher temperatures (10 ºC). However. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). Shewanella putrefaciens and Ph. both organisms are capable of growth at chill temperatures. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15). followed by moisture loss.10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. in numerical terms. but is not limited to. Vibrionaceae will succeed (17 ).1 Chilled storage For chilled products or chilled components of frozen products. The fish component of recipe dishes. by a loss of texture. Spoilage of this material is not expected to differ greatly from the processes described elsewhere. At refrigeration temperatures (5 . phosphoreum. as this organism is a common contaminant of freshwater fish species (19). This is also true of chill storage and the distribution chain. advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’.
freezing also has a protective role in improving the safety of raw materials by. within dairy products are well known.FISH AND SEAFOOD 2.5. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. botulinum. By retaining the intrinsic spoilage flora of seafood. auto-oxidation of fats/oils).2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage.3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. this should also take into account shelf life of product components used in the assembly of the final product. However. producing characteristic off-flavours.g. Bacillus spp. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e.. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme.5. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. The type and nature of the enzymes and their activity will depend on the bacterial species. Still. 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. the shelf life of products is limited by the natural spoilage process described earlier. Serratia spp. This reinforces the need for prompt processing and temperature control of ingredients and finished products. eliminating nematodes in products that do not have a heat treatment step. Pseudomonas spp. 44 . Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24). As a note of interest. for example. The conditions and freezing process most favourable for good-quality products will. 2.. where the variety of components and ingredients used in manufacture is varied and complex.. For example. enzymic processes are often ignored or overlooked as insignificant or unimportant because of the rapidity of fish spoilage. also be those that have the least effect on the viability of microorganisms (21). which would warn the consumer of the potentially unsafe nature of the product. and Aerobacter spp.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. however. These have been proved to impart off-flavours. Products that are developed with an extended shelf life beyond the 6 . released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15).
Counts of between 103 . any risk of C. perfringens contaminating seafood will depend on the source of raw material. or a heat shock followed by holding at a temperature suitable for growth. farmed or free-living crustacea (30). histamine intoxication.. Cases quoting contamination of seafood with organisms such as S. are dealt with elsewhere in this book. Inappropriate holding times and temperatures will. A number of other Bacillus spp. Incidents in such diverse materials as cream. 2. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. and many raw or processed foods (26). perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). typhi.3 Clostridium perfringens A study examining C. Hazards attributable to the non-seafood components are listed below and this information. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. 27).6.6. botulinum type E and Vibrio parahaemolyticus (25).104/ g should be treated with some concern (26). Germination of spores requires favourable conditions. with pathogenic microorganisms or viruses. and this issue may be of greater consequence for warmwater.2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. air. low oxygen or anaerobic conditions. 45 . Additional hazards from marine bio-toxins. permit rapid growth of B. spices and herbs and vegetable shoots have been reported (12.6 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. and some effects related to product type and time to toxin production have been noted. 2. potato. In addition.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. 13. along with previous discussion. These are C. cereus. botulinum when held in favourable conditions for growth (28) i. in some cases.6.1 Bacillus cereus This is a common contaminant of dust. agricultural run-off or sewage can contaminate in-shore species. may be of benefit within any risk assessment for chilled and frozen prepared fish products. water. Therefore. soil. have been linked with food poisoning and should also be considered. and Campylobacter are recorded in filter feeding molluscs such as mussels. etc. Vibrio spp.e. 2.
but under the appropriate conditions.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). An acceptance has now developed in the UK of the presence at low levels of L. Rates of contamination with smoked seafood are much higher than those with unsmoked products. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. infected food handlers. 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). but. 2. 2. growth may occur in foods (26). 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. at the present time.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). which is not affected by any subsequent cooking process. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32).6. if any other potentially pathogenic organisms. It is unclear. L. Shigella spp. and inadequate cooking or holding within the temperature growth range (27). will colonise factory environments in a similar fashion except for possibly Vibrio spp. 2. 46 . Sources of contamination are many because of the widespread distribution of this organism in the environment. These include inadequate refrigeration and preparing foods far in advance of planned service. indigenous or other.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. aureus enterotoxins in foods.6. when introduced or inoculated into sterile components of chilled prepared recipe dishes. monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. and this is believed to be due to a tolerance to brine used in the smoking process (32). Chilling may help Shigella to survive.5 Shigella spp. aureus (35). the restrictions on growth rate seen with seafood will not apply. monocytogenes as a contaminant in foods (31). These conditions will allow the production of the toxin.6. L. However. and will therefore cause “food poisoning” even in a product that is cooked before consumption.
are not mandatory. Microbiological specifications do not necessarily relate to product safety. 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. with respect to the number and types of ingredients used. . etc. such as batter mixes. Microbial guidelines. it is not surprising that Microbiological Standards (defined as mandatory compliance required (37)) are limited to those raw materials in which specific hazards occur. and are a recognised means of providing data on manufacturing standards. cereus emetic toxin are often confused owing to similar symptoms. It should be remembered that 47 . (36). as published by for example. The following tables (Table 2. pastry.7 Published Microbiological Criteria Due to the complexity of many processed food products. 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). and not to final products. Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems. 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.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. although due to long analysis times required. 2. 2. but these may be eliminated by a subsequent heat-treatment stage. aureus enterotoxins and B.CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts.6. pasta.III based on IFST Guidelines (39)) provide example guidelines for seafood products. and as such the monitoring is for verification purposes rather than control. unless the criteria is for a pathogenic organisms where only non detection is acceptable. For instance. but provide the industry with an indication of the microbial level at which action is required to ensure product safety. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). 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. and their association with similar materials. the fresh product is often consumed before the results are obtained.II and 2.
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. fish fingers. pasta. aureus V. 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. monocytogenes B. coated fish.g.II IFST Guidelines for part-cooked foods – e. batter etc) should also be consulted when establishing the specifications for raw materials and final product. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp. cereus Staph. TABLE 2.FISH AND SEAFOOD the microbial criteria for other products (rice.fish products. L.III IFST Guidelines for processed foods . coli GMP* <102 Maximum 104 TABLE 2. 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.
Powers E.M. Mejholm O. London.. 2. 1997. 1997. 1997. 96. 233-7. Bacterial histamine production as a function of temperature and time of incubation.E. 14.C. Journal of Milk Food Technology. Pathogenic microorganisms associated with fresh produce. 38 (11). Journal of Food Protection.R. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. Washington D. Incidence and levels of Bacillus cereus in processed spices.. Fish: microbiological spoilage and safety. Christiansen T.M. Journal of Food Science. Masuoka Y. Ward D. in Fish Inspection Quality Control and HACCP: A Global Focus. Behling A.K. Wienke A.. Journal of Hygiene. Tu A. Recent Advances in Food Science. Oxford. Collette R... Hoshino T..A.. 166-73. 10.. Food and Drug Administration.J. 2. 49 4.. London.J. Kluwer Academic. Powers E. Hackney C. 1991. 47. International Commission on Microbiological Specifications for Foods. 1. Woolaway M.C.. Cambridge. 24 (5).R. Toxin Related Diseases: Poisons Originating from Plants. Aureli P. Communicable Disease Report. 11. 1982. Letters in Applied Microbiology. Microbiology of processed spices.C.L.W. 15. Huss H... 1311-7. Gilbert R. 6. 204-16.H. 1993. Microorganisms in Foods 6: Microbial Ecology of Food Commodities.. New York. 373-8. 668 -70. Van Nostrand Reinhold. 167-93. Yumoto I. Proper processing and handling of raw tuna. IBH Publishing Co. Ohgiya S.8 1. Dalgaard P. 2001. 8. 1976. Bartlett C. Letters in Applied Microbiology. 7. Microbiology of Marine Food Products. Communicable Disease Report.L. Bartram P. Journal of Milk Food Technology.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). 5. 2005. Fish and Fisheries Products: Hazards and Controls Guidance. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature.R. 13.R.. Matsuyama H.. Importance of Photobacterium phosphoreum in relation to spoilage. Pvt.L. 1991. Outbreaks of foodborne illness in humans.G. Hobbs G. 59 (2). 67-73. Martin R.. Taylor S.R. 215.. 39 (10). England and Wales Quarterly Report. References US Food and Drug Administration. 1962. Lawyer R. 7 (24). 25 (1).M. Slavin J. Animals and Spoilage. Latt T. The bacteriology of fresh and spoiling fish and some related chemical changes.. 1997. Shewan J. 70-2. Eds. 663-87. Brown T. Murrel H. 1976. Taylor and Francis. Ishizaki K. Kumeta H. 9.. 5 (3). 12. Sakamoto T. Ltd.. Food Science and Technology Today. . 1986.. Beuchat L. 3.T..
655-60.. Halligan A. 41-3.. London. Gould G. 4. 1990. Gram L.K. Trolle G. 1974.E. Is Aeromonas spp. Blackie Academic Press. 1992. Lund B. Effects of freezing and storage on microorganisms in frozen foods: a review. Leistner L. 2000. 26. Walker S. Blackwell Scientific Publications. Granum P. Anon.A. Andersson A. 11. Speck M. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures.. 27. International Journal of Food Microbiology.. PHLS Microbiology Digest. 32.M. Fresh and processed fish and shellfish .. . Golden D. 1986. 28. Eds. Ray B.H.. Matches J.. in Anaerobic Bacteria in Habitats Other than Man.. 13 (1). 34-40.W. 274-86. Clostridium perfringens in the environment. Food preservation by combined methods... Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens. perfringens? International Journal of Food Microbiology.H. 1996.J. Huss H. 1994. Liston J. 1979. 1996. Keusch A.. 23. 1995. New York. 309-16. Wood L.J. Jemmi T. Ronner U. Huss H. Loessner M.J. Leatherhead Food Publications. 13. 20. 19. Gram L. 1992. Jay M. 1987. Glasgow. Journal of Food Safety. The Emerging pathogens . Brooks J.A Literature Survey. 22. 28. Blackie Academic and Professional. Oxford.Yersinia. 65. Leatherhead.L. 59-72. Eds. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service. Aspen Publishers. 21..C. 31. Food Control. 30. 4 (1).M. 351-72. Microbiology in fishery science.J. Springer. Liston J. in The Microbiological Safety and Quality of Food. 472-506. Ed. Lund B. 333-6. Food Research International. Food Microbiology. Journal of Food Protection. Mead G. Barnes E. Report on Vacuum Packing and Associated Processes. in Advances in Fish Science and Technology. 18.FISH AND SEAFOOD 16. Applied Microbiology. Connel J. 1993. U. 1977. a foodborne pathogen? Review of the clinical data. 50 17. Her Majesty’s Stationery Office.. Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants. International Commission on Microbiological Specifications for Foods. 2005.. 29. Modern Food Microbiology. 24.R. London. 40 (5). 1988.. 1996. Survey of the incidence of Aeromonas and Yersinia species in retail foods. Gaithersburg.C. What problems does the food industry have with spore forming pathogens B. cereus and C.V. Food Taints and Off-flavours. coli (VTEC) . Baird-Parker T. 25 (2). 51-158 Saxby M.. Aeromonas and Verotoxigenic E. Fishing News Books Ltd. Curran D. 28 (4). 9.J. SAB Symposium Series No.C. 25. 155-9.R. Morgan D. Advisory Committee on the Microbiological Safety of Foods. Anaerobes in relation to foods of plant origin.
Institute of Food Science and Technology. Shigella as a foodborne pathogen.A. 35. Foodborne Pathogens an Illustrated Text. 41. Varnam A. Chilled Food Association. Psychrotrophic Bacteria in Foods: Disease and Spoilage.G. 38. 788-801. 1999. Campden & Chorleywood Food Research Association.europa. 40.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. Phil Voysey. 34. 36. Cambridge. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs. http://www. Journal of Food Protection. Principles and Practices for the Safe Processing of Foods. Smith J.co.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology. Kraft A. 50 (9). Chilled Food Association. Woodhead Publishing Ltd. 39. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. 60 (4).chilledfood. Wolfe Publishing.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33. London.uk/cday/posters2008/npd6. Development and Use of Microbiological Criteria for Foods. 1998. U.K.L. CRC Press. http://eur-lex. http://www..F.campden. 51 .A. 1991. 37. Gloucestershire. Microbiological Criteria. 2006. 1992. 372-6.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1.eu/LexUriServ/ LexUriServ. Shapton N.K. Journal of Food Protection.A. 1997. McCarthy S.H. British Retail Consortium.2.pdf Shapton D. London.. 1987. Evans M. U.
The class having species of economic and public health importance is the Pelecypoda. mussels. Many microorganisms ingested by shellfish survive the digestive process (3. chitons. Plankton and algae form the bulk of their diet from seawater. Other species of shellfish such as crabs.1 Definitions General features of the mollusc The phylum Mollusca.1 3. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). and scallops. In the process of filtering water. scallops. which includes oysters. Special tracts of cilia move this mucus toward the mouth. snails. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells. The term shellfish in this context is limited to oysters. Gastropoda. octopi. cockles.3. clams. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. respectively. clams.6 litres. chemical contaminants and other impurities on the mucus of the gills. S. Pacific Regional Laboratory Northwest U. The phylum consists of five classes: Amphineura. while 53 . they also trap bacteria. the adductors. composed of two valves hinged together dorsally. For example. O.7 litres of water per hour. 4).S. and 2. and shrimp. Box 3012 Bothell WA 98041-4421 United States of America 3. The class Pelecypoda includes all of the bivalves such as clams. Scaphopoda. oysters and clams can process about 3. whelks. abalones. In these forms. Jr. lobster. completely encloses the body. viruses. Strong muscles. Pelecypoda. Food and Drug Administration 22201 23rd Dr. where it is eventually swallowed and digested. the shell.E. P. cockles and scallops. Pelecypods have little need for locomotion. lying well within the mantle cavity. slugs. and Cephalopoda. mussels. can hold the shells tightly shut against enemies (1). mussels.1. squid. oysters. MOLLUSCAN SHELLFISH Carlos Abeyta. cockles. In most species.
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. Whatever contaminants are in the water will eventually get into the shellfish.North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp. but maintain contact with the water by means of a siphon tube (5).I). Mercenaria mercenaria Venus mercenaria Mya arenaria Country Scientific nomenclature Hard clam (also used for Quahaug/quahog) Pacific . dwelling on the bottom or attached to structures in the water column.2 International commercial products Globally. many species of bivalves are edible (Table 3. Clams are benthic and live burrowed in the mud. 3. Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 . are not as vulnerable to contamination through pollution of their beds as the bivalves.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 . The habitat of oysters and mussels are sessile. and usually at higher concentrations than found in their habitat.1.FISH AND SEAFOOD commercially valuable.
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 .
FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. extended above the surface to maintain water currents needed for respiration and feeding. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint. but 56 .1 Clam Clam is the common name of a large group of often edible. The muscular foot is used to burrow in mud or sand. Most clams are only a few centimetres in their maximum dimensions. mostly marine bivalve molluscs. littleneck. geoduck. or siphons.2. Jacques Iceland scallop Bay scallop Weathervane scallop (also called Alaska scallop) Atlantic North East Atlantic North Atlantic West Atlantic Pacific – North America Pacific – North America Pacific – Japan Common scallop Sea scallop (also called giant smooth) Japan Atlantic – North America Pecten varius Pecten maximus Chlamys islandica Argopecten irradians Pecten caurinus Pecten aequisulcatus Pecten laquaetus Pecten yessoensis Penten magellanicus or Placopecten magellanicus Pecten meridionalis Pecten novaezealandiae Pecten jacobaeus Aequipecten gibbus Chlamys opercularis Chlamys varius Amusium balloti Mediterranean – South East Europe New Zealand Australia Mytilus galloprovincialis Mytilus canaliculus Mytilus planulatus Commercial scallop Australia New Zealand Great scallop Calico scallop Queen scallop Variegated scallop Scallop saucer Atlantic/Mediterranean Atlantic – North American Atlantic Atlantic/Mediterranean Australia 3.1. laterally compressed. Buried clams leave tubes. cherrystone. A mantle tissue encloses the body of the clam. and other names. Different species are variously called quahog. hard-shell or soft-shell clam.
Oysters lack a foot or have a reduced foot. Live: in shell Fresh: in shell or shelled meats. uncooked. uncooked meats packed unprocessed. Frozen: shelled meats. skewered with bamboo sticks. then heat-processed. Fresh: in shell. sun-dried. Live: in shells. in cans with either brine or clam liquor. either whole or minced. Soups: oyster stew. or shelled meats. Canned soups include clam chowder. hermetically sealed. or shelled meats. 57 . Frozen: in shell. smoked meats are also canned. Dried: shelled meats. oyster soup. oyster bisque. boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil. all in cans.1. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. Smoked: meats. uncooked (shucked). These molluscs feed on minute plants and animals carried to them by the current. the meats are smoked and packed in oil in cans or jars. Dried: Hoshigai (Japan) shelled meat. after precooking and brining. packed in weak brine.2 Oyster Oysters are marine bivalve molluscs.MOLLUSCAN SHELLFISH the giant clam of the Indian and Pacific oceans may be nearly 130 cm long and weigh 227 kg. and no siphon. Semi-preserved: meats cooked and packed in spiced vinegar. trimmed and packed. 3. having rough and irregularly shaped shells that are closed by a single adductor muscle. clam madrilène and clam liquor. Canned: clams are steamed open and the meats removed from the shells. tidal waters or in brackish water in the mouths of rivers. washed. Canned: meats removed from the shell by steaming.2.
with the posterior ends of the shells (valves) broad and rounded. 58 . They have filibranch gills. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. Live: whole with shells. or packed with jelly. The familiar marine mussel is asymmetrical. and held apart. vinegar solution. The posterior halves of the valves contain a large adductor muscle that holds the valves closed. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). or filaments. Fresh: shelled meats (often only the adductor muscle and the roe are eaten. The adductor muscle is the only edible part of the animal. sauces.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.1. mussel paste.1. in which the individual branches. boiled. in mussel liquor. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves.FISH AND SEAFOOD 3. Fresh: cooked meats. scallops can swim or leap by a series of rapid closings and openings of the valves. also with other ingredients.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. by interlocking tufts of hair-like cilia. Semi-preserves: cooked meats with vinegar-acidified brine and spices. Unlike other sessile bivalve molluscs. brine. smouldered and afterwards dried (Japan).2. are united.2. The shells are fluted and round with a broad. cooked meats packed in dry salt for transport. scallops possess a single large adductor muscle. gutted. Salted: cooked meats bottled in brine.) Dried: peeled. flattened ear or wing at the hinge area. Canned: cooked meats. 3. The common scallop is about 5 cm long. Smoked: meats may be canned in edible oil. and the anterior end smaller and pointed. Bait: used live extensively for baiting fish hooks for line fishing. Internally. mussel in butter sauce. In some species.
Whatever contaminants are in the water will eventually get into the shellfish (7). depending on length of journey. Cockles range from about 1 to 15 cm in length and have two valves of equal size. Raw and partially cooked shellfish. 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. clams and mussels. Canned: in brine. mostly in oysters.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. The microflora are dependent upon a number of factors such as season. also bottled in brine.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves. they accumulate pathogenic microorganisms from polluted waters.1. Fresh: meats removed from the shell by boiling.2. Live: in shell. 3. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding. 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 . Because molluscan shellfish are filter feeders. Vinegar cured: bottled in malt vinegar after brining. are vectors of disease transmission (8). which affects the temperature. in sauce. 3. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms. Canned: in own juice. and the environment. pH. nutrient concentration and pollutants of the water column. and butter.MOLLUSCAN SHELLFISH Frozen: shelled meats. salinity. Salted: meats either lightly or heavily dry salted.
15).3. and shucked shellfish that is shellfish. spirochetes of the genus Cristispira and Saprospira. 10. dredges.. rakes. and yeast cells of Rhodotorula rubra and Trichosporon spp.1 Shellstock 3. or by 60 .1. Bacillus spp. Campylobacter coli Bacillus cereus Hepatitis non-A. the initial flora composition and concentration of bivalves are highly variable. Enterococci spp.. from which one or both shells have been removed. responding to the environment in which they are harvested. The largest groups found are the Pseudomonads.3. whole or in part.. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish.. Moraxella spp.. Temperature. They are termed viable non-culturable microorganisms.. Flow diagrams of the major processing steps of molluscs are presented in Figures 3.3 3.1. Alcaligenes spp. There are microorganisms. and cockles The processing of bivalves takes two forms. observed microscopically.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp. 12.1 Processing and its Effects on the Microflora Oysters. In summary. 11. salinity. mussels.. molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. Acinetobacter spp. shellstock (live) which is shellfish in the shell.. Micrococcus spp. For example. At harvest. The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora.000 /g. clams. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. and Aeromonads. nutrients.. Water temperatures affect the concentrations of microflora in the water. that have never been cultured on laboratory media.1 and 3. but have never been cultured (14.2.1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. 3.3. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. 3. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp.1. Other microorganisms found in lesser numbers are the Achromobacter spp. 13). Vibrios. Pseudomonas spp. (9. mechanical harvesters. Flavobacterium spp.
usually in open-mesh containers and/or burlap sacks. Shellstock are placed in drycooler storage containers in the facility. Secondly. or immediately placed in refrigerated trucks. after harvesting.1). or from other sources approved by the State Shellfish Control Agency (SSCA). wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. Generic flow diagram of live shellfish from harvester to point of sale 61 . Shellstock may be wet-stored at the processing facility in tanks of clean seawater. for transportation to the point of sale. in containers from which they can be retrieved easily. The shellstock is transported dry.MOLLUSCAN SHELLFISH hand (Figure 3. or in bay-approved shellfish-growing waters prior to sale or processing. by truck or boat.3.1. usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock. Thirdly. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. Fig. 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. Water used for shellstock washing is from approved growing areas. If by boat. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. There are three primary reasons for wet storage: first.
22). involving 691 cases was attributed to oysters contaminated in a wet storage area (27). including V. 16. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. 26). 24. Furthermore. Harvest conditions are another factor that influences the growth of bacteria in shellstock. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. however. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. shellstock are placed in refrigerated trucks for transport or placed in wet storage. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. Generally. when it reaches the processing facility. The washing of sediment and detritus soon after harvesting will prevent further contamination. Generally. Klebsiella. metabolic waste accumulates in the shell serving as nutrients for bacterial growth. 25).FISH AND SEAFOOD 3. 21. bivalves tightly close their shells thus trapping water and microflora. Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. and thus contaminating the shucked meat at the point of sale. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF).2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. synthetic seawater for the purpose of de-sanding and salt uptake. without adequate refrigeration. Enterobacter. shellstock are not refrigerated on the harvest boat. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . handling. An outbreak of infectious hepatitis in Sweden in 1956. and the sanitary practices applied during harvesting and shipping. Water used for shellstock washing should be of good sanitary quality. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. coli. the bacterial counts in the meats will increase.2 ºC (45 ºF) or below (19). to avoid possible contamination of the shellstock. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. The most important factor in controlling bacterial growth in shellstock is temperature. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. there is evidence that in the absence of adequate refrigeration. At this temperature the multiplication of faecal coliforms (E. At harvest. for a considerable length of time. Over a period.1. hydrophila (18). 23. with prolonged storage the microbiological quality of the bivalve decreases. and temperature storage of shellstock to point of sale. If long delays occur before reaching the processing facility and/or refrigeration. Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7.1.3. 17). and Citrobacter species) and Vibrios. with the exception of A. are prevented. some pathogenic organisms will survive and multiply in shellfish (20. vulnificus.
Meats are collected in either perforated or non-perforated buckets for delivery to the washing station. 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. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab. thus contributing to high bacterial counts in the final product (28).2 Shucked shellfish 3.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. are applied to the shucking operation of shellfish. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. washing. through which the washed meats are discharged for rinsing and then packed in containers. or subjected to splash from foot traffic. whereas cotton become wet and 63 . and fish.1 Processing description The processing of shucked shellfish is limited to washing of shellstock.3. Due to the nature of the shucking operation. shellstock is subjected to contamination when stored on the floor in standing water. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations. removal of shellfish (shucking). lobsters. If shellfish are not reasonably clean at the time of processing.1. clothing becomes very soiled.2.1. washing and culling shellfish prior to storage is essential. the tank is opened through a drain gate and chute. Containers are placed in packing boxes for shipping at refrigerated temperatures. To remove the shellfish attached to the remaining valve.1. In the blower-washer. Studies have shown the use of rubber gloves reduces the contamination level. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation. 3. Under these conditions if the shuckers enter the packing room. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). 3. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. Shuckers routinely use either cotton or rubber gloves during the shucking operation.2). Thus.3.2 Effects of microflora during processing The same considerations as previously discussed. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported. and packing shellfish into containers (Figure 3. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank. In addition. the finished product may become contaminated. for the effects of microflora during the processing of shellstock. After washing of meats.3.2. 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).
2 ºC or less. Shucking buckets and storage containers are kept at adequate height above the floor to prevent contamination from floor splash. which may lead to contamination of product.2.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). and sanitised before each filling. Fig 3. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. 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. 3. 2. Separate rooms or lockers for storage of clothing. aprons. 64 . Shucking containers are rinsed clean with running water. Shucked meats are thoroughly drained. There is a tendency to store such articles on the shucking benches or in packing rooms. and gloves help prevent the contamination of shellfish. within one hour. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker. 4.
Micrococcus. As described previously. 3. at the turn of the century.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. 65 .2 ºC. a carbohydrate. rough. anywhere during the process of shellfish from harvesting to the point of sale. 35. This process changes the bacterial profile to microorganisms favouring low pH. Earlier studies. The spoilage process in shellfish may be divided into three stages: increase of acidity. particularly oysters stored at 7 ºC. clean gloves. and yeasts. Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. 34. and packing the shellfish in a timely manner. these include lactobacilli. Shellfish. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow.2 ºC or less. Much of the work has been done with oysters and clams. 36).8% (37). Shucked shellfish are held and transported at temperatures of 7. 32). These organisms may play a significant role in the spoilage processes. Flavobacterium.47 to 6. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. shucking pails. are an excellent medium for growth of bacteria. Proteus. Controls that should be applied include using clean packing equipment. Glycogen. streptococci.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. thus producing acid and gas (38). particularly shucked meats. skimmers. is stored in the bivalve as a source of energy. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. whereas bacterium such as Pseudomonas. Alcaligenes. Pseudomonas. Temperature abuse will result in high bacterial counts. spoilage will occur. All equipment coming in contact with the shellfish such as colanders. showed that organisms such as Achromobacter. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10. Shucked shellfish are an excellent medium for the growth of bacteria. Glycogen is hydrolysed by the initial flora found in shellfish. In summary. The environment has an effect on the bacterial population found in shellfish. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. 33. glycogen levels range from 0. and proteolysis (10). Lactobacilli seem to be the predominant microflora in shellfish. 31. where there is a breakdown of temperature control or prolonged storage of shellfish. For example. abundant gas production. blowers and other utensils that have cracked. The initial flora found in shellfish is the major factor of spoilage in bivalves. in oysters. Achromobacter and Flavobacterium levels decrease during storage. 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.
It is a common marine bacterium. These organisms can be present in the shellfish from the point of harvesting. diarrhoea. from harvesting to finished product (i.1. prevention of cross-contamination. ranging from epidemic cholera to sporadic cases of diarrhoea. nausea. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. 41).5. shellstock and packing of shellfish in jars) (29). approximately 23 cfu/g. only a small percentage of strains in the marine environment are potentially pathogenic (45). headache and fever. lot and retail outlet. 3.e. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. The incidence of V. coastal areas). Diseases caused by Vibrios are enteric in nature. Products involved were from the same processor. Sporadic cases are frequent along the coast of the United States.FISH AND SEAFOOD 3.5 cases/100. and harvesting of shellfish in the cooler months.1 Vibrio parahaemolyticus V. 66 . The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). of V. parahaemolyticus causes gastroenteritis lasting 24 .48 hours with abdominal pain. V. 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. primarily in brackish or saltwater. parahaemolyticus.5. during processing. parahaemolyticus from shellfish is low. there is a recent report of low levels of V. Between May and September 1997. V. however. They can be introduced to the shellfish from the environment or added during processing. 44). where outbreaks occur regularly. Fortunately. The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. and estimates based from a hospital study show less than 0. This bacterium is present in all the major processing steps of a shellfish plant. Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. parahaemolyticus possibly producing illnesses (40). widespread in both polluted and unpolluted waters. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). Major outbreaks are associated with the warmer months. One of 250 cases reported involved shucked products with low levels. 3. although non-hemolytic strains have been associated with illness (43. and very common in Japan.1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries.000 population/year (46).5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8.
primarily in immunocompromised individuals with a mortality rate exceeding 50% (50).3 Vibrio cholerae non-01 Illness caused by V. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52. these individuals are advised not to eat raw shellfish. 56). the organism has been found in the Gulf of Mexico. fever.0. chills. parahaemolyticus growth in shellfish meats is temperature dependent. cholerae non-01 has been associated with septicaemia. abdominal cramps. which have been implicated in shellfish-related disease outbreaks (31). lasting 6 . 58).48 hours of onset. and rapidly become inactive at low storage temperatures. All isolates are considered pathogenic.8 case/100. cholerae non-01 is ubiquitous in the estuarine environment.7 days.MOLLUSCAN SHELLFISH Control of V.1. 3.000 population/year (8. Inadequate refrigeration will accelerate the growth of V. It causes diarrhoea. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. 57. 48). 46. Onset of symptoms occurs in 48 hours and can be severe. or primary septicaemia. or immunosuppression. The organism attaches to the intestine and release of a toxin is suspected. like V. Control of oyster-associated 67 . cholerae non-01 is generally a less severe gastroenteritis than that caused by V. Oysters are the most hazardous food associated with V. In one study. and nausea within 24 . V. gastroenteritis. and preventing cross contamination once the product is cooked. vulnificus. 53. Again. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47).II (8. 49). V. cirrhosis. 23 of 24 major USA-West Coast estuaries were positive for V. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. cholerae. High-risk individuals. parahaemolyticus. It is one of the most severe foodborne infectious diseases. those who have liver disease. in the range of 0. vulnificus are fever. 46. Death has been reported to occur within 36 hours from onset. 54. nausea. cholerae non-01 (51). The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. The bacterium is an estuarine species that causes wound infections. V.1.5. vulnificus is by rapid refrigeration during warm weather months.5. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. Healthy individuals are susceptible to gastroenteritis. 3. Primarily. The occurrence of V. They are sensitive to cold. parahaemolyticus the estimated annual incidence rate is very low. leukaemia.4 . and vomiting. diabetes. Control of V. Hazards from this bacterium can be controlled by thorough cooking of shellfish. are particularly susceptible to primary septicaemia.2 Vibrio vulnificus The symptoms of V. however. 55. with a fatality rate of 50% for individuals with septicaemia.
3.5. respectively.1. vulnificus. vomiting. fluvialis.g. these organisms cause gastroenteritis lasting 24 .S. cholerae 01 is transmitted by faecal contamination of food or water. 62. 61. however.10% carbon dioxide for optimal growth conditions. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. In addition. There are several other Vibrio spp. and shock. Illness can include abdominal cramps. preventing crosscontamination of cooked seafood. Illnesses are associated with the warmer months. and illness lasts 7 . cholerae 01.5%) and 2 . 3. and southern Asia including India and Bangladesh.1. and the transmission of the disease by consumption of raw. V.2 Campylobacter jejuni C.5 Other Vibrios spp. 68 . Gulf coast environment. salinity. The control of V. abdominal pain. diarrhoea. imported product from a country experiencing an epidemic. mimicus. 3. including V. (8.4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. 63). It causes diarrhoea. acidity and heat. headache and fever. dehydration. Control of infection from these organisms is no different from the other Vibrios. processors should know the product source. The presence of these strains appears to be in low numbers and non-virulent (59). Onset of illness is from 3 to 5 days after ingestion of contaminated food. V.10 days. cholerae 01 establishing in the U. whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. parahaemolyticus and V. Onset of disease is 6 hours to 5 days. associated with shellfish-borne illness outbreaks. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). 60).48 hours with abdominal pain. There is evidence of strains of V. cholerae non-01 illnesses is by cooking seafood thoroughly. Poor sanitation and contaminated water supplies will spread the disease. cholerae 01 in molluscan shellfish. and V. In general. and nausea. particularly in oysters.5. fever. low pH.FISH AND SEAFOOD V. or cross-contaminated shellfish (57.5. undercooked. jejuni requires reduced levels of oxygen (3 . nausea. widespread in both polluted and unpolluted waters. V. The frequency of infection of these organisms is no different from other Vibrios. Death may occur after severe fluid and electrolyte loss. e. drying. is the same as previously discussed for the non-01 strains. hollisae. nausea. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. In developing countries. They are common marine bacteria. the pathogenic severity is lower than V.
mimicus. 71). 3.3. however. V.5. have been isolated from shellfish beds throughout the world (67.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. of Cases 4 362 0 1 0 a Includes V. they survive well at refrigerated temperatures.II Molluscan shellfish associated outbreaks caused by Vibrio spp. 81. vulnificus. from Pacific shellstock oysters purchased from a retail grocery store (74). hydrophila group (75. They have been implicated in limited shellfishassociated gastroenteritis cases (8. In one case. V. shigelloides (79. ranging from 0.3 Other organisms of concern 3. control of C. 80) are ubiquitous in the marine environment. Campylobacter appear to survive well in oysters during storage. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. jejuni were recovered. Recently. although their role as a pathogen is highly controversial and circumstantial. 73). and their immediate proper refrigeration. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. 68. C. The seasonality of 69 . 76. To reduce the risk of shellfish-borne campylobacteriosis. C. 82. and V. hollisae Infective dose varies with host susceptibility and virulence of the strain.4 to 114 cells. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. 72.500 cells/gram of Campylobacter may cause illness (64. Although. 83. jejuni begins in the harvest of shellstock from approved waters. Campylobacter spp.5. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC. however. 66). This finding is significant in that C. infection occurred among participants of a firemen's banquet. of Incidents 4 279 0 1 0 No.MOLLUSCAN SHELLFISH TABLE 3. fluviallis.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). 69. 85). jejuni does not multiply in shellstock at refrigerated temperatures. 65. 70. Human feeding studies suggest that about 400 . whereas at 10 ºC Campylobacter survive only for 5 days (71). 77. 84. cholerae 01 and non-01. 78) and P. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish. parahaemolyticus. V. V.
Similar results were seen by other investigators (86. soy products. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. Caution should be exercised by individuals with haematological malignancies (88). coli Enterotoxigenic E. They are easily destroyed with mild cooking temperatures.000 to 3. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. dried milk/whey drink powders. FDA/ Administrative Guidelines. FDA/Administrative Guidelines 3 Source (91). aureus C. 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 <10.6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. parahaemolyticus Staph. Note: Although molluscan shellfish are not included in the selected high risk food.000/gram (29).000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures. cholerae V. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V.600. being isolated more often during the warmer month.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. Test Standard Plate Counts E. coli Salmonella V. 3. and where indicated are intended to be used for official control purposes. 2 Source (90). non-dairy frozen desserts. prepared sandwiches. 70 . 87) of the outgrowth of A.000 cfu/g1 <230/100 g2 <1000 per gram. vulnificus in molluscan shellfish. Control of these organisms in shellfish products is limited. In one study. jejuni V. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation.0001 negative for the presence3 Under review4 Source (89). Pathogen monitoring of Selected High Risk Foods Domestic including: milk. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. hydrophila group stored at refrigeration temperatures. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish.
Nauman R.. 105-8. Applied Microbiology. in Microbiology of Marine Food Products.E. Liston J.7 1. Jr. A typhoid fever epidemic caused by carrier bootlegging oysters.. Galtsoff P. Lee J. 36. 10. 4.. Smith F. 14. 44. 2. Bacterial flora of the market oyster. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters.R. Colwell R. 1936. 1982. 19-34. Journal of Milk and Food Technology.R. Microbiological characteristics of Dungeness crab (Cancer magister). Gill S. Clinical Microbiology Review. Effects of harvesting waters and storage conditions on yeast population in shellfish.W.J. 30. Rippey S. Microbiology of bivalve molluscan shellfish. 1960. 71 .D. 13. 104-9. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves. Eliot C. Fleet G. 71-6. 1984. U.S. 3. 8. 1940. 64. 30.. 1983.M. 231-51. Jr. Vanderzant C. Applied and Environmental Microbiology. 9. Organisation for Economic Co-operation and Development. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). 33643.. 755-76. 11. Cook D. Van Nostrand Reinhold. 1968. Viability and release Salmonella charity and Escherichia coli from oyster faeces. Journal of the Mississippi Academy of Sciences.. 6. 12. Thompson C. 1975. 33-6. Schmidt C. 82-7. in Multilingual Dictionary of Fish and Fish Products.R. Lovelace T. Villee C. Government Printing Office. Bernard F. 1994. 5. in General Zoology. Cook D. Ed. American Journal of Public Health. 419-25.E.MOLLUSCAN SHELLFISH 3. Proceedings of the National Shellfisheries Association. 13. 42. 245-7. 1926.C.K. USA.B. Tubiash H.R. Colwell R. The American Oyster. 15. American Journal of Hygiene. Fate of enteric bacteria in estuarine sediments and oyster faeces. 7. 447-52. W. Ray S.A.A. Infectious diseases associated with molluscan shellfish consumption.C.L. Washington D. New York. 1995..S. 1970. 1991.W.K..R. Fish and Wildlife Service.N. Eds. Wiley-Blackwell. Hood M. 1981. Philadelphia. Saunders Company. 1973. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay.H. Eds..F. 544-8... in Fishery Bulletin 1964 No. Ward D.S.A. Veliger. Applied and Environmental Microbiology. water and sediment from Galveston Bay. Hackney C.S. Galtsoff P. Applied Microbiology. 16. 1964. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). Tall B.R. Old H. 6. 46. 8. 58. References The Phylum Mollusca. 633-40. 1963. Biology of the oyster in relation to sanitation. American Journal of Public Health.. 72-8. Walker W. 17. 7. Pfeifer D.. 29. 26. Rowse A. Journal of Food Protection.
Liston J. Journal of Food Science. Timetemperature effects on the bacteriological quality of stored shellfish. Rodrick G. 5. 50. Stott R. Native Littleneck clams (Protothaca staminea). Maryland.W..W. 52. Eds. Abeyta C. Public Health Report. 1967. 1989. Bacteriological studies of commercial shellfish operations on the Gulf Coast. re-laying. 31. 123-8. 1490-5. 1205-10. Kelly C. Svenska Lakartodningen. 1954.H. 27. 1971..J. P-1. Olympia oysters (Ostrea lurida). 28. Temporal occurrence of Vibrio species and Aeromonas hydrophila in estuarine sediments. 4. Ruple A.. Jones W.FISH AND SEAFOOD 18. Saikon Publishing Co. 1221-8. 21. Food and Drug Administration. 23.A. Establishment of critical control points in west coast oyster processing plants. in International Symposium Vibrio parahaemolyticus... Bacteriological changes in live shellfish: Pacific oysters (Crassostrea gigas). Son N. U. Fraiser M.M. Ltd. 167-71. and storage.B... U.E. in National Shellfish Sanitation Program Manual of Operations. Vaughn M..B. 1984. Chesapeake Science. 1985. 1964.W.S. 26.S. 1983.. Special Report U.W. Blake N.D. Journal of Food Protection. 69.J. Part II Harvesting. Effects of storage on microbial loads of two commercially important shellfish species. Journal of Food Protection.H. Influence of U. 22. 1992. Beck W. 989-1003..A.B. LaRock P.C. Washington D. Arcisz W..S. crabs and mullet.A. American Society for Microbiology. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. 47 (5). Applied and Environmental Microbiology.. Koburger J. Applied and Environmental Microbiology. 1980. Takeda Y. Hood M. Sadaguchi G. Kelly C.T. Colburn K. 1961. Ness G. Ericksen T.G. Tokyo. U. 1973. Roos B.. Crassostrea virginica and Mercenaria campechiensis..W. FDA Gulf Coast Technical Services Unit. Handling and Shipping Shellfish. Food and Drug Administration. Hoff J.C. 343-9. 30. 72 . 19. Behaviour of pathogenic bacteria in the oyster. Fleet G. 343-5.A.J.F... 1995.. seafood-handling procedures on Vibrio parahaemolyticus. Presnell M.. 45. Crassostrea commercialis. Bacteriological survey of an oyster bed in Tangier Sound.. Williams L. Applied and Environmental Microbiology. Fujino T. during depuration. 32. 994-1002.S.. Hepatitis epidemic conveyed by oysters. F61-9. Ed. Public Health Service.. Cook D.. I. Jr. 1214. 20. Public Health Service Technical Report.A. Survival of enteric organisms in shellfish. Sakazaki R. and Manillal clams (Venerupis japonica). The influence of time and temperature on the bacteriological quality of shell oysters during processing and shipping.S. 25. Trost P. 29. 24.A.E. Presnell M. 53. Vasconcelos G. Incidence of Samonellae in clams. 40. oysters. 324. 1956. Wekell M. Food and Drug Administration. Kaysner C.
A. kanagawa-negative Vibrio parahaemolyticus from patients and the environment in the Pacific Northwest. Sidwell V. Hlady W. 204-5.C.. Kelly M. Bacteriological examination of oysters stored at low temperatures. Stott R. Thompson C. 41. Journal of Infectious Disease. 25. 35. DePaola A. Journal of Food Science. 1937.M... Jr. 46.B.A. Vibrio parahaemolyticus illnesses. 1349-51. 97.M. Morris J. 107-8. Volume 1. Novak A. 1989.. Yokohama Medical Bulletin. Joseph M.L. Stroh E.T. 1926. Geiger J. Journal of Infectious Disease. Peeler R. Borgstrom G.M. 1989. Joseph S. 1954.S. 1961.W..L.. 73 . 1958.A.. Journal of Clinical Microbiology. Epidemiology and spectrum of Vibrio infections in a Chesapeake Bay community. Grodner R. 1989. Kaysner C. Bulletin of the Johns Hopkins Hospital. Geographic and monthly variation in composition of oysters. Abbott S. 44. Fieger E. In vitro haemolytic characteristics of Vibrio parahaemolyticus: Its close correlation with human pathogenicity.. 69. Janda J. Libonati J.. 133-47... Obara Y. 41.. U. 1987. 1976... 38. Urease-positive. Bacteriological findings in Baltimore oysters. Hoge C.. Microbiology of shellfish decomposition. Kelly C. 1997.M... Arcisz W. Ed.G. New York.. Watsky D... The bacterial content of market oysters. 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. U.. 33. Kaysner C. Journal of Bacteriology. 40. Bothell. Tanikawa E. Jacobson M. 1969. Wekell M. 27. Takeda Y. 1. 2820-2.. Applied Environmental Microbiology. 353-7. West Coast States and British Columbia.D.. The bacterial flora of market oysters. Studies on pathogenic halophilic bacteria. Academic Press.A. 34. 37. 1916. 36. Powers C. 13-7. Washington Report October 14... 313-22. 47. Takizawa K. Shellfish Specialist. Parasitenkunde. 1914. in Fish as Food. Public Health Reports. Akiyama S. 128-31. Bacteriological control of oysters during processing and marketing. 43. Loomis A. Food and Drug Administration. Kato T. Crassostrea virginica. 160. Sample T.. Takikawa I..P. Journal of Bacteriology. 41. Israel E.N.. Infectionskrankheiten und Hygien.MOLLUSCAN SHELLFISH 32. 273-80. (2). Abeyta C. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States west coast. Vibrio infections associated with raw oyster consumption in Florida.W. 1979. 53.E. Marine Fisheries Review. 1147-9. Yamai S. Zentralblatt für Bakteriologie.F. 2..M. 100.G. Jr. 38. Serological and hemolytic characteristics of Vibrio parahaemolyticus from marine sources. Miyamoto Y. Leitch J. 2891-3. 39. Journal of Clinical Microbiology.S. 60. Vanderzant C. 27.D.. 45. Ishibashi M. Berry F. 1997. Journal of Food Protection. 985-93. Ward W. 42.
1982. 1988..D.T.A.G. Wilson R. Characterisation of Vibrios cholerae isolated from oysters...M.M.. 1983.B. 60. Kaper J. Morris J. Lancet.W. 1987. Abeyta C. Ecological relationship between Vibrio cholerae and planktonic crustacean copepods. Colwell R. 41. Twedt R.J. Hebert S. 1984.B. 1979. Journal of Food Protection...E. McFarland L... 37. Jr.. Hickman F..G. Wilson T.. Morris J. Riddle C. 10.G.F.A. Maden J.M.J.O. An epidemiological study of V. Safrin S. Adams M. 52. 802-6.L. Blake P. 1982. McCay C.B.A. Jr.T. 656-8. Gulf Coast. Hunt J. Annals of Internal Medicine. McFarland L. Tacket C. 1981.. 61..K.. 305-9. Clinical features and an epidemiological study of Vibrio vulnificus infections.R. 53. 74 . Applied and Environmental Microbiology. 50. 49. Davis B. Stott R.....A. Weaver R.A possible endemic focus in the United States.. Becker S.M.I. Non-01 Vibrio cholerae bacteraemia: Case report and review. 53...N. and enterotoxin production of Vibrio cholerae in Chesapeake Bay. Barrett T.. Wachsmuth I. Wathen H. Non-O group 1 Vibrio cholerae gastroenteritis in the United States: Clinical.G.E.. 8.A. Journal of the American Medical Association. Feldman R. Lockman H.R. Blake P. R..G.G. Cholera and other vibrioses in the United States. Jr. Motes M. Blake P. Ecology. Jr.... Hug M.W. Black R. 2050-3. Hollis D.P. Brenner F.. 1981. Puhr N.. Ed. 56.. 1294-7. Roberts N... Becker R. Non-01 Vibrio cholerae in shellfish..M. Feeley J.G. Colwell R.. 1012-7. Colwell R. cholerae in Louisiana..FISH AND SEAFOOD 48.M. Blake P.M..A. Jr.C.. in Vibrios in the Environment.. Conte J. 558-61. Kaysner C. Jr..S.A. Roderick G. 51. Jacobs R.V. Small P... New England Journal of Medicine. Francis D. and laboratory characteristics of sporadic cases.T.. Wekell M.. Applied and Environmental Microbiology.W. 58. Cholera ....M..F. 1980.. Hug A.. Applied and Environmental Microbiology. Johnston J. sediment and waters of the U. epidemiologic.. 149. 45. Applied and Environmental Microbiology.. 312. Duran W. 1984. Wazenski T. New York. Bradford H. Incidence of Vibrio cholerae from estuaries of the United States West coast. Morris J. DePaola A. McPhearson R. Peeler J. Vibrio vulnificus: Man and the sea. Craig J... West P.G. Twedt R.A. Presnell M. Rahman R. 305.D... Kaper J. 59. 1. Bradford H. 343-50. Vibrios in the Louisiana Gulf Coast environment.T. Plaard R. Siebeling. New England Journal of Medicine.A.T.. Lee J.. Morris J.. Spite G. 1344-8..J. Journal of Infectious Disease.. 1985. Joseph S. Pons V.. 1985. 5972. Snyder J. Zywno S.M.E.E.. 1983.W.. Jr. John Wiley & Sons Inc. DePaola A. serology. 299-312. Illness caused by Vibrio damsela and Vibrio hollisae.. 55.R. Tacket C. Review of Infectious Disease. Caraway C. 253. Allegra D. 46.R. Miller H.O. 57.. Microbial Ecology. 54. 94. 91-103. 275-83. 1475-8.O.F.C.. Leitch J.
infections in man.A. Fitzpatrick M. Infective dose of Campylobacter jejuni in milk.M.. 472-9. Pearson A. Experimental Campylobacter jejuni infection in humans.. Davis B. Robinson D.S.A... Moore J. Blaser M.A. 169-71. Clements M. Hunt J.I.J. Kaysner. Rowe B. 1987.. Plenum Press. 72.M. Wekell M.G.R. 65.X.R. Levine M.G. 56. 70.. a newly recognised Vibrio species.. Black R. 73.. Huq M.L. Proudford R. Moore J. 257. Brussels. International Journal of Food Microbiology.. Ireland. Wilson I.G.E. 4. Wilson T. Feldman R.D.F.H. Alam A.. 67. Public Health Laboratory Service. Davis J.P. 68. Eds. from patients with diarrhoea. Shandera W. Journal of Rapid Methods and Automation in Microbiology.S. 1980. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness. Wareing D. Helicobacters and Related Organisms.J. Jr. 1991. 157. Skirrows M. 621-4.. 1996. Clements M... in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections. 4.R. Wekell M. 1995. New York.. 6-9 September 1983. in Campylobacters.M. Microbial Ecology in Health and Disease. British Medical Journal. 11. New York. 1584. Blake P. Journal of the Medical Society of New Jersey. Hughes T.K. Black R. Matsuda M. Wilson I. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N.G. Newell D. EF-6. Abeyta C.. Eds...E.W.J. Newell D. 64. Isolation of vibrio-like group. Wareing D. Journal of Clinical Microbiology.. 13.M. Jr. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation. Morris G. 1983.. Griffin M. Plenum Press.. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp.A. 75 ..M. Arumugaswamy R. Huges T. (jejuni. 66. 607-9. Journal of Infectious Disease. Feldman R. 1981. 282. 63. Eds.E. Annals of Internal Medicine.. coli and lari) from shellfish and the marine environment. 80. 99. Ketley J.G.A.K..M. S41.. Stott R.K. 4. Blaser M.. Abeyta C. 69... Campylobacter gastroenteritis associated with raw clams. Murphy P. Jr. 71.A.A. C.L. in Campylobacters.. Studies of Campylobacter jejuni infection in volunteers.... Kaysner C. A method for isolation of Campylobacter spp.A.B. Kaysner C..MOLLUSCAN SHELLFISH 62. 1996..R. Wilson T... Ketley J. Brenner D. 51-64.A. Jones D.J. 1993. Deeter F. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster. Journal of Food Protection. 256. 1983. 323-5. Abeyta C. Dalley E.. Austin S. Disease from infection with Vibrio mimicus.. Helicobacters and Related Organisms.G. 1980. Levine M.M. Murphy P. 101-4..D.M. London.G.. 1988.E...M..P.R. Johnston J.
Annals of Internal Medicine. 998-1001. Hirsch R.. In vitro and in vivo pathogenicity of Plesiomonas shigelloides. Applied and Environmental Microbiology. 87.. Effects of processing and storing oyster meats on concentrations of indicator bacteria...R. 223-47. 79.. Hickman-Brenner F. 849-55. 1997.A. Gracey M.A..C. 343-9. 1988.H. 81. 48. Seidler R. Jr. 598-601. Applied and Environmental Microbiology. Hazan T.. 52..A.. Bark D.M.. Journal of Food Protection. Department of Health and Human Services/Public Health Service/Food and Drug Administration... 1986. Wekell M. 76...A. III. Farmer J. Stelma G. Kelly M.. Beaman J. 449-57.. 76 . 80.L. 85. Stott R..M. Herrington D. Herrington T. Esch G. Koburger J. 84.M.. 55. Fla. Ruple A.A. Peterson D. 1989. Abeyta C.C.. epidemiology. 1984.. Colwell R. Sullivan J. 979-85. Journal of Food Protection. Vandepitte J. Jr. 1985. 39. 105.N. Incidence of motile aeromonads from United States west coast shellfish growing estuaries. 5.. Tzipori S. Oyster borne disease outbreaks in St. Kaysner C. Clinical features.M. 1146-9. Jr. Singelton F. Hood M.. 1981.W. Partridge K. 83. Prevalence and distribution of Aeromonas hydrophila in the United States.. Trost P. 75.L. Kaysner C. 36. Kaysner C... Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species.. Robins-Browne R. GA.J.J. Abeyta C. Journal of Food Protection. Lockman H.. Report (1/13/84). Hunt J. Infection and Immunity.FISH AND SEAFOOD 74. Miller M.B. 1987. Van Damme L. 27. Levine M. 52. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness. 1989... The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish. 731-8.F.D. Burke V.D. Journal of Applied Bacteriology.. 7-12.A. 47.L.. 1978.M. Plesiomonas shigelloides: An opportunistic food and waterborne pathogen. Abeyta C..R. Wekell M. Cook D. Aeromonas hydrophila: ecology and toxigenicity of isolates from an estuary.A. Petersburg. Journal of Food Protection.J. Frequent isolation of Edwardsiella tarda and Plesiomonas shigelloides from healthy Zairese freshwater fish: A possible source of sporadic diarrhoea in the tropics. 690-4. Journal of Clinical Microbiology.P.W. Plesiomonas enteric infections in the United States. Kaper J.. 77. 475-9. Tall B. Blake P.. Wachsmuth I.. Fliermans C.T.K. and treatment of Plesiomonas shigelloides diarrhoea. vibrios and Aeromonas hydrophila. Robinson J. 47..M. Atlanta. 50. Wekell M.. Tenge B. Cooper M. 359-77. 1980. 53. Applied and Environmental Microbiology. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters.W. 1984.A.. Journal of Food Protection.M. Kain K.J. Journal of Rapid Methods and Automation in Microbiology... Holmerg S.D. 1990. Baker R. 78. 82. 86.
Food and Drug Administration. Martin R. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease.S. 1995. Gaithersburg. Government Printing Office . U. Slavin J. 89. in U.H. Food and Drug Administration..12). 2000. Ed. Fish Inspection.. Ed.E. U. Food and Drug Administration Compliance Policy Guides. Sherlock C. Ed.. Technomic Publishing Co. 63-8. Washington D..S.8 Further Reading Ahmed F. 1991. Food and Drug Administration Compliance Program Guidance Manual. 90.C. Part I Sanitation of Shellfish Growing Areas. Food and Drug Administration. Food and Drug Administration.. Food and Drug Administration.E. Seafood Safety. Collette R. 560. National Academy Press.MOLLUSCAN SHELLFISH 88.W.A. Chapter 5: Foods. 1987. Gould G. Burdge D.S. 3.L. 91. U.R.. 7. Food and Drug Administration.030.1987. Lancaster. U. 77 .. mussels.Pathogen monitoring of selected high risk foods (FY 88/89).S. Pittsurgh. Program 7303.W.S. The Microbiological Safety and Quality of Food.600 (CPG 7119. fresh or frozenadulteration by bacteriological contamination Sec. Aspen Publishers Inc. Smith J. oysters. Quality Control. and HACCP: A Global Focus.M. U. Washington D.. Washington D. Clams. in National Shellfish Sanitation Program Manual of Operations..C. 1995.S. 1997.C. U.S. Food and Drug Administration. Baird-Parker T. Food and Drug Administration.S.C. in U. Lund B. Inc.
79 .0 kg polythene bags. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp.5 kilo pack (boxes or bags).5 and 1. and Trachypenaeus spp. CRUSTACEAN SHELLFISH Linda Nicolaides. and available in 0.Ph. e. M.. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. Parapeneopsis spp. crabs.5 and 1. packed and frozen in 0. the front pair ends in claws. Pandalus borealis (deep-water shrimp).g. Peeled ready-to-cook shrimp are raw. lobsters and crabs have ten legs. (deep-water shrimp). with additional water to form a glaze. Pandalus montagui and Crangon crangon (inshore shrimp).0 kg polythene bags. They possess a segmented body. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. Such products are usually only partially cooked before freezing. peeled product.4.. jointed limbs and a chitinous exoskeleton. Shrimps.. Peeled and cooked shrimp are ready-to-eat after controlled thawing.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. with shells and heads on or off are usually packed. crayfish and shrimp (1). and are usually aquatic. Species eaten from tropical waters include Penaeus spp. lobsters. Crayfish. and frozen in 1 and 2.
boiled to kill the crustaceans and cook their meat prior to consumption. traditionally. or frozen crab. which include the environment from which they have been captured or harvested. only live carcasses are processed.g. the geography of the area they are captured from or are grown in. stored and transported as frozen product. lobster tails and cooked crabmeat presented decoratively in their shells. 4. However. during the peeling of cooked shrimp. their feeding and living habits. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. For example. tropical waters. The structure of the circulatory system of crabs is not closed.2 Initial Microflora As with finfish. Pasteurised crabmeat is edible meat removed from the exoskeleton by hand. Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function.g. lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption. spoilage bacteria are of little importance as. e. storage above the recommended storage temperature. for crabs and lobsters. 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. and the temperature and quality of the waters in which they exist.. If these products are subjected to temperature abuse. Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. packed into polythene bags. Shewanella putrefaciens and 80 . so that spoilage may occur during landing and transportation to the processing plant. shrimp die immediately after capture. with the increasing demand for ready-to-eat/cook crustacean products. which means that the haemolymph can be a reservoir of bacteria.FISH AND SEAFOOD Crabs. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp. Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP). Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance. the season. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. As with finfish. Cooked and chilled. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. In comparison. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed. particularly for members of the genus Vibrio (2). the potential exists for crosscontamination from both the processor and from the environment. Following capture or harvest. particularly after it has been cooked. hence.
Alcaligenes.. The rate of spoilage will be a function of time and temperature. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . When the shrimp arrive at the processing plant they are washed and then sorted into size. glazed and frozen (see Figure 4. corynebacteria. or transported directly to processing plants. In contrast. After capture and transport to the processing plant.for example. or cooked. 4. A large proportion of shrimps are produced by aquaculture. lobsters and crayfish are harvested from cages or pots containing bait. however. they are washed. up to 16 days for tropical species compared with 8 . and Achromobacter spp. Shrimp will then be glazed and frozen in a raw state. Shrimp might be further processed with the shell on. peeled and cooked. the rate of deterioration of the crustacean shellfish by these organisms is reduced. 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. with “head on” or “head off” whereby the edible tail is removed from the head. so they are either sold on the local market or are processed in local processing plants.1.3 Processing and its Effects on the Microflora Crabs.. Pseudomonas spp. or are found in brackish water around the shores. The claws are usually bound together to prevent the animals fighting and causing damage to each other. especially if they are stored in ice or iced water. 4. uncooked prawns and shrimp is presented in Figure 4. Micrococcus spp. gills and thorax.1). chilled 81 . Vibrio spp. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected.10 days for temperate water species (3). Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. bacterial contamination and spoilage are inevitable. Shrimps can be processed whole. As with finfish.. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed.3. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. or with the shell and central vein removed. from the time of capture and processing. graded according to size. In comparison shrimps die quickly after capture. thus. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots.1 Prawns and shrimps An example of the process used for preparing frozen. Hence. These animals can survive out of water if the surrounding environment is kept cool and damp.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. the genera associated with tropical species include Enterobacteriaceae. They are then taken to market alive in either holding tanks. chilled sea water. whether GHP are in place.
1.frozen uncooked prawns 82 . and packed to meet the customer’s specification. 4. Flow diagram . should be followed to ensure that the product is not re-contaminated with microorganisms present in the processing environment or from the workers. spices and cooking batter. including GHP. After cooking. Good Manufacturing Practice (GMP). The cooking stage will eliminate the intrinsic microbial flora of the shrimp as well as any contaminants that have been added during handling and transportation to the processing plant. Breaded products introduce an added risk of spore forming bacteria. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour. Fig.FISH AND SEAFOOD frozen.
contaminating microflora may include enteric bacteria.4. If boiled. they include Sh.CRUSTACEAN SHELLFISH 4. dull and sticky. slightly iodine smell. If hygienic procedures are not followed. Once removed and cleaned. bacteria of public health significance. the meat is either repacked and sold in the shells. and have a sweetish. since crabs and lobsters are traditionally consumed immediately after cooking. putrefaciens. 83 . whilst boiled crustaceans have a pleasant slightly sweet smell. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. and a strong ammoniacal odour is produced. viruses or parasites. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. An offensive odour is also produced. putrefaciens and Pseudomonas spp.1 Prawns and shrimps Freshness indicators for prawns and shrimps are for the shells to be crisp and dry and feel sharp and cool to the touch.2 Cooked crabs and lobsters As indicated earlier. the joint between the tail and carapace is open and the limbs hang loosely. there is no indication of life. immediately prior to boiling.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. 4. The shells have a crisp. 4. spoiling prawns and shrimps have shells that are soft. However. 4.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold. With spoiled crabs and lobsters. Heat is produced if the shrimps decompose in a confined space. with the growing demand for ready-to-eat products from crab and lobster meat. If the tail drops. bright and dry appearance.3. there is no associated spoilage problem. The shell will be soft. wet and ‘soapy’ or ‘jammy’ to the touch. In comparison. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. for crabs. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step.4. or packaged and processed. indicative of protein degradation and the growth of the principal spoilage bacterium Sh. or in hermatically sealed cans that have undergone an appropriate heat treatment.
5. 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. in domestic/imported shrimps (15).5.2 Listeria spp. 16). 6. as well as for Salmonella. which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). Aquatic birds and the hands of workers have also been identified as sources of the pathogen. 4.1 Pathogens: Growth and Survival Salmonella spp. monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 .5% of frozen raw shrimps from different countries (13). Although foodborne illness had not been associated with the product. although efficient cooking will quickly eliminate it (6). The presence of L. Following this. monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. later surveys in 1990 recovered the pathogen from retail raw. animal manure. fish fingers. in 24. often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. nearly 30 years elapsed before the potential hazard of L.cooked prior to consumption. shrimps and cockles sold (17). Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. The above results prompted the FDA to develop a compliance programme for the pathogen. One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). cooked and smoked fish. even if GMP is followed. An import alert was put in place in June 1987. shrimp and shellfish (11). Salmonella cannot be completely removed from raw shrimp during processing. and usually for both. monocytogenes was absent from 40 samples of cooked prawns. Further surveys identified Listeria spp. a common pond fertiliser used in aquaculture. 7). Hence. Moreover. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. 5. However. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14.5 4. monocytogenes in cooked ready-to-eat foods.FISH AND SEAFOOD 4.
about 50 . no incidence of foodborne illness caused by L. whilst boils and cuts are also common sources of the pathogen. 24. Once the competing microflora has been removed by a heat treatment. However. is widely distributed in aquatic environments. Crabs are covered in a chitin exoskeleton. Control should be achieved by promoting GHP and staff training programmes. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. the causative agent of cholera. therefore. 4.5. 85 . as part of the natural bacterial flora. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. botulinum type E is commonly associated with fish and fishery products. The main source of contamination is the hands of processing personnel. was associated with seafood consumption. Researchers have demonstrated that inoculated shrimp supported toxin production by C.70% of healthy individuals carry toxin-producing strains of Staph.3 Staphylococcus aureus Staph. Crustaceans have been linked with food intoxications attributed to Staph.5. which can become colonised by V.5 Vibrio spp. To date. although contamination by types A and B may occur from the environment. e. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. The pathogen has resistant spores. monocytogenes has been attributed to crustaceans. particularly crabmeat (18. The former. pregnant women and the immuno-compromised. Vibrio cholerae. so is able to survive heat treatments that eliminate vegetative cells. V. the pathogen can grow at temperatures as low as 3 °C.CRUSTACEAN SHELLFISH old. V. An epidemic that began in South America in 1991. which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. cholerae O1 and V. cholerae non-O1. combined with the degree of handling of these types of product. 4.5. cholerae. especially in products that have been vacuum packaged. botulinum type E at 10 °C but not at 4 °C (20). 19). 4. aureus because of the ubiquity of this organism in the processing environment.4 Clostridium botulinum C. aureus on their hands. pasteurisation and cooking. 22) as well as from undercooked crustacea when harvested from contaminated waters (23.g. 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. 25). cholerae is divided into two groups. aureus is not a member of the intrinsic flora associated with crustacean shellfish.
4. However.7 Aeromonas hydrophila and Aeromonas sobria A. although A.5. they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). 32).8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans. whilst 5. vulnificus is an invasive and lethal pathogen associated with wound infections. V. coli. 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. Parasitic infections are controlled by either cooking infected material prior to consumption. hydrophila and A.5. 31. in a survey carried out in the United States between 1977 and 86 .5.6 Campylobacter jejuni and Campylobacter coli C.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A. particularly trematodes (6). However. It is more commonly implicated in countries where crustacean shellfish are eaten raw. Caliciviruses.5. Hepatitis A virus presents the most serious hazard. 4.2 °C) so that GHP coupled with controlled low temperature storage is essential to prevent the pathogen from reaching levels that may present a hazard. non-B Hepatitis viruses (31). to date. V. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. jejuni has not. 4.8% of samples contained C. Of these.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. A. 4. Quantitative levels were below limits of detection in all cases (<0. Norovirus. South America. a marine halophile. occurs in crustaceans from warm environments. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates. hydrophila has been isolated from prawns (29). Astroviruses and non-A. however. or freezing at a temperature of -23 °C for 7 days (30.g. hydrophila is able to grow well at refrigeration temperatures (0 . but has the potential to cause fatal foodborne illness (26). Japan. e.30 MPN/g) (27). been implicated in a case of foodborne infection associated with crustaceans. parahaemolyticus. the potential hazard exists as C.
which results in a persistent and apparently permanent loss of short-term memory (36). 39).1 EU Legislation Microbiological standards for cooked crustaceans and shellfish. although it is one of the European Union (EU) legislated allergens. from farm to fork (37. 4. consistent with that in the stomach. They can survive low pH levels. produced or imported into the European Union (EU). 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. 2073/2005. which could be transferred to the meat during unskilled removal from the shell (35).5. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. only 0. or chlorine concentrations similar to those found in drinking water.6. A summary of microbiological criteria for crustacean shellfish is listed below. 4. Provisions laid down concerning the use of additives would also apply to these types of products.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33).6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. The toxin causes amnesic shellfish poisoning (ASP) when consumed. Selected parts from this regulation is presented below. 4.CRUSTACEAN SHELLFISH 1981. The risk of such hazards is far higher with fin fish and molluscan shellfish. 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. Norwalk viruses can also withstand heating to 60 °C. 38. 87 . Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34).5. are included in the Commission Regulation (EC) No.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. 4.
FISH AND SEAFOOD
220.127.116.11 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 18.104.22.168.1 Chapter 1 - Food safety criteria
L. monocytogenes Prepared crustacean products fall into the category described in the regulation as “Ready-to-eat foods available to support the growth of L. monocytogenes other than those intended for infants and for special medical purposes”. During capture and preparation there is a zero tolerance for the presence of L. monocytogenes on the product. However, to take into account the issues of natural habitats being contaminated with low levels of this pathogen there is a slight loosening of the regulation at the point of sale and for the duration of the products shelf life (refer to Table 9.I, Food Category 1.2). If the level of L. monocytogenes should be above this level then the consignment is considered to be unsatisfactory. Salmonella All cooked crustaceans and molluscan shellfish should be free from Salmonella at the point of sale and for the duration of their shelf life (refer to Table 9.I, Food Category 1.16). Should any sample test positive for Salmonella then the product is declared to be unsatisfactory. 22.214.171.124.2 Chapter 2 - Process hygiene criteria
Process hygiene criteria for E. coli and coagulase-positive staphylococci in shellfish and shucked products of cooked crustaceans and molluscan shellfish are laid down (refer to Table 9.II, Food Category 2.4.1). 4.6.2 International requirements
International requirements are based upon having a risk-based, preventive management system in place at all stages of the supply chain. The requirements for this preventive approach is described as follows (37): • Fish and fishery products should be prepared in plants certified by the local competent authority. All certified plants should comply with the GHP (39); • The fisheries industry should take responsibility, implement and maintain safety management systems based upon HACCP; • The national competent authority is responsible for the certification of fish processing and manufacturing plants, verification of effective systems, and
issuing of certificates of compliance for export products. This includes auditing and inspection programmes; • National surveillance and monitoring programmes should be in place to demonstrate that all identified hazards are under control, e.g. biotoxins, and to identify potentially emerging hazards; Within this requirement is the flexibility for countries and trading blocks to set specific standards required to deliver an equivalent requirement with those at national level. Such standards need to be science-based, especially if they are more stringent that those recommended by the Codex Alimentarius Commission. Verification that GHP have been followed during the handling and preparation of crustacean shellfish for consumption is usually done by assessing the level of E. coli and/or coagulase-positive staphylococci in the products. Legal requirements generally require that these groups of indicator organisms are present in levels <100 cells/gram of product. Pathogenic bacteria such as Salmonella spp. should be absence is 25 grams multiplied by the number of subsamples tested. 126.96.36.199 United States The FDA brought into force a Federal Mandate Seafood Rule in 1995 which forms the foundation for good practices required for the handling and processing of imported fish and fishery products into the U.S. The best practices include GHP and HACCP (37). All fish and fishery products produced in and imported into the U.S. after 17 December 1997 are required to demonstrate that HACCP principles were applied to their handling and processing. The USFDA has a zero tolerance for Salmonella in fish and fishery products; this includes farmed shrimp. The US FDA considers that the presence of pathogenic microorganisms in seafood is an indicator of adulteration (32). 4.7
Encarta 1997. Encarta 97 Encyclopedia World English Edition. Microsoft. The International Commission for Microbiological Specifications for Food. Fish and Fish Products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. The International Commission for Microbiological Specifications for Food. New York, Kluwer Academic/Plenum Publishers. 2005, 174-249. Cann D.C. Bacteriology of shellfish with reference to international trade, in Handling, Processing and Marketing of Tropical Fish. Ed. Tropical Products Institute. London, Ministry of Overseas Development. 1976, 377-94. D’Aoust J.Y., Gelinas R., Maishment C. Presence of indicator organisms and the recovery of Salmonella in fish and shellfish. Journal of Food Protection, 1980, 43, 679-82.
FISH AND SEAFOOD 5. 6. 7. 8. 9. 10. 11. Fraiser M.B., Koburger J.A. Incidence of salmonellae in clams, oysters, crabs and mullet. Journal of Food Protection, 47, 343-5. Reilly P.J.A., Twiddy D.R., Fuchs R.S. Review of the occurrence of Salmonella in tropical shrimp. FAO Fisheries Circular No. 851, 1992, (FIIU/C851). Reilly P.J.A. Salmonella and Vibrio cholerae in brackishwater cultured tropical prawns. International Journal of Food Microbiology, 1992, 16, 293-301. Twiddy D.R. Antibiotic-resistant human pathogens in integrated fish farms. ASEAN Food Journal, 1995, 10, 22-9. Olsufjew N.G., Petrow V.G. Detection of Erysipelothrix and Listeria in stream water. Zhurnal Mikrobiologii Epidemiologii i Immunobiologii, 1959, 30, 89-94. Anon. First Listeria finding in crabmeat confirmed by FDA. Food Chemistry News, 1987, 29 (14), 38. Ryser E.T., Marth E.H. Incidence and behaviour of Listeria monocytogenes in fish and seafood, in Listeria, Listeriosis and Food Safety. Eds. Ryser E.T., Marth E.H. New York, CRC Press. 2007, 617-54. Anon. Mexican crabmeat, Greek pasta automatically detained. Food Chemistry News, 29 (19), 43. Anon. FDA checking imported domestic shrimp, crabmeat for Listeria. Food Chemistry News, 1987, 29 (24) 15-7. Archer D.L. Review of the latest FDA information on the presence of Listeria in foods. WHO Working Group on Foodborne Listeriosis, Geneva, Switzerland. 1988, 15-9. Anon. FDA to sample shrimp for Salmonella, Listeria. Food Chemistry News, 1988, 30 (36), 9-10. Anon. Monitoring of changes to listeriosis problem urged. Food Chemistry News, 1989, 31 (20), 13-7. Richmond M. Report of the Committee on the Microbiological Safety of Food. London, HMSO. 1990, 133-7. Swartzentruber A., Schwab A.H., Duran A.P., Wentz B.A., Read R.B. Jr. Microbiological quality of frozen shrimp and lobster tail in the retail market. Journal of Food Protection, 1980, 40, 765-9. Wentz B.A., Duran A.P., Swaartzentruber A., Schwab A.H., McClure F.D., Archer D., Read R.B. Jr. Microbiological quality of crabmeat during processing. Journal of Food Protection, 1985, 48, 44-9. Garren D.M., Harrison M.A., Huang Y-W. Clostridium botulinum type E outgrowth and toxin production in vacuum-skin packaged shrimp. Food Microbiology, 1994, 11, 467-72. Molitoris E., Joseph S.E., Krichevsky M.I., Sindhuhardja W., Colwell R.R. Characterisation and distribution of Vibrio alginolyticus and Vibrio parahaemolyticus isolated from Indonesia. Applied and Environmental Microbiology, 1985, 50, 1388-94.
12. 13. 14.
15. 16. 17. 18.
CRUSTACEAN SHELLFISH 22. Varma P.R.G., Iver T.S.G., Joseph M.A., Zacharia S. Studies on the incidence of Vibrio cholera in fishery products. Journal of Food Science and Technology, 1989, 26, 341-2. Madden J.M., McCardell B.A., Morris J.G. Jr. Vibrio cholerae, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 525-42. Twedt R.M. Vibrio parahaemolyticus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 543-68. Oliver J.D. Vibrio vulnificus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 569-600. Pearson A.M., Dutson T.R. Quality Attributes and Their Measurement in Meat, Poultry and Fish Products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994. Reinhard R.G., McAdam T.J., Flick G.J., Croonenberghs R.E., Wittman R.F., Diallo A.A., Fernandes C. Analysis of Campylobacter jejuni, Campylobacter coli, Salmonella, Klebsiella pneumoniae and Escherichia coli 0157:H7 in fresh handpicked blue crab (Callinectes sapidus) meat. Journal of Food Protection, 1996, 59 (8), 803-7. Buchanan R.L., Palumbo S.A. Aeromonas hydrophila and Aeromonas sobria as potential food poisoning species: a review. Journal of Food Safety, 1985, 7, 15-29. Palumbo S.A., Maxino F. Williams A.C., Buchanan R.L., Thayer D.W. Starch ampicillin agar for the quantitive detection of Aeromonas hydrophila in foods. Applied and Environmental Microbiology, 1985, 50, 1027-30. Wekell M.M., Manger R., Colburn K., Adams A., Hill W. Microbiological quality of seafoods: viruses, bacteria and parasites, in Seafoods: Chemistry, Processing Technology and Quality. Eds. Shahidi F., Botta J.R. Glasgow, Blackie Academic and Professional. 1994, 196-219. Schantz J., McKerrow J.H. The dangers of eating raw fish. New England Journal of Medicine, 1989, 320, 1143-5. Ahmed F.E. Seafood Safety. Seafood Safety. Committee on Evaluation of the Safety of Fisheries Products, Food and Nutrition Board, Institute of Medicine. Washington D.C., National Academy Press. 1991. Blackwell J.H., Cliver D.O., Callis J.J., Heidelbaugh N.D., Larkin E.P., McKercher P.D., Tayer D.W. Foodborne viruses: their importance and need for research. Journal of Food Protection, 1985, 48, 717-23. Kapikian A.Z., Chanock R.M. Norwalk group of viruses, in Virology. Eds. Fields D.M., Knipe D.M. New York, Raven Press. 1990, 359-77. Horner R.A., Postel J.R. Toxic diatoms in western Eashington waters (US west coast). Hydrobiologia, 1993, 269/270, 197-205. Perl T.M., Bedard M.S.M., Kosatsky T., Hockin J.C., Todd E.C.D., Remis R.S. An outbreak of toxic encephalopathy caused by eating mussels contaminated with domoic acid. The New England Medical Journal, 1990, 322, 1775-80.
23. 24. 25. 26.
34. 35. 36.
FISH AND SEAFOOD 37. Ababouch L., Gandini G., Ryder J. Causes in Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. Australian Government. Australian Quarantine and Inspection Service Hazard Analysis and Critical Control Point (HACCP). A Guide to Compliance with Export Control (Fish and Fish Products) Orders 2005. FAO/WHO. General Hygiene Requirements. Rome, Food and Agriculture Organisation of the United Nations. 2003.
Further Reading Spoilage and pathogens
Gram L., Huss H.H. Fresh and Processed Fish and Shellfish, in Microbiological Safety and Quality of Food Volume 1. Eds. Lund B.M., Baird-Parker A.C., Gould G.W. Maryland, Aspen Publishers Inc. 2000, 472-506. Ababouch L., Gandini G., Ryder J. Causes of Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. International Commission on Microbiological Specifications for Foods. Fish and fish products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1998, 130-89. Sutcliffe P., Disney J. Handling, Processing and Marketing of Tropical Fish. London, Tropical Products Institute. 1977. Connell J.J. Control of Fish Quality. Oxford, Fishing News Books. 1995 Ward D.R., Hackney C. Microbiology of Marine Food Products. New York, Van Nostrand Reinhold AVI. 1991 Pearson A.M., Dutson T.R. Quality Attributes and their Measurement in Meat, Poultry and Fish products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994.
4.8.2 Pathogens - general
International Commission on Microbiological Specifications for Foods. Microorganisms in Foods 5. Characteristics of Microbial Pathogens. Eds. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1996. Varnam A.H, Evans M.G. Foodborne Pathogens - An Illustrated Text. London, Wolfe Publishing Ltd. 1991. Doyle M.P. Foodborne Bacterial Pathogens. New York, Marcel Dekker. 1989.
where the salt replaces moisture in the fish flesh. split and opened out flat. and is sometimes known as ‘pickling’. and the extracted moisture is allowed to drain away. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. and before significant spoilage takes place.5. alternating fish layers with a layer of salt. rather than for preservation itself. The processes take anywhere from a few hours to a few days or even months. Fish can be salted in three different ways or their combination: Dry salting. drying. their combinations and/or variations. and marinating processes. It includes the salting. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. drying and smoking processes. Salting is both a method of preserving fish. The fish is headed.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. Usually. both to preserve and to create a variety of new products. This can be called ‘kench’ curing and is mainly used for non-fatty white fish. and salting is done by stacking the fish in layers. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. Fish curing is practised worldwide. and a preliminary operation to some smoking. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. dependent on the country or the type of product. fish is immersed for a few hours in a relatively low concentration brine. SMOKED AND DRIED FISH Dr. CURED. 93 . Wet salting or brining.
thus delaying spoilage for a few days. prior to or during the boiling process. Fish in contact with refrigerated plates freeze. The fish is mixed with dry salt but in a watertight container. are added to the marinade. Longer shelf life is achieved in the products when salt is added. It has found limited application with fish. When the process is running in a closed system. as concentrated brines are formed. Salt-boiling. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. Vacuum drying. evaporated water is removed by vacuum pump. this ‘brine’ can be collected and placed again over the fish several times. Salt. it can also be named ‘pickling’ or ‘blood pickling’. Salted-boiled fish are the products obtained on either boiling fish in a brine. Included in this group are some lightly preserved products where no acid is added to the marinade. spices). Freeze drying. This is the common method of drying fish. depending on the type of product. usually vinegar. The products are usually consumed raw. This is a mixture of a dry and a wet salting process. Heat is transferred to fish from heated air or a heated surface. Usually. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms. the ice sublimes and the vapour is removed from the fish by vacuum pumps. Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. or boiling it with dry salt and water. As the water is drawn out of the flesh by the salt a brine is formed. such as ‘gravad’ fish. Thus. the preservative effect is due to the low pH used in the process. thus reducing the water activity of the product. as well as other flavourings (sugar. the product rehydrates quickly but does not retain reabsorbed moisture on cooking. After the freezedrying process. 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 . Smoking is an ancient process for preserving and flavouring food. is used as a preservative instead of salt. thus making the product unsuitable for microbial growth. under normal pressure. Marinating or pickling. and the fish can be immersed in such brines for long periods. Food products can be dried in three different ways: Air or contact drying. and the air movement removes the vapour. this was done naturally by exposure of the fish to the air. The product is dried by contact with heated surfaces or by radiation.FISH AND SEAFOOD Dry / wet salting. Marinades are acidified brines. without other preservation methods applied after boiling. It is not commonly used for fish products. Traditionally. where an organic acid.
Cold-smoking. Usually. the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. In this process. 95 . or by means of smoke-flavouring processes with liquid smoke. Smoking can be either traditional. temperatures below 30 °C are used. It can be smoked either traditionally. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke. SMOKED AND DRIED FISH of smoke. and the preservative effect is only slight. The process was designed so that the smoke is filtered to remove all smoke particles. It is commonly used in place of traditional smoke.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. 5. smoking is now used mainly to give the fish a characteristic flavour. leaving only elevated levels of carbon monoxide. including pathogens. The different spices used in the immersion brines are also a potential source of microorganisms. the fish is subjected to heat for a period that does not coagulate protein. After the salting step. However. Usually. by exposure to the wood or sawdust smoke. or liquid smoke. or added to the brine with other flavourings. by 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. With the exception of some Asian countries. Smoked fish is prepared by subjecting the fish to the direct action of smoke from wood or wood sawdust burning. fish can be smoked by means other than the direct action of smoke. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. process temperatures are over 60 °C. sporeforming bacteria or osmophilic moulds. Clear smoke. Smoked-flavoured fish. It can be injected into the product. In this process. Hot-smoking. Also. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour. such as injection or dipping in a solution of liquid smoke. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. and filtered to remove tars and particulate matter.CURED. which impart the flavour of smoke to fish. The smoke itself may contain spores of moulds. or by means of smoke flavouring processes. 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. the salt used in the processes may contain halophilic bacteria.
only the most common processes can be described here. a 0º brine saturation means pure water.I). Therefore. the fish is usually salted with a 70 . For smoking salmon. Thus. % 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.6 132. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat.8 211.2 237.4 184. or salt in the water phase. it causes swelling because the fish absorbs water from the salt solution (1). This should be distinguished from the salt content. processes and producers’ private recipes. Salt in the water phase links the two conditions of moisture and saltiness. all using salting steps.8 211. When a 100º brine is used. The dissolution of salt in water is dependent on the temperature. and salt content. it is important to understand the concept of salt concentration.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) 26.0 g of salt per litre of brine). it is impossible to establish standardised processing stages in the curing of fish.8 79. sometimes the salt crystallises onto the surface of the fish. TABLE 5. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.0 158. a 90º or above brine (237. The strength of the solution is described as degrees of saturation (brineometer degrees). Different brine strengths are required according to the curing processes and/or the type of product.2 105. it is important to state the temperature. At 18 °C a 100º brine is produced when the solution contains 26.0 .3 Processing and its Effects on the Microflora Owing to the wide variety of products.6 g of salt or more per litre of brine) may cause salt burn in this way. etc.4% salt (Table 5.4 52.FISH AND SEAFOOD 5.80º brine (184. When working with salting. A 100º brine solution means that the solution is saturated and no more salt can be dissolved.6 264. Salt concentration means the percentage of salt in the water contained in the fish. is employed. smoking or marinating processes.2 g of salt per litre of brine). When a milder brine. about 50º (132.
the fish is mixed with one part of salt per eight parts fish.1. The calcium and magnesium impurities.3. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. The most common fish used in the northern hemisphere is cod (Gadus morhua). and for a heavy salting a ratio of 1:3 salt:fish is used (1). can be dried further to produce ‘klipfish’ or ‘bacalão’.1.CURED. and essentially free from iron and copper (3). It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. are likely to cause hydroscopicity of the product and. producing characteristic pink patches. respectively.3. for which there is a high demand in Mediterranean countries. This is because fatty fish turn rancid quickly. ‘Salt burn’ might occur when the surface of the fish dries out too quickly as a result of the small grain size of the salt. however. The salt used should be of food-grade quality. outgrowth of halophilic bacteria may occur. which include calcium and magnesium. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1).1 Salting Salting of fish for preservation purposes has been used since pre-history. low in calcium and magnesium. On the other hand.35% and 0. lean white fish is used for this purpose. Also. are recommended to help make the product whiter. 97 . having a short section at the end of the tail. However. and such delays.15% calcium and magnesium. This condition is known as ‘putty fish’ (4). and salt can speed this oxidation process. the fish is restacked with fresh salt after about a week. and a ‘putty’ fish product is also obtained (1). SMOKED AND DRIED FISH 5.1 Dry salting / Kench curing Usually.1 ppm and 10 ppm. Salting can be done in a number of different ways. caught in North Atlantic waters.15 .0. which consequently inhibits the diffusion of the water from the bulk of the product to the surface. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. See Figure 5. respectively (1). mainly in tropical conditions. For a light salting. if very pure sodium chloride is used for curing purposes. When pure salt is used. 5. addition of levels of 0. but the concentration of copper and iron must be kept under 0. A ‘green cured’ product is obtained when the water content has been reduced to about two-thirds of the initial amount. 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. Usually it is followed by other preservation techniques such as drying and/or smoking. when the air humidity is high. ‘Green cured’ split cod with about 60% of the backbone removed. and the salt has penetrated throughout the fish (3). The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. it tends to lead to a slight yellowing and softening of the product.
FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat.1. when a light brine is used.5.2 Wet salting or brining As referred to above. although toxin production would be unlikely (4). such as Staphylococcus aureus could survive and grow at such water activities in the product. such as 98 . Fig. Only small fish can be salted whole.1. so that the risk of spoilage is high.90 are rarely achieved. it appears that water activities much less than 0. Food-poisoning organisms. Where fish are salted in their whole form.3. eviscerated and/or split). and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. For large fish it is advisable to split it before salting. Processing of dried salted cod 5. The greatest barriers to salt penetration are the skin and scales. the process is mainly for flavouring purposes prior to the use of other preservation processes.
the fish enzymes play an important role by maturing or ripening the product.03 2.56 6.33 5.41 5. but as it is placed in containers.5 3.74 9. a saturated brine is used.23 3.98 6.II.06 4.44 2.03 4. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained.76 4. For preservation purposes.16 7.0 3.10 1.76 4.88 5. thus.23 2. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.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) 2.26 7.84 1. TABLE 5.44 2.70 6.76 4.25 5.61 3.90 (1).5 10. and can be collected and replaced again over the fish.76 4.51 3.76 4.99 2.91 2.98 5.66 5. Histamineforming bacteria do not generally grow at water activities below 0.61 3.09 8.76 99 .60 2.41 4.26 1. if split fish is dry-salted.23 2. Anchovies are the main fish used for these purposes.17 5.3. can be cured in a saturated brine solution for 6 days and. headed and gutted.41 6.CURED. the liquid that comes out of the fish forms a brine. herrings and other fatty fish can be used.22 6. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 . 5. The main objective of the brining method of salting fish is to limit air penetration during processing.23 8.49 4. limiting the amount of rancidity that can develop.54 5.30 2. after being drained and pressed at 465 kg/m2 for 15 hours. sardines. with pressure being applied to the top of the container to help the brine to drain away. SMOKED AND DRIED FISH smoking.51 5.95 3.35 4. the fish is dry-salted. and a red burgundy colour will develop in the product.66 5.09 8.80 5.85 3. There are several variations of this process.5 3.0 2.64 4.70 3.00 3. with a shelf life of about 4 weeks (water activity (aw) about 0.78) (1). but mackerel. Here.11 4.5 4.26 2.1.78 6.65 2.0 4.86 2. See Table 5.04 5.78 2.46 4. Fresh sardines. Histamine formation is of importance when using scombroid and some clupeidean species.98 6.17 4.26 4. This is why it is the preferred method for fatty fish. The fish are kept immersed in their own brine near saturation point for several days.96 1. the brine is then replaced over the fish.45 3.47 6.3 days.25 5.19 3. the product obtained is called gaspé-cured (5).05 7.85 3. they can be stored at 28 °C.11 3.11 9.41 3.3 Dry / wet salting In this process.73 1. For example.26 3.
The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. up to 1 . The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. Another variation uses sour cream instead of the second brine step. They will have a salt content >6% (w/w) in the water phase.FISH AND SEAFOOD 5. Traditional jellied eels are a product where pieces of fish are cooked in boiling water. and packed (3). only the nonspore-forming organisms are eliminated in the process and. until the flesh is soft. they are packed for consumption within 2 weeks at chill temperature (6). After this. There is epidemiological evidence that these products have been the cause of some 100 . 5.3. herring or other fatty species are used. has a shelf life of several years. usually requiring refrigeration below 10 °C for longer-term storage. and lasts for 3 . salt. ‘ikan kaju’. but it has further smoking. a Peruvian product. then packed in barrels with a 70º brine. The fish can be soaked in a high concentration of vinegar (about 4 . Other preservation techniques can be used for extending the shelf life of these products. and spices) is added to the brine. and 6% salt.4% of salt) for distribution and sale (1). However. or can be boiled by being placed in a basket. A much firmer product is obtained when the fish is salted prior to acidification. drying and salt-boiling steps until a hard translucent block is obtained (4).7%) for as long as 3 weeks. and even death. containing 1 kg of salt for every 25 kg of fish. they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. Ceviche. Usually. placed in a marinade of lime juice. because of the salt content of the product. As referred to above. or a pH <5 and they are stabilised for a limited period.30 minutes. it can be repacked in milder brine (1 . flavouring (sugar.6 days without refrigeration (1). The lime juice plays the role of the vinegar. whereas the latter method preserves the fish for only a few days.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. Once cooled.3.3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. the occasional implication of pindang in cases of foodborne illness. In a second step. which is dipped into a boiling brine for 10 . Another salt-boiled Indonesian product. usually flavoured with onions and hot chillies. The first process can give a longer shelf life. is made of different varieties of fish.2 months. even if the product is hermetically sealed. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. Herring can also be cured for several days in an 80 to 90º brine with 2. outgrowth of bacterial spores after processing may occur (4).5% 120grain distilled vinegar.2% acetic acid and 2 . giving a cooked appearance to the product. underlines the need for careful control (4). All these products are ‘semi-preserves’. Afterwards.
are sundried and. bacterial toxins and parasites (7). and the flesh will break down. are commonly dried in Asian countries. It is important to assure the quality of the raw material. under a light pressure. depending on the thickness. such as squid. etc.There are several products named marinades (matjes. sugar and spices. and the stacks are placed in the refrigerator. and a fine product is obtained. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival.5. but sometimes it is dried without prior salting. 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). it will last for many years (5). Stockfish products have a golden brown colour and are very hard. Some bacteria and enzymes are still active in the presence of acid and salt. the fish is dried naturally in a relatively cold environment. air flows naturally. Shark pectoral fins. Gravad fish is a Scandinavian product made of salmon fillets. Dried shrimps are widely used in Asia. jellyfish. split and dried gadoid species) are traditionally made in cold climates. Italy and Spain (5). the temperature and the amount of pressure applied (5). 101 . by using either solar heaters or mechanical dryers. used as an appetiser. gutted. mixed with salt. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures.CURED. SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. They are an important dried product. Provided the product is kept in dry and fairly cool conditions.) where no acid is added to the process. Stockfish is extensively used in Portugal. are likely to survive in these mild-processed products. Matjes are made of fat young herring. which are placed in a mixture of salt. Other products. after frying.4 Air drying This is probably the oldest method of fish preservation. It takes about 2 days to marinate completely. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). marinated products will keep for several months at a temperature of 4 °C. Fresh dill is placed between the layers of fish. Usually. such as Listeria monocytogenes and Clostridium botulinum. are generally air-dried at sea. 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. In some coastal areas of Portugal. At pH levels below 4. South America and the Caribbean. In Scandinavian countries. Stockfish (dressed. 5. In the first case. sea cucumbers and seaweeds. some fish species such as moray eel and varieties of dog fish. The most common method used for fish products is the air-drying process.0) (7). Dried octopus is also a very common dish in the interior of the country. still made without prior salting on a large scale.3. Foodborne pathogens. gravad fish. the fish is dried after salting or brining.
It is important to balance the drying rates and quality/safety issues.83 (9)). aureus) or spores (e. making a water barrier and consequently. Whilst the low aw of the final product will inhibit microbial growth. the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or. more commonly nowadays. botulinum growth might occur in the bulk of the product. and increasing amounts of such products are making their way into the market place. neither it. Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. the protein must be set or denatured with low-temperature drying before applying higher temperatures. However. that may have contaminated the product during the drying process. and it is exported to Portugal. dried salted cod (bacalão) is now mainly produced in Norway. Thus. but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). spoilage and possibly C. Dried salted products must have a water activity below 0. Whilst the production of dried fish products is tightly controlled in industrialised countries. the product can be held for months at room temperature. or wooden drying rack to dry in the sun. toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0. 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. some of which are listed in Table 5. if the rate of water removal is too high in comparison with the diffusion rate. tarpaulin.the lower the humidity. In conclusion. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain. In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. Staph. The product is dried to a final water content of about 25 to 38% (3). the quality is more difficult to control. Italy and South America. To prevent this. France. skin up. When no more water can be removed from the fish. The fish must be stored in reasonably dry conditions to avoid moisture absorption.g. Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s. the fish must be dried slowly enough to avoid case-hardening. After dry salting. the diffusion of the bulk water to the surface of the fish is blocked. and the water must be changed several times before cooking.III.g. nor subsequent cooking operations will affect the presence of toxins (e. 102 . Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources.75 (1). botulinum). by warm air circulating in a drying chamber. and has the primary objective of creating a product with specific taste and textural characteristics. Iceland and Canada. the skin will form a hard case (case-hardening). typically in stews and soups. the better the moisture transfer. C. which leads to mould growth (aflatoxins. They must be soaked in water.FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. Spain. 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 . there is an increasing demand from people that have emigrated to industrialised nations for traditional products.
It has been reported that the smoke from mahogany. and the use of smoke generated by superheated steam or liquid smoke. aureus (toxin) C. swordfish. using electrostatic filtration of the smoke. eel. It can be used for cold or hot smoking. haddock. Maintaining the temperatures of pyrolysis between 425 and 200 °C. oysters. sprats. is mainly a process for flavouring fish rather than a preservation technique. Hardwoods are considered better because of the milder flavour they impart to the food. In the mechanical kiln. There are many fish species that are used for smoking purposes .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. mussels and many others from tropical countries. Top-quality fish must always be used. as referred to above. The smoking procedures vary widely. Staph. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10).5 Smoking Smoking today. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. softwood colours the product quickly. making full description impossible here. aureus and Saccharomyces cerevisiae (10). However. clams.3. white mangrove and abura has an inhibitory effect on some microorganisms. cod. mackerel. The quality of the smoke is related to the type of wood used. botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. cod roes. Aroma and flavour are a blend of smoke components (3). SMOKED AND DRIED FISH TABLE 5.salmon. 5. such as Escherichia coli. Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . tuna. Blends of various woods can be formulated to produce sawdust with specific flavours. herring. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish. trout.CURED.
they can be vacuum packaged.10% of the weight of the fish. the excess salt is removed with water. canned or bottled. skin off or on. The maintenance of temperatures below 30 °C avoids protein coagulation. Brining at 10 15 °C is recommended (10). but this concentration will again depend on the type of product and the length of the brining stage.5.and hot-smoking is the temperature of the process. as it retains more moisture. usually dry-salted and cold-smoked. permitted colourings or smoke flavours can also be added to the product in the brining step. Also. Whether the product is prepared headed. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. The salting period is dependent on the size of the fish and its presentation (whole. which is important for the appearance of these products (4). Cod roes. whether cold. Sugar can be added together with the salt. but a ‘safety’ (i. thus allowing the deposit of smoke on the fish surface. spices. the following step is salting. 70 – 80 º brine is commonly used. thawing must be done slowly for 12 . wooden boxes or master cartons (10). Drying should be minimal at the start.18 hours at 4 °C. skin on or off). Depending on the type of product. Wet salting or brining is preferable to dry-salting when smoking fillets because. gutted or filleted. The length of the smoking process for such products is much greater than for the hotsmoked fish. it will reduce the salty flavour. are boiled before smoking (12). Sugar. and the cold smoke is denser. This difference is crucial for safety issues because the flesh is not cooked. However. giving the product the desired appearance and flavour (4).FISH AND SEAFOOD water flow system with chlorinated water (25 . When frozen/thawed fish is used. If wet salting is chosen. All the smoked products. smoked cod fillets and finnan haddock are usually cooked before consumption. it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11). after being smoked need to be chilled immediately to 4 °C before being packed. In cold-smoking. tuna and swordfish. covering the fish surface completely (10). low temperatures will slow the uptake of salt. shrink packaged. as well as many other products prepared in the same way. or quick-frozen and stored at -18 °C (2). the highly desirable cold-smoked salmon. all cold-smoked products should be cooked before consumption.e. it will take up salt twice as fast as fresh fish (10). If frozen fish is used. but it will increase towards the end of the process (10). temperatures must be kept under 30 °C throughout the process. 104 . Thus.50 ppm) should always be done just before process initiation (3). however.or hot-smoked. an attractive glossy pellicle on the fish fillet surface is achieved after draining.1 Cold-smoking The major difference between cold. are not cooked prior to consumption. owing to the elution of soluble proteins. filleted.3. such as smoked salmon-trout. The fish can be dry-salted using a weight of salt 5 . pasteurisation) temperature is not achieved in any stage of the process. Thus. or packed in retortable foil pouches. Some herring products. 5.
and their sensitivity to salt (13). On the other hand. the products are cooled. a drying period at 30 °C. SMOKED AND DRIED FISH During cold-smoking. gravad salmon (also called gravad lax or gravlax). thirdly. refrigerated storage and vacuum packaging still allow its growth (12). This temperature should be maintained during storage and distribution (3). a final cooking period at 80 °C (12). See Figure 5. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). During smoking. The exception is vacuum packed fish. which underlines the danger of storage at abuse temperatures of >10 °C (13).3. Pathogenic vibrios. such as Yersinia enterocolitica. a partial cooking period at 50 °C.5. Smoked salmon fillets are usually vacuum packed whole or after slicing. as. Smoked products can be also packed under controlled-atmosphere packaging. but the proteolytic types A and B. in order to produce toxin. The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. and hotsmoked salmon (kippered) (3). further cooled to 3 °C within 12 hours and then packed. can be isolated from seafood. If the refrigeration chain is maintained. Hot-smoked products require a longer cooling stage. stored at 4 °C and packed. condensation will occur inside the pack. Vacuum packing is recommended only for coldsmoked salmon. To avoid growth of non-proteolytic C. 5. botulinum type E the product must have at least 3% salt in the water phase and be stored below 5 °C. if a 105 . can grow with salt levels of 10% in the water phase. Furthermore. These products should be vacuum packed at room temperature. typically mesophilic. Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). Although other human pathogens that can grow at refrigeration temperature. A typical temperature set is as follows: firstly. Length of the smoking process at each stage will depend on the species and the type of product. secondly. there is no point in the process that can fully assure the absence of L. thus allowing any psychrotrophic C. 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. if they are taken from a chill room at 3 °C and packed. are of concern when smoking fish from tropical waters (13). botulinum type E present an even longer time than before to grow and produce toxin (14). Neither the smoking temperature nor the salt content is enough to kill the organism. and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). monocytogenes.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. if present. but they should be cooled to a temperature of 10 °C within 3 hours.CURED. Afterwards.2.
it will have a reduced shelf life. 5. Process of cold-smoked sliced salmon 106 . and moulds will grow on the product (12).FISH AND SEAFOOD product is packed too warm without being cooled to room temperature.2. Fig.
In terms of safety.6 hours and smoked for 3 . The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. to have no further advantage over other packing methods for other fish (12). A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean. white fish about 8 days. resulting in a characteristic flavour (20. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. including ‘umami’ (16. and many closely related Aspergilli (Aspergillus ochraceus.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. fatty fish will remain in good condition for 6 days. Natural smoke extracts. They are generally not cooked prior to consumption. 22. cooking in brine. 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). Hot-smoked products made from white fish will usually keep better than those from fatty fish. rewetted and remoulded to give the desired shape and consistency. Katsuobushi. The classical liquid smoke is 107 . but seems. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). are usually 80º brined for 5 . synthetic smoke flavours.3. including Eurotium repens and Eurotium rubrum. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). 21. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). such as yeast derivatives. Aspergillus ostianus.CURED. apart from the improved appearance.3½ hours using the typical temperature set for hot-smoking referred to above (12). 18). When refrigerated at 3 °C.5. Aspergillus oryzae. similar to masmeen. Growth of these moulds on the tuna results in the extraction of moisture. moulding it into long thick sections. Vacuum packaging retards the onset of rancidity when fatty fish is used. are used. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. 17. 5. oysters and mussels are usually wet-salted in 50º brine for 5 minutes and hot-smoked at 80 °C for less than 1 hour (12). a very expensive and fashionable smoked product in the Netherlands. Mackerel. (Penicillum cyclopium and Penicillium puterellii) (19). but mechanical drying is used instead of sun drying. Boiled or steamed clams. 23). Eels. Aspergillus tamarii) and several Penicillium spp. break down of fats and the moderation of smoke flavour. then hot smoking and finally sun drying it. Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. chilled or freeze-thawed. SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). The production method is similar to masmeen. The product is removed from the drying cycle. and substances unconnected with smoke but with a smoky flavour or smell. They will keep well frozen.
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). botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). nutritional neutrality and antioxidant activity (27). Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. The European Scientific Commitee for Food. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. In experiments comparing traditionally smoked fish with liquid smoke. the salt level could effectively be reduced from 4.8% and still provide protection for 7 days (25). According to some recent research. or applying it as an atomised spray within a modern automatic smoke kiln. It has been shown that liquid smoke can reduce the formation of PAHs (3). smoking.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. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. both of which are considered potential carcinogens. 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. 5. applying it as a dip after brining. When liquid smoke was used. drying.06 μg/kg of benzoanthracene in the final foodstuff (28). spraying is preferable because it is easiest to control flavour and acceptable with this method (3).4 Spoilage Brining. Fresh fish spoilage may be measured by 108 . so as to impart an unnatural red colouration to the flesh.3. consistency of flavour and colour. or smoke extracts in organic solvents. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish.5. 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.6 to 2. 5. injecting it into the fish flesh.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. 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.03 μg/kg of benzopyrene. Liquid smoke was considered to provide advantages of safety.
however. both of these will affect the spoilage patterns. contact with the bacteria 109 . the fish are cut and splayed to allow greater surface contact with the salt.g. When acid marinating or smoking. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). or extended refrigerated storage. SMOKED AND DRIED FISH grading schemes or quality indices. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime. a product may become undesirable for consumption as a result of changes in its sensory characteristics.. Spoilage needs to be carefully defined for each product.. When heavy brining or salting is important for product shelf life. Upon salt packing. marination or drying). which are found in the salt. a process similar in microbiological effect to that of saturated brining. The use of drying and/or salt to lower the water activity. the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. the consequent reduction in the pH will help to inhibit spoilage organisms. Once these changes are defined. and of smoke to add chemical preservatives. which may also be produced to describe the quality attributes and their loss over time for processed fish. Often. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt. the bulk of the microflora comprises Bacillus spp. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure). limits the types of microorganism capable of growing. Within the limits of safety. Brining. and. and high concentrations are usually required to inhibit their growth fully. Even saturated brines allow some halophilic organisms to grow. these bacteria tend to continue to dominate the flora in the liquid. contaminant bacteria from other materials used to produce the product. In salts. but. For some products. Once placed into brines. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. Some brines are re-used over a period of days and a load of 10. for some. also. Usually. the production process will modify the microbial flora present on the product. chemical oxidation. a very dry product is required to produce 'shelf-stability'. otherwise. Micrococcus spp. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). pickling and dry-salting all introduce bacteria to the fish. the shelf life may be estimated and investigation made into the causes of the loss of sensory quality. Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2. these halophilic organisms may grow and cause spoilage. therefore.000 cfu/ml of brine may build up.3%). the fish flora are transferred to the brine mix. and coryneform bacteria. Most marinated and smoked fish products are first brined or salt-pickled. rock and manufactured salts did not usually contain halophilic bacteria (29). Stronger and longer brining has an effect like that of pickling. or their products require refrigerated storage. spoilage is due to endogenous microorganisms and enzymes of the fish. for others. The microflora attached to the fish is relatively unaffected by light brines used before smoking.CURED. as the fish are added.
as little if any growth will occur below 5 °C on salted fish (29). A. 34) have shown that P. yeasts and fungi. powdery growth. W. Aspergillus sydowii. 33).. The microbiology of heavily salted products is dominated by Gram-positive. E. (syn. Sporendonema sebi)). as the water activity of saturated brine solutions is low enough to inhibit most bacteria. The bacteria are osmosensitive and will lyse in water. sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). flavus. yeasts. and still produce a reasonable dried product.FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. Only halophilic microbes may grow. Unlike ‘pink’. and Aspergillus penicilloides. Several Penicillium spp. halotolerant or halophilic micrococci. may also be found in the salt used in production. Studies conducted on dried and salted fish from Indonesia (31. the best way of controlling the organisms is to refrigerate the fish. or after drying if the fish is stored in excessive humidity. rancidity due to lipolysis. spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. protein breakdown. gaspé). pisce was the main fungus isolated. most notably Penicillium citrinum and Penicillium thomii. lactic acid bacteria (LAB) and moulds. The bacteria form coloured colonies on the surface of salted fish prior to drying. if not too contaminated. although it may occur in other salted fish. The metabolism of the organisms will produce off-odours. were also isolated. 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. including hydrogen sulphide (26). halophilica was isolated 110 . Therefore. spore-forming bacteria.g. sometimes covering the whole of the fish with a whitish. though only the latter two species were considered likely to have grown on the fish (32). At the pre-drying stage. ‘pink’ and ‘dun’. This condition is often associated with case-hardening. 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). which. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage. Basipetospora halophila. rubrum are also common isolates from dried fish. The condition describes the growth of the halophilic moulds (Wallemia sebi. Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. niger. Eurotium amstelodami. it was found in about 50% of samples examined. including A. Aspergillus wentii. Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration. or fat breakdown) caused by enzymes rather than by microbial growth. (syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. repens and E. like the ‘pink’ organisms. together with a range of Aspergilli. 32. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. B. and Halobacterium spp. it is possible to wash off the infected parts. ‘dun’ is not characterised by the production of offensive odours.
and microbial spoilage is more likely. It has been reported that the type of salting (dry. it was. able to grow on dried salted fish. has been isolated from dried fish (35). SMOKED AND DRIED FISH infrequently at low levels. where a high internal temperature is reached. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. but this will not prevent the growth of spoilage bacteria. Cold-smoked fish will contain microbial flora representative of that of the raw fish. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). 36). Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. Marine vibrios were the dominant flora in smoked salmon that was drysalted. LAB. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. around the water activity of saturated sodium chloride. Smoked fish products tend to have a light brine treatment. flavus. not the growth of spoilage organisms. which does not create the salty conditions required for spoilage by those halophilic microorganisms. botulinum type E. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. so the bacteria spoiling these products will be psychrotrophic or psychrophilic. Fish processed by cold-smoking. including A.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). For more information on spoilage see Table 5. Refrigeration is necessary. will have a different residual post-smoking flora from 'hot-smoked' fish. however. but. with a lower aw limit for growth of 0.5% sodium chloride in the water phase will inhibit some bacteria. Table 5. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. Vacuum packing limits chemical oxidation. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage.747. as already stated. most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations.IV. Pseudomonadaceae. 33. 111 .or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. The heat during hot-smoking will kill vegetative microorganisms but not all spores. The preservative effect of the smoke will inhibit some spoilage microorganisms. The salt in the water phase must be high enough to inhibit the growth of C. Aspergillus spp. The brining to 3 . Aspergillus clavatus and Aspergillus niger. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life.CURED. In Vietnam..
Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad.not used/unchanged. halophila Halophilic spoilage organisms Halophilic spoilage organisms.IV Spoilage of salted.variable.FISH AND SEAFOOD TABLE 5. 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. boiled. smoked and dried Citrus marinade Dried Brined or salted then liquid or cold smoked Brined or salted then liquid or hot smoked 1 2 +++ Jellied eels + Ikan kaju Ceviche Stockfish Coldsmoked fish Hotsmoked fish +++ +++ ++ +/++ + + ++ ++ - ++ + + + ++ ++ ++ ++ + ++ + ++ Antimicrobial compounds added Refrigeration applied 3 v. + to +++ strength of preservative effect 4 Post-process contaminants 112 . . and Halobacterium spp. sebi and B. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted.W. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. Dun. LAB.
However.0-9.5-8. with this change. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). Since neither the smoking nor the drying parts of this curing process are particularly severe. the fermented products being the most dangerous group (7). but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. In the United States.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. botulinum.8 0. botulinum type E and production of toxin. botulinum Staph.97 0. when present in the raw material.86 0. The concentration required to prevent growth at room temperature can vary from as low as 3. The presence of salt in cured products has a great effect on the growth of the bacteria. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13). aureus Yeasts Moulds Lactic acid bacteria pH 5. smoked and dried. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C.5 5. under optimal conditions.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). SMOKED AND DRIED FISH TABLE 5. and as botulism is a serious hazard in foods. the products were heavily salted. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E. botulinum type E. It has been shown that. could not grow and produce toxin. the non-proteolytic type E strains can grow in up to 5% sodium chloride. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). although the combination of the two factors. types A and B are not affected by 3 . salted and pickled products accounted for 10.61 Organisms Most Pseudomonadaceae C. the importance of cold-chain integrity must be emphasised.5-10 4. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. pressure on the traditional methods of curing.5-10.75 0.5.5 4. 9 and 8 of the outbreaks. Before. respectively.97 0.5 5.5 0-11 3.5% to 5%. popular demand for healthy foods has led to a change in the tastes preferred by the public and. but if the fish products are stored at temperatures <10 °C. and C. sodium 113 .5-8.CURED.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades.8 1.9 0. botulinum type E Staph.
The FDA established that hot.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. are particularly susceptible to contamination and. they will grow rapidly (7). gills and guts as well as in several lightly preserved fish 114 . as they require more handling. but. coli all occur on fish products as a result of contamination from animal/human reservoirs.4 Listeria monocytogenes The psychrotrophic L. throat and skin. but requires higher temperatures for toxin production (>10 °C). Infection by Shigella sonnei has increased in the UK. It is a halotolerant bacterium able to grow at water activities as low as 0.5. the human nose. if the person handling the product is a carrier of Staph. if storage temperatures are over 10 °C. these parameters change as a result of the interference of food components and other limiting factors that can act in an additive or synergistic way to prevent Staph.30% of the population positive for enterotoxin-producing strains (41). monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. has been reported (42). 5. if recontamination occurs and storage conditions are favourable. growth and toxin formation may occur. Shigella and E. In foodstuffs. Sliced smoked fish products. The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). it should contain not less than 3. when the bacterial load of the product is lower.0 .5. they do not grow well in the raw material. more importantly. However.or cold-smoked fish should have at least 3. As with other organisms. and the minimum pH for growth is 4.5% salt in the water phase.0% salt in the water phase (30).2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and.94 (7). Once the product is contaminated.5. 5.4. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0. if it contains sodium nitrite ≥100 ppm. but. As they are poor competitors. the organism may contaminate the product. 5. Usually. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . aureus. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene.5. Thus. aureus growth. the risk of infection can be eliminated by proper cooking. An outbreak of Shigella in an Italian cruise ship. related to the consumption of a raw smoked fish meal. the organism can survive most of the light curing processes.3 Enterobacteriaceae The pathogens Salmonella. after processing.86 and 10% sodium chloride. The organism is mesophilic with a minimum growth temperature of 7 °C.
which. the USA established ‘zero tolerance’ for the organism. The round worms. monocytogenes in levels at or above 102 per gram (46). a number of fish products. there were no reported outbreaks related to cured fish. and the tape worm Diphyllobothrium spp. gravad fish. This includes matjes herring. and other local traditional products (7).5. widely used as starter cultures in food processes. most of them are of little concern to public health. the CT 3162-FAIR program.5 Parasites Despite the presence of parasites in fish being very common. the only concern to public health is when eating raw or uncooked fish products.2. SMOKED AND DRIED FISH products such as cold-smoked fish. do not survive 1 min at 60 °C.9. has also been detected in cod (7).514. Until 1997. cold-smoked fish.95 (45).CURED. Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. salt levels of 8 .the round worm Anisakis spp. marinades (ceviche) (43) and gravad fish (44).6 and water activities below 0. a round worm. Of microbiological concern in the northern hemisphere are two parasites . Its limiting conditions for growth are 0 . researchers are looking for other ways of preventing the pathogen’s occurrence or growth.45 °C. from seawater fish. which include intermediate hosts. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. Thus. Two outbreaks in the United States have been reported to be linked to salmon (10). 5. which considered L. Pseudoterranova dicipiens. such as ‘lightly preserved’ fish products. As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism. Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. a short period of freezing either of the raw material or of 115 . monocytogenes’ in 25 g of the finished product. in the EU Fisheries R&D-FLAIR program. which means that the process must conform to a level of ‘no detectable L.0. Some LAB. Unlike some European countries. The use of LAB in fish preservation is a topic that has been studied in projects (UP. have been shown to have an anti-listerial activity in many food products.8 . are considered unsafe. lightly salted caviar. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). The Health Protection Agency (HPA) in the UK considered that it was unacceptable for readyto-eat foods (including all the products that are consumed without prior cooking) to contain any serogroup of L. and in CT95-1207FAIR). when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). monocytogenes pathogenic for only specific segments at risk. As they are heat-sensitive. both being detected in cold-smoked salmon. They have complicated life cycles. in combination with curing agents and refrigerated storage.12%. favours growth of psychrotrophic LAB.92 . or nematodes. pH 4. from freshwater fish.
cholerae Vibrio parahaemolyticus L.<104 20 .<100 20 . † Guidelines for aerobic colony counts may not apply to certain fermented foods. coli (total) Listeria spp. cereus and Bacillus subtilis# <20 <20** <20 <10 20 . 5. salami.<100 10 .<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . E. monocytogenes) (cfu/g unless stated) <106 106 . monocytogenes Staph.VI Guidelines for the microbiological quality of ready-to-eat smoked fish.<107 ≥107 N/A <100 <20 100 . ‡ On occasions some strains may be pathogenic.6 Published Microbiological Criteria TABLE 5. aureus C.100 >100 N/A Pathogens Salmonella spp. and the levels or absence of indicator organisms or pathogens.<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. the organism should be identified # N/A denotes not applicable 116 .<103 N/A 100 .<104 100 . for example. 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E. and unpasteurised yoghurt. Acceptability is based on appearance.FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). soft cheese. smell.<100 200 . texture. (not L. If the Bacillus counts exceed 104/g.<104 104 .<100 20 . perfringens B. These foods fall into category 5. coli O157 & other VTEC Vibrio.<100 >104 >100 N/A N/A <20 20 . not detected in 25 g Campylobacter spp. taramasalata and cooked shellfish sampled at the point of sale (adapted from (11)) Criterion Satisfactory Borderline limit of acceptability (cfu/g unless stated) Unsatisfactory Unacceptable/ potential hazard* (cfu/g unless stated) (cfu/g unless stated) Aerobic plate count† (30 °C.
Horner W. botulinum * Limit of tolerance 1x 103 ETEC/g.7 1. or presence of toxin 0.A. Fernandes R. Ed. Assurance of Seafood Quality. * Staph. 8. 7. 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. 3. Eds. Torry Advisory Note No. monocytogenes Salmonella spp. 37(revised). Preservation of fish by curing (drying.. 1993. Moody M.M. Smoked. Food spoilage fungi. 4.W. Doré I. Huss H. Hall G. Flick G. Horne J. USA.. 120-38. 334. aureus * Vibrio cholerae V. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. Regenstein C. salting and smoking). Martin R. New York. Bannerman A. 1990. 82 (revised). Introduction to Fish Technology. 117 .H. Marinades. 3273. 1984. in Fish Processing Technology. parahaemolyticus Vibrio vulnificus C. 1994. References Regenstein J. SMOKED AND DRIED FISH TABLE 5.CURED. Van Nostrand Reinhold. 56.F.F. Flick G. 2. 1997. Chapman & Hall. 381-406. Urner Barry Publications Inc. coli (ETEC) L.M.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. 1991. Eds. except in the viscera of dungeness crab.E. Torry Advisory Note No. Wareing P. 2007. Torry Advisory Note No. Birnie K. in The Seafood Industry. McLay R... Leatherhead. 9. cured and dried fish. New York. Fernandes R. London. Catching. Wareing P.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.. 254-82 5.. Van Nostrand Reinhold. Leatherhead Publishing. 6. in Micro-Facts: The Working Companion for Food Microbiologists. Handling and Processing Eels. 1982. The Smoked and Cured Seafood Guide. 1979. Hot Smoking of Fish. FAO Fisheries Technical Paper.
118 .. Kinoshita Y. and Biochemistry. 2324-5. 553-72..A.E.C. 19. Doe P. 51-81. Shuto Y.. 508. Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish. 840-6.. Teeny F. 1981. 1051-5. O-methylation of 2. Horne J. Torry Advisory Note No. 335-40.. Ogiwara Y. 24. London.. Doi M. Sakakibara H. 61 (8).M.B.. 843.M..H. Bioscience.. Torry Advisory Note No. 22. Embarek P. 1989.. Microbiological control for fish smoking operations. 2002. 1977. Fish Drying and Smoking: Production and Quality. Perusing Asian soup options. Ninomiya Y. 12. 59 (12). Kinoshita Y. 1992. Agricultural and Biological Chemistry. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour. Biotechnology. Patel T. Flavour constituents of dried bonito (Katsuobushi). in Fisheries Processing. 16. Lindemann B.. 45 (10). 53 (4)..C. Matsui M. Druschitz A. Packing Fish in a Modified Atmosphere. Biotechnology and Biochemistry. Huss H. Ed. 22 (revised). Bioscience. Torry Advisory Note No.. 17 (9). 1994. The discovery of umami. 1984. 1997.. Aoshima H. Peterson M. 2008. Food Chemistry. Journal of Food Protection. Jeppesen V.. 56 (6). Shuto Y. 1386-7.. 11.P.. 21. Food Product Design. 935-41. 5 (revised). Lancaster. Doi M. Yamauchi H. Halpern B.. Martin A. Botulism and Fishery Products. Technomic Publishers. 1980. 18. Biotechnological Applications. 14. Bannerman A. 25. Food Control. Yajima I. 20. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor. Dillon R.. 27 (9). 108 (3). 18.FISH AND SEAFOOD 10.. Cann D. Martin A.. 1998. Biotechnology and Biochemistry. 6 (4). Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species.R. Pelroy G. 6 (6). 1995. Doi M. Mitou M.. Kanayama T. 1990. Hosokawa M. Recommendations for the Preparation of Smoked Salmon. Shuto Y. 23.W..E. 88. 17.. Processing Cod Roes.K. Bioscience. 1995.M. 2007. 15. Food Reviews International. Hayashi K. Control of biological hazards in cold smoked salmon production...F. Matsui M. Doi M. 958-60. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors. Paranjpye R. 13. Cann D. Yokayama S... Torry Advisory Note No. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour. Shiemori Y. Chapman & Hall. Ninomiya M. Eklund M. Bannerman A. What’s in a name? Are MSG and umami the same? Chemical Senses. 1982.
119 . Cox J. 1971.. 1985. Nahrung. 88-90. 1996. 43. 2005.D..A. Hess E. Hocking A. Food Microbiology. 29 (2). 38..H. 12.E.J. Lancet. 3 (4).S...R. Food and Drug Administration. Cann D.V. Hocking A. Ed. 36. Pszczola D.M. in Reports of the Scientific Committee for Food (34th Series). 197-208. 31.I.A.D. The control of botulism in hot-smoked trout and mackerel. 27. 74.I. Institute of Medicine. in Smoke in Food Processing. 1997. Journal of Food Technology. Incidence of Listeria monocytogenes in an acidified product. Hocking A. Importance of autolysis and microbiological activity on quality of cold-smoked salmon.. Fungi associated with Indonesian dried fish.CURED. Ed. 348. Fuchs R. Simini B. Modern Food Microbiology. Ed.. 311-7. Smoke flavourings. 218-25. Letters in Applied Microbiology. Wheeler K. Food Technology.Y. Townsend J. 34. Frank M. 74 (2). 742. 29.I. Sprouse R. Ed. 14.C. 22. Gill T. Luxembourg. Commission of the European Communities. Hocking A. 1995.. Maga J. 164-9. Taylor L. The microbiology of sea-water fish. Oxford. Røntved S. Boca Raton. Academic Press. 1994.B.T. 28.M. Washington DC. 40. Springer Science. CRC. Lucas F. 49 (1). Press Inc. Scientific Committee for Food..A. Hansen L. Pitt J.. Pitt J. Mycotaxon.. Studies on Sporendonema epizoum from ‘dun’ salt fish. Interactions among xerophilic fungi associated with dried salted fish. 107-11. Jay J.D. Blackie. Sirvas S.. Food Research International. Golden D.A. Food and Drug Administration. in Fish as Food. Journal of Applied Bacteriology. 1996. Antimicrobial properties of smoke. Shewan J. Journal of the Fisheries Research Board of Canada.. 123-9. Smoked and smoke-flavoured fishery products. 1995.F.O. Huss H. 1993. New York..K. 35.. An outbreak of Shigella. in Fish and Fishery Products Hazards and Controls Guide. 33. 181-8. Tour highlights production and uses of smoke-based flavours. Fungi and Food Spoilage. SMOKED AND DRIED FISH 26.. Committee on Evaluation of the Safety of Fishery Products.D. 32. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. 30. 1991. London. 70-2. Rockville. Studies on salt fish. 1941. 1994. 351-7. 42. CEC. Journal of Tropical Medicine and Hygiene. 41. 39. 38 (3).E. New species of fungi from Indonesian dried fish. Anggawati A. Diyaolu S.. Loessner M. Borgstrøm G.F. 1991. US Food and Drug Administration.. V. ceviche. 37. 1961. 1986. Fungal flora of South Vietnamese fish and rice. 98100. Ahmed F.M. 5. Adebajo L. 27686. Seafood Safety. 74. Pitt J.1988. 1979.. National Academic Press. Wheeler K.
Tham W.. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale. Harrison M. 1982.L. Fletcher R. 120 . 5 (1). 13-8. 1980. Amsterdam. Round Worms in Fish. and smoked fish establishments good manufacturing practices. Association of Food and Drug Officials. Moral A. Danielsson-Tham M. 46. Richards J.H.gov/~comm/haccp4x5.E. 1995. 188 (3). 833-41.. Advances in Food Research. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions.D. Elsevier Science Publishers B.. Hughes A. 3.. 22. 1989. Appendix 5: FDA and EPA Guidance Levels.S. http://www. Canadian Journal of Microbiology.. An outbreak of listeriosis suspected to have been caused by rainbow trout. Persson I.. 45. 55 (4). Eds. Roberts D. Journal of Food Protection. Mentzing L-O.A. 1992.cfsan. Eklund M. 197-209..html Gilbert R. 58 (8). 48. 1991. 1996. 507-10. Prevalence of Listeria monocytogenes and other Listeria spp. 863-6. 80. 1994. Liston J. 232-6. Little C. Davies A.. Eklow A.. in smoked and "gravad" fish. Wooten R.. 135-85. in Fish and Fishery Products Hazards and Controls Guide. 35 (11) 2904-7. 37 (1). Huang Y. Pelroy G. Peterson M. Journal of Clinical Microbiology.FISH AND SEAFOOD 44. Food and Drug Administration. Journal of the Association of Food and Drug Officials.W.J.. Danielsson-Tham M-L. Pathogenic microorganisms of importance in seafood..C.. Loncarevic S. Ericsson H.. Zeitschrift für Lebensmittel-Untersuchung und –Forschung. 49-59. Ribeiro C.C. 163-7. Journal of Food Protection. Donovan T.. 1997... Paranjpye R. 1976. Cann D. Tham W.. 26 (4). Communicable Disease and Public Health. 5-7. Beltran A.8 Further Reading Gibson D. 47..J. Torry Advisory Note No. salted.. US FDA Centre for Food Safety & Applied Nutrition. US FDA Centre for Food Safety & Applied Nutrition. 2000. Huss H. Pelaez C. Bolton F. 1980. 45 (9). Loncarevic S. Hurst A.. Cured.. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects. Nye K.W.V. in Quality Assurance in the Fish Industry. Hobbs G.. Susuki E. Clostridium botulinum and its importance in fishery products.R...A. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon. Gibbs P..N. Acta Veterinaria Scandinavia.M. 5.M. Unnerstad H. de Louvois J.. 2001. Ed. Jacobsen M..A. Incidence and growth of psychrotrophic Clostridium botulinum in food. Garren D. Washington D.. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus.fda. Food Control..
Hitchins A. International Journal of Food Microbiology. 1974.J.. Yndestad M. Contamination pattern of Listeria monocytogenes and other Listeria spp. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. Incidence of Listeria in hot and cold-smoked fish. Caugant D.. 55 (11). Kramer D. Peterson M. 502-8. 14. van Spreckens K. 61 (12). Beuchat L. 94-8.C. Combined effects of water activity. 394-6. Journal of Food Safety. Nicolaides L. Rørvik L.. Journal of Food Protection. 309-16. Poysky F.E. 1995.CURED. 1994. 121 . 58 (5). 1987. Elsevier. 1992.R. Hudson J.W. SMOKED AND DRIED FISH Sutherland J. Eklund M.A. Letters in Applied Microbiology.K. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes.R.. 1994. 844-6. 19-27. 42. Taylor S. Pelroy G.. Food Microbiology. Davies A.. Histamine production by psychrophilic flora. 309-18. detection and growth of Listeria monocytogenes in seafoods: a review.A. 293301.. Applied Microbiology. solute. New York.N. Letters in Applied Microbiology. and salt concentration on growth and survival of Vibrio parahaemolyticus. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. Marine toxins of microbial origin. 866-70. 11. Prevalence of Listeria in smoked fish. 1992. Patel T. 1995. 1973. 1075-80.. International Journal of Food Microbiology. 21 (3). in a salmon slaughterhouse and smoked salmon processing plant. 19. in Seafood Quality Determination. 11.... Jemmi T. Lashbrook L.S.P.T. 1988. Dillon R.. Beuchat L. 1995. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. Tran T..A. Slade A. Poindexter C. Letters in Applied Microbiology. Applied Microbiology.. Liston J. Interacting effects of pH. International Journal of Food Microbiology. 217-36. Keusch A. 4389-95. Bayliss A.. Ratnam S. 27 (6).178-80. 1994. 1991. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants. Competition of food bacteria with Listeria monocytogenes during enrichment culture. Presence. Dallas H.. Ben Embarek P. 25. and temperature on the growth of Vibrio parahaemolyticus.R. pH and sodium chloride. 23 (1). 25 (5). temperature. Eds. Roberts T. 1734. Paranjpye R.. Food Technology. Fuchs R. 1994. Journal of Food Protection..
though some are produced elsewhere. Fermented fish products are largely confined to east and south-east Asia. producing the fish sauces and pastes. Those to which only salt is added tend to be mainly autolytic. 6. As with most traditional products that are still produced principally on a cottage industry or domestic scale. The fish sauces are produced on the largest scale. they are exported to European. koji.6. The relative importance of these two activities depends on the formulation of the product. whereas in those where a carbohydrate source such as rice or sugar is also added.I. 123 . added during preparation. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6. For example. Carbohydrates are added to some fish sauces during their preparation. the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. lactic fermentation can play a significant role.25% and are used mainly as a condiment.1. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. in economic terms. typically in the range 15 . there are numerous variants of some common themes and a host of local names used to describe them. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. Essentially. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. North American and other markets and are. the most important of all fermented fish products.1 Definitions As applied to fish products. FERMENTED FISH Prof. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt.
to products analogous to other lactic fermented foods such as salami.1. a variety of products can be produced within a single country. although garos. where the bacterial production of lactic acid is a major feature. These have long since fallen from use. salty taste. 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. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. 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.III. who had adopted them from the Greeks.II. and the more widely known Worcestershire sauce a very distant relative. and pissala. In Europe.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. made from whole fish in France. 6. may be direct descendants. A selection of Asian products is presented in Table 6. 124 .2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. made from fish livers in Greece.FISH AND SEAFOOD TABLE 6.
pa-chao. funa-suchi. crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 . shrimps.g.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 .FERMENTED FISH TABLE 6. e.III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. pla-som. 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. pla-chao. salt/fish ratio Production period Type of use Hoi-dong Molluscs Whole or without shells Whole - Main dish Kapi (fish or shrimp paste) Nam-pla (fish sauce) Nam-budu Small fish. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps. mam-pa-kor. som-pa. cencalok burong-isda. som-fak/ som-fug 125 Malaysia Philippines Thailand .8 months Main dish TABLE 6. burong-bangus pla-ra. burong-dalag.12 months 10% raw sugar added. som-pa-keng pekasam.g. burong-ayungi.3 months Main dish Tai-pla Bowels 1:3 6 . mam-chao. e. pa-khem.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. ayu-sushi. tai-suchi phaak.
TABLE 6. which employ lower levels of salt than Asian products. in Norway and surstrømming. and their microflora tends to reflect their local environment more than that of marine species. and counts ranging from below 104 /g to in excess of 107 have been reported. made from herring. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. which has been increasing in popularity over the last 50 years. malt or flour (1). rakefisk or surfisk. some fermented fish products remain popular. paddy ponds beside rice fields used for collecting fish when the field has dried up. in Sweden. or buried salmon. fermented trout. and the initial microflora is likely to be considerably larger in these cases.the flooded rice fields. and a large local freshwater reservoir (2). For example. 6.FISH AND SEAFOOD In Northern Europe. A modern account of their production. is a traditional. the most popular varieties of which are rakørret. in a survey of fermented fish products in north-east Thailand. Norwegian gravlaks. and old descriptions of their production often refer to the addition of whey. freshwater fish are the usual raw material. for example. The diversity of fish used in fermented products (see. 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. especially in fish/salt products. relatively mild-tasting product. 126 . Penaeus indicua Many of the fish used in fermentation are uneviscerated. These are often described as fish/salt products. Here.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. it was found that several different sources were used for the fish . however. There are also more heavily fermented products. Lactic fermented fish products are often associated with inland areas such as the Central Luzon region of the Philippines and the north-east of Thailand.IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. Table 6.
Bacillus spp. principally species of the genera Rhizopus. 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. the Philippine fish sauce. This.FERMENTED FISH Where other ingredients are used. Mucor.74 or below. sufficient to saturate the water phase present and produce an aw of 0. Similar results were seen with patis. and studies 127 . traditional starter culture preparations such as koji (Japan). in many cases.1 – 6. in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter.3 6. and the anaerobic conditions that prevail in the bulk of the material. More recently.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. work with nam-pla in Thailand has isolated Halobacterium and Halococcus spp. It also demonstrated their significant proteolytic activity during the first month of fermentation. mean that microorganisms have been generally held to play little or no role in the process that follows. were found to be the predominant isolates. angkak (Philippines and Thailand) and look-pang (Thailand) are used. either directly or as the partially saccharified rice they produce. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). however. It has also been shown that a perfectly acceptable patis could be produced from fish that had been sterilised with ionising radiation (6). the product contained about 500 cfu/g comprising mainly Micrococcus and Bacillus spp. 4).3. they can contribute to the initial microflora. after one month. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. Other halophilic species have been identified from fish/salt products. The amount of salt used is often only vaguely defined but is generally sufficient to reduce the aw below levels at which most bacteria associated with the raw material will grow and. In another microbiological study of four fish sauces. In some cases.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. which included nam-pla and patis. however. Hansenula. Generally. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. Endomycopsis and Saccharomyces . Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). When this is a simple carbohydrate source such as sugar.4. These preparations contain a mixed microflora of moulds and yeasts. suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. or boiled or roasted rice. (3. 6.
2. Patis (Condiment) 128 . 6. 6.12). Belacan (Condiment) Fig.1.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 . Fig.
This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues.3. 14).4. To produce semi-solid fish pastes as opposed to fish sauces. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are. the 129 .6. In fish sauce production. 6. thought to be largely the result of autolytic breakdown of the fish tissues. eviscerated fish are sometimes used to slow the autolytic process.FERMENTED FISH Fig. however. Ika-Shiokara (side dish) Fig.
and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens. In contrast to free-swimming fish. Hoi-Malangpu-Dong (side dish.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. snack) 130 .5.2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. which is eventually separated from the residual fish tissues. 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. for example. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. particularly glutamate. 6. Fig.6 – 6.5). Some examples of products and their formulation are presented in Table 6.8.3. volatile fatty acids and nucleotides. Figure 6. filtered and bottled. 6.V and Figures 6.
V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C. In these cases. Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus.25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0.6. central. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. Fig.03 Preparation time 6 .10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 .7 days 5 . 3:1:0.12 months Main dish 2 Pla-jao C.FERMENTED FISH TABLE 6. the prefix 'pla' is replaced by 'kung'. C NE.20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE. N. C. N S 5 .4 are also produced using shrimps. south of Thailand.12 months Shelf life Use 1 . N. NE Formulation Fish (whole or pieces): salt: roasted rice. NE. north.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0.6. Kung-Chom (staple food.5-1. north-east. N. C NE. side dish) 131 . NE 10 .12 days 3 .2-4 Fish (pieces): salt:khaomark (fermented rice).3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 . S.
If acidity is a desired sensory property of the product. giving a reduced rate of acid generation. Phillipines Journal of Food Science and Technology. 106) Fig. (from Arroyo et al. while they are able to grow at high salt levels. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6. Generally. This can be clearly seen from the data on reported shelf lives of a number of fish salt carbohydrate products and their respective pH ranges and salt contents (Table 6. the salt is of far greater importance in determining the product’s long-term stability. The LAB isolated from these products are those with more marked salt tolerance but.3. This is readily explained by the susceptibility of LAB to elevated salt concentrations. typically in the range 2 . their growth and metabolism are nonetheless slowed. then this will take longer to develop than if lower salt levels are used. Red burong isda 132 . 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.6.10%.salt and lactic acid..7. these products are salted at lower levels than the fish sauces and pastes. 2.VI).FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content.2. but there is considerable variation in this. It also indicates that.
9. Philippines Journal of Food Science and Technology.1 4.1 .17.4.8 2.9 .6.12 months 1 .9 4.FERMENTED FISH (from Arroyo et al.5 .5 3.8 .3 5.8 .5.5.4 . 133 .2 7. 106) Fig.3 .9 4.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).5.0 Shelf life 3 weeks 6 . White balao-balao TABLE 6.0 .2 4.8 pH 4.0 .5. 2.8..3 years 1 .7 .188.8.131.52 3.0 .9.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.2 4.5 .6.
Addition of carbohydrate will. In som-fak 9% of LAB isolated were able to ferment rice whereas 19% could ferment garlic and omission of garlic from the formulation resulted in lower acid production during fermentation (20.VII). although the final population of LAB may be similar. 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 . Where they are used. such as salt content. efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. inulin.4. Rice is the most common carbohydrate source used.0 . alternatively. When added as boiled or roasted rice alone.3.5) despite other factors. 21). a number of amylolytic species have been described and an amylolytic strain of Lactobacillus plantarum has been isolated from burong bangus in the Philippines (17). a common ingredient in these products. also serves as a valuable source of the more readily fermented carbohydrate.9) compared with the white product (pH 4.2. For example. being similar (18.FISH AND SEAFOOD 6.3. the rice may be added in a partially saccharified form such as kao-mark (Thailand). a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. Although the ability to ferment starch is not widespread in the LAB. Garlic. of course.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. 19). This results in a more active lactic fermentation and a lower pH. The effect of this will be to decrease the product’s final pH and increase the amount of total acidity. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3.2 Inclusion of more carbohydrate gives a faster fermentation and a stronger acid taste Increasing the proportion of carbohydrate in the formulation decreases the product’s buffering capacity and provides the LAB with more substrate on which to act. TABLE 6. In several products.1 . also lower the unit cost of the product and this is probably a significant factor in many cases.
6.17. the pH drops from 6. Traditional packing techniques used with lactic fermented fish products. Clearly. reporting the product’s pH to range from 4. There is little published information on the factors that limit the shelf life of the low-salt products. Results of a product survey were consistent with this.5. The mixture is allowed to ferment for 2 . This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). In a successful fermentation. 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.5 in the first 2 . is something of an exception since millet is used along with garlic as the carbohydrate source. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. but they are likely to behave similarly to lactic fermented products produced from other commodities.3 days.9%. thereafter declining to about 4.5:9:3. The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. markedly extending the shelf life (25).2 at the end of fermentation. Generally. The growth of yeasts and moulds could.3 weeks at around 20 °C. A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak.8 . and there is a marked regional variation in the preferred type. Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. allowing other spoilage organisms to grow. which ranged from 7. It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24). red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. 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). the Korean fermented fish product. Excessive growth and acid production by the LAB could render products too sour for acceptability. presumably correlating with the salt content. 135 .FERMENTED FISH Sikhae. in turn. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage.7 to below 4.7 up to 6.4 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). Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage.2. lead to an increase in pH. the level of salt employed is on the high side for fish/salt/carbohydrate products. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. such as wrapping in banana leaves. will not reliably exclude air. around 12%.
and consumed without cooking. fish products are the most common source of botulism. There is little in the way of documented information associating fermented fish products with foodborne illness. 6.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. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. In the fish sauces and pastes. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. but we are unaware of any reports of this happening. such as soaking the fish at temperatures below 5 °C before fermentation and the use of starter cultures to ensure a rapid and reliable decrease in pH. viable numbers will probably decline quite rapidly. are Vibrio parahaemolyticus and Clostridium botulinum. outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. V. During fermentation. 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. Several measures have been suggested for reducing the risk of botulism. Because of the severity of the illness it causes.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. As in Norway. however. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety. 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 . outbreaks of 136 . Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process.87. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. The bacterial pathogens most commonly associated with the living environment of fish. under conditions inimical to growth and high ambient storage temperatures. although these limits apply when all other factors are optimal (26). In Japan. parahaemolyticus has been reported growing down to a pH of 4. typically 30 °C. After capture.8 and at salt levels as high 10%.
. Novus Press. Greenland and Alaska. walrus nose. 1966. Dollar A. Yhoung-Aree J. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. Liston J.O. One study with the trematode Haplorchis taichui. Foodborne trematode infections are emerging as a major public health problem. Microbiology and chemistry of fermented fish. Saisithi P. Raw materials subject to this process include seal flipper. affecting an estimated 40 million people. botulinum and the production of toxin.6 1. Dhanamitta S. 6. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. 137 . Rakefisk and Herring in the Norwegian Tradition.. 1989. which will persist into the product. Mahidol University. whale meat. 2. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy.. though 27% survival was still recorded (33). A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31).M. probably reflecting greater awareness of the critical factors that determine the product’s safety (27). Freshwater fish are the most important intermediate reservoir hosts of foodborne trematodes. Where these products are consumed without final cooking. botulinum. and salmon heads and eggs. which is generally consumed raw (29). Kasemsarn B. Oslo. particularly in south-east Asia and the western Pacific regions (30). and these are often used in the production of fermented fish products. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. beaver tail and paw. giving an alkaline fermentation. Many of these products are described as fermented. References Riddervold A. Studies to determine the effect of traditional processing techniques on the infectivity of metacercariae was identified as a priority research area by a WHO study group (30). and warm weather can result in rapid growth of C. salt content was found to have the greatest impact on the viability of larvae. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). Salting levels are often insufficiently low to prevent growth of C.FERMENTED FISH botulism have become less frequent in recent years. although the absence of any source of fermentable carbohydrate means that the process is putrefactive. 31. Journal of Food Science. Fermented Fish Products Study (Thailand) Phase 1. 1990.. Baseline Study (1987-1988). 3. they could act as an important vehicle of transmission. Puwastien P. while increasing the acetic acid content of the formulation had little or no effect (32).. Institute of Nutrition. With this particular organism. 105-10. Thailand. Lutefisk.
Microflora of four fermented fish sauces.. Kyong-Haeng L. Reddi P. Applied Microbiology. 70. Rattagool P. Takano M.. Itoh T. 185-92.. 14.K.. 6338.H. Tsunemi M. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries.. Dymsza H.T. 80.. Biotechnology and Biochemistry. 16. 1979. 12. International Journal of Systematic and Evolutionary Microbiology..H. 1986. 29.. 57. Moonmangmee S. 25. in General and Applied Aspects of Halophilic Microorganisms. from fermented shrimp paste in Thailand. Journal of Food Science and Technology. 124-30.J. Some chemical changes in fish silage.R. Fukuda H. Lentibacillus kapialis spp. Government of Australia.S. 1995. 1957. International Journal of Systematic and Evolutionary Microbiology.A.. Plenum Press. Halophiles in Thai fish sauce (Nam pla). Rodriguez-Valera F. Tropical Science. Cooke R.. Backhoff P. Sands A. 61-73.. Salinococcus jeotgali spp. 141-2. Crisan E. 138 . Production and characterisation of NaCl-activated proteinases from Virgibacillus spp. 1991. 2007... 37. Journal of Food Technology. 1985. Dhamcharee B. Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme. 11.T. 17-30. 9. Canberra.R. Nov. Journal of Fermentation and Bioengineering. a traditional Korean fermented seafood. Catheptic activity of fish muscle. Yongsawatdigul J. Thongthai C.. Agents of proteolysis and its inhibition in patis (fish sauce).. Ahn H. Saono S. 15. Namwong S. A halophilic serine proteinase from Halobacillus spp. Thawai C. microbiological and chemical properties of salted and fermented squid. Pakdeeto A. Fermented fish products of south east Asia. 6. Lee S. Nov. New York.. Rodtong S.R. Liston J.. 2008. 1395-401. Chung Y. SK33 isolated from fish sauce fermentation.. Journal of Food Protection. Dong-Ho K.. Yook H. isolated from jeotgal. Orejana F.. 643-7. 2007. 381-8. 106-8. Journal of Scientific and Industrial Research (India).D. Effects of gamma radiation on sensory qualities. 3. Phil. Takada K. 43. Ed. 1972.M.. Oda K. 63. 13. 11. Hull R. Bioscience. Protein hydrolysate from fish. Sinsuwan S.. Tanasupawat S. SR5-3 isolated from fish sauce: purification and characterisation.. Hiraga K. 17.. Kaneko Y. 2000.V... Ono H...K. Journal of Food Science. 5.. Aslam Z. 10. 1976.. 3649. Kudo T. Process Biochemistry... Olympia M. Yasir M. Lim J.W.. 8.FISH AND SEAFOOD 4. Jae-Hun K. 934-9.. 353-63. Constantinides S. 2006. 16A.M.. Suntinanalert P. Velankar N. 1975. Adams M. Im W. Byun M. 57. 7... Tanasupawat S.
20.W. in the Philippines.H. Gram L. 38. Gram L. Valyasevi R. New York. Ed. Journal of Food Science and Technology. Journal of Agricultural Science (Tokyo). Eds. Tropical Science. Wedell-Neergaard C... a Thai low-salt fish product. ASEAN Food Journal. International Commission on Microbiological Specifications for Foods. Food Chemistry. World Health Organisation Technical Report Series.. 1983. Yeast-flora in red burong-isda a fermented fish food from the Philippines. Avurhi J. 1992. 1987.. Two outbreaks of botulism associated with fermented salmon roe – British Columbia.H.H. Erikson U. 28 (6). Paludan-Müller C. Isaac-Renton J.. 1-4. 426-35. 2002. Huss H. 21. Control of foodborne trematode infections. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation. Characterisation of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation. Benjakul S. New York. Ben Embarek P. 139 ..H.L. Kozaki M. Hazards from northern native foods. 28. The fermented fish food. International Journal of Food Microbiology. Dawar M. 29. Journal of Applied Microbiology..H.W.. in Microorganisms in Foods.. Canada Communicable Disease Report. Moody L. 1983. Kozaki M. a Thai low-salt fermented fish product. 27. 25. 69-104. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper. Paludan-Müller C. Dodds K. 1815.. 1990. Gram L. 24. Fung C.. Journal of Agricultural Science (Tokyo).. Marcel Dekker. Eds. 1998.H. Rustad T.. in Clostridium botulinum: Ecology and Control in Foods. 1999.. 219-29..W. Yamprayoon J. Dodds K.D. 28. 3. 22. 3-10.L.. Vibrio parahaemolyticus.A. 92. Owens J. Huss H. 305-22. 46.K.. Volume 5: Microbiological Specifications of Food Pathogens. burong-isda.D. 1996. 307-14. Riebroy S.FERMENTED FISH 18. Sakai H. 23.. International Commission on Microbiological Specifications for Foods. Hauschild A. Hauschild A. Marcel Dekker.B.. 2002.A. London. Fermentation and spoilage of som fak. Blackie... in Clostridium botulinum: Ecology and Control in Foods. Tanaka M. Ostergaard A. 27. 274-86..M. 26. WHO Study Group. 105-12. Caldo G. 2007.. Hauschild A. Huss H. 30. 1992. Epidemiology of human foodborne botulism. Saisithi P. 103. Caldo G. Patrick D. Traditional fermented fish products with special reference to Thai products... 104-6. Wainwright R. 138-44.B. Martin J.. Sakai H. 19. 28. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak.. Visessanguan W. 1995.
32. 6..R. Sripa B. 23. Deerasamee S. Fermented flesh foods. 110-1. Adams M.. Honjo S. Wongsawad C.J. 33.. 2005. Aspen Publishers Inc. Bloemsma B.H. Survival of Haplorchis taichui metacercariae in lab-pla. Nout M... Fermentation and Food Safety. Huss H. 117. Kikukawa H. Sriplung H.. Todoroki T. 140 . Hanai S.R... Ruamsuk A.. 1986.FISH AND SEAFOOD 31. Miwa M.. 854-60.. International Journal of Cancer.. Progress in Industrial Microbiology. 36. Chuboon S. Gaithersburg.. Uchida K. 29.R.7 Further Reading Adams M.. International Journal of Food Science & Technology. Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom. Nithikathkul C. Thai traditional food preparation.. Kittiwatanachot P.. Southeast Asian Journal of Tropical Medicine and Public Health. Srivatanakul P. Horst K. 2005. Jedpiyawongse A. Survival of Anisakis larvae in marinated herring fillets. 1995. 661-70. northeast Thailand... 2001. Roepstorff A. 15998.
Nevertheless. It remains to be seen if this approach gains momentum in the future.g.C. FISH AND SHELLFISH TOXINS Jeffrey L. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins. Wright. Since the original chapter in this series was published (6). and a second updated edition dealing with the pharmacology. Increasingly sophisticated instrumentation (e.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world.D.7. The manual on harmful marine algae and their products. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue.. While some of these toxins only occur in seafood from a restricted geographical region. Ph. C. several scientific reports have described their efficacy and potential. physiology and detection of marine and freshwater toxins has also appeared (8). the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7. produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity.3) and some practical applications for detecting shellfish toxins have been reported (4.M. Although most of these antibody-based test kits have yet to receive international regulatory approval. known and unknown. including the well known marine toxins 141 . more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins.. most notably the azaspiracids and the cyclic imines as well as some additional polyethers. 5). An emerging technology is the use of immunosensors (1 . Another book dealing with the clinical effects of a variety of poisons and toxins. Scientific and Cultural Organisation) has been updated (7). a number of new toxins and toxin families have been identified.
2. Around twenty PSP toxins are known. There is a recommendation from a joint FAO (Food and Agriculture Organisation of the United Nations)/IOC/WHO (World Health Organisation) report that marine biotoxins be classified according to their structural chemistry group rather than the symptoms they induce (e. Saxitoxin Group instead of the Paralytic Shellfish Poisons). The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13).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. 7. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11).FISH AND SEAFOOD has been produced (9). Consequently. 7. 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. all of which contain a tetrahydropurine skeleton (Figure 7. Gymnodinium catenatum. Not all the toxins found in shellfish extracts are microalgal products. 142 . neoSTX and Gonyautoxins (GTX) 1-4. Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic. Many of these species are extremely common and are found throughout the world in a variety of cold. can become contaminated with these toxins.g. In addition.1). a great variety of shellfish from all latitudes. even crustaceans such as lobsters. dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). and a tropical species Pyrodinium bahamense (16). The Ncarbamoyl sulfates are readily hydrolysed to STX. and a review on the risk assessment of marine toxins has also been produced (14). PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. some are the result of biotransformation processes by the filter-feeding host. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). and sub-tropical waters and habitats (15). All PSPs degrade rapidly in alkaline conditions. and another. temperate. with intoxications occurring in many countries even today (15).
30 minutes of consumption. and humans seem to be particularly susceptible. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. 143 . 7. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water. Neurophysiological studies have shown that PSP toxins all possess the same mode of action though some derivatives such as STX and neoSTX are considerably more potent than the C-toxins (15). PSPs bind to site 1 of sodium channels reversibly blocking sodium conductance in nerve and muscle membranes by binding to a specific receptor site located on the outside surface of the membrane close to the ion channel opening.2 Toxicology PSP toxins affect the central nervous system of mammals.2. However. this is never sufficient to render the seafood safe to consume. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis.FISH AND SHELLFISH TOXINS STX GTX2 GTX3 GTX5 (B1) C1 C2 NEO GTX1 GTX4 GTX6 (B2) C3 C4 R1 H H H H H H OH OH OH OH OH OH R2 H H OSO3 H H OSO3 H H OSO3 H H OSO3 - R3 H OSO3 H H OSO3 H H OSO3 H H OSO3 H R4 H H H SO3 SO3 SO3 H H H SO3 SO3 SO3 - Fig. and fingers within 5 . tongue. Human symptoms begin with tingling and numbness of the lips.1.
The mouse bioassay has been in place for decades and the method has been standardised. Thailand. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). While the chemical detection methods are most promising for the future. TTX most commonly occurs in many species of puffer fish. and over ten analogs are known (22. as well as other parts. China. and has also been evaluated by a number of EU laboratories.FISH AND SEAFOOD 7. After some development and various trials. other derivatives have been reported. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). specifically 144 . One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. Two approaches have been taken here.3. Most intoxication events have occurred in South East Asia. and while simple and relatively inexpensive to operate. 7. the detection limit (4 µg/g) of the method is close to the safe level and does not provide any information on the profile of PSP toxins present in the extract. and islands of the South Pacific.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0.2. 15. 7.2). Alternative HPLC methods have subsequently reduced this to a single HPLC run. a criticism has been that such methods are time consuming. Since its discovery. and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. 17). Taiwan.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). a pre-column method has received Association of Analytical Communities (AOAC) approval (18). However it remains as the monitoring method of choice in many countries.8 µg/g) of shellfish or seafood tissue. Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. 24) (Figure 7. 23. including Japan. 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.
preventing depolarisation and propagation of action potentials in nerve cells. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. 7. Chemical methods for TTX detection have also been investigated and are mainly based on its smooth conversion by heating in an alkaline solution to a detectable fluorescent (C9) base.2 mg of purified toxin can be lethal. respiratory failure and coma. The onset of symptoms usually occurs rapidly.3. where puffer fish of the genus Fugu are a delicacy. molluscs. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. 26). raising speculation that TTX is produced by a bacterial source. and even newts. and once again the mouse bioassay has been used to detect the presence of the toxin. and blocks diffusion of sodium through the sodium channel. Typically this has been accomplished using HPLC followed by post 145 . it is served in speciality restaurants with qualified chefs to properly prepare the fish. Structure of tetrodotoxin 7.2. and more recently by an actinomycete Norcardiopsis dassonvillei (26). In Japan. All of the observed toxicity is secondary to the action potential blockade.3. An oral dose of 1 . However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. nausea and vomiting. horseshoe crabs. Severe poisoning causes paralysis and respiratory failure. 23. 25. A review of TTX detection methods has been published (12. gonads and roe (24). and in mild cases results in distal muscle weakness. hypotension. 7. and ingestion of larger amounts leads to cardiovascular effects including bradycardia. the blue-ringed octopus. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera.FISH AND SHELLFISH TOXINS in the liver. 27). Fig. Since then.2 Toxicology Like the PSP toxins.
but it is known that such congeners are less toxic than DA itself (30). 7.4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA).1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28). These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. Fig. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA.3 for examples). need further development (24).4. a polar glutamate agonist. Immunoassay methods have also been investigated. However this method has been criticised for being complex.3. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. Currently 12 isomers of DA have been reported (see Figure 7. and more rapid LC-MS methods [(M+H)+ m/z 320] have been reported for the detection and identification of TTX and its various derivatives (27). and while promising. but all of them are considerably less toxic than DA itself (29). 7. Structure of domoic acid and three naturally occurring photoisomers 7. but 146 .FISH AND SEAFOOD column oxidation to the fluorescent derivatives.
and review of the toxicologic pathology of DA has been published (36). 147 . and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). has been reported as a medium-level producer of DA (33). southern and western portions of Ireland. 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. 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.. the Philippines. and in locations around New Zealand (32. disorientation and coma. which has been well documented in studies along the west coast of the US.39). Penetration of the blood brain barrier is poor in healthy subjects. and sea animals such as sea lions and even whales (35). 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.4. seizures. To a great extent. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. Recently. multiseries. Another factor is the movement of the toxin through the food chain. scallops. found in the Pacific around Vietnam. and is mainly distributed in the blood. Victims display a variety of symptoms including headache. For example. Pseudonitzschia australis. Okinawa and more northerly parts of Japan.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. as does impaired renal function which increases serum concentration and residence time of the toxin. including persistent short term memory deficits (36 . the Mediterranean. Levels of DA in the other strains are generally <1 pg/cell. but any impairment of the blood brain barrier results in additional risk. 7. and this has led to ASP toxicity in birds. 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. Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits.10% of the administered dose. cormorants and pelicans (34). 33). and clams. Indeed.2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. The oral absorption of DA is around 5 . herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. nausea. another diatom species Nitzschia navisvaringica.
4).4.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. and consequently were often overlooked as the real source of the toxins. and dinophysistoxin-2 (DTX-2) (Figure 7. To further complicate matters. dinophysistoxin-1 (DTX-1). Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. the heterotrophic dinoflagellates Dinophysis spp. 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. DTX-2 was discovered in contaminated Irish shellfish (46). In the context of this chapter. Since then. 7. 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.. 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. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue).3 Detection methods DA is water soluble and can be extracted from shellfish tissue by the acidic PSP extraction method although this may result in some decomposition. The benthic Prorocentrum spp. 48. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present.FISH AND SEAFOOD 7. 41). However. are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. and this is typically the method of choice. produce a variety of other 148 . An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. as can LC/MS. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). 49). Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). California. Alternatively. Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). though this method is most commonly used to analyse plankton and seawater (42). and an immunosensor method has also been reported (4).. in Monterey Bay. although in the latter study the probes for Pseudonitzschia pungens. and these methods have been reviewed (40. Dinophysis spp. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47. australis. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp.
Lately these compounds.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay.4. While these esters are not toxic in vitro. which include the pectenotoxins and the yessotoxins. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. Dinophysis acuta.. In comparison. DTX-5. The parent acids are the only DSP congeners to be found in shellfish tissue.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. DSP toxins produced by benthic Prorocentrum spp. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). Prorocentrum maculosum. Today over ten Dinophysis spp. are recognised as DSP toxin producers. 7. 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. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. and include Dinophysis acuminata. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. although occasionally fatty acyl esters linked through the 7-OH group (referred to as DTX-3 compounds) are also found and are likely a result of bioconversion in shellfish tissue.5. are also recognised as a threat to human health. In this case Dinophysis was suspected as the culprit organism. 149 . such as Prorocentrum lima. This was probably not the first DSP episode. and Prorocentrum belizeanum. Dinophysis fortii. 49). Structure of okadaic acid and the two related DSP toxins 7. and Dinophysis norvegica among others (47. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. Fig. Dinophysis caudata.
This regulatory level includes the possibility of LSPs in the DSP fraction being tested. and DTX-2. various European countries. Since the late 1970s. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 .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. it does not provide information on the DSP toxin profile. even with the minimal of clean up steps (48).5. 7.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. Human symptoms include diarrhoea. DSP-Check is a commercially 150 . DSP episodes have been reported from Japan. nausea. New Zealand and Chile. which can lead to an underestimation of the total DSP content in a sample. Due to the lipophilic nature of the toxins. and a detection limit of 100 µg/kg has been reported (12). 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. DSP episodes have been reported worldwide and are included in many monitoring programs (47). and while this provides critical seafood safety information. vomiting. and abdominal pain.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. most progress in DSP toxin detection has been using new or improved LC/MS methods and a detection limit of 10 µg/kg has been reported. Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. The 7-O acyl derivatives (DTX-3) of OA that can form in shellfish also show poor cross reactivity and this further exacerbates the problem. or indeed on the presence of other associated LSTs that can occur with DSP toxins and which result in a positive mouse bioassay result (vide infra). 7. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). In recent years. A regulatory level of 80 . Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996. 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. 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. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method.3 days.
they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness.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).FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. 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. and another rapid antibodybased test kit for DSP toxins has also been reported (53). Within this group. fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays.5 and 7. Some representative examples of both groups of toxins are shown in Figures 7. for many years these compounds were included in the DSP toxin complex.6. 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. with a claimed detection limit of 20 µg/kg. 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. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. In yet another approach. 58). Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . 55). 7. However. Indeed. and the latter group are usually found to be responsible for any reported shellfish toxicity (59). with good agreement with the mouse bioassay and LC–based methods (54. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). Fig.
YTX and homo-YTX 7. 7.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59). 58).FISH AND SEAFOOD Fig.6. As is typical for many algal species. eastern Canada. 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. the toxin content and profile may vary among different strains. Structure of the two parent yessotoxins. While 152 . PTX-1 and PTX-2 are frequently the most common derivatives to occur and PTX-2 is the most potent following intraperitoneal (IP) injection (230 µg/kg body weight).6. New Zealand and South America (49.6. 7.1 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). 57. The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. which may co-occur in blooms with other organisms. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. Japan. many parts of Europe and the Mediterranean.
a regulatory level of 1 mg/kg shellfish tissue has been set. though its cell effects may be linked to modulation of calcium channels (62). and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. chemical analysis is best carried out by LC/MS and several approaches have been described (57.3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue. 58). There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7.7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. The regulatory level of 160 µg/kg shellfish tissue has been established and the mouse bioassay is often the primary detection method although LC/MS methods are preferred and this requires some simple clean-up of the crude extract prior to chemical analysis. The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. but various congeners are acutely toxic in the mouse bioassay following IP injection. The structures and chemical data including the molecular weight of each member have been collated (56). the PTXs are susceptible to bioconversion in shellfish tissue and consequently it is often desirable to determine the profile of toxins present. but less so following oral dosing (61). Some of the PTXs are potent cytotoxins in vitro (60).7). 7. it may be compromised by other biological materials in the matrix. The crude extract may be used directly in the mouse bioassay. While LC/UV detection is possible for PTXs. 7. Estimates of acute toxicity of the parent compound following IP injection vary considerably. which is further compounded by differences in the experimental design. The YTX group has not been reported to cause human illness. Because of the number and complexity of the YTX group.FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. There are no clear cases of human illness resulting from ingestion of PTXs (59). which aids detection by LC/UV. 64). Many YTX toxins contain a conjugated diene system.6. some of which are found as glycosides and sulfates. The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds. However. the PTXs show considerably less toxicity when administered orally. Their toxicology and mechanism of action remain uncertain. For many years it was believed that 153 . Although there is no evidence that yessotoxins are responsible for human illness. but a fluorescent derivative has been prepared for others.
They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. PbTx-1 and PbTx-2. 66).FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida. 7. 66). and Heretosigma akashiwo. the same toxins have been reported to be produced by three genera of Raphidophytes.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. Structure of the two parent brevetoxins. Fig. These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). 154 . In the Gulf of Mexico. namely Chatonella marina. and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf.S.7. A complicating factor both in terms of establishing the profile and toxicity of brevetoxin-contaminated shellfish has been the discovery of the so-called “brevetoxin metabolites”. and cockles (65. and one of the brevetoxin metabolites BTX-B2 7. Interestingly. oysters. and off the coast of New Zealand (65). Fibrocapsa japonica. Many of these species are associated with massive fish kills. whelks.7. brevetoxins are found in mussels. was chemically characterised as PbTx-2 (67). which are found in Japanese waters (66). but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65. verruculosa off the mid Atlantic coast of the U.
The mouse bioassay is used as the primary detection method.7. this family of toxins has proven to be a challenge both to identify and monitor. 66. with hot spots in the coral reef areas of the Caribbean and French Polynesia.FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68). A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. with a mean duration of symptoms of around 1 day. In this application. 7. 69). the competitive ELISA method correlated most closely with the mouse bioassay results. and mammalian body fluids without pretreatment or dilution (72). 7. shellfish homogenates. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 .3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. and an action level has been set at 800 µg/kg (or 80 µg PbTx-2 equivalents/100 g or 20 MU/100 g or 4 µg/mouse) of shellfish tissue to trigger regulatory action. and in severe cases seizure.7. vertigo. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. which can result in a cascade of associated events in a cell (66). nausea.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels. malaise. While the mouse bioassay continues to be used. Ingestion of brevetoxins results in a variety of symptoms including parathesia.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). The brevetoxin metabolites are reported to be less toxic than the parent compound (70). and diarrhoea. The human health implications following exposure to NSPs have been reviewed (69). 7. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. The mean time to onset of symptoms is around 3 hours.
Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin. and only a few structures have been identified. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish. green and brown seaweeds and also occur in sediments and coral debris. lipid soluble polyethers.8). These include species belonging to the genera Ostreopsis and Coolia.1 Origins and distribution It is recognised that several species of benthic dinoflagellates can produce CTXs and the gambiertoxins.FISH AND SEAFOOD transferred through the food chain to the larger carnivores. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins. complex. A detailed comprehensive review of ciguatoxins has recently been published (74). most likely a consequence of biotransformation of the parent toxins as they pass up the food chain. The diversity of benthic phytoplankton that exist on reefs further complicates the picture. but more are suspected to exist. Fig. 7. a group of closely related polyethers that are considered the source of ciguatoxins. CTXs are large.8. Distribution through the food chain begins with 156 .8.
although not all CTXs are equipotent. the neurological symptoms can persist for days.1 µg of ciguatoxin can result in human illness (74.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. Like the brevetoxins. Other symptoms include a metallic taste. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. and there are suggestions that such incidents are increasing and spreading worldwide (74).8. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. Globally. Even when suspect samples are available.76). where cold objects feel hot and vice versa. all of which can be linked to the effect of CTX on excitable membranes. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. Patients have reported symptoms lasting weeks and even months. Local island populations attempt to avoid poisoning by catching only smaller fish. Another symptom that can develop over time is extreme fatigue. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . myalgia. 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. These include paresthesias (numbness and tingling of the extremities). weeks or even months (74 . 7. which in turn are consumed by carnivores such as snappers. pruritus. which can be confused with chronic fatigue syndrome. 78). but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. only trace amounts of CTXs can cause illness following consumption of contaminated fish. arthralgia.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. groupers. and temperature sensation reversal. Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. sharks. barracudas. 75. and the sensation of loose teeth (77). and it is estimated that ingestion of as little as 0. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. 7.3 Detection methods As a consequence of their potency. 75). and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. and in fact CTX poisoning is often only identified after consumption of contaminated fish. there are neither standards nor an official testing program. The onset of symptoms can occur rapidly within 24 hours.8. and although the gastronomical problems may dissipate within a day or so. which can return following the consumption of alcohol and certain foods such as oily fish (74. and moray eels.
has been developed. The original structure proposed revealed an entirely new class of shellfish toxin. In this way Caribbean and Pacific CTX-1 compounds can be distinguished (74). and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. followed by clean-up on silica and aminopropyl cartridges. A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine. 158 . at C-3 and C-23).5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. almost 20 additional derivatives have been reported (83 . although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). 84). Immunoassays may be an alternative detection method (79). Acetone extracts of fish tissue are subjected to solvent partitioning. high-throughput screen (74).80%).9). minor revisions of a few stereocenters were made. Chemical detection methods are severely compromised by the lack of standards. and another dealing with the chemistry and detection methods has been published (85).g. and following synthesis of the molecule some time later. Recent reviews of this new group of toxins have appeared (83. following consumption of contaminated mussels Mytelius edulis (82). another in vitro sodium channel-based assay.30%) and AZA3 (5 . specific for CTX activity. only a few laboratories are able to undertake such analyses. Usually the major AZA derivatives found in shellfish are AZA1 (50 . The majority of these have been proposed based on LC/MS analytical data. which features characteristic fragment ions corresponding to sequential losses of three molecules of water.FISH AND SEAFOOD Lethal Dose (MLD) 0. This is then subjected to LCMS/MS analysis. which are based on an LC-MS/MS approach.25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. 7. and only a few structures (AZA 1 . Since the original discovery and identification of the toxin. but this only has application in a clinical setting (81). Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e.9). with AZA2 (10 . and the corrected structure was published in 2004 (86) (Figure 7. Consequently. Recently. and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue.85).20%) present in lesser amounts (Figure 7.9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland.
1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. diarrhoea and stomach cramps. cassipes produces AZAs in culture. In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown. Donax spp. 89). Portugal. cassipes (83).2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. Scandinavia. these molluscs were never considered to be the biogenetic source. and most recently in a crustacean (Cancer pagurus). and to date this has been the only source from which these toxins have been found at toxic levels. 7. While this result is not necessarily in debate.9.FISH AND SHELLFISH TOXINS Fig. However. Ostrea edulis). but current data indicates that unlike the DSP compounds. Structure of the three principal azaspiracids found in shellfish 7. Mytilus galloprovincialis).9. which can persist for 2 . it has been impossible to establish that P. vomiting. These symptoms are not unlike those experienced following the consumption of DSP toxins. it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). The bivalves affected include mussels (Mytilus edulis.3 days. and some time after the original episode in Ireland. oysters (Crassostrea gigas. Following the original episode. cockles (Cerastoderma edule). and to date no long-term effects have been recorded (83. scallops (Pecten maximus). Ensis siliqua. 90). 84.). and off the coast of Morocco (83. 88. AZA toxins do 159 . and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. clams (Tapes philipinarum.9. which are ingested by P. 7. as in other cases of shellfish toxins. AZA occurrence has been reported by several European countries including France. There has been considerable debate over the source of azaspiracids.
Using the same approach. the pinnatoxins. 7. though the possibility exists that they may inhibit other phosphatases. 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. and closely related ptierotoxins (Figures 7.12). 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. 92). Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. which are particularly amenable to detection by positive ion electrospray mass spectrometry. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. 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). 7. Based on IP minimum lethal dose data. it is suggested that the order of toxicity is AZA2 (110 µg/kg) >AZA3 (140 µg/kg) >AZA (150 µg/kg).9. but all contain a spirocyclic or cyclic imine function within their structure. AZAs also display cytotoxicity towards various mammalian cell lines (83. Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. However.10 – 7. Once again. abnormalities in the thymus and spleen. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 . parenchymal cell pyknosis in the pancreas. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. Importantly these pathological outcomes were reported as being different from those caused by DSP.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. Oral administration of AZAs at substantially higher doses than those used in IP injection studies did not result in any mouse mortalities after 24 hours. The mouse bioassay has been used to monitor shellfish extracts. 91. PSP. and necrosis of epithelial cells on the microvilli (91). 85).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 spirolides. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. and erosion and bleeding in the stomach.10 Cyclic imines These toxins are represented by several distinct structural types. and ASP. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic.
In only two cases (gymnodimines and spirolides) has a planktonic source been identified. selliformis (97). gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). it was proposed that a dinoflagellate identified as Gymnodinium spp. 95). Pacific oyster. they are sometimes referred to as emerging toxins of uncertain human health concern..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. 7.10). The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7.1 Origins and distribution The original observation of toxicity in oysters led to a wider survey of other shellfish species in New Zealand when it was discovered that the toxin was present in a variety of bivalve molluscs including mussels. a species closely related to Gymnodinium mikimoitoi was the planktonic source of the toxin. The gymnodimines are the smallest members of the cyclic imine group with a molecular weight range of 500 . 95).10. Some time later. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94.10. two additional congeners.1. cockles. scallops.FISH AND SHELLFISH TOXINS health. Subsequent taxonomic studies revised this again to a new dinoflagellate K.520 Da (94. At the time of the initial observation of gymnodimine. 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. surf clam and New Zealand abalone. 7. 161 . Fig.10. Structure of the cyclic imine gymnodimine 7. but they are included here for completeness.
Reduction of the imine group eradicated the toxicity. 50%) of 96 µg/kg. paralysis of the hind quarters and eventually complete immobility. together with a lack of evidence of human intoxication.10. and so spectroscopic detection by UV methods is not favoured. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction. and this is the detection method of choice. finishing with 100% methanol. There is a rapid onset of symptoms including respiratory distress. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds.1. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine. the first step in the process is typically an initial clean-up step using a short C18 cartridge and elution with water containing increasing amounts of methanol. 7. A later study using purified toxin established an LD50 dose (Lethal Dose. 95). Based on this diminished oral toxicity. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay.3 Detection methods The characteristic “fast-acting.10.11). However. Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. it has been stated that regulation against gymnodimine is not required (98). Most cyclic imines lack an extended chromophore. 95) (Figure 7. Since their original discovery which resulted in the characterisation of spirolides A-F (99.FISH AND SEAFOOD 7.10. abdominal breathing. They were originally discovered in extracts of mussels (M. The time lapse between ingestion and death is between 5 and 15 minutes.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). Oral administration of the toxin to mice by gavage resulted in about an 8. In particular. almost a dozen derivatives in total have been identified to date (94. extracts exhibited the fast acting symptoms that became associated with cyclic imines following IP injection. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay. Whether isolated from dinoflagellate cells directly or from shellfish tissue.1. and lack of responsiveness to stimuli.900 µg/kg).100).to10-fold reduction of toxicity (LD50 600 . all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach. 7.
707 µg/kg. Interestingly. the western coast of Scotland. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). the gulf of Maine.10. by gavage (87 .1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. While oral toxicity is considerably less than by IP injection.11.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra). their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. Fig.11. fasted mice). the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102). southern Ireland. 7. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98).2. All data reported here are for desmethyl spirolide C. in fjords in Denmark and Norway. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94.2 Toxicology The spirolides are the most studied group of the cyclic imines. or by oral ingestion (381 . 7.10. 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.2.166 µg/kg. 95). Structure of four spirolide toxins A-D 7. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . fasted mice).8 µg/kg). those spirolides possessing the vicinal dimethyl grouping (see Figure 7. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 .
However. The compounds are zwitterionic and therefore somewhat more polar than the spirolides. 7. 7.10.12). the mouse bioassay can be used to detect these fast acting toxins. To date. Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. and in fact these compounds share a number of other structural features. which includes two diastereomers. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7. 95.FISH AND SEAFOOD (100).12. experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C.3 Detection methods As discussed for gymnodimine.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. 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. Fig. A number of methods following this procedure have been described (94.10.2. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . 102). Three members of this group have been reported. While the mechanism of action remains to be clearly defined. 7. consistent with modulation of acetyl cholinergic receptors (103).3 Pinnatoxins and pteriatoxins Pinnatoxins A .
There is strong circumstantial evidence to suspect that these toxins. The LD99 dose for the pteriatoxins is reported to be in the range (8 .1 Origins and distribution Shellfish of the genus Pinna.3 Detection methods No formal detection methods for these compounds have been reported. 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. While this is recognised as a real possibility for a wide range of toxins. once again reflecting improvements in purification and spectroscopic methods permitting structural identification of new and complex structures.3.10. 99%) values for pinnatoxins A-C are in the range (22 .10. like the gymnodimines and spirolides. to date action or 165 .10. Despite this progress.180 µg/kg). Coupled with this. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). However. 7.2 Toxicology There is little information describing the toxicology of these compounds (98). This is generally a result of increased monitoring and continually improving detection technologies and methods.FISH AND SHELLFISH TOXINS 7. have long been suspected of causing human illness after ingestion.most likely a dinoflagellate. but this remains to be conclusively established (106). 7. 7. The LD99 (Lethal Dose. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other. which occupy tropical waters around Okinawa and the south east coast of China. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds. arise from an algal source . new toxin groups such as the azaspiracids and cyclic imines have also been reported. and more advanced means of detection.100 µg/kg). which in many cases are now found to be more widespread than previously believed.3. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins.11 Conclusion There has been considerable progress in the detection of known marine toxins. while pinnatoxin D was essentially inactive. Only the acute toxicity in mice following IP injection has been reported. there is no compelling evidence to suggest that they have resulted in human toxicity (98). With increasing knowledge of the chemistry of toxins.3. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104).
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Bean J. Environmental Health Perspectives.. 2002.. Botana L... Toxicon. Poli M... Baden D. Abraham W. 456-79. Butera R. Schrank K.E. Kubanek J. Freidman M.M.M. Neurologic Shellfish Poisoning (NSP). 2008. Wolny J. Kirkpatrick B. Bourdelais A... 131. 2004. Naar J.A.A. Rome.S.. Bossart G. Ed..D. Naar J. 2008.G.. 2007.D..G.. like ciguatoxins. Ciguatera fish poisoning: treatment prevention and management. 551-8.. are potent ichthyotoxic neurotoxins that accumulate in fish..M. Ed. Bourdelais A.. 171 .. Detection of brevetoxins in the twenty-first century.A. 68. Bienfang P. Weisman R. Locatelli C... Dickey R.S... 67. Marine Biotoxins.. 179-85.. Buonocore M. Backer L. Botana L. Food and Agriculture Organisation.J.P. Boca Raton.. Pierce R..M. Baden D. toxicology. Henry M. Mild ciguatera poisoning: case reports with neurophysiological evaluations. Flewelling L. Backer L. 707-23. Dechraoui M-Y. Watkins S. Zaias J. 519-50. Reich A. Pierce R.FISH AND SHELLFISH TOXINS 65. 66. Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2. 2007. in Seafood and Freshwater Toxins.. Chest.. 76.. Fleming L. New fish-killing alga in coastal Delaware produces neurotoxins. 220-6. 110... Whitney P.G.. Granholm A.S. Harmful Algae. Benson J. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. Chen A. Steidinger K. Granade R. 825-34. Baden D.. 465-70..E.C. Toxicon. Dickey R. Ramsdell J. Reich A. Gannon D. Baden D.. Abbott J.G.. Flewelling L. Kubanek J. Ayyar R. Manzo L. Food and Agriculture Organisation of the United Nations. Cheng Y. in FAO Food and Nutrition Paper 80. Pierce R.... Ramsdell J..N. 50.. Landsberg J.. 71. 2004.. Fleming L. 99-115. 50. 2004. Prockop L. Naar J. 73. 117-71. Clark R. Ed. Fernandez M.H.. Botana L. West Indian Medical Journal.. Tomas C. 479-516.G. Poon-King C..A. CRC Press. Squicciarini D. 2008. 2007.... Lenzi A.. Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater. Abraham W. 2000...C. Tomas C..J. Wang Z. Baden D. Boca Raton.. 2008. 53. Boca Raton. 110. Marine Drugs. Ed. 187-94.. Backer L.E. 74.R.E. Kirkpatrick B.. Naar J. Wanner A. 69. Bottein M-Y. 1598-603. Jacocks H. The molecular and integrative basis to brevetoxin toxicity. 77. Aerosolised red tide toxins (Brevetoxins) and asthma.. 72. 3. in Seafood and Freshwater Toxins. Poon-King T. Johnson D. in Seafood and Freshwater Toxins.. Brevetoxins.S.M. and detection.B. 75. 70. 2002..P. Food and Agriculture Organisation.P.. CRC Press.W. Gandini C.. 23.... shellfish and mammalian body fluid. CRC Press.E. Ciguatera toxins: Chemistry. 6. Zaias J. Lancaster J.. Literature review of Florida red tide: implications for human health... Environmental Health Perspectives. Muscle Nerve. Fleming L. Cheng Y..
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.
ingredient and packaging buyers. 3. 2. Figure 8. Determine the CCPs. Establish the corrective action to be taken when monitoring shows that a CCP is not under control. Establish a system to monitor control of the CCP.1 Assemble HACCP team HACCP requires management commitment of resources to the process. Each stage is discussed below in detail. Conduct a hazard analysis. 8. 7. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available.1 is a flow diagram illustrating this 12-stage logic sequence. quality assurance or control.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. hygiene managers. Verification . tests and other evaluations. 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. operation or stage in the food chain including raw materials. Establish documentation concerning all procedures and records appropriate to these principles and their application. procedure. procedures.The application of methods. Establish the critical limit(s). 6.3.A point. in addition to monitoring to determine compliance with the HACCP plan. food microbiology. 5. It is recommended by the Codex Alimentarius that the practical application of the HACCP principles be approached by breaking the seven principles down into a 12-stage logic sequence. Establish procedures for verification to confirm that the HACCP system is working effectively. The team should also include 177 . Validation . from primary production to final consumption. food chemistry or engineering.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . 4.Obtaining evidence that the elements of the HACCP plan are effective. 8.
FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations. chemical and/or physical). The scope of the plan should be determined by defining the extent of the production process to be considered and the categories of hazard to be addressed (e.g. Fig. If expert advice is not available on-site.1. 8. it may be obtained from external sources. Logic sequence for application of HACCP 178 . biological.
it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments.3. It is also important to identify the consumer target groups. shelf life. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. there may be species-specific hazards related to certain fish and shellfish. 8. is a cooking process required?). 8.3. such as enteric viruses in shellfish. contaminated fish and shellfish should not be used. freezing.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora. etc. may need to be considered specifically.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. The description should include information such as the product name. which should be described in detail. 8. In addition. pH.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries. processing conditions (e.g. composition. or in aquaculture operations.3.2 Describe the product It is important to have a complete understanding of the product. aquaculture drug residues and potentially pathogenic Vibrio spp. storage and distribution conditions and instructions for use.1.3.3. physical and chemical structure (including water activity (aw). challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. Expertise on such inherent hazards. fermentation. Vulnerable groups of the population. etc. such as children or the elderly. including practices onboard fishing vessels. packaging.g.). These factors are likely to have a significant effect on some potential hazards.). naturally present on fish from warm waters (> 15 °C). marine toxins associated with particular fish and shellfish species. 8. heat treatment.3. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . If the processing does not include a kill step. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation.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. However.
and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. For example. ingredient specifications. It is essential that the diagram is accurate.4. and therefore a detailed description of the procedure applied must be available. the qualitative and/or quantitative evaluation of the presence of hazards. 180 ● ● . If HACCP studies are applied to proposed new process lines or products. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. conduct a hazard analysis. cleaning and hygiene procedures and storage conditions. The operation should be observed at all stages.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. because the hazard analysis and decisions regarding CCPs are based on these data. and identify any measures to control identified hazards The HACCP team should list all hazards that may reasonably be expected to occur at each step in the production process. or circumstances of any reclaim or rework activity.3. Ideally. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working.FISH AND SEAFOOD product.3. However. 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. 8. 8. time/temperature data. It may include details of the factory and equipment layout. once determined. it is essential that the details of each step are fully appreciated and recorded. features of equipment design.3. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised.6 List all potential hazards associated with each step. adequate chilling of finfish immediately after capture is vital for the control of several potential hazards.5 On-site confirmation of the flow diagram The HACCP team should confirm that the flow diagram matches the process that is actually being carried out. survival or multiplication of pathogenic microorganisms. it should also include details of CCP steps. 8.
181 . For example. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual. where a hazard is repeated. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards.). since this may interfere with the analysis. One control measure may be relevant to several process steps. Salmonella spp. Hazards introduced during processing of fish products depend very much on the characteristics of the process. and there are an equally varied range of potential hazards associated with them. any food safety objectives or manufacturer’s food safety requirements. botulinum. and viruses). Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. Note: it is important at this stage that no attempt is made to identify CCPs. Vibrio spp. 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. Pathogens may be part of the resident microflora of the living animal (e.6. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage.3. The HACCP team should then determine what control measures exist that can be applied for each hazard.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products. or desirable. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin. and the potential growth of Listeria monocytogenes on smoked fish. The incidence of psychrotrophic. or may originate from polluted water or from post-capture contamination (e. For example. Therefore it is not possible.g. the hurdle effect. the number of consumers potentially exposed and their vulnerability. 8. 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.g.
If a significant hazard has been identified at a step where control is required for safety. biological or sensory parameters. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. However.FISH AND SEAFOOD 8. chemical.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. given the enormous variety of fish products and processes in use. then the process must be modified to include a control measure. 8.7.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits.3.3. application of the decision tree should be flexible. 8.1 Fish and seafood Again. and the reader is referred to the appropriate product chapter. However.3.8. More than one critical limit may be defined for a single step. For example.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. Criteria used to set critical limits must be measurable and may include physical. and its use may not always be appropriate. it is not possible to generalise on likely CCPs. 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 . it is necessary to specify both time and temperature for a thermal process.2) to provide a logical. 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. Where possible.3. but for which no control exists at that step or any other. critical limits should be specified and validated for each CCP. structured approach to decision making. it can be said that effective control measures are likely to include the following: • • • • • 8.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE
• • • 8.3.9
Water activity (aw) values for dried fish pH values in fermented fish Microbiological quality of water in shellfish production areas Establish a monitoring system for each CCP
Monitoring involves planned measurement or observation of a CCP relative to its critical limits. Monitoring procedures for target levels must be able to detect loss of control of the CCP, and should provide this information with sufficient speed to allow adjustments to be made to the control of the process before the critical limits are violated. Monitoring at critical limits should be able to rapidly detect when the critical limit has been exceeded. Monitoring should therefore either be continuous, or carried out sufficiently frequently to ensure control at the CCP. Therefore, physical and chemical on-line measurements are usually preferred to lengthy microbiological testing. However, certain rapid methods, such as ATP assay by bioluminescence, may be useful for assessment of adequate cleaning, which could be a critical limit for some CCPs, for example pre-start-up hygiene. Persons engaged in monitoring activities must have sufficient knowledge, training and authority to act effectively on the basis of the data collected. These data should also be properly recorded. 8.3.10 Establish corrective actions
For each CCP in the HACCP plan, there must be specified corrective actions to be applied if the CCP is not under control. If monitoring indicates a deviation from the critical limit for a CCP, action must be taken that will bring it back under control. Actions taken should also include proper isolation of the affected product, repair of defective equipment, and an investigation into why the deviation occurred. A further set of corrective actions should relate to the target levels, if process drift is occurring. In this case, only repair of the process defect and investigation of the fault are required. All corrective actions should be properly recorded. 8.3.11 Establish verification procedures
Verification usually involves auditing and testing procedures. Auditing methods, procedures and tests should be used frequently enough to determine whether the HACCP system is being followed, and is effective at controlling the hazards. These may include random sampling and analysis, including microbiological testing. Although microbiological analysis is generally too slow for monitoring purposes, it can be of great value in verification, since many of the identified hazards are likely to be microbiological. In addition, reviews of HACCP records are important for verification purposes. These should confirm that CCPs are under control and should indicate the nature
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of any deviations and the actions that were taken in each case. It is also useful to review customer returns and complaints regularly. 8.3.12 Establish documentation and record keeping
Efficient and accurate record keeping is an essential element of a HACCP system. The procedures in the HACCP system should be documented. ● ● Determination of critical limits The completed HACCP plan
Examples of documented procedures include: ● ● The hazard analysis Determination of CCPs
Examples of recorded data include: ● ● ● Results of monitoring procedures Deviations from critical limits and corrective actions Records of certain verification activities, e.g. observations of monitoring activities, and calibration of equipment.
The degree of documentation required will depend partly on the size and complexity of the operation, but it is unlikely to be possible to demonstrate that an effective HACCP system is present without adequate documentation and records. The length of time that records are kept will be as per company policy, but should not be less than one year beyond the shelf life of the product. Three to five years is typical for many food companies. 8.4 Implementation and review of the HACCP plan
The completed plan can only be implemented successfully with the full support and co-operation of management and the workforce. Adequate training is essential and the responsibilities and tasks of the operating personnel at each CCP must be clearly defined.
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Fig. 8.2. CCP Decision Tree (Adapted from Codex Alimentarius Commission, 2003) Finally, it is essential that the HACCP plan be reviewed following any changes to the process, including changes to raw materials, processing conditions or equipment, packaging, cleaning procedures and any other factor that may have an effect on product safety. Even a small alteration to the product or process may invalidate the HACCP plan and introduce potential hazards. Therefore, the implications of any changes to the overall HACCP system must be fully
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considered and documented, and adjustments made to the procedures as necessary. Triggered reviews/audits should occur as a result of changes, whereas a scheduled review/audit should be annual, as a minimum. 8.5 References
Wareing P.W., Carnell A.C. HACCP – A Toolkit for Implementation. Leatherhead, Leatherhead Food International. 2007. Bernard D., Scott V. Hazard Analysis and Critical Control Point System: use in controlling microbiological hazards, in Food Microbiology: Fundamentals and Frontiers. Eds. Doyle M.P., Beuchat L.R. Washington D.C., ASM Press. 2007, 971-86. Su Y-C., Daeschel M.A. Sanitation and HACCP, in Surimi and Surimi Seafood. Ed. Park J.W. Boca Raton, CRC Press. 2005, 141-62. Huss H.H. Considerations in the application of the HACCP principles to seafood production, in Assessment and Management of Seafood Safety and Quality. Eds. Food and Agricultural Organisation, Huss H.H., Ababouch L., Gram L. Rome, FAO. 2004, 153-77. Chen H-C., Chang P.C-M. Frozen seafood safety and HACCP, in Handbook of Frozen Foods. Ed. Hui Y.H., Cornillon P., Legaretta I.G., Lim M.H., Murrell K.D., Nip WK. New York, Marcel Dekker. 2004, 339-63. Mortimore S., Mayes T. The effective implementation of HACCP systems in food processing, in Foodborne Pathogens: Hazards, Risk Analysis and Control. Eds. Blackburn C.deW., McClure P.J. Cambridge, Woodhead Publishing Ltd. 2002, 229-56. Mayes T., Mortimore C.A. Making the Most of HACCP: Learning from Other’s Experience. Cambridge, Woodhead Publishing. 2001. Mortimore S.E., Wallace C., Cassianos C. HACCP (Executive Briefing). London, Blackwell Science Ltd. 2001. Chartered Institute of Environmental Health. HACCP in Practice. London, Chadwick House Group Ltd. 2000. Jouve J.L. Good manufacturing practice, HACCP, and quality systems, in The Microbiological Safety and Quality of Food, Volume 2. Eds. Lund B.M., BairdParker T.C., Gould G.W. Gaithersburg, Aspen Publishers. 2000, 1627-55. Stevenson K.E., Bernard D.T. HACCP: A Systematic Approach to Food Safety. Washington D.C., Food Processors Institute. 1999. National Seafood HACCP Alliance. IQF cooked shrimp processing HACCP model, in HACCP User's Manual. Ed. Corlett D.A. Gaithersburg, Aspen Publishers. 1998, 454-73. Mortimore S., Wallace C. HACCP: A Practical Approach. Gaithersburg, Aspen Publishers. 1998.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Khandke S.S., Mayes T. HACCP implementation: a practical guide to the implementation of the HACCP plan. Food Control, 1998, 9 (2-3), 103-9. Forsythe S.J., Hayes P.R. Food Hygiene, Microbiology and HACCP. Gaithersburg. Aspen Publishers. 1998. Food and Agriculture Organisation. Food Quality and Safety Systems: A Training Manual on Food Hygiene and the Hazard Analysis and Critical Control Point (HACCP) System. Rome, FAO. 1998. US Food and Drug Administration. Fish and Fishery Products Hazards and Controls Guide. Rockville, Food Drug Administration. 1998. National Advisory Committee on Microbiological Criteria for Foods. Hazard Analysis and Critical Control Point Principles and Application Guidelines. 1997. Gardner L.A. Testing to fulfil HACCP (Hazard Analysis Critical Control Points) requirements: principles and examples. Journal of Dairy Science, 1997, 80 (12), 3453-7. Martin R.E., Collette R.L, Slavin J.W. Fish Inspection, Quality Control and HACCP: A Global Focus. Proceedings of a Conference, Arlington, May 1996. Lancaster P.A., Technomic Publishers. 1997. Codex Alimentarius Commission. Hazard Analysis Critical Control Point (HACCP) System and guidelines for its application, in Food Hygiene: Basic Texts. Ed. Codex Alimentarius Commission. Rome, FAO. 2003, 21-31. Garrett E.S., Jahncke M.l., Tennyson J.M. Microbiological hazards and emerging food safety issues associated with seafoods. Journal of Food Protection, 1997, 60 (11), 1409-15. National Seafood HACCP Alliance. HACCP: Hazard Analysis and Critical Control Point Training Curriculum. Raleigh, North Carolina State University. 1997. Dillon M., McEachern V. HACCP and quality assurance of seafood, in Fish Processing Technology. Ed. Hall G.M. London, Blackie. 1997, 224-48. Savage R.A. Hazard Analysis Critical Control Point: a review. Food Reviews International, 1995, 11 (4), 575-95. Pearson A.M., Dutson T.R. HACCP in Meat, Poultry and Fish Processing. Glasgow, Blackie. 1995. Huss H.H., Food and Agriculture Organisation. Assurance of Seafood Quality. FAO Fisheries Technical Paper No. 334. Rome, FAO. 1994. Bonnell A.D. Process control, in Quality Assurance in Seafood Processing: A Practical Guide. Ed. Bonnell A.D. London, Chapman & Hall. 1994, 39-53. Pierson M.D., Corlett D.A., Institute of Food Technologists. HACCP: Principles and Applications. New York, Van Nostrand Reinhold. 1992. Hudak-Roos M., Garrett E.S. Monitoring, corrective actions and record keeping in HACCP, in Quality Assurance in the Fish Industry: Proceedings of an International Conference, Copenhagen, August 1991. Eds. Huss H.H., Jakobsen M., Liston J. Amsterdam, Elsevier. 1992, 521-31.
Food Control. 33-44. 1992. International Journal of Food Microbiology. 15 (1/2). 188 . International Commission on Microbiological Specifications for Foods. Microorganisms in Foods. Mayes T. WHO. Blackwell Scientific Publications. Bryan F. Geneva.L.. Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality.H. 1992. Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety. Oxford. Development and use of the HACCP concept in fish processing. 1988. World Health Organisation.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H. Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. 3 (1). 1992. 14-9.
Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials. final product specifications. however. A number of standards are provided in food legislation. to personnel requirements. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. from structural conditions in the factory or process facility. have been replaced by several linked measures on food safety rules and associated animal health controls. 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. and conditions of raw materials. transport and delivery vehicles requirements. under which food is produced to optimise public health and to prevent. EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr.1 Introduction Hygiene is an important aspect of ensuring food safety. eliminate or acceptably control pathogen contamination of food. the existence of microbiological standards alone cannot protect consumer health. and one that plays an important role in most countries’ food legislation.9. with the intention of introducing consistency and clarity throughout the food production chain from primary production. including microbiological criteria. Hygiene is a general concept that covers a wide subject area. to sale or supply to the final consumer. 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. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. The new legislation was designed to establish conditions. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 .
handling and offering for sale or supply to the consumer. and sets out what those enforcing the provisions have to do. packaging.15). Regulation 854/2004 relates to the organisation of official controls on products of animal origin.1991. p. Council Directive 92/48/EEC of 16 June 1992 laying down the minimum hygiene rules applicable to fishery products caught on board certain vessels in accordance with Article 3(1)(a)(i) of Directive 91/493/EEC (OJ L 187. 41). European Communities L 268. 9. distribution. N. transportation. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No.9. p. processing. 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. manufacturing.1992. Subsequently. storing.9.B. 190 • • . A number of more detailed implementing and transitional measures have been adopted at EC level.1).FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No.7. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin.1991. as last amended by Regulation (EC) No 806/2003. 24. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. Council Directive 91/493/EEC of 22 July 1991 laying down the health conditions for the production and the placing on the market of fishery products (OJ L 268. p. 7.2 Legislative Structure From 1 January 2006. as last amended by Regulation (EC) No 806/2003. 24. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation. 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. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No. J.
national legislation in the form of a Statutory Instrument (S. domestic preparation or handling. p. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. for example.1993. Wales and Northern Ireland).11) The EU hygiene regulations apply to all stages of food production including primary production. The Food Hygiene (England) Regulations 2006 (S.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175.I. right through to the implementation of procedures based on HACCP principles. Wales and Northern Ireland. etc. with primary production. and equivalent legislation in Scotland. 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. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. 191 . Additionally the requirements of EC Regulation 853/2004 must be complied with. food storage that is for private or domestic consumption. 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. and binding in its entirety on all member states from the date of entry into force. penalties and powers of entry. There are some exemptions. of stock farming. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. setting offences. and of hunting and fishing. 852/2004. the legislation is directly applicable law. The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No.I. e. Although the regulations have the force of law. As regulations. 2073/2005.g. which will be covered later in this chapter. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. 1 . 9. 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.3 Regulation (EC) No. 2006 No. is required to give effect to the EU regulations. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs.) in England. revocation of existing implementing legislation. 19.7.14. Primary products are defined as products of primary production including products of the soil.
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Annex I – Primary production
Annex I (Part A) relates to general hygiene provisions for primary production and associated operations covering: (a) the transport, storage and handling of primary products at the place of production, (b) the transport of live animals, (c) for fishery products - transport operations to deliver primary products (which have not been substantially altered) from the place of production to an establishment Food business operators have the responsibility to ensure primary products are protected against contamination. Any community and national legislation relating to the control of hazards in primary production such as measures to control contamination resulting from surroundings e.g. air, soil, water etc., and measures relating to animal health and welfare, and plant health that may impact on human health should be complied with. Requirements for record keeping are also laid down. This relates to animal feed (nature and origin), veterinary medicines administered to animals (date given and withdrawal periods), any diseases, analysis of samples from other animals that might impact on human health as well as reports on animal checks performed. Part B of Annex I contains recommendations for guides to good hygiene practice. 9.3.2 Annex II – Stages other than primary production
Annex II of the regulation lays down additional general hygiene requirements that must be met by food business operators carrying out production, processing and distribution of food at all stages following primary production. A summary of Chapters I to IV of Annex II is provided as follows: 184.108.40.206 Chapter I Chapter I applies to all food premises, except premises to which Chapter III applies. Food premises must be kept clean and maintained in good repair and condition. Their layout should allow for this. The environment should allow good hygiene practices and provide temperature-controlled handling and storage conditions where necessary ,and enable foods to be kept at correct temperatures and be monitored.
Additionally, there are requirements for provision of adequate lavatories, basins, ventilation, lighting and drainage.
220.127.116.11 Chapter II Chapter II applies to all rooms where food is prepared, treated or processed, except dining areas and premises to which Chapter III applies. The design and layout of rooms should allow for good hygiene practices between and during operations. Therefore floor and wall surfaces, ceilings and windows should be constructed to prevent dirt accumulating. Surfaces where food is handled must be well-maintained and allow easy cleaning and disinfection, preferably using smooth, washable corrosionresistant and non-toxic materials. There should be facilities for cleaning or disinfecting, and for storing working utensils or equipment. Clean potable water and adequate provision for washing food is needed.
18.104.22.168 Chapter III Chapter III applies to temporary premises (e.g. marquees, market stalls, and mobile sales vehicles), premises used primarily as a private dwelling-house but where foods are regularly prepared for placing on the market, and vending machines. Premises and vending machines should be practically sited, designed, constructed and kept clean and maintained in good repair and condition so as to avoid the risk of contamination, in particular by animals and pests. Facilities should allow adequate personal hygiene, and surfaces in contact with food should be easy to clean. Enough potable water and storage arrangements for hazardous or inedible substances are required as well as adherence to food safety requirements.
22.214.171.124 Chapter IV Chapter IV applies to all transportation. Conveyances and/or containers used for transporting foodstuffs are to be kept clean and maintained in good repair and condition to protect foodstuffs from contamination and are, where necessary, to be designed and constructed to permit adequate cleaning and/or disinfection. Food should be maintained at appropriate temperatures.
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126.96.36.199 Chapter V Chapter V refers to equipment requirements. Adequate cleaning and disinfection is to be carried out frequently for articles, fittings and equipment in contact with food where contamination needs to be avoided. Equipment should be installed to allow adequate cleaning, and be fitted with the required control device.
188.8.131.52 Chapter VI Chapter VI refers to food waste. Food waste, non-edible by-products and other refuse is to be removed from rooms where food is present as quickly as possible to avoid accumulation. Such waste is to be deposited in closable containers, to allow easy cleaning. Refuse stores should allow easy cleaning and be free of pests. Waste must be eliminated hygienically in accordance with European Community legislation.
184.108.40.206 Chapter VII Chapter VII refers to the water supply. There must be an adequate supply of potable water, which should be used whenever necessary to ensure foodstuffs are not contaminated. There are also requirements for recycled water, ice in contact with food, use of steam and for the water used in the cooling process for heat-treated foods in hermetically sealed containers. Specific requirements are that clean water may be used with whole fishery products. Clean seawater, may be used for live bivalve molluscs, echinoderms, tunicates and marine gastropods; and for external washing. If it is used, there must be adequate facilities to ensure the supply does not create a source of contamination for the foodstuff. Recycled water used in processing or as an ingredient must not present a contamination risk. It must be of the same standard as potable water, unless the competent authority is satisfied that the quality of water cannot affect the wholesomeness of food in the finished form.
Additionally, ice that comes into contact with food or which could possibly contaminate it needs to be made from potable water or, in the case of chilling whole fishery products using clean water. Ice must be made, handled and stored under conditions that protect it from contamination.
220.127.116.11 Chapter VIII Chapter VIII sets out personal hygiene requirements for those working in a food handling area including clean protective clothing and the requirement that those carrying or suffering from a disease are not permitted to handle food. 18.104.22.168 Chapter IX Chapter IX covers provisions applicable to foodstuffs. A food business operator should not accept raw materials or ingredients, other than live animals, or any other material used in processing products, if they are known to be contaminated with parasites, pathogenic microorganisms or foreign substances, to be toxic or decomposed to such an extent that, even after the business operator applied normal hygienic processing , the product would be inedible. Raw materials must be kept under appropriate conditions throughout production, processing and distribution. In particular, temperature control (i.e. cold chain and food thawing) requirements are laid down.
22.214.171.124 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 126.96.36.199 Chapter XI Chapter XI lays down heat treatment requirements for food that is placed on the market in hermetically sealed containers. The main relevant process parameters (particularly temperature, pressure, sealing and microbiology) must be checked by the food business operator to ensure that the required heat treatment has been achieved. Also, the process used should comply with an internationally recognised standard (e.g. pasteurisation, ultra high temperature or sterilisation). 188.8.131.52 Chapter XII Chapter XII states training requirements for food business operators to ensure that food handlers are trained in food hygiene matters and in the application of HACCP principles.
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The Regulation requires that food business operators must notify the relevant competent authority of each establishment under their control that carries out any of the stages of production, processing and distribution of food, with a view to the registration of each establishment. Food business operators must also ensure that the competent authority always has up-to-date information on establishments, including the notification of significant changes in activity, and closure of an existing establishment. Food business operators must ensure that establishments are approved by the competent authority, following at least one on-site visit, when approval is required by the national law of the Member State in which the establishment is located, or under Regulation (EC) No. 853/2004, or by a separate decision adopted. Separate rules apply for businesses producing products of animal origin. 9.3.4 HACCP
Food business operators, other than at the level of primary production, and associated operations must put in place, implement and maintain a permanent procedure or procedures based on principles of the system of hazard analysis and critical control points (HACCP). Emphasis is placed on risk-related control, with responsibility placed on the proprietor of the food business to ensure that potential hazards are identified and systems are developed to control them. Under HACCP, food business operators must amongst other considerations, identify hazards to be prevented or eliminated or reduced to acceptable levels, identify critical control points (CCP) at which control is essential to prevent, eliminate or reduce hazards and establish critical limits at these points, implement effective monitoring procedures, and establish corrective actions in the case where a CCP is out of control. Procedures must be followed to confirm the above is in place and up-todate, as well as providing the competent authority with documents and records as evidence, when required. 9.4 Regulation (EC) No. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin
Regulation (EC) No. 853/2004 lays down hygiene rules for products of animal origin that apply in addition to the general hygiene rules of Regulation (EC) No. 852/2004. 9.4.1 Definitions
The following definitions apply for fishery products and live bivalve molluscs:
and chopping. slime or sand. live tunicates and live marine gastropods. heading. slicing. where appropriate. to remove mud. shelling. reptiles and frogs) whether wild or farmed and including all edible forms. such as gutting. skinning. if necessary: chilling or freezing: filleting. filleting. slicing. or in natural sites. followed by wrapping or packaging. shucking. after preparatory work such as bleeding. live echinoderms. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. purification centres or dispatch centres in tanks or any other installation containing clean seawater. Fresh fishery products means unprocessed fishery products. Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. 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.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. in particular as a result of feeding on plankton containing toxins. and all mammals. where necessary. Conditioning means the storage of live bivalve molluscs coming from class A production areas. including products packaged under vacuum or in a modifiedatmosphere that have not undergone any treatment to ensure preservation other than chilling. 197 . mincing or processing. to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing. heading. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. gutting and removal of fins and. 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. whether whole or prepared. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness. parts and products of such animals.
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. 9. the code would be UK. please refer to Regulation (EC) 853/2004. (iii) Food business operators must have in place systems and procedures to identify food business operators from whom they have received and to whom they have delivered products of animal origin.4. 198 .1. EÜ.4. when a product's packaging and/or wrapping is removed or is further processed in another establishment.2. EK. EY.2 Form of the identification mark (i) The mark must be legible and indelible.2. Section I of Regulation (EC) 853/2004 are given as follows: 9. (iv) If the identification mark is applied in an establishment within the Community. EF. and the characters easily decipherable and be clearly displayed. (ii) However. the food business operator may apply the same identification mark to both types of food.1.4. a new mark must be applied to the product. the mark must show the approval number of the establishment.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. In this case. for other codes. the new mark must show the approval number of the establishment where this occurred. Section I of Regulation (EC) 853/2004. EO.4.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. (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. ES. the mark must be oval shaped and include the abbreviation CE.2 Fishery product requirements 9.2. Extracts of the requirements of Annex II. For the UK. EB or WE. (iii) Additionally.FISH AND SEAFOOD 9. EK. EG. EC.
and fishery products carried in bulk. C and D. as amended.2. involved in primary production and such operations accompany the needs of Annex I of Regulation (EC) 852/2004 b) for other establishments including vessels. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . a) requirements for establishments. tunicates and marine gastropods when placed on the market live. Chapter III.4. including vessels.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. specifically relating to fishery products are given below. Extracts of the requirements of Annex III. for full requirements. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. and are intended for further handling. (iii) In the case of liquid.3 Method of marking (i) The mark is to be applied directly to the product. the mark may be applied to the outer surface of the container or packaging. processing.4. Part A and Chapter V apply to retail. reference should be made to the actual Regulation. With the exception of Chapters I and II. wrapping or packaging in another establishment. the colourings used must be EC authorised colours. its wrapping or the packaging. (ii) For products of animal origin that are placed in transport containers or large packages. 9.1. (v) When the mark is applied directly to products of animal origin. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. the wrapping or the packaging. Section VIII of Regulation 853/2004. it is sufficient to apply the mark to the exterior of that package only. Therefore. The requirements of this section accompany those in Regulation (EC) 852/2004.2. This section does not apply to bivalve molluscs.LEGISLATION 9. they would need to be obtained following Section VII on live bivalve molluscs. it applies to those animals when not placed on the market live. echinoderms. or be printed on a label affixed to the product. i. Parts A. granulate and powdered products of animal origin carried in bulk.e. the approval number of the establishment and the abbreviation for the community where appropriate. Chapter IV.
There is a derogation from point a.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. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking.2. 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. if a vessel has water coming in that is used with fishery products. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products.2. fishing and collection of live fishery products with a view to their being placed on the market.1 Chapter I .e. bleeding.. Structural and equipment requirements refer to the following points 1. gutting. 2.) 9. A. removing fins.4. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. from the place of production to the first establishment of destination. Chapter VII of Regulation 852/2004. it must be placed somewhere that will not contaminate the water supply. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. heading. surfaces that contact fishery products need to be made from cleanable corrosion-resistant material. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 . refrigeration and wrapping. (a) primary production covers the farming. or to handle or process them after harvesting. The definition of fishery products covers primary production as well as its associated operations. this supplements the requirements of Annex II.FISH AND SEAFOOD c) for the water supply. clean seawater may be used for the handling and washing of fishery products. i.
(iii) In vessels equipped for chilling fishery products in cooled clean seawater.LEGISLATION (i) Such vessels need to have holds. Also. and not more than 0 °C after 16 hours. (ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. any waste processing unit needs to have a separate hold to store the waste. (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. A temperature recording device must be present and placed where the temperature in the hold is the highest. (iii) The above requirements for holds in vessels designed and equipped to preserve fresh fishery products for more than 24 hours must be met. 3. Storage areas for the finished products also need to be sufficiently large and cleanable. tanks must incorporate devices that allow a uniform temperature throughout the tanks. and allow the monitoring and. Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas. where necessary. Requirements for factory vessels a) Factory vessels (i) These must have at a receiving area reserved for taking fishery products on board so each catch can be separated (ii) There must be a hygienic system for conveying fishery products from the receiving area to the work area (iii) Large work areas are needed to allow fishery products to be prepared and processed hygienically. 201 . Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. to ensure that melt water does not stay in contact with the products. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. recording of temperatures. where necessary. Such devices must achieve a chilling rate that ensures that the mix of fish and clean seawater does not exceed 3 °C six hours after loading. 4.
b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. Handlers may use spiked instruments to move large fish or fish that might injure them. provided no damage occurs to the flesh. and products washed thoroughly. c) Fishery products. e) In cases where whole fish intended for canning is frozen in brine. must be chilled or.Requirements during and after landing 9. other than those kept alive. the product must be kept at no more than -9 °C. 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. landed as soon as possible.2. where fishery products are displayed for sale. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. 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 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. The brine must not be a source of contamination. if this is not possible.FISH AND SEAFOOD (vi) A water tank needs to be placed where there is no contamination of the water supply and hand washing taps should be designed to prevent contamination. B.4. (ii) In the case of food business operators responsible for auction and wholesale markets etc.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. b) Factory vessels on board which crustaceans and molluscs are cooked. chilled and wrapped.2. Chapter II . Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen.
They should be wrapped. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . part I.LEGISLATION consumption. points 1 and 2. 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. Chapter I.3 Chapter III . C. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels.Requirements for establishments. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. C. Fillets and slices must then be removed from work tables as soon as possible after preparation. packaged and chilled as quickly as possible after preparation.2. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. 9. Requirements for frozen products An establishment on land that freezes fishery products needs to have equipment that meets that for freezer vessels as stated in Section VIII. (ii) Heading and gutting must be carried out hygienically. (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices. Food business operators need to co-operate with the relevant competent authorities so as to allow them to carry out official controls as laid down in Regulation (EC) No. (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. dispatched. they must be stored under ice in appropriate facilities.2. including vessels. B. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed. 854/2004. and where necessary. prepared or processed immediately after arrival at an establishment on land.4. Additionally.
(iii) sprat. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. 9.2. 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. (iii) A document from the manufacturer. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . must accompany fishery products referred to above when placed on the market. (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. (iv) (wild) Atlantic and Pacific salmon.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. if the processing is insufficient to destroy nematode larvae.4. D.2. or a stabilising treatment. raw materials must have no guts. (ii) Compliance with the above is not necessary if epidemiological data indicates that the fishing grounds of origin do not present a health hazard in relation to the presence of parasites and this has been authorised by the competent authority.4 Chapter IV . stating the type of process they have undergone. Once produced. If no other method of preservation is used. (b) fishery products from the following species. they must be cooled rapidly. and (c) marinated and/or salted fishery products. (ii) mackerel. cooling must continue until a temperature 204 . and whole fish must be gutted and washed beforehand.Requirements for certain processed fishery products A. except when supplied to the final consumer.
the following conditions need to be met: A. 2074/2005). cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. 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. and the raw material is processed within 36 hours after loading.Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. However. 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. Shelling/shucking is to be done hygienically. 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. C. pressing. 205 . B. Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. 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).2. separation. raw materials need to be chilled as soon as possible and should meet the temperature requirements in Chapter VII. 2073/2005. 2074/2005.5 Chapter V .4. 2073/2005). (ii) The production process for fish oil must ensure that all raw material intended for the production of crude fish oil is subject to a treatment including (where relevant) heating.2. 9. refining and purification steps before being placed on the market for the final consumer. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No.LEGISLATION approaching that of melting ice is reached. and after this. processing. B. centrifugation.
Fishery products contaminated with parasites may not be placed on the market for human consumption.6 Chapter VI . Diodontidae and Canthigasteridae. Fresh.FISH AND SEAFOOD D. it should be kept at no more than -9 °C. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. 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. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption. and when fishery products are wrapped on board fishing vessels. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market. In the case of whole frozen fish in brine intended for the manufacture of canned food. 9.7 Chapter VII . E. thawed unprocessed fishery products.2.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. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. 206 . (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product.Storage of fishery products (i) Fresh fishery products.2. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum.2. may only be placed on the market in wrapped/packaged form and must be appropriately labeled to provide information to the consumer on preparation/cooking methods and on the risk related to the presence of substances with adverse gastrointestinal effects. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae.4. 9.4. The scientific name must accompany the common name on the label. prepared and processed fishery products belonging to the family Gempylidae. Molidae.2.
melt water must not remain in contact with the products. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. 9. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability.Transport of fishery products The above points for Chapter VII apply in relation to transport. species belonging to the Gadidae family. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar.4. Helicolenus dactylopterus.8 Chapter VIII .4. (ii) If fishery products are kept under ice. Note: (i) Frozen fishery products do not need to be kept at a temperature of -18 °C if they are being transported from a cold store to an approved establishment to be thawed on arrival for the purposes of preparation and/or processing. species belonging to the Merlucciidae family. The temperature of frozen fishery products must not fluctuate above -15°C.2. 9. 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.2. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them. provided that the journey is short and the competent authority allows this.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.). 2074/2005. (d) 60 mg of nitrogen/100 g of whole fishery products used directly for the preparation of fish oil for human consumption as referred to in the second sub-paragraph of Part B(1) of Chapter IV of Section VIII of Annex III to 207 .2. Sebastichthys capensis.
Chapters V. taken from at least three different points and mixed together by grinding. please refer directly to the regulation. Apart from the provisions on purification.4. (b) In the case of other operations. The sample must consist of about 100 g of flesh.3. 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. 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. they supplement the requirements of Annex II to that Regulation. VIII and IX. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. 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.FISH AND SEAFOOD Regulation (EC) No 853/2004. (b) and (c) of Part B(1) of that Chapter. however. as amended. and point 3 of Chapter VII applies to retail. specifically relating to bivalve molluscs are given below. Extracts of the requirements of Annex III. tunicates and marine gastropods.3 The regulation details specific hygiene requirements for bivalve molluscs. they supplement the requirements of Annex I to that Regulation. Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation. VI.4. Section VII of Regulation 853/2004. The copy of 208 . 9.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. reference should be made to the actual regulation. Chapter IX applies to pectinidae harvested outside those areas. this Section also applies to live echinoderms. The information required in the registration document is given in point 4 and includes the health status of the production area. shellfish species etc. Live bivalve mollusc requirements - 9. for full requirements.1 Chapter I . where the raw material complies with points (a).
4.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority. C.Hygiene requirements for the production and harvesting of live bivalve molluscs A.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market . processing or relaying. B. (ii) Provided the requirements of chapter V are met. poles or other fixed means are to identify the boundaries of the sites. 9. 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. 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.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. 209 . B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with. collection from: . Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. Requirements for production areas (i) The competent authority classes (classed A. they need to be sent to a processing establishment and undergo specified treatments such as sterilisation in hermetically sealed containers or specific heat treatments such as immersion in boiled water so the internal temperature of the mollusc flesh reaches at least 90 ºC and this temperature is maintained for at least 90 seconds. Transport needs to allow sufficient drainage for best survival conditions.Class A means they can be placed directly on the market .3.2 Chapter II .
shells of live bivalve molluscs are to be washed throughout with clean water and the molluscs are to come from one of the following: (i) Class A production area (ii) Relaying area (iii) Purification centre (iv) Another dispatch centre 9. the purification system to allow them to resume /maintain filter-feeding activity. 1 milligram of yessotoxin equivalent per kilogram.4.Hygiene requirements for purification and dispatch centres A. B.5 Chapter V .4. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. and there are requirements for tanks and water storage containers.Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc.4. 160 micrograms of okadaic acid equivalents per kilogram. containers and a label to identify purification.3. Before dispatch. 800 micrograms per kilogram. (ii) amnesic shellfish poison (ASP).3 Chapter III . 210 .3. (iv) yessotoxins. the levels of marine biotoxins should not exceed the following limits. Requirements for purification centres There are requirements for live bivalve molluscs to be washed. requirements if there are many batches of live bivalve molluscs. Requirements for dispatch centres Handling should not result in contamination to any degree. dinophysistoxins and pectenotoxins together.4 Chapter IV .Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. 9.3. for: (i) paralytic shellfish poison (PSP).FISH AND SEAFOOD 9.
dispatch centre or processing establishment.4.Other requirements (i) The temperature at which live bivalve molluscs are stored/ transported in should be at one that does not adversely affect food safety or their viability. must be waterproof.Specific requirements for pectinidae harvested outside classified production areas Requirements for harvesting and handling following harvesting and health standards for live bivalve molluscs must be met.4. Finally.3.4. by way of derogation from directive 2000/13/EC.1). including the identification mark. Registration document requirements also apply. Also.3. the species of bivalve mollusc and date of packaging (at least the date and month) are required.6 Chapter VI .2. 211 . (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.4.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.Identification marking and labelling In addition to general identification marking (contained in 9. 160 micrograms of azaspiracid equivalents per kilogram.LEGISLATION (v) azaspiracids. 9. the label. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’. Requirements of Chapters III and IV are to be complied with. (ii) Individual consumer-size packages of live bivalve molluscs must be closed and remain closed after leaving the dispatch centre and until presented for sale to the final consumer.4.8 Chapter VIII .3. 9.7 Chapter VII . 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.9 Chapter IX .3. (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. they cannot be re-immersed in or sprayed with water.
1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. Chapter V of Regulation 853/2004 apply. 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. chemical. they derive from: (i) poisonous fish. Section VIII. 9. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption Regulation (EC) 854/2004 gives requirements for official controls on products of animal origin and states requirements for those enforcing the provisions.5 Regulation (EC) No. or (iii) bivalve molluscs. Fishery products in particular must comply with the requirements of Annex III of the regulation. (ii) fishery products not complying with the requirement concerning biotoxins. or - - 212 . This refers to official controls or production and placing on the market of fishery products. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures.5. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. echinoderms.FISH AND SEAFOOD 9. 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. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004. Therefore the health standards for fishery products as previously stated in Annex III. In this regulation. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given.
Sampling plans and frequency are required. - - - - B. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. Live bivalve molluscs from Class B areas must not exceed.2 This states official controls for the production and placing on the market of live bivalve molluscs. Live bivalve molluscs from class A areas may be collected for direct human consumption provided health standards are met. Live bivalve molluscs from Class C areas must not exceed 46. as previously stated. chapter V is reinforced. section VII.2. Also. 213 . in 90% of the samples. live echinoderms.5. In the remaining 10 % of samples. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3.1 Chapter II . A summary is given as follows: 9. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. coli per 100 g of flesh and intravalvular liquid. 4. coli per 100 g of flesh and intravalvular liquid. three dilutions MPN test specified in ISO 16649-3. live tunicates and live marine gastropods.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.000 E. The reference method for this analysis is the five-tube. that microbiological criteria and chemical contaminants amongst other requirements are met. live bivalve molluscs must not exceed 46.Official controls concerning live bivalve molluscs from classified production areas A. 000 E. Classification of production and relaying areas: This section reinforces that specified in Annex III. The reference method for this analysis is the five-tube. coli per 100 g of flesh and intravalvular liquid.600 E. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated. Regulation (EC) 854/2004.5. Annex II 9.
boundaries or class of a production area or its closure.5. Procedures for re-opening a closed production area are stated D. 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. 214 . 2073/2005.5. Additional monitoring requirements This concerns the competent authority in relation to monitoring of classified production areas where bivalve mollusc harvesting is forbidden and of relaying and production zones to ensure there is no malpractice etc.6 Regulation (EC) No. the competent authority is to close the production area. 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. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. 9. E. 9.2 Chapter III . which has applied since 1 January 2006. establishes microbiological criteria for a range of foods.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. F. opening or closing a production area. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying.FISH AND SEAFOOD C. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions.2. 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. 9.
ensuring that foods being placed on the market do not pose a risk to human health.6. 9.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria.6. 215 . and/or on the quantity of their toxins or metabolites. with chapter 2. Two kinds of criteria have been established: food safety criteria.4 referring to fishery products. volume.II. a batch of foodstuffs or a process. based on the absence. 9. area or batch. The relevant criteria are outlined in Table 9. The criteria are outlined in Table 9. applying to products placed on the market and process hygiene criteria that are applied during the manufacturing process.I. presence or number of microorganisms. per unit(s) of mass.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.LEGISLATION The aim of this legislation is to complement food hygiene requirements. and the legislation applies to all businesses involved in food production and handling. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product.
monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1.I Food safety criteria Sampling plan(1) n 10 5 Absence in 25 g EN/ISO 11290-1 c m M Products placed on the market during their shelf life Limit(2) Analytical reference method(3) Stage where the criterion applies Food category Microorganisms 1.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life . monocytogenes 1. other than those intended for infants and for special medical purposes(4) (8) L.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L. monocytogenes. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1.TABLE 9.2 Ready-to-eat foods able to support the growth of L.3 Ready-to-eat foods unable to support the growth of L. monocytogenes L. other than those intended for infants and for special medical purposes EN/ISO 11290-1 Products placed on the market during their shelf life Before the food has left the immediate control of the food business operator. monocytogenes.
coli (15) 1. tunicates and gastropods 9 (18) 2 E.17 Live bivalve molluscs and live echinoderms.25 Live bivalve molluscs and live echinoderms.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. 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. manufactured from fish species associated with a high amount of histidine (17) Histamine .26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1.27 Fishery products which have undergone enzyme maturation treatment in brine.
excluding sprouted seeds. Engraulidae.. sugar. The most recent edition of the standard shall be used. Malle P. Other categories of products can also belong to this category. when he is not able to demonstrate. wine. products heat treated in their final package). c = number of sample units giving values between m and M For point 1. 43-9. 5 This criterion shall apply if the manufacturer is able to demonstrate.. A pooled sample comprising a minimum of 10 individual animals. soft drinks.1 . biscuits and similar products.94. including cocoa and chocolate products. Assay of biogenic amines involved in fish decomposition. when recontamination is not possible after this treatment (for example. AOAC Internat. to the satisfaction of the competent authority. 2. cider. products with pH ≤ 5. coli is used here as an indicator of faecal contamination. References: 1. spirits and similar products. — bread. J. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. that the product will not exceed the limit 100 cfu/g throughout the shelf life. m = M.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. E. Valle M. according to which the whole batch is to be deemed unsafe. monocytogenes. 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. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . live bivalve molluscs. Bouquelet S. 1996. beer.0 and aw ≤ 0.92.1.. Bouquelet S.. Malle P. Clupeidae. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. 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. Dervin C. Single samples may be taken at retail level. 79. Products with pH ≤ 4.25.. honey and confectionery. — — bottled or packed waters. Coryfenidae. uncut and unprocessed vegetables and fruits. Particularly fish species of the families: Scombridae. — fresh.4 or aw ≤ 0. Duflos G. This criterion shall apply to products before they have left the immediate control of the producing food business operator. Scombresosidae. to the satisfaction of the competent authority. subject to scientific justification. shall not apply. Pomatomidae.
monocytogenes in ready-to-eat foods able to support the growth of L. monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory. if the following requirements are fulfilled: 1. tunicates and gastropods in relation to testing E. — unsatisfactory. if the presence of the bacterium is detected in any of the sample units. 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 are ≤ the limit. AOAC Internat. if all the values observed indicate the absence of the bacterium. 1999. L. if the presence of the bacterium is detected in any of the sample units. 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. Histamine in fishery products from fish species associated with a high amount of histidine: — satisfactory. (1) 219 . Interpretation of the test results relating to Table 9. if all the values observed indicate the absence of the bacterium. 82. J. L. where the limit refers to a pooled sample. 1097-101. if all the values observed indicate the absence of the bacterium. a maximum of c/n values observed are between m and M 3. — unsatisfactory. Salmonella in different food categories: — satisfactory. coli. no values observed exceed the limit of M. 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. L. the mean value observed is ≤ m 2. — unsatisfactory. excluding live bivalve molluscs and live echinoderms. — unsatisfactory.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus.I The limits given refer to each sample unit tested. if any of the values are > the limit. The test results demonstrate the microbiological quality of the batch tested(1). — unsatisfactory. if the presence of the bacterium is detected in any of the sample units.
II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2.TABLE 9. coli 5 shucked products of cooked crustaceans and molluscan shellfish 2 100 cfu/g 1000 cfu/g EN/ISO 6888-1 or 2 FISH AND SEAFOOD Coagulasepositive staphylococci 5 Improvements in production hygiene 220 .1 Shelled and E.4.
if a maximum of c/n values are between m and M. 14 (Hygiene requirements specific to the U. and the rest of the values observed are ≤ m. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. c = number of sample units giving values between m and M The most recent edition of the standard shall be used.7. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health. S.1 In the U. S. 2006 No. — acceptable.7 Food Hygiene (England) Regulations 2006.II The limits given refer to each sample unit tested. and the rest of the values observed are ≤ m. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. 2006 No. — unsatisfactory.LEGISLATION 1 n = number of units comprising the sample. if one or more of the values observed are > M or more than c/n values are between m and M. 14 details temperature control requirements for foods in general. For example. Schedule 4 of the Food Hygiene (England) Regulations 2006. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. the defendant will need to prove that the food was on service or display. if a maximum of c/n values are between m and M. — acceptable. 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 . 9. The regulations prescribe a chilled food holding temperature of 8 °C or less. The regulations provide for defences in relation to upward variations of the 8 °C temperature. with chill holding tolerance periods.I. The defendant may be required to produce well-founded scientific proof to support his claims. which overrides the 8 °C requirement. if all the values observed are ≤ m. 2 Interpretation of the test results relating to Table 9. The test results demonstrate the microbiological quality of the process tested. E.K. — unsatisfactory. if all the values observed are ≤ m. tolerance periods for chill-holding of foods and hot-holding variations.I.K) Temperature control requirements 9. if one or more of the values observed are > M or more than c/n values are between m and M.
or.food.gov. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. 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. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature. was kept at above 8 °C or the recommended temperature for an unavoidable reason. for a limited period consistent with food safety. 3). Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing.I. 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. 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. The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment. 9. 2006 No. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions. such as that below.K. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. it would need to be proved that the food was being transferred to or from a vehicle used for the activities of a food business.I. 2006 No. S.8 Guidance In the U.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours.S. and was kept at above 8 °C or the recommended temperature. 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. Alternatively.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . 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. For other food.
12. The revised text of Regulation (EC) No.4. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41.2004.2004. 852/2004 is now set out in a Corrigendum (OJ No.6. 9. 22.2005. L338.2004. these are not be discussed here. L139. 3) as amended by Regulation (EC) No. 25. 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.LEGISLATION 9.4.2004. 30. as amended. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 25. Regulation (EC) No. 1. Regulation (EC) No. 2076/2005. Regulation (EEC) 2136/89. 853/2004 is now set out in a Corrigendum (OJ No. 30. L226. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito.2004.2006. 2074/2005. 1688/2005. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. 1441/2007. L226. 854/2004 is now set out in a Corrigendum (OJ No. 2076/2005. 62) as amended by Regulation (EC) No. 223 . 2073/2005 on microbiological criteria for foodstuffs (OJ No. as read with the corrigenda at OJ No. 2074/2005 and Regulation (EC) No. 55).6. L139. 2076/2005. 2075/2005 and Regulation (EC) No.10. Regulation(EC) No. namely. Regulation (EC) No. Commission Regulation (EC) No.4. 206).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.2004. 22) as amended by Regulation (EC) Nos.6.10 Further Reading Regulation (EC) No. 1663/2006. laying down common marketing standards for preserved sardines. The revised text of Regulation (EC) No. 1662/2006. 2074/2005. Regulation (EC) No. 30. 83) as amended by Regulation Nos 882/2004. 2074/2005. 2076/2005. 1). The revised text of Regulation (EC) No. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 14. Regulation (EC) No. L283. 1019/2008 and as read with Regulation 2073/2005. L226. L139. 25.
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. 22.10. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338. 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. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004.2001. 22.2005.2005.1. (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. 6) 224 . 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. 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.12. 23.12. 21.2000.
The minimum pH value for growth is thought to be pH 5.3 . 10.5. Some death will occur at this pH if the temperature is 4 °C.45 °C (1). At pH 4. no growth is observed (1).5 minutes. Morphology Gram-negative rods.000-fold increase in number is observed after one week in food systems (1).3 Temperature Aeromonas grows at temperatures of 2 .5.0 . 225 . and 2.1 10. At 5 °C. Aeromonas has the potential to grow over a pH range of ca 4 .1.1.3 minutes.2 Oxygen requirements Facultative anaerobe.5 µm 10.0 x 1.1.1. 10. PATHOGEN PROFILES 10.4 Heat resistance Aeromonads are sensitive to moderately high temperatures.3.1. a 10 to 1.10 (3).1 Aeromonas spp. under optimal incubation temperatures. respectively (1).10. 10. 0.5 pH Under otherwise optimal conditions. and D-values at 45 and 51 °C have been reported as 29.1. The optimum growth temperature is 28 °C (2).
5 . E and. forming at least seven different toxins.2% sodium chloride (NaCl) (1). A to G. B. but the spores of this organism are more so. F are the types that are responsible for most cases of human botulism (5. botulinum is prevented.0 µm.1 Morphology Gram-positive.22. at 85 ºC for 5 minutes.2. which may be inactivated by heating at 80 ºC for 20 . 10. 10. botulinum types produce heat labile toxins. Group I will not grow at temperatures of 10 ºC or less.1.3 Temperature All strains of C. botulinum are not particularly heat resistant.even as low as 3 ºC (7).4 x 1. and groups I and II are those responsible for the botulism cases. 10. but the interior of the food may do so. spore-forming rods. 0.5% NaCl (4). but Group II strains are psychrotrophic. 10.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 .2.2 Clostridium botulinum Seven different types of C.7 . All C. 226 . many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. botulinum grow reasonably well in the temperature range of 20 45 ºC.2 Oxygen requirements Although C. and is sensitive to > 4. being capable of slow growth and toxin production at low temperatures . All type A strains are proteolytic and type E strains are usually non-proteolytic. botulinum is an obligate anaerobe. 6). It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C.2.2. or at 90 ºC for a few seconds.2. to a lesser extent.30 minutes.4 Heat resistance The vegetative cells of C. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different. Types A. 10.FISH AND SEAFOOD 10. botulinum are known. There are four main groupings of the organism. Thus an aerobically packed product may not support the growth of the organism on the surface. types B and F can be either.
21 minutes) (9).2 Oxygen requirements C. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0. These concentrations correspond to limiting aw of 0.3 .0 . pH and temperature.3.like other clostridia .3 10.97) (10.1 Clostridium perfringens Morphology Gram-positive spore-forming rods. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. the minimum aw for growth may be lower. Note also that if humectants other than salt are used.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. they may survive mild heat treatments (70 . In commercial situations. 11)). These values have been established under carefully controlled laboratory conditions.10. 0.0 µm.3.g. 10. therefore. perfringens .9 x 2. where oxygen levels are reduced. It will not.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e.94 for Group I and 0. respectively (8.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. 10. The organism will grow well in the centre of meat or poultry dishes.97 for Group II (5). 10. parcooked meals with pH value > 5. botulinum spores The most heat resistant spores of Group I C.0 and aw > 0.2. particularly by cooking. botulinum depends on solute.0.2. safety margins must be introduced to allow for process variability. However.1.PATHOGEN PROFILES 10.7 Characteristics of C. 10.is an obligate anaerobe.6 and pH 5. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. 227 . 9).2. vacuum packed. grow on the surface of foods unless they are vacuum or gas-packed.
15. 13).3 Temperature The most significant characteristic of C. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. 10. As in the case of other factors limiting the growth/survival of this organism. Its optimum temperature for growth is 43 . very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. perfringens appears to be 0.50 ºC. The lowest aw recorded to support the growth of C.3. 16). It grows best at pH values between 6 and 7 (the same pH as most meats). vegetative cells die rapidly (12). depending on strain and other conditions. etc. 10. In addition. type of solute.3. 10.45 ºC. perfringens spores The spores of C.3. Some studies indicate that the presence of 3% NaCl delays growth of C. pH. the limits for aw are affected by temperature. Salt concentrations of 6 . Under otherwise ideal conditions.10 ºC. perfringens is not a tolerant organism with respect to pH.3.5 minutes (17). 13).3. perfringens has the potential ability to grow within the temperature range 15 .93 and 0. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. Recorded heat 228 . 18).8% inhibit growth of most C. although C.7 Characteristics of C. will survive greater extremes of pH (and aw) (12. While some growth can occur at 50 ºC. death of the vegetative cells of this organism usually occurs rapidly above this temperature (12. perfringens is not tolerant of low water activities.3.5 pH C. (It is unusual for spores to be formed in foods after the growth of this organism (14)). lower concentrations may be effective in combination with other factors. and their heat resistance is also affected by the heating substrate. perfringens in roast beef at 60 ºC is 14.it is destroyed by heating at 60 ºC for 10 minutes (14.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. At cold temperatures. perfringens strains. 10. The D-value for C. however. Spores. perfringens. the enterotoxin is not heat resistant . 0 .6 Aw/Sodium chloride C. perfringens in vacuum packed beef (18). perfringens can vary quite considerably in their heat resistance.FISH AND SEAFOOD 10.
98 and 0. pH < 5. respectively (21). requiring days or weeks to reach maximum numbers. it is not a sporeformer. Morphology Gram-positive short rods. 10.0 µm. so can be destroyed by pasteurisation.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17.at refrigeration temperatures down to.albeit slowly .3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. monocytogenes is not a particularly heat resistant organism.3 . The upper temperature limit for the growth of L.5 . Under ideal conditions.3 is the lowest value where growth has been recorded. -0. Its optimum growth temperature. 60 and 65 ºC were reported as 10. using hydrochloric acid as an acidulant).6 . monocytogenes in crawfish tail meat at 55. or even below 0 ºC. the organism is able to grow at pH values well below pH 5 (pH 4.2.5 x 0. and where acetic acid is used as an acidulant.0 minutes for heat resistant spores to 1. 229 . 10. D-values for L. 10.8 minutes for heat sensitive spores (12).64. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.184.108.40.206. growth at low temperatures can be very slow. the lowest limit for growth is likely to be considerably higher .4.4 ºC is probably a more likely minimum for growth in foods (19).4 Heat resistance L.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.4 10.4. However.19 minutes. monocytogenes is reported to be 45 ºC (20). being potentially capable of growing . 0.1 Listeria spp. 1. 10. however. 10.2 has been suggested as the lowest working limit (22).4 . In foods.especially at refrigeration temperatures. is between 30 and 37 ºC.4. and temperature.2 Oxygen requirements Aerobe or microaerophilic. however.0.
monocytogenes is quite tolerant of high NaCl/low water activities.5 pH Plesiomonas has a pH range for growth of pH 4. Morphology Gram-negative short rods.FISH AND SEAFOOD 10.5. 10.1 Plesiomonas Morphology Gram-negative.5.6. It grows best at aw of ≥ 0. 25).7 x 1.6 Aw/Sodium chloride L.5 .5 10.0 x 1 .5 (24.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.6 10.0 µm. 10.8 .8.1 Salmonella spp. at salt levels found in foods (10 . The bacterium may survive for long periods at aw values as low as 0.3.83 (20).3 µm.4. 25). peritrichous flagella. 10. 10.5.5.12% NaCl or more).0 .5.5. but has been shown to be able to grow at an aw level of 0. 10.90.3 Temperature Plesiomonas can grow in the temperature range of 8 . 10.45 ºC. 230 .5 . It is likely to survive. 0. or even grow.0. 0.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24). 10. the optimum being 30 ºC (23).2 Oxygen requirements Facultative anaerobe.1.97.
5.2 Oxygen requirements Facultative anaerobe. 10. or possibly 0.6. substrate.6. and appropriate heat treatments must be determined experimentally for low-aw foods.3 Temperature Salmonellae can grow in the temperature range of 7 .8 . some strains are able to grow at temperatures as low as 4 °C (26).5 .93 (as reported in dried meat and dehydrated soup).48 ºC. The optimal pH for Salmonella growth is between 220.127.116.115.4% NaCl. 231 . the optimum being 35 . a heat resistant organism.37 ºC. with a z-value of 4 . therefore.20 times more heat resistant than the average strain of Salmonella at high aw (27). temperature and pH. pasteurisation and equivalent treatments will destroy the organism under normal circumstances.6.6. D60 ºC values normally range from about 1 to 10 minutes.5 pH Salmonella has a pH range for growth of pH 3. 10. Some death will occur at pH values of less than about 4.5. Salmonellae are quite resistant to freezing. under otherwise ideal conditions. However. 10. and with an appropriate acid. Salmonellae are quite resistant to drying.PATHOGEN PROFILES 10. 10.9. depending on the type of acid and temperature.6. depending on serotype. although salt tolerance increases with increasing temperature (28). Salmonella has the potential to grow at aw levels as low as 0. strains vary in their ability to withstand heating. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. However. Salmonella senftenberg 775W is about 10 . The growth of Salmonella is generally inhibited by the presence of 3 .5 ºC. It is not.6 Aw/Sodium chloride Where all other conditions are favourable. Growth is slow at temperatures below about 10 ºC. Furthermore.4 Heat resistance Salmonella is not a spore-forming organism.
7.7. aureus is more limited under anaerobic than aerobic conditions. aw. stressed cells can also be less tolerant of heating. Under most circumstances.7 10. 0. Most strains of staphylococci can grow within the pH range 4. 10. 10. depending on aw. with an optimum of 35 . The following relate to limits for growth only.7.5). 10.5 pH The pH at which a staphylococcal strain will grow is dependent upon the type of acid (acetic acid is more effective at destroying Staph.7. 10. Staph.).3 Temperature Under otherwise ideal conditions. aureus can grow within the temperature range 7 – 48. etc.2 to 9. 232 .9 µm diameter.7.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH.2 Oxygen requirements Facultative anaerobe. 31). NB: The growth of Staph. aureus than citric acid).3 (optimum 7.20 minutes. The limits for toxin production are also narrower than for growth. It can survive well at low temperatures. however. aw and temperature (sensitivity to acid increases with temperature). In meat macerate. but may cause some cell damage (30).FISH AND SEAFOOD 10. aureus viability.1 Staphylococcus aureus Morphology Gram-positive cocci.0. the D60 ºC value is 2 .0 . fat content.7. As is the case with other bacteria.5 ºC. under otherwise ideal conditions (29. Freezing and thawing have little effect on Staph.37 ºC (29).7 . the organism is heat sensitive and will be destroyed by pasteurisation.
8.2.8 x 1. 10.99 little or no toxin production in anaerobically packed foods. an aw of 0. 10.5) (29. 34).1 Vibrio cholerae Morphology Gram-negative curved rods.PATHOGEN PROFILES 10.8 10.3 Temperature The optimum temperature for growth has been reported as 30 ºC.4 .9. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31.10% NaCl. pH. 233 . atmosphere.6 µm 10. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant. aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.2 .7. but certain strains can grow in 20% NaCl.5 . * dependent on temperature.0. aureus can grow in 7 . In general.0 (optimum 7.7. It can grow over the aw range 0. strain. with a temperature range for growth of 10 . An effect of increasing salt concentration is to raise the minimum pH of growth.86 is the generally recognised minimum in foods (32). 10.6 Aw/Sodium chloride Staph.43 ºC (35). 31) between 0.87 and > 0.2 Oxygen requirements Facultative anaerobe.8.7 Limits permitting toxin production 10 .0 .8. Staphylococci are more resistant to salt present in foods than other organisms.7.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. 0. However.>0.83 .99 aerobically under otherwise optimal conditions. Staph. and solute. In general.
1 Vibrio parahaemolyticus Morphology Gram-negative curved rods.5 ºC).65 minutes (35). with an optimum of 0.9.9.5 pH This organism is tolerant of high pH values. but not acid conditions. 10.998.6 Aw/Sodium chloride V. cholerae is sensitive to heat at temperatures of >45 ºC. 10. are 5 and 43 ºC. declining in number during storage. and it is sensitive to refrigeration temperatures (0 .1 . It can grow in the presence of 0. it has a pH range for growth of pH 5 .97 .9. 10. 10. 38).8.FISH AND SEAFOOD Also.3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.4.6.0. 37.9.6 (35. 10. under otherwise favourable conditions. cholerae can grow over the aw range 0. with an optimum of 37 ºC (39).8.0% NaCl (35). there is evidence that V.8.4 Heat resistance V. with optimum at 7. The D-value for V. respectively. parahaemolyticus is easily destroyed by drying. but only moderately sensitive to freezing (40).984 (35). cholerae in crab meat homogenate at 60 ºC is 2.2 Oxygen requirements Facultative anaerobe.9 10. 234 . 10. V. cholerae may persist on frozen fish for >180 days (36).
4% (39. 10. The optimum concentration of NaCl equates to aw of approximately 0. 10.11.8 .5% NaCl.5 pH The pH range for growth is 4.0. 41).8. 10. with an optimum pH of 7.10. parahaemolyticus are not heat resistant. vulnificus is heat sensitive. 235 .PATHOGEN PROFILES 10.02 . respectively. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.9. 10.4 Heat resistance V. V.9.10.6 Aw/Sodium chloride V. and are readily destroyed by cooking.9. The organism can survive for long periods at freezer temperatures (42). with a temperature range for growth of 8 . 10.5 minutes. 10. parahaemolyticus will grow at aw levels as low as 0.6 (41).43 ºC (42). 41). respectively (39).10.10 Vibrio vulnificus 10. and 2.3 Temperature The optimum temperature for growth has been reported at 37 ºC. but the optimum concentration is 2 .29 minutes.1 Morphology Gram-negative curved rods. depending on solute.922. parahaemolyticus is unable to grow in the absence of salt.10.2 Oxygen requirements Facultative anaerobe. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43). The D-values at 55 ºC for clam and crab homogenate are 0.6 .98 (35.4 Heat resistance V.
. Clostridium botulinum. Potential hazards associated with REPFEDS. Hauschild A.. 4.M. Hauschild A. Peck M. Palumbo S. 11. Murrell K. Diseases Caused by Bacteria. Doyle M.H. in Foodborne Bacterial Pathogens. Ed. Aeromonas. ASM Press.M..R. New York. Lund B.A. in Foodborne Disease Handbook. New York.10. 1993.W...5 .L. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’. 1996. Dodds K. 97-131. Johnson E.M.. 9.. Clostridium botulinum. Betts G. Great Britain. Volume 1: Diseases Caused by Bacteria. Volume 5: Microbiological Specifications of Food Pathogens.P.N. Faris A. Austin J..E.. Doyle M.FISH AND SEAFOOD 10. 5. Gorham J.P.P. 57-63. Bhunia A. 10. 236 . Beuchat L. New York. Eds. 7.N. Gorman J.D. Aeromonas hydrophila. 1994.H.. Marcel Dekker.. New York. 6. 2001. 279-303. Ed.O...D. 383-408. 8. Aeromonas species.J.D. 3. Cliver D.0% NaCl. 357-67. Eds.10..L. Smith J. Marcel Dekker. vulnificus grows over the range 0. Washington D. in Microorganisms in Foods. International Commission on Microbiological Specifications for Foods. Doyle M. Eds.6 Aw/Sodium chloride V.L. in Foodborne Bacterial Pathogens. with an optimum of 2.. Hui Y. Hauschild A. Kovacek K.H.R.. Marcel Dekker. Aeromonas hydrophila group. 1989.H. 2.R. Microbiology and Molecular Biology. in Foodborne Pathogens. Fratamico P. in Food Microbiology: Fundamentals and Frontiers. Clostridium botulinum and Clostridium perfringens. New York. Marcel Dekker. Volume 1. Blackie. 1-27. Bier J. in Clostridium botulinum: Ecology and Control in Foods.. 121-76. Kim J. Stelma G.K. Dodds K. Eds. Eds. Abeyta C. Hui Y. Eds. 111-89. in Foodborne Disease Handbook.5 pH The pH range for growth is 5 . 5-19.C.W. 2005. Juneja V. 1989. Marcel Dekker.10.5% (42). 10. 2003. 1993. 1995. 1994... Foegeding P.W. Novak J. Gaze J. Murrell K. Cliver D. References International Commission on Microbiological Specifications for Foods. 1-19.D. Eds. Clostridium botulinum. 329-49. Food Control.5. Montville T.. Marcel Dekker Inc. Stelma G. New York.11 1.. Principles of control. London.L.H. 6 (1). Miliotis M.. Notermans S. New York.W.W. 10.O. Ed. with an optimum pH of 7. in International Handbook of Foodborne Pathogens. in Clostridium botulinum: Ecology and Control in Foods. Marcel Dekker.H..8 (42). Caister Academic Press. Dodds K.
. Johnson E. Clostridium perfringens. Listeria. 26. in Food Microbiology . Listeria monocytogenes. 201-8. Evans M. 229-40.. Wrigley D. 237 .H. Marcel Dekker. 735-42. Volume 5: Microbiological Specifications of Food Pathogens. Lund B. 137-64. Varnam A. in Foodborne Diseases.. Kim C. London. Clostridium perfringens. 1996.G. 20. Ed. 1991. Washington D.G. Ryser E.V.O. Eds. 18.. Volume 1. Doyle M. Marcel Dekker. Labbe R.. Blackie.P.. 1990. Washington D. Murrell K. Rinke H.O. Evans M. Marcel Dekker. Plesiomonas shigelloides. Matthews K. 17.M. Doyle M.A. 1994. 2005. Blackie. 191-243..R. 15. 129-38.. 13. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs. Doyle M. Plesiomonas shigelloides. New York. in Food Microbiology: Fundamentals and Frontiers. 311-25. in Microorganisms in Foods.H. London. Emery D. London. International Commission on Microbiological Specifications for Foods. Growth of Listeria monocytogenes at refrigeration temperatures. Varnam A.P. 2006. Plesiomonas. 157-62.T. 68 (2). Academic Press. Walker S. Cliver D. 1996. Clostridium perfringens. Ed.. Beuchat L. Diseases Caused by Bacteria. New York. 1990. in Foodborne Pathogens: An Illustrated Text.J. Marth E. Volume 5: Microbiological Specifications of Food Pathogens. International Commission on Microbiological Specifications for Foods. ASM Press. 2007. Koburger J. Montville T. ASM Press.A.PATHOGEN PROFILES 12.. London. Hui Y.H. Clostridium perfringens food poisoning. 1989. International Commission on Microbiological Specifications for Foods. Ed.J. 982-6. Riemann H. in Foodborne Bacterial Pathogens. Blackie. New York. 1989.. 1996. 19. Volume 5: Microbiological Specifications of Food Pathogens. Gorham J. 14.An Introduction.O. in Foodborne Infection and Intoxication. 22.R. Ed. Ed. 33. Banks J. Listeriosis and Food Safety. 24. 133-67. Montville T.. Cliver D.P. Archer P. 1989.. in Foodborne Disease Handbook. Eds. 336 (8721). McClane B. Labbe R.J. 208-13. International Commission on Microbiological Specifications for Foods.R. 351-82.. 25. Lancet.D. 141-82. Eds. Clostridium perfringens gastroenteritis.. 23. International Commission on Microbiological Specifications for Foods. 112-25. Journal of Applied Bacteriology.P..K.H. 1990. Halvorson D. 16. in Microorganisms in Foods.. Clostridium perfringens. in Foodborne Bacterial Pathogens. in Microorganisms in Foods. Juneja V.A. London.C. Avian Disease. International Commission on Microbiological Specifications for Foods. Eds.J. Wolfe Publishing Ltd. Nagaraja K. 2001.M.A.. Ed.C.A. Elsevier. Marcel Dekker. London. Eds. 21.G. Cliver D. New York. Foodborne disease due to Bacillus and Clostridium species. Listeria monocytogenes.
Golden D. Stress Response and Resistance. 34. International Commission on Microbiological Specifications for Foods.D. Blackie. 414-25. 523.H. 13 (11). 35.K.62. Springer Science. Vibrio parahaemolyticus. 85-106. in Foodborne Pathogens: Microbiology and Molecular Biology. Volume 5: Microbiological Specifications of Food Pathogens. 2005. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems.FISH AND SEAFOOD 27. in Foodborne Infections and Intoxications. 543-68. Gustafson J. 1994. Volume 5. 1983. in Foodborne Pathogens. International Commission on Microbiological Specifications for Foods. Warner R.. Ed. 45 (3). Cleland D. 2005.A. Twedt R. in Microorganisms in Foods. 1996. 1993. 642. Ed. 273-84. Stewart G. 31. Bergdoll M. Jay J.O. Ed. 39.. Reed G. Vibrio parahaemolyticus. in Foodborne Disease.O. Salmonella. Loessner M.P. in Foodborne Bacterial Pathogens. Loessner M. Food and Environmental Sanitation. 985-98. 327-52. Microbiological Specifications of Food Pathogens. 28. Vibrio cholerae. Eds. 238 . Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning.. Bhunia A. Marcel Dekker. 29. Doyle M. CDR Review. Woodhead Publishing Ltd. London. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. Eds. Marcel Dekker. Ed. in Modern Food Microbiology. Ed. Crowcroft N.A. 327-445. 40. Doyle M. 1989. New York.S. in Understanding Pathogen Behaviour Virulence.C. New York.S. R15764. London. 33.. London. 331-57. Academic Press. Cambridge. Microbiological Specifications of Food Pathogens. Lee-Wong A.J..A. 1996. Blackie. Oliver J. Golden D. Wymondham. London. Griffiths M. in Microorganisms in Foods. Tohyama Y. 545-66. Staphylococcal Food Poisoning. 4 (13). Salmonellae. Applied and Environmental Microbiology. 32. Staphylococcal gastroenteritis.L. in Microorganisms in Foods. Academic Press. 8. Cholera: current epidemiology. Volume 5. Dairy. Staphylococcus aureus. Ed. International Commission on Microbiological Specifications for Foods.M. 1930. 1989.M. D’Aoust J-Y.P. Ed. 1990. Smith J.P. International Commission on Microbiological Specifications for Foods. Cliver D.M.C. London. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures.R. Caister Academic Press. Bergdoll M. Blackie. 30. 38. Jay J. International Commission on Microbiological Specifications for Foods.M. Wilkinson B. Japanese Journal of Experimental Medicine... 37. 2005. Cliver D. Staphylococcal intoxications.. 217-64.S. Fratamico P. 2005. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). 36. 1996.. New York. Riemann H. Eds..J. 426-35. International Commission on Microbiological Specifications for Foods.
and Campylobacter species. Loessner M. 2005..A.. Springer Science. in Modern Food Microbiology. Yersinia. Ruple A..M. International Commission on Microbiological Specifications for Foods. 657-78. Jay J.J. Golden D. New York.W.D. 43.M. 436-9. Jay J. Foodborne gastroenteritis caused by Vibrio. 239 . Cook D. Ed. Blackie. 55 (12). 1992.PATHOGEN PROFILES 41. London. Microbiological Specifications of Food Pathogens. Volume 5. 985-9.A.. Journal of Food Protection.. International Commission on Microbiological Specifications for Foods. Vibrio vulnificus. 1996. in Microorganisms in Foods. Golden D. 42.J. Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters. Loessner M.
FISH AND SEAFOOD 240 .
defra.org.uk/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: firstname.lastname@example.org@ifm.uk Institute of Fisheries Management (IFM) 22 Rushworth Avenue West Bridgford Nottingham NG2 7LF United Kingdom Tel: + 44 (0)115 9822317 Fax: + 44 (0)115 9826150 Email: email@example.com/ .uk Web site: www.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: firstname.lastname@example.org Web site: www.uk Web site: www.seafish.uk or v.org.ifm.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.uk Prepared Fish Products Association (PFPA) Federation House 6 Catherine Street London WC2B 5JJ United Kingdom Tel: + 44 (0) 208 78362460 The Scottish Government Fisheries Group Environment and Rural Affairs Department Pentland House 47 Robb's Loan Edinburgh EH14 1TY United Kingdom Tel: + 44 (0) 8457 741741 or + 44 (0)131 5568400 Fax: + 44 (0)1397 795001 Email: ceu@scotland.
C.gov.ie/templates/homepage.matis.dardni.uk/index/fisheriesfarming-and-food.fao.dk/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 .fiskehandlerne.bim.suite.org Web site: www.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: email@example.com/fishery/ European Association of Fish Producers’ Organisations H.gov. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.no Web site: www.com/ Danish Institute for Fisheries Technology and Aquaculture (DIFTA) illemoesvej The North Sea Centre DK-9850 Hirtshals Denmark Tel: + 45 98 944300 Fax: + 45 98 942226 Email: difta@difta. Andersens Boulevard 37.dk Web site: www.com 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 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.is/ 242 .uk Web site: www.difta.eapo.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.dk Danmarks Fiskehandlere H.asp Food and Agriculture Organization of the United Nations Fisheries and Aquaculture Department Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57053152 Email: FAO-HQ@fao. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@email@example.com Web site: www.FISH AND SEAFOOD Department of Agriculture and Rural Development.fiskeriforskning.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: firstname.lastname@example.org Web site: www. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: info@eapo.
uk Web site: www.ifst.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.org.co.gov.ifr.hpa.fdf.gsi.cmar.ac.seafood.uk Web site: www.au New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: email@example.com Web site: www.com Web site: www.htm Food and Drink Federation (FDF) 6 Catherine Street London WC2B SJJ United Kingdom TeI: + 44 (0) 207 8362460 Fax: + 44 (0) 207 8360580 Email: firstname.lastname@example.org/ 243 .bsi-global.au Web site: www.org Other Sources of Information British Standards Institution 389 Chiswick High Road London W4 4AL United Kingdom Tel: + 44 (0) 208 9969001 Fax: + 44 (0) 208 9967001 Email: email@example.com/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.gov.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: dhmail@dh.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: firstname.lastname@example.org Web site: www.org CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: email@example.com 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/en/index.csiro.
Web site: www. jsp Food Safety 10 .nhs.scot.euro.org.who.int.net/web/indexen.uk/scieh.int Web site: www.euro.who.sofht.uk Web site: 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 .00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: firstname.lastname@example.org.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.php Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: email@example.com Web site: http://www.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: firstname.lastname@example.org Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.uk Web site: www.se Web site: www.org Web site: www.int/foodsafety World Health Organization (WHO) Headquarters CH .who.co.hps.co.scot.int/en/ WHO Regional Office for Europe Scherfigsvej 8 DK-2100 Copenhagen Ø Denmark Tel: + 45 39 171717 Fax: + 45 39 171818 Email: email@example.com/ 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 244 .who.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: email@example.com/index.sik.
fpa-food. Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: info@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.fda.org.sofht.au/ International Association for Food Protection http://www.co.org/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.gov/foodsafety/ Food Safety Authority of Ireland http://www.gov/ Institute of Food Technologists 525 W.org/ Royal Institute of Public Health (UK) http://www.ift.riph.uk/ Society of Food Hygiene and Technology (UK) http://www.FoodSafety.uk/ 245 .org/ The Association of Food.eu/pol/food/index_en. Beverage and Consumer Products Companies http://www.foodprotection.cdc.csiro.cfsan.org/ Institute of Food Technologists http://www.foodscience.who.CONTACTS US FDA .int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa.fsai.gov/ World Health Organization (WHO): Food safety programmes and projects http://www.ie/ Food Science Australia http://www.org Web site: http://www.htm Centre for Disease Control and Prevention (CDC) (US) http://www.ift.
FISH AND SEAFOOD 246 .
to test microbiological criteria 19 Aeromonadaceae. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. as 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 in fish sauce 127 Bacillus cereus. causing spoilage of dried fish 110. preserved by dry/wet salting 99 Anisakis spp. source of PSP toxins 142 Alexandrium ostenfeldii. in crustacean shellfish 87 Alteromonas spp.INDEX Achromobacter spp. growth and survival in crustacean shellfish 86 Aeromonas sobria. pathogenic microorganisms in molluscan shellfish 59-60 as contaminant in enrobing process 37 as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45 growth in vegetable and rice 247 . growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. planktonic source of spirolides in shellfish 163 Allergens. 111 Astroviruses. in crustacean shellfish 86-7 ATP. 102 Bacillus spp. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. as autochthonous bacterial flora of fish 3 as spoilage organisms in oysters 65 as spoilage organisms in temperate crustacean shellfish 80 as spoilage organisms in tropical crustacean shellfish 81 in molluscan shellfish 60 Aerobic plate counts. 69-70 Aeromonas hydrophila. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. 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 gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. preserved by marinating 100 Chemical detection methods. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. fish sauce 123 Basipetospora halophila. 98-9 Brochothrix thermosphacta. 131 Carbon dioxide. of fresh fish spoilage 11 Chemical pollutants. 68-9 Capture and handling. 136. as autochthonous bacterial flora of fish 3 Clostridium botulinum. 137. in crustacean shellfish 86-7 Campylobacter coli. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. associated with consumption of fish/fish products 16. 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. smoked and dried fish 113-6 hazard in mild-processed marinated products 101 hazard in sun-dried products 102 non-proteolytic – risk of growth in fish recipe meals 44 248 . in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. as cause of spoilage of MAP fish 15 Calciviruses. of fish – effects on microflora 5-6 Carbohydrate. for PSP toxins 144 Chemical indicators. of recipe fish meals 43 Cholera. importance in lactic acid fermentation in fish products 130. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. 226 Breading process 36-8 Brevetoxins. caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. 113. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang.
to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. to determine fresh fish spoilage 11 Eels. 37 as spoilage organisms in chilled products 43 as spoilage organisms in tropical crustacean shellfish 81 as spoilage organisms in vacuum-packed dried fish 111 growth and survival in cured. as contaminants in scampi 36. of fish – different processes 94 Dun. smoked and dried fish 114 Enzymic spoilage. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . to determine fresh fish spoilage 11 Corynebacteria. 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. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. 149-50. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). as autochthonous bacterial flora of fish 3 Crabs.INDEX pathogen profile 226-7 Clostridium perfringens. of prepared fish products 44 Escherichia coli. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. to detect NSP toxins 155 ELISA kit (Biosense). hot-smoking 107 ELISA assay. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp. source of azaspiracids 159 source of DSP toxins 148-9. for detection of DA 148 ELISA kit (DSP-Check). 152 Diphyllobothrium. as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45-6 pathogen profile 227-9 Coated fish products. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. growth and survival in crabmeat 84 growth and survival in cured. adverse condition on poorly preserved fish 110 EC grading scheme. processing 334 Cockles. 106 Contacts 241-5 Conway method. 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.
130-5 Fish-spoilage bacteria 12 Fishtester. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. of fish – effects on microflora 5-6 Hepatitis (non-A. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. fish sauce 123 HACCP. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . of fish at sea 5 Fatty fish. as autochthonous bacterial flora of fish 3 in molluscan shellfish 60 spoilage organism in oysters 65 Food hygiene legislation 189-224 Food of animal origin. as source of inulin in lactic acid fermentation in fish products 134 Garos. non-B) viruses. fermented fish product 126 Green cured split cod 97. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. preserved by marinating 101 Gravlaks.FISH AND SEAFOOD EU legislation. member of cyclic imine group 161 Gymnodinium catenatum. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes. concerning crustacean shellfish 87-8 European Grading Scheme. specific EU hygiene rules 196-224 Fresh fish. source of PSP toxins 142 Gyoshoyu. for evaluation of fish quality 9 Eurotium. as source of ciguatoxins 156 Garlic. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. of prepared fish products 44 Gambiertoxins. of fresh fish spoilage 911 Evisceration. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. to measure fish skin resistance 10 Flavobacterium. 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. 102 Gymnodimines. causing pink condition on fish surface 110 in fish sauce 127 Handling. mould causing spoilage of dried fish 110 Evaluation. processing 35 Fish component.
growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. for fresh and frozen fish (Table) 20 Micrococcus spp. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149. 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. 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. hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. enabled by carbohydrate source in fish products 130. food hygiene 189-224 Lingulodinium polyedrum. in fish fillet 1 Lipid oxidation. definition 1 Legislation.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. to reduce rate of fish spoilage 12 Ikan kaju. 135 I-zushi. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. preserved by marinating 101 Microbiological criteria. for microbial inactivation in fish products 127. as autochthonous bacterial flora of fish 3 Lean fish. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. as source of botulism 136 International legislation. preserved by salt-boiling 100 Karenia brevis. smoked and dried fish 114-5 hazard in cold-smoking 105 hazard in mild-processed marinated products 101 Listeriosis. 100-1 Masmeen 107 Matjes. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. for chilled and frozen prepared fish products 47-9 for chilled and frozen raw fish 19-20 for crustacean shellfish 87-8 for cured. concerning crustacean shellfis 88 Jellied eels. preserved by salt-boiling 100 Ionising radiation. outbreak caused by gravid 115 Lobster. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits.
pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. shigelloides. fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. of crustacean shellfish – effects of processing 81-3 of cured. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . caused by lipid oxidation 11 Oysters. for detection of azaspiracids 160 for detection of ciguatoxins 157 for detection of DA 148 for detection of DSP toxins 150. in crustacean shellfish 867 Norwalk virus. 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. 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. reduction in smoking process 108 Norcardiopsis dassonvillei. 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. 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. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu.
81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 . planktonic source of azaspiracids 159 Pseudomonadaceae. for detection of PSP toxins 144. 225-39 Pathogens. growth and survival in chilled and frozen raw fish 19 growth and survival in crustacean shellfish 86 growth and survival in cured. 145-6 definitions 28-30 non-seafood ingredients 32 Processing. contact details 241-2 Prorocentrum spp. fermented fish product 124 Plankton. reduction in smoking process 103. detection methods 165 origins and distribution 165 toxicology 165 Pissala. as cause of histamine fish poisoning 15 as spoilage organism in chilled products 43 as spoilage organism in fresh fish 14-5 Pickling 94. smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164. spoilage organism in oysters 65 Protoceratium reticulatum. 197 Plesiomonas. 161. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation. preserved by salt-boiling 100 Pink. 100-1 Pindang. causing spoilage of dried fish 110-1 Photobacterium phosphoreum. as source of toxins 159. as source of yessotoxins 152 Protoperidinium cassipes. smoked and dried fish 115 Pasta. causing spoilage in vacuum-packed dried fish 111 Pseudomonads. mould causing spoilage of dried fish 110 Potato. 163. pathogen profile 230 in molluscan shellfish 5960. 69-70 Polycyclic aromatic hydrocarbons. 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.INDEX Parasites. as source of DSP toxins 148-9 Protein content. in fish fillet 1 Proteus. as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. as component for seafood ready meals 40 Pathogen profiles. effects on microflora of fish 5-9 Professional bodies. in molluscan shellfish 60 Pseudomonas spp. 152-3 Penicillium spp. 108 Polypaecilum pisce. as component for seafood ready meals 40 Prawns.
for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops. processing 34. fermented fish product 126 as source of botulism 136 Rakørret. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova. 100 Salting 97-9 definitions 93-4 Sardines.FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries. for microbial inactivation in fish products 127. as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. description and uses 58-9 Scampi. assembly and packaging. as source of tetrotoxin 144-5 Putty fish. detection methods 165 member of cyclic imine group 164 origins and distribution 165 toxicology 165 Puffer fish. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 growth and survival in cured. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products. for evaluation of fish quality 9 Radiation. as carbohydrate source in lactic acid fermentation in fish products 134 as component for seafood ready meals 40 RIDASCREEN. 36 Seafood. hazard in salting of fish 97 Salt concentration.) 41 ingredients preparation. detected in cod 115 Pteriatoxins. fermented fish product 126 Raphidophytes. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. antibody-based kit to detect STX 144 RT Freshmeter. importance in lactic acid fermentation in fish products 130 importance in salting. smoking and marinating of fish 96 Salt-boiling 94. to measure fish skin resistance 10 Salmon. producers of brevetoxins 154 Raw fish. processing and effects on microflora 60-1 Shewanella. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. 83 254 . flow diagram (Fig. in molluscan shellfish 60 Rice. resulting from poorly salted cod 110 Pyrodinium.) 41 Recipe fish products 38-40 Rhodotorula rubra. 135 Rakefisk. growth and survival in chilled and frozen raw fish 19 Pseudoterranova dicipiens. chilled and frozen 1-25 Recipe dish. preserved in saturated brine 99 Sauce. processing 35 Shellstock. vacuum packing 105 Salmonella spp. source of PSP toxins 142 Quality Index Method. flow diagram (Fig.
role in fish degradation 10.INDEX Shigella spp. fermented fish product 126 Taste panel evaluation. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. 13. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. of fish after capture 5 Surstrømming. fish sauce 123 Shrimps. for evaluation of fish quality 9 Torrymeter. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO. processing and effects on microflora 63-4 Skin resistance. grading of 31 hot-smoking 107 US legislation. concerning crustacean shellfish 89 Vacuum packaging. 14 Torry Scale. as indicator of fresh fish spoilage 11 Toxins. contact details 241-2 Trematode infections foodborne 137 Trichosporon. in molluscan shellfish 60 Tuna. 101 Storage. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. of crustacean shellfish 80. fish and shellfish 141-74 Trade associations. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. as contaminant in scampi 36 as contaminant in enrobing process 37 as hazard in dry salting of fish 89 growth and survival in chilled and frozen prepared fish products 46-7 growth and survival in crustacean shellfish 84 growth and survival in cured. smoked and dried fish 114 pathogen profile 232-3 pathogenic microorganisms in molluscan shellfish 59-60 potential for growth in pasta component of seafood ready meals 40 Stockfish. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 . as measure of fish degradation 10 Sliming.
processing for seafood recipe meal centres 39 Vibrio spp. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01. 62 Yersinia spp. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80. as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. 68 Vibrio cholerae non-01. 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. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. of shellstock 61. 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. 152-3 256 . 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi.
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