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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
published by Leatherhead Food International and RSC Publishing. and a detailed list of contacts for various food authorities. and growth characteristics for key pathogens associated with the product. The books are intended as a source of information for microbiologists and food scientists working in the food industry and responsible for food safety. Each book provides a brief overview of the processing factors that determine the nature and extent of microbial growth and survival in the product. both in the UK and elsewhere. I also acknowledge and thank Jackie Apps. and Ann Pernet for the work put into typesetting and indexing. The books in the series are designed as easy-to-use guides to the microorganisms found in foods. I thank the contributing authors for their collaboration. which has been the chief factor in ensuring the completion of this book. Training Manager. Technical Team Leader for her valuable editorial contribution. My gratitude is extended to Dr Peter Wareing. Rhea Fernandes Leatherhead Food International v . thank you for all the support. Acknowledgements All the authors involved in writing the first edition of this book are gratefully acknowledged.Goldwyn. for his scientific input and Victoria Emerton. Fish and Seafood. and Meat Products. All handbooks also contain a review of the related legislation in Europe and UK. potential hazards associated with the consumption of a range of products. guides to HACCP.FOREWORD The Microbiology Handbook series includes Dairy Products. Alison Turner. I dedicate this book to my husband .
7 References 3.8 References MOLLUSCAN SHELLFISH 3.1 Definitions 1.8 References 1.1 Definitions 3.3 Initial Microflora 2.5 Pathogens: Growth and Survival 3.5 Factors Affecting Fresh Fish Spoilage 1.6 Published Microbiological Criteria 4.8 Further Reading iii v xi 1 1 3 5 9 12 15 19 20 24 27 27 28 31 32 43 45 47 49 53 53 59 60 65 66 70 71 77 79 79 80 81 83 84 87 89 92 2.2 Initial Microflora 1.2 Initial Microflora 3.4 Spoilage 3.8 Further Reading CRUSTACEAN SHELLFISH 4.5 Pathogens: Growth and Survival 4.4 Spoilage 1.5 Spoilage 2.1 Definitions 4.6 Published Microbiological Criteria 3.3 Processing and its Effects on the Microflora 4. CHILLED AND FROZEN RAW FISH 1.1 Introduction 2.7 Published Microbiological Criteria 1.2 Initial Microflora 4. vii .CONTENTS CONTRIBUTORS FOREWORD INTRODUCTION 1.6 Pathogens: Growth and Survival 1. 3.6 Pathogens: Growth and Survival 2.3 Processing and its Effects on the Microflora 1.4 Processing and its Effects on the Microflora 2.9 Further Reading CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.2 Definitions 2. 4.7 Published Microbiological Criteria 2.4 Spoilage 4.3 Processing and its Effects on the Microflora 3.7 References 4.
6 References 6. 854/2004 of the European Parliament and of the Council Laying Down Specific Rules for the Organisation of Official Controls on Products of Animal Origin Intended for Human Consumption 212 9.8 Guidance 222 9. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin 196 9.5 Pathogens: Growth and Survival 6.7 Food Hygiene (England) Regulations 2006.2 Paralytic Shellfish Poisoning (PSP) Toxins 7.12 References HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8. 2073/2005 on Microbiological Criteria for Foodstuffs 214 9.2 Initial Microflora 5.7 Further Reading FISH AND SHELLFISH TOXINS 7.4 Amnesic Shellfish Poisoning (ASP) Toxins 7. 9. 7.10 Cyclic Imines 7.11 Conclusion 7.1 Introduction 189 9.1 Definitions 5.6 Published Microbiological Criteria 5.9 Azaspiracids 7.6 Regulation (EC) No.4 Spoilage 6.4 Spoilage 5.4 Implementation and Review of the HACCP Plan 8.2 Definitions 8.8 Ciguatoxins 7.9 Other Relevant Legislation 223 9. 14 (Hygiene requirements specific to the UK) 221 9.5 Regulation (EC) No. CURED.8 Further Reading FERMENTED FISH 6.6 Lipophilic Shellfish Toxins (LST) 7.2 Legislative Structure 190 9.3 Processing and its Effects on the Microflora 5.5. S.5 Pathogens: Growth and Survival 5.1 Definitions 6. 2006 No.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 6.5 Diarrhetic Shellfish Poisoning (DSP) Toxins 7.1 Introduction 7.2 Initial Microflora 6.3 Processing and its Effects on the Microflora 6.7 Neurotoxin Shellfish Poisoning (NSP) Toxins 7.1 Introduction 8. SMOKED AND DRIED FISH 5.10 Further Reading 223 viii .7 References 5.3 Regulation (EC) No.3 Tetrodotoxin 7.3 Stages of a HACCP Study 8.4 Regulation (EC) No. 852/2004 on the General Hygiene of Foodstuffs 191 9.I. EU FOOD HYGIENE LEGISLATION 189 9. 8.
5 Plesiomonas 10.11 References 225 225 226 227 229 230 230 232 233 234 235 236 241 247 CONTACTS INDEX ix .10 PATHOGEN PROFILES 10.1 Aeromonas spp.10 Vibrio vulnificus 10.2 Clostridium botulinum 10.3 Clostridium perfringens 10. 10.8 Vibrio cholerae 10. 10. 10.7 Staphylococcus aureus 10.9 Vibrio parahaemolyticus 10.6 Salmonella spp.4 Listeria spp.
Poultry. the consumption of fish and seafood has increased. Chandan R. Jacobsen C. cured. xi . chilled and frozen prepared fish.. Wageningen Academic Publishers. Saebo A.M. fermented fish. Wageningen. Oehlenschlager J. crustacean shellfish. Bekaert K. 2006. and. 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. fish and seafood can be involved in the transmission of pathogenic microorganisms and toxins. Safety and Processing of Wild and Farmed Fish.A... smoked and dried fish. rivers and lakes whose water can range from pristine to contaminated. season. The second edition of this handbook is a review of the entire book for currency of information. Catches can be gathered from seas.INTRODUCTION Fish and seafood are a main source of animal protein in the diet. Eds. Cross N.. molluscan shellfish. Hoboken. 94767. Seafood Research from Fish to Dish: Quality. Geographical region. Wiley. Often contamination is from human and animal sources.B. 2007. Key changes in this edition are the recent regulatory changes pertaining to food hygiene and microbiological criteria for foodstuffs. Clark S. Hui Y. The Microbiology Handbook. thus. in Handbook of Food Products Manufacturing: Health. for fish. Milk..H. and fish and shellfish toxins. Seafood and Vegetables. Luten J. Seafood processing: basic sanitation practices.Fish and Seafood consists of the microbiology of seven different product categories: chilled and frozen raw fish.. Nollet L.. Meat. Because of their health advantages over red meats. Further Reading General Fish/Seafood Microbiology Cross N..
in Minimal Processing Technologies in the Food Industry.A.L. Marcel Dekker. The incidence of Listeria spp.M.. The fish industry... 459-72. Johnson J.International Commission on Microbiological Specifications for Foods. New York. Noordwijkerhout. November 1995. Hui Y. Journal of Food Protection. Eds. Gram L. Bengtsson N. Kluwer Academic / Plenum Publishers.. 8 (2). Biogenic amines in fish and shellfish. 200-23.M. Proceedings of the International Seafood Conference. Huss H. and Clostridium botulinum in smoked fish and shellfish. xii . Ed. in Food Processing: Principles and Applications. Huss H. 2004. London.. 1997. Eds. Tong P. Eds. Oxford.. 121-37. Blech Z. 431-43. 33 (1). Cambridge. Blech Z. Gudmundsson M. International Commission on Microbiological Specifications for Foods. 87121. in Fish Processing Technology...L. 413-30. Heinitz M. Blackwell Publishing. Blackwell Publishing. November 1995.Y. Ohlsson T. Ventresca P. International Journal of Food Microbiology. 1996. Martin R. Elsevier.A.J.. Davies A.P..D. Hall G. Flick G. Proceedings of the International Seafood Conference. Safety and Quality Issues in Fish Processing. Ed.S.L. Modified-atmosphere packaging of fish and fish products.H. 60 (4).A.S... in Microorganisms in Foods 6: Microbial Ecology of Food Commodities.H. 2004. Stanfield P. Bruinsma B. in Seafood from Producer to Consumer: Integrated Approach to Quality. Elsevier. Luten J. Amsterdam.N. 36 (1-2). Academic Press.B. Gorham J. Minimal processing in practice: seafood. Stanfield P. Oxford. Control of indigenous pathogenic bacteria in seafood. Dalgaard P. Gram L. in Advances in Food and Nutrition Research. Ed. 543-61.Y. 329-65. Ed. New York. Microbiological spoilage of fish and fish products. Simpson B. 1996. Amsterdam. 174-249.H. Woodhead Publishing Ltd. Noordwijkerhout.H. in Seafood from Producer to Consumer: Integrated Approach to Quality. Luten J.B. 1998..R.H. Vo1ume 39. Nip W-K. 167-77. 2002. 1997. 2002. Ed.. in Kosher Food Production. 318-23. Huss H. 2003.E.. Hafsteinsson H. Ashie I. Blackie. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products... Microbiology of fish and fish products. Food Control. Seafood: processing. Journal of Food Protection. Cambridge. 1997. 2005. Salmonella spp. 1996. Predictive microbiological modelling and seafood quality. 91-8. Dalgaard P. CRC Critical Reviews in Food Science and Nutrition. Woodhead Publishing Ltd.K. basic sanitation practices. in Food Plant Sanitation. 245-66 Bremner H. Smith J. Seafood processing: basic sanitation practices.R. Ed. Hui Y. London. Taylor S.. Rawles D. Spoilage and shelf-life extension of fresh fish and shellfish. Smith J. 372-6. 1997. 61 (3). Fish and fish products (Microorganisms in fish products). McCarthy S. 1997.
44 (12). Eds. Glasgow. Jakobsen M. Ed. International Journal of Food Microbiology. Bacterial pathogens transmitted by seafood. Liston J. 1992. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. 93-105.International Commission on Microbiological Specifications for Foods. Volume 5: Microbiological Specifications of Food Pathogens. Institut fur Veterinarmedizin. International Journal of Food Microbiology.H. Pearson A.M.H. Berlin. Garrett E. Ed. Dalgaard P.R. 269-82. Blackie.D.R. 1990.. Copenhagen. 46 (1). Jakobsen M. Ed. Sterling Publications Ltd. Microbiological indicators and seafood processing. Eds. Evaluation of the bacteriological quality of seafood. 166-73. 121-8. Application of predictive microbiology to assure the quality and safety of fish and fish products. Ed. Blackie. Liston J. 1994. Liston J. in Food and Beverage Technology International: USA.. 1996. 2257. 26 (3). Copenhagen. Amsterdam. xiii . London. 5662. Eds. Chapman and Hall.. August 1991. Ward D. McMeekin T. 1989. 319-33. Ed. in Fisheries Processing: Biotechnological Applications. August 1991.M. Gram L. Seafood toxins of microbiological origin. 1994.A. Fish: microbiological spoilage and safety.R.H. 1994. in Microorganisms in Foods. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. in Quality Assurance in Seafood Processing: A Practical Guide.A. Bernoth E-M. Hobbs G. 1-17.. London. Ross T. Sofos J. Liston J.. 54-61. 5 (3). 16 (1). Bonnell A. 90-106. and specifications for seafood products. A new look at microbiological standards. PHLS Microbiology Digest. Amsterdam. Modelling of microbial activity and prediction of shelf-life for packed fish. poultry and fish. Martin A. 265-79.D. in Public Health Aspects of Seafood-borne Zoonotic Diseases. 25-39. Chapman & Hall. 1995. 1994. Olley J. Bacterial spoilage of seafood. in Quality Attributes and Their Measurement in Meat. Food Technology. Gibson D. 26 (3). Activities Report of the R&D Associates. Twigg B. Microbial growth and its control in meat. Amsterdam. The microbiology of seafood products.S.J. Huss H. Eds. August 1991. Microbiological quality of fishery products. Poultry and Fish Products. 459-78. Evaluation of the bacteriological quality of seafood. Bonnell A. 305-17. 118-20. International Commission on Microbiological Specifications for Foods. London.M. Liston J. 1992. in Quality Assurance in the Fish Industry: Proceedings of an International Conference. Elsevier. 1994. Microbial hazards of seafood consumption. 11 (2).. Copenhagen. 359-403. Dutson T. 1991. Fish spoilage and safety.. 1992. Ward D. Elsevier. International Journal of Food Microbiology.. Food Science and Technology Today. guidelines. 1990. Qualitative and quantitative characterization of spoilage bacteria from packed fish. 1995. Huss H. Elsevier.N. Dalgaard P. 1992. Jakobsen M. London. Huss H.. Gilbert R. Gram L.
Applied Science Publishers. 1986. Hobbs G. Elsevier Applied Science Publishers.J. in Microbiology of Frozen Foods.M.K. Microbiological hazards associated with fishery products. Wiley. 103.R. Microbiological standards. Garrett E. Microbiology of frozen fish and related products. 1982. Dicharry A. Davies R. 169-208. 46 (1). Chichester... 104-9. Regenstein J.a review. Shelf-life extension of fresh fish .C. London. Hobbs G. guidelines. 42 (3). Hodgkiss W. Ed. 117-27.. Volume 1. 1988.spoilage of fish. Lamprecht E. 1985. 1988. 71-117.R. 42 (3). Ed. 1988. 11 (2). Part I .K. Microbiology of fish. Journal of Food Quality. Robinson R. in Developments in Food Microbiology. The bacteriology of fish handling and processing. Hackney C. Norris J. Food Technology. xiv . 199-226. 8-16.Pedrosa-Menabrito A. 90-3. and specifications and inspection of seafood products. 1987. Ed.S.. Food Technology. Simmonds C. London. Eyles M. in Essays in Agricultural and Food Microbiology. Seafood-borne bacterial pathogens of marine origin. CSIRO Food Research Quarterly.
Fish are an important part of a healthy diet since they contain high quality protein. 1 . and is mostly interspersed between the muscle fibres. The connective tissue content of fish is also lower than that found in meat.21%. giving it its characteristically flaky structure. crustaceans.5% (1). respectively (1). The lipid content. fins and a streamline body. “fish” will be used for fresh and seawater finfish.1. Although fish is broadly similar in composition and structure to meat there are a number of distinctive features. lean fish are those fish that store most of their fat in the liver.1 Definitions Fish are classified as any of the cold-blooded aquatic vertebrates of the super class Pisces typically showing gills. and excludes molluscs and Crustacea. ‘fish’ also refers to the flesh of such animals used as food. migratory patterns. which can be up to 67%. However. offering less than 0.2 . Fish fillet composition can vary significantly within the same species due to feed intake. Muscle composition and structure of fish also differ from those found in other meat. Covadonga Arias Department of Fisheries and Allied Aquacultures Auburn University Auburn Alabama 36849 United States of America 1. In addition. and spawning season. most fish contain omega 3-fatty acids and other essential nutrients. some regulatory agencies such as the US Food and Drug Administration (FDA) will include molluscan shellfish. typically fluctuates between 0. In this chapter. This super class of vertebrates includes all the bony and cartilaginous finfish. In addition. Fish fillets are a poor source of carbohydrates. Protein content in fish fillet varies typically from 16 . CHILLED AND FROZEN RAW FISH Associate Prof. but typically present a low fat percent when compared to other meats. and other forms of aquatic animal as part of their ‘fish’ definition. while fatty fish have fat cells distributed along their bodies. Fish can be divided into fatty and lean fish. typically 3 and 15% of total weight.20%. it shows a typical season pattern especially in migratory species such as herring or mackerel. Fish flesh is dominated by the abundance of white muscle in relatively short segments. The lipid fraction is the component showing the greatest variation.
Frozen fish is fish that has been cooled to. Modified-atmosphere packaging (MAP) refers to packaging systems in which the natural gaseous environment around the product is intentionally replaced by other gases. during storage. juiciness. transportation and sale. and texture of the fish. transportation and sale. Spoilage describes changes that render fish objectionable to consumers. 2 . and maintained at temperatures just below the freezing point. usually carbon dioxide (CO2). hence. Shelf life is defined as the time of storage before microbial spoilage of a fish is evident. and flavour based on the maturity. Quality refers to palatability and organoleptic characteristics such as tenderness. nitrogen (N2) and oxygen (O2). DMA is dimethyl amine. Spoilage potential is a measure of the propensity of microorganisms to render fish objectionable to consumers through the production of offensive metabolic byproducts. odour and texture. and maintained at or below -2 °C (normally below -12 °C) during storage. firmness. but not below 3 °C during storage. but no further control is exercised during storage.FISH AND SEAFOOD Chilled fish is fish that has been cooled to. renders that fish objectionable to consumers. colour. This technique is primarily used for bulk storage. Raw fish refers to fish that has not been cooked but excludes fish treated with curing salts and/or subjected to fermentation. Fresh fish is raw fish that has not been processed. transportation and sale. The proportion of each component is fixed when the mixture is introduced. colour. Organoleptic refers to qualities such as appearance. marbling. at -2 to -4 °C. spoilage microflora describes an association of microorganisms that. 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. through their development on fish. and maintained at or below 7 °C. Superchilled fish is fish that has been cooled to. frozen or preserved.
) are also common aquatic bacteria. Members of the families Vibrionaceae (Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp. several culture media containing sodium chloride and more than one incubation temperature should be used. In fact. More accurate. microorganisms colonise the skin. molecular-based methods have not yet been used to address this issue. The number and diversity of microbes associated with fish depend on the geographical location. It must be noted that mesophilic bacteria can rapidly overgrow psychrotropic organisms. Moraxella. The autochthonous bacterial flora of fish is dominated by Gram-negative genera including: Acinetobacter. Flavobacterium. Gastrointestinal tracts and gills typically yield high bacteria numbers. and typical of the fish flora. However. the natural fish microflora tends to reflect the microbial communities of the surrounding waters. 3 . For example. posing a concern for seafoodborne illnesses (2). healthy fish is considered sterile and should not present any bacteria or other microorganisms.CHILLED AND FROZEN RAW FISH Vacuum packaging (VP) refers to packaging systems in which the air is evacuated and the package sealed. are present there as result of contamination (anthropomorphic origin or other) or are introduced to the fish during harvest. Lactobacillus and coryneforms can also be found in varying proportions (1). standard culture-dependent methods can only recover between 1 to 10% of total bacteria present in any given sample.I).2 Initial Microflora The subsurface flesh of live. 1. potential human pathogens may be present in both scenarios.II) (3). It is crucial to mimic the environmental physicochemical parameters when isolating bacteria from fish. In general. Clostridium. processing or storage (Table 1. as with other vertebrates. and those that although not autochthonous to the aquatic environment. the season and the method of harvest. Micrococcus. Fish harvested from clean and cold waters will present lower bacterial numbers than fish from eutrophic and/or warm waters. Human pathogenic bacteria can be part of the initial microflora of fish. some species (most Vibrios) require sodium chloride for growth. gills and the gastrointestinal tract of fish. although these are influenced by water quality and feed. 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. whenever possible. Shewanella and Pseudomonas. Gram-positive organisms such as Bacillus. since they heavily depend on the type of sample analysed and the protocol used for isolation.
>106 Salmonella spp.1 µg toxin lethal dose Clostridium botulinum non-proteolytic type E Pathogenic Vibrio spp.35 °C 8 . 4). sewage.106 cells/g.I Indigenous bacterial pathogens typically present in fish (3) Organism Temperature range Estimated Minimum Infective Dose 00.106 cells/g (exception: V. and mud Ubiquitous.102 101 .1 . 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. 105 .II Non-indigenous bacterial pathogens frequently present in fish (3) Organism Listeria monocytogenes* Primary habitat Soil. In addition. human origin Minimum Infective Dose Variable depending on the strain (>102 cells/g) Staphylococcus aureus 105 . Toxin levels 0.19 µg/kg bodyweight from < 102 . and risk associated with the consumption of seafood is low (2. birds. Escherichia coli Yersinia enterocolitica 101 .109 cells/g) * Some sources considered L.0.26 °C 10 .FISH AND SEAFOOD TABLE 1. stream water. However. vulnificus) 5 . numbers of pathogenic bacteria in raw fish tend to be low.37 °C High for most species. Plesiomonas shigelloides 3 . 4 . Intestinal track of terrestrial vertebrates Human origin Fecal contamination Ubiquitous in environment Shigella spp. estuarine environments. Vibrio cholerae Vibrio parahaemolyticus Vibrio vulnificus other vibrios Aeromonas spp.37 °C Unknown Unknown TABLE 1.103 High (107 . during storage indigenous spoilage bacteria tend to outgrow potential pathogenic bacteria.14 .
After capture. Inadequate circulation of chilled brines may result in localised anaerobic growth of some microorganisms. and during harvest. mackerel or herring. or traps. bruising and bursting of the gut will increase the number of sites open for bacterial attack and spread. and conditions of storage.a practice that may have both advantages and disadvantages. potential contaminants added during handling and processing. in which the health of each animal can be assessed prior to slaughter. Of particular concern is the use of wooden or soiled plastic containers for storage and unloading at the quayside. In addition. The action of intestinal enzymes and activity of the gut bacteria on the flesh around the belly cavity may produce discolouration. Used refrigerated brines can be contaminated with high numbers of psychrotrophic spoilage bacteria. This contrasts greatly with the meat industry. and the killing process is designed to minimise stress. The length of time that set nets have been in the water or the time trawlers’ nets are towed. freezing facilities (-18 °C) may be used to prevent the catch from deteriorating. Fish may be eviscerated prior to storage at sea .3. and especially when fish is stored on board for longer periods. However. decks. Physical damage such as loss of scales. chilled brine or refrigerated seawater at -2 °C. the fish may be stored in the vessel for periods ranging from just a few hours to several weeks in melting ice. digestion and off-flavours in uneviscerated fish. The decision to eviscerate the catch at sea will depend greatly on the size of the fish and the duration of storage at sea. has an effect on the amount of stress and physical damage that the fish will suffer during capture. the cuttings provide areas of exposed flesh that are open to microbial attack. in recent decades. and their re-use will increase the crosscontamination of other fish with such microorganisms. 1. icing or freezing. birds and soil. and spoilage.1 Processing and its Effects on the Microflora Capture. ropes. however. cortisone levels increase during prolonged stress and can alter the fillet quality.CHILLED AND FROZEN RAW FISH Shelf life depends on the initial microflora on the fish. In eviscerated fish. hook and line. handling and processing Wild finfish are usually caught by net. offering better control of fish health prior to. human hands and clothing. with fish such as tuna and cod being more commonly eviscerated than sardines. If evisceration is carried out at sea. These contacts will not only increase the bacterial cross-contamination between fish batches but will introduce microorganisms from other sources such as humans. with very little control over the condition of the fish at the time of death or the duration of the killing process. in which the bacterial load 5 . finfish will almost invariably come into contact with nets. care should be taken in removing all the gut contents and washing the carcass thoroughly prior to refrigerating. aquaculture practices have been expanding worldwide. with the production of off-odours. During capture and storage.3 1. Increasingly. boxes and/or baskets.
By vacuum packing foods. Anaerobic atmospheres have less effect on fresh fish shelf life. allowing more growth of spoilage organisms. Bacterial composition under MAP shifts from mostly Gram-negative to predominantly Gram-positive (lactic) bacteria. 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. a reduction in the oxygen tension is achieved which. Low oxygen levels have been shown to substantially prolong the freshness and quality life of refrigerated seafood products. However. and to limit the loss of quality (5). 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. fish have a higher post mortem pH. however. potential spoilage bacteria are among the psychrophilic and psychrotrophic flora present on temperate-water fish before death (9).3. but isolate the food in an environment different from the bulk atmosphere. In MAP. These changes can be deleterious. These containers are also used for displaying the fish during auction at the quayside. from water activity (aw) through to physical damage. if there is some oxygen demand from the product. by actively altering the composition of the 6 . and of the quality of the water used for ice and to wash fish. use of effective detergents and disinfectants. MAP extends the shelf life of most fishery products by inhibiting bacterial growth and autoxidation. 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. and minimising handling will all reduce microbial cross-contamination. ferric ion (Fe3+)). not part of the normal flora of the meat animals (8). partial fermentation of sugars occur leading to lower pH. The atmospheric conditions surrounding a product may be passively or actively altered.FISH AND SEAFOOD can be substantial. In the absence of oxygen. for example. will result in fully anoxic conditions. What is more. in time. As with all foods. but if applied properly should extend the life of the product. Brochothrix thermosphacta and psychrotrophic lactic acid bacteria (LAB) can produce spoilage characteristics. 7). Carbon dioxide is the most important gas in MAP of fish because of its bacteriostatic and fungistatic properties. they are usually process contaminants. Packaging changes the intrinsic and extrinsic parameters affecting a product. nitrogen.).2 Modified-atmosphere packaging A natural atmosphere rich in oxygen (21%) is responsible for oxidative processes and for all aerobic respiratory life. the natural atmosphere is replaced with a controlled gas mixture (carbon dioxide. cleaning of the work environment. Both carbon dioxide and low pH inhibit the growth of the typical spoilage bacteria such as Pseudomonas and Shewanella. oxygen etc. 1. often in the absence of adequate refrigeration.
but only in recent years has it seen a marked expansion in use and market share. slower-growing flora (11).3 °C and produce toxins. The products. after additive application. although less information on their effectiveness is available. but can dissolve into fish liquids and deform packages. Modified-atmosphere preservation of fish was first reported in the 1930s. the pathogens that may be able to grow before spoilage occurs. Of particular concern are the psychrotrophic type E and non-proteolytic type B and F strains. The single most important concern with respect to the use of MAP is the potential for outgrowth and toxin production by C. botulinum. including salt. which are able to grow rapidly at low temperature. High concentrations of carbon dioxide have the most pronounced microbial effect. Micrococcus and Moraxella). Growth and toxin production have been detected in artificially contaminated packs of whole trout after 1 week’s incubation at 10 °C (13). sorbates and chelating agents such as ethylenediaminetetraacetic acid (EDTA) (11). Sulphur dioxide. MAP has been applied to fresh meat and fish with a resulting commercially viable extension in shelf life (10). The principal effect of raised carbon dioxide-MAP is an extension of the 'lag' phase of the growth of the bacteria on the fish. Replacement of oxygen with nitrogen. and the problems of muscle structure damage by the modified-atmosphere (11). as they are able to grow at temperatures as low as 3. which provides guidance relating to vacuum or modified-atmosphere packaging for specific fish products (14). and increase in-pack drip (12). may not be considered ‘fresh’ fish. This has been driven partly by increased consumer demand for fresh and chilled convenience foods containing fewer chemical preservatives. In the United States. VP is one of the oldest forms of altering the interior gaseous environment of a pack. The microflora of meat is not the same as that of whole. discolour pigmented fish (11). the inhibition of common 'spoilage' bacteria (Pseudomonas. but residual oxygen and other electron acceptors may be sufficient to allow oxidative spoilage of fish (12). nitrous oxide and carbon monoxide have also been suggested as possible replacement gases in trace amounts for MAP/CAP. does inhibit some aerobic bacteria and reduce the rate of oxidative rancidity. Gases used in MAP of fish most commonly include carbon dioxide and nitrogen. Many additives have been tried in conjunction with changed atmosphere. the higher pH and reduction potential (Eh) of the fish muscle. a modified-atmosphere may contain any gas necessary for the desired effect. gutted or filleted fish. an inert and odourless gas. and the promotion of a predominantly Gram-positive.CHILLED AND FROZEN RAW FISH surrounding gas. and the MAP of fish has more challenges to overcome as a result of: a comparatively large initial load of bacteria present. Flavobacterium. phosphates. without overt signs of spoilage. the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) has published a number of recommendations on 7 . The (International) Codex Committee has published a Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003).
Only the highest-quality fish. this problem occurs to a lesser extent with meat. 4. leading to the recommendation that the sale of MAP/VP raw fishery products be allowed only when certain conditions are met. should be used. It was also noted that organoleptic spoilage and rejection by the consumer should occur before the possibility of toxin production. The only processing dip to be used is 5% potassium sorbate and 10% sodium tripolyphosphate. oxidative rancidity is a significant source of spoilage and hence the complete removal of oxygen and the use of oxygen-impermeable films are preferred. These conditions include storage of the product at ≤ 3. The growth of these bacteria is reduced by high carbon dioxide concentrations but. Freshwater fish have different intrinsic bacterial flora and also do not contain such high concentrations of amines (e. Storage studies have shown that the extension in shelf life of MAP cod fillets was proportional to carbon dioxide concentration up to around 50% (V/V). It highlights temperature control as the primary preventive measure against the possible hazard of toxin production by C. from where drip loss and gaping of the muscle meant that organoleptically classified rejection occurred sooner than with no carbon dioxide addition (12). however. but is recognised as a drawback of carbon dioxide application. owing to the dissolution of cellular structures. rejection due to alteration in fish texture with greater carbon dioxide makes this unsuitable (13). The replacement gas should contain at least 40% carbon dioxide. from both a microbiological and a chemical standpoint.3 °C at all points from packaging onwards. The problem of water loss in MAP fish is common. In the United Kingdom. at all times. For fatty fish. greater advantages are seen. For other less fatty marine fish. as already stated. adequate shelf life and cooking requirements. The 1985 British Guidelines on MAP/CAP fish from the National Fisheries Institute state that: 1. Although all fish species need to be treated separately. 2. The addition of high levels of carbon dioxide to these fish. Subsequent storage must be at or below 3 °C.FISH AND SEAFOOD the safety of MAP and VP refrigerated raw fishery products (15). Sensitive and accurate time-temperature indicators must be used to ensure that the product has not been temperature-abused. and the use of a HACCP plan. 5. 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.g.g. 3. however. may result in unacceptable colour and textural change and hence little shelf life extension (11). grouping of fish types is possible to estimate the best carbon dioxide level to be used in MAP. use of high-quality raw fish. the Sea Fish Industry Authority has published guidelines for the handling of fish packaged in a controlled-atmosphere (16). botulinum. TMAO) that can be reduced to produce 8 . trimethylamine and sulfides). adequate product labelling in respect of storage temperature.
TABLE 1. 17. Loss of freshness and spoilage cannot be separated as processes. Spoilage of fish starts upon death due to autoxidation (oxidation of unsaturated lipids). evaluation of taste is unavoidable as the visual pattern of degradation of the eyes and gills is unavailable for analysis. with no smell. loss of the fresh characteristics may be simply measured by comparative visual and smell analysis. Taste panel evaluation of cooked fish may be used to score or grade colour. 1. rejection of whole fresh fish for human consumption may be made without chemical or microbiological evaluation. the skin turns from iridescent to dull and bleached with bacterial slime.60% carbon dioxide with a balance of nitrogen for fatty fish have been recommended (16. Degradation of whole fresh fish stored in ice generally follows a set pattern. or for that matter evaluation of the taste of the fish.4 1.1 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). these are summarised in Table 1. texture. Gas mixtures of 40% CO2/30% N2/30% O2 for white fish and 40 . smell and 9 . The eyes turn from convex and clear to concave and opaque. and metabolic activities of microorganisms present in the fish. B. Various methods for whole fish freshness evaluation by a trained panel have been proposed.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.4. and the flesh turns from bright and elastic to dull and soft (1). as traditional spoilage patterns of the external organs are typically very distinct.III.CHILLED AND FROZEN RAW FISH off-odours. the gills from pink and shiny. reactions caused by activities of the fish’s own enzymes. C (C=reject) Significantly. but it is a commonly held view that loss of freshness is related to autolytic degradation and spoilage is more microbial in origin (1). 18) and are probably the most widely used. so lower carbon dioxide concentrations may have better preservative effects. and this pattern is the basis of freshness grading schemes. When working with fillets. Over time. to brown and slimy with an intense off-odour. A.
as little endogenous reduction of TMAO occurs (18). an intense odour compound.8 to 6. an osmoregulator. the muscles harden as residual adenosine triphosphate (ATP) is reduced and the myosin and actin filaments bind to form actomyosin (18). sea-weedy taste. These processes are strongly influenced by the physical conditions of storage. sulphurous and/or rancid smells. the Torrymeter. However.5 (18. is significant and may even be responsible for the ultimate sensory rejection of fresh cod and other fish with high initial TMAO content (20.g. By measuring the concentrations of the six components. the Eh of fish muscle remains relatively high (20). an anti-freeze. which increases from 0% towards 100% with time. Because fish tend to have relatively little residual glycogen compared with mammals. development of sickly sweet. Changes in the resistance of the fish skin after death are used as the basis for tests that employ an electrode measurement of skin resistance (e. or its remaining shelf life (1). During the initial storage. or simply a waste product present due to bioaccumulation (22). To a varying extent. post-rigor values are typically in the range of pH 5. this will generally correlate with the length and severity of rigor. possibly slightly metallic. rigor resolves. ii) Neutral taste. The process of ATP breakdown is used as an indicator of fish freshness (24). spoiled and putrid. the muscles relax again and the fish returns to a flaccid state. iii) Traces of sour. TMAO permits a high Eh to remain in the muscle tissue.FISH AND SEAFOOD taste of cooked fish. RT Freshmeter or Fishtester). texture becomes soft and watery or hard and dry. 19). After some time. cabbage-like. After death. The processes of degradation being analysed by the quality scoring methods above are a complex mix of physical. fruity and/or bitter off-flavours. After death. From being flaccid. %). little flavour. Generally. HxR and Hx) gives a quantity (K-value. 23). 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. As the fish degrades. IMP. ammoniacal. bacterial reduction of TMAO to TMA. biochemical and microbiological actions. 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. rigor mortis is the first noticeable change in the fish. thus. all marine fish use/have TMAO (21). iv) Enhancement of the spoilage characteristic of phase iii. chemical. which has been ascribed a number of possible functions as: a trimethylamine (TMA) detoxified waste product. the pH drop of the muscle is correspondingly less. AMP. ADP. reduction in ATP and increase 10 . the cooked analysis of fish passes through four phases of spoilage: i) Delicate sweet. the conductance generally increases. a ratio of concentration of the hypoxanthine (Hx) and inosine (HxR) to total concentration (ATP.
the importance of microbial action increases with time (5. and are especially important in the spoilage of frozen fish. The Conway method (and variations of it) uses a strong inorganic base to volatilise the bases in the fish sample. This measure is highly dependent upon the temperature of storage and may not reflect the rate of loss of quality equally at different temperatures. 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). Chemical degradation continues after the initial post mortem phase. as significant proteolysis is observed only during the latest stages of spoilage and after rejection (30. hypoxanthine. Oxidative rancidity is known to reduce the quality of fatty fish in particular (19. a correlation can be made between the spoilage of the fresh fish and the production of TVB-N. 26). Volatile bases are the best-characterised chemical indicators of fresh fish spoilage. Lipid oxidation can be a result of enzymic action or a cascade reaction initiated by free radicals (26. However. The major spoilage odours and flavours of fresh fish are undoubtedly principally microbial in origin. For some fish species. and a segregated weak acid to absorb them. as microbial spoilage is limited by the low storage temperature (1. 27) produced by aerobic respiration and other forms of metal ion reduction (28). these measurements are usually used to quantify the amount of spoilage. therefore. Evaluation of Total Volatile Base-Nitrogen (TVB-N. which may also be formed by bacteria (25). or a specific fraction of the volatile bases. Quality indices based upon the products of microbial metabolism do not explain changes in quality until microbial growth produces measurable changes in the fish. 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. 18). Lipid oxidation and other oxidative changes lead to oxidative rancidity. is later significantly reduced by bacterial action. 31). 7). colour changes. also termed TVN). using Conway diffusion chambers (29) allows determination of changes of mg-N/100 g fish. so the measurement is effective only during the initial loss of freshness. not to describe freshness. does not show correlation between muscle pH and the amount of volatile bases contained within the fish at rejection. for example the TMA fraction. Comparing results for different fish species. 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. 11 . however. however.CHILLED AND FROZEN RAW FISH in hypoxanthine by endogenous enzymes allow this measurement to be used for determining freshness (19). Neither does the change in pH during storage correlate well with the production of TVB-N. During spoilage. the residual acid is then titrated.
including the ability of the endogenous bacteria to grow at low temperatures and lower endogenous enzyme activity. For fresh fish. 1. by altering the gas atmosphere (extrinsic).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. The reasons for the differences between tropical. 5).1 Factors Affecting Fresh Fish Spoilage Temperature By far the most effective method of reducing the rate of whole fresh fish spoilage is temperature control (1. For fresh fish iced immediately after capture and continually stored in ice.and temperate-water fish spoilage rates may be many. For example. Fish spoil as a result of the chemical. This increase has been reported to be moderately dependent on the temperature of the sea from which the fish are taken (33.5. a minority are often associated with the majority of the spoilage.5 1. biochemical and microbiological reactions taking place within and on the fish. The application of ice storage increases the shelf life of fresh fish from a matter of hours at ambient temperatures to days or weeks.FISH AND SEAFOOD 1. yoghurt spoilage by yeasts and clostridial spoilage of cheese (5) are examples where it has been recognised for many years that a particular minority of the microbial flora present in the product is responsible for its spoilage. The factors are interactive and cannot be completely isolated. Also. 1. the lower the temperature of storage. Not all the bacteria growing on a food will lead to the production of objectionable characteristics.5. but also crucial for the assurance of consumer safety. realisation that the bulk of the microbial population on newly caught fish does not cause off-flavours and off-odours stems from work 12 .5. 34). careful temperature control during storage is not only important in terms of quality loss. The concept of specific spoilage organisms (SSOs) is not new.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 results of altering one parameter may have far-reaching consequences on others. the pH of the fish muscle may change (intrinsic) and a different microbial population may develop due to the change in atmosphere and pH. The sum of these changes may result in a different spoilage profile. All chemical reaction rate kinetics (and thereby microbial growth) are temperature-dependent. the slower the spoilage processes proceed (within limits).
on fish are diverse and in many cases species-specific. its ability to reduce TMAO to TMA. more especially. or faster than the rest of the flora of ice-stored fresh marine fish (9).1 Pseudomonas spp. Pseudomonas spp. The Pseudomonadaceae family represent a large and poorly defined group of microorganisms.3. and proteolytic and lipolytic enzymes that may affect the quality of fresh and. It is able to grow as fast as. During the 1970s. putrefaciens is considered an SSO of temperate-water marine fish species stored in ice. As previously noted. Sh.5. obligate respiratory bacteria. putrefaciens to the spoilage of fresh marine fish has been recognised since the 1940s.2 Shewanella putrefaciens Sh. ‘oniony’ and ‘faecal’ odours from the production of ketones. 37). It is thus an important spoilage organism of gadoid fish such as cod. motile with polar flagella. Members of the genus are able to produce pigments. processed (e. putrefaciens growth may be used as indicators of spoilage (30). plus other characteristic compounds of the species responsible for spoilage. catalase-positive. 35). the characterisation of spoilage of each product must be made before the identification of the responsible agent(s) can proceed. although only since the late 1960s has it been realised that specific metabolites of Sh. form methylmercaptane (CH3SH) and dimethylsuphide ((CH3)2S) from methionine and produce hypoxanthine (Hx) from inosine monophosphate (IMP) or inosine. work with inoculated sterile fish blocks resulted in identification of a specific minority of microorganisms. Bacteria identified as being associated with the spoilage process of fresh fish are as follows: 1. They are generally characterised as Gram-negative rods. It is also present in fresh water. aldehydes.g. therefore. These organisms are described as potential spoilers. 13 . which produced the characteristic spoilage compounds of the fish (30. 35. and may play some role in the spoilage of freshwater fish.3.5.e. 1. esters and nonhydrogen sulphide sulphur-containing compounds such as methyl sulphide (36. 36). it is often isolated as about 1 . oxidase-positive. The spoilage compounds associated with the growth of psychrotrophic Pseudomonas spp. 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. The spoilage of freshwater fish is generally ascribed to the growth of Pseudomonas spp.CHILLED AND FROZEN RAW FISH started in the 1940s. i.10% of the total flora of fresh fish from temperate marine waters. The importance of Sh. mediated spoilage is characterised by ‘fruity’. produce hydrogen sulphide (H2S) from cysteine. the spoilage of a product may be strongly influenced by the conditions under which the product is held. putrefaciens spoilage of fish is due to its biochemical action on muscle. (9) and they are considered SSO of iced freshwater fish (35). frozen) fish products.
Ph.how to reduce a molecule that they are not able to take into their cells efficiently. In fact. Initially. putrefaciens may grow in the absence of oxygen using alternative terminal electron acceptors (ATECs). and its metal ion reduction ability has led to intensive investigation into its role in iron and sulphur cycles. 1. 38). This wide-ranging respiratory capability is thought to be unique (39). phosphoreum has also been identified as responsible for histamine fish poisoning. Ph. mesophilic strains of Sh. Shewanella oneidensis. Another distinct feature of the physiology of Sh. Shewanella affinis. the bacteria have a significant problem to overcome . Sh. baltica has been identified as the main H2S producer in cod during cold storing (37). Shewanella livingstonensis. phosphoreum increased in notoriety when it was proposed that reduction of TMAO to TMA limited the shelf life of MAP cod fillets (12). Shewanella gelidimarina. putrefaciens were required. owing to its cell size and activity. putrefaciens lies in its chemotaxis: the bacterium. Because no other TMAO-reducing bacteria were present in sufficient numbers to produce the quantities of TMA that were related to rejection. putrefaciens has been studied. 41). they are heterogeneous. but does show strong chemotaxis up gradients of most of their alternative electron acceptors (39). putrefaciens growth has been collected (12. phosphoreum were required for 50% of taste panellists to reject a sample. Shewanella frigidimarina. unlike many other motile bacteria. at the time of rejection. In terms of its taxonomy. and Shewanella aquimarina. with several new species being described. putrefaciens were identified as Shewanella algae. In 50% N2/50% CO2 MAP cod. putrefaciens strains are characterised as microaerophiles or anaerobes. Later. like all members of the Pseudomonadaceae. Sh. 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. as the ferric iron is insoluble. putrefaciens was not present in such numbers (23. whereas >108 cfu/g Sh. The localisation of the reductase on the outer cellular membrane offers the bacteria a solution to this problem (40). but Sh. it is strictly respiratory. a population of Ph. Research has shown that approximately 107 cfu/g of Ph. phosphoreum sufficient to cause spoilage was found. although. does not show carbon-source chemotaxis.5.3 Photobacterium phosphoreum Recognised for some time as being present on spoiling fish (30). Shewanella waksmanii.FISH AND SEAFOOD for which the most compelling evidence showing spoilage as a result of Sh. Sh. The prevalence of the bacterium in so many environments. and recent taxonomic description using modern molecular methods bifurcated this species. 41). Shewanella denitrificans. Shewanella olleyana. additional psychrotrophic species such as Shewanalla baltica. it was proposed that this organism. The ferric reductase activity of Sh. It is also able to produce hydrogen sulphide and a range of other off-odour compounds.3. This type of intoxication occurs when bacteria convert the histidine 14 . and Shewanella profunda were described.
botulinum recorded up to 1963.3 °C (Group II) and these are non-proteolytic (i. (and very rarely F and G) have been reported to cause human disease. algal toxins. presents a potential risk. MAP studies have also demonstrated this organism’s sensitivity to oxygen and have shown that they are also inhibited by high CO2 concentration (43).e. Review of outbreak data suggests fresh fish to be safe. Types A. thermosphacta is a well-characterised psychrophilic spoilage organism of meat. 5) reports that fresh fish consumption has never been shown to cause human botulism. E. Of 404 intoxication outbreaks of type E C. bacteria and higher parasites all present some risk associated with fish consumption (1. human viruses. especially type E botulinum intoxication. previous exposure and access to supportive treatment. where a particular 15 . including antisera. Of these. Huss (1. 5).6 Pathogens: Growth and Survival Endogenous chemicals. D.CHILLED AND FROZEN RAW FISH present in fish into histamine. it is bacterial risks that increase after capture of the fish. Adult disease is caused by the production of the botulinum toxin.6. B. thermosphacta did not produce off-flavours until 2 . 6). F and G) is well documented (44). 1. E. a neurotoxin that causes flaccid paralysis and is associated with a variable (44) mortality depending upon dose. botulinum. Acetate production has been reported as a good indicator of spoilage by this organism. 1. The heterogeneity of the types (A. this is probably due to spoilage occurring before toxin elaboration. 75% (with a 35% mortality rate) occurred in Japan. There are few human bacterial pathogens that can cause primary infections or disease and are capable of persisting in the aquatic environment.5. 1. thermosphacta in the spoilage of some MAP fish. E and F include bacteria with minimum growth temperatures of approximately 3.3 days after having reached 108 cfu/g (42). However. these risks are substantially reduced (1. but with proper cooking. C2.1 Clostridium botulinum C. a convenient but diverse species of bacterium with a number of different types. age. B.4 Brochothrix thermosphacta and lactic acid bacteria B. 5. Fewer still are capable of growing on fresh fish. they do not produce significant product spoilage). Growing evidence suggests a role for B. thermosphacta on spoiling fish in a 40% CO2/30% N2/30% O2 MAP (42). B. types B. The remaining few bacteria present a major risk involved with the consumption of raw seafood such as sushi or oysters. Some Ph. C1. lightly preserved fish products are associated with botulism. phosphoreum strains have great capacity as histamine producers even under refrigeration conditions. When inoculated into VP corned beef and sliced ham. Recent studies have investigated the dominance of B.3.
parahaemolyticus cases have increased across the world. 1. Human disease caused by the organism has only been observed in conjunction with marine bivalve (mostly oysters) and some crustacean consumption. Despite the ubiquitous nature of type E C. 1978 . but as fresh fish are often cooked before consumption and the botulinum toxin is heat-labile. The bacterium causes primary septicaemia. Southeast Asia. which is believed to be required for virulence. use of post harvest treatments and/or cooking of fresh seafood will reduce risks. V. salted or pickled) fish have also been associated with botulism. In Japan. and proper cooking (boiling for 1 minute. 5). Only about 1% of marine isolates produce a thermostable haemolysin. USA. smoked. including epidemics in India. sushi) (5). 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. accounted for the majority of cases (1.g. 11).2005). V. which was probably contaminated from an uncooked food source or an excreting foodhandler. The first report of the organism being a foodborne agent was with shirasu. Generally. there remains only an ‘extremely small’ risk of intoxication if reasonable precautions are taken during handling. Japan. I-sushi. vulnificus is associated with warm marine and estuarine waters.g. again. Russia. parahaemolyticus O3:K6 is responsible for many of the recent V. France.FISH AND SEAFOOD fermented raw fish product that is consumed uncooked. there is no specific guideline that describes a minimum level of V. which can cause human gastroenteritis. especially in individuals with underlying diseases (patients suffer from immune and liver diseases or blood 16 . parahaemolyticus outbreaks. parahaemolyticus infections since 1996 and is referred to as the pandemic strain (47). were associated with finfish between 1978 and 1998 (46). Since 1996. and when in contact with contaminated water (no CDC reported cases from fish. Other lightly or semi-preserved (e. is small. whether stored aerobically or in modified-atmospheres. Proper temperature control (<3. or cooking at 80 °C for 5 minutes) (44) will substantially reduce risk.6. storage and preparation (11). parahaemolyticus in sea water fish and shellfish that could potentially be hazardous to humans. the risk posed to the consumer of fresh fish. This strain has been responsible for 50 to 80% of all V. The risk of toxin formation before apparent spoilage has been studied for some MAP fish (45).2 Vibrio parahaemolyticus and other vibrios V. parahaemolyticus is a marine organism. Proper chilling. Currently. and North America. due to the cultural preference for raw fish dishes. A unique clone of V. only shellfish are associated with the disease and no reported cases by the Center for Diseases Control. botulinum in the marine environment (isolated from around 90% of marine and environmental samples from northern European waters and its consequent presence in seafood (a surveyed incidence of as high as 65%) (2.3 °C) will eliminate all risk. the majority of outbreaks are caused by consumption of raw fish products (e. the majority are associated with products not cooked immediately before consumption. a Japanese boiled and semi-dried sardine dish.
Again. 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. shigelloides is similar to Aeromonas spp. infection. 1. L. and especially estuarine environment. but proper cooking should substantially reduce risk of infection. Some isolates are true psychrophiles. Similar problems as those encountered for Aeromonas spp. There is no risk associated with the consumption of properly chilled and cooked fresh fish.4 Listeria monocytogenes L. They are aquatic organisms with an epidemiology that is yet to be fully understood. their potential to produce disease-causing exotoxin in food has not been fully examined (48). sodium channel blocking and haemolytic activity (48).6. monocytogenes incidence has been between 0. no more than circumstantial evidence exists linking the consumption of seafood to Aeromonas spp. possibly due to under-reporting because of their likeness to E. is historically associated with faecally contaminated water. It is known that Aeromonas spp.26 to 0. and proper cooking will substantially reduce risk. 1. 5. Chill storage may not eliminate growth. listeriosis cases have been reportable to the CDC. P. there is no risk from the consumption of properly handled and cooked fresh fish. Since 2000. the causative agent of cholera. are associated with human diarrhoeal illnesses. though it is a true mesophile with a minimum growth temperature of 8 °C and shows consequential seasonal variation in its environmental isolation. The major virulence factor appears to be the production of toxins with enterotoxic. Most cases have been sporadic rather than associated to large outbreaks.55 cases per 100. Although the psychrophiles are unable to grow at body temperature. 17). monocytogenes has been well documented as a foodborne human pathogen. with minimum growth temperatures around 0 °C and optima of 15 . Despite this. In the US.CHILLED AND FROZEN RAW FISH disorders). proper chill storage will eliminate any risk associated with this organism. The bacterium has been documented as being responsible for a few fishborne outbreaks of gastroenteritis..20 °C. which is often fatal (>50%).6. others are psychrotrophic mesophiles (49). however. coli on isolation media (50). and especially Aeromonas hydrophila. cholerae O1. and fish and shellfish are probably the major reservoirs for the organism (50). It is particularly associated with disease following consumption of raw oysters from warm sewagepolluted waters. cytotoxic. but the bacterium is known to survive and grow in the shallow marine. in assessing the risk of fishborne infection by P. shigelloides are encountered. are present on fish and can grow to significant numbers during storage (1.000 17 . V.3 Aeromonas Aeromonas spp. in its habitat.
Of fish-transmitted human diseases scombroid poisoning is commonly reported in the US (118 outbreaks with 463 cases reported by CDC 1998 . The organism poses a serious risk in chilled products not cooked before consumption. full risk analysis is not possible (52). Lightly preserved products. 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. some Vibrio spp. Low-temperature storage of potentially toxigenic (<5 °C) fish at all times is the most effective way to control histamine production (6). leads to the production of histamine.48 minutes in salmon have been reported (53). neonates.6. It is environmentally ubiquitous but its true frequency in the marine environment is not well studied.FISH AND SEAFOOD persons. 1. AIDS patients and patients undergoing immunosuppressive therapy) and a lack of data covering infectious doses. Owing to the dependence of the virulence of this organism on the immune state of the host (usually requiring lowered cellular immunity. Klebsiella pneumoniae and Hafnia alvei. scombroid poisoning is usually a mild intoxication and it is not a significant cause of death.. there is a possibility for growth of the organism on chill-stored fish. It is regularly isolated from seafoods (51). Photobacterium spp. especially pickled fish.pregnant women.2002 (55)) and is also common worldwide. 1. foetuses. Decarboxylation of histidine by a wide range of bacteria including Enterobacteriacae.6. 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). at risk groups . Like other toxins. it is not apparent to the consumer and it cannot be destroyed by cooking. D60 of 1.5 Scombroid fish poisoning The production of histamine and other biogenic amines with human immunological activity. are exceedingly difficult to produce within the legal limits of histamine as the temperatures used during production can lead to rapid growth of histidine decarboxylating bacteria. which has a maximum permissible level (for fish products from fish species associated with high amount of histidine) of 200 mg/kg fish in the EC.98 minutes in cod and D60 of 4. alcoholics. but proper cooking will reduce any risk. but cooking will significantly reduce the risk of infection. there is a far greater risk of unacceptable histamine levels being attained before sensory rejection. but more especially by Morganella morganii. Fortunately. Being a psychrotroph. If fish are not properly refrigerated. Clostridium spp. and Lactobacillus spp. 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...6 Parasites Helminthic parasites can occur extensively in finfish but very few are capable of infecting humans.
Cooking properly immediately before consumption is also an effective way of reducing or eliminating risk of fresh fishborne disease. Testing methods should be practical and the enforcement of such criteria should translate as a reduction of potential microbiological risks to consumers. Ababouch L.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.IV) (57). 1997) Huss H. 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. Faecal coliform counts may be used instead of E. 444.H. coli proposed by the International Commission on Microbiological Specifications for Foods (ICMSF)..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. including parasites and/or quantity of their toxins / metabolites per unit of mass. No. 1. especially in warm-water areas and where fish are to be consumed raw. coli counts where this method is preferred. Assessment and management of seafood safety and quality FAO Fisheries Technical Paper. Gram L.. parahaemolyticus Staph. In summary. long storage under refrigeration or one of the former prior to freezing. 2003. For fish from inshore or inland waters of doubtful microbiological quality. FAO. it may be desirable to test for Salmonella and V. risk of human disease caused by natural bacterial contamination (not through faecal pollution) of fresh fish is extremely low. The most effective control of all bacterial risks is continual chill storage in ice (0 °C) or freezing. An increase of APC to levels in excess of 106 cfu/g is usually indicative of inadequate refrigeration. EC. Criteria should set standards that are attainable by the currently accepted GMP and applied only when there is absolute need for it. Rome. volume. A limited number of nematodes and trematodes found in finfish have also been identified as a cause of human disease (56). 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. TABLE1.CHILLED AND FROZEN RAW FISH mildly processed fish such as sushi. area or lot (CAC. parahaemolyticus (see Table 1.1997. aureus 10 7 7 2 3 3 19 5 5 5 0 2 2 0 102 103 103 104 . which in many cases will eliminate the risk.
1992. 8 (2)..N..R.K.T. in FAO Fisheries Technical Paper No. 1992. Food and Agriculture Organisation of the United Nations. 67-71. 2001. Simpson B.A.E.K. Incidence of foodborne pathogens in European fish. Ed. 5 (1). Rosnes J.. Gibbs P. Huss H. 1997. Food and Agriculture Organisation.H. Assurance of seafood quality.H. Kirby R. in FAO Fisheries Technical Paper No. 36.J.. 334. International Journal of Food Microbiology. 1996. Rhodehamel E. 11. Huss H. Control of indigenous pathogenic bacterial in seafood. Feldhusen F.. Theoretical analysis of two hot water cabinet systems for decontamination of sides of beef. 107-27. Food Control. Ahmed F.A. 13. Davies A.. 348. International Journal of Food Sciences and Techonology. Fletcher R. 12. Capell C.. 12.. 4. 5.8 1. Ashie I. 10. 24. 1994. International Journal of Food Science and Technology. Davies A. Nychas G. M.R. 1994. 1651-60. Codex Alimentarius Commission. 1989. Code of Practice for Fish and Fishery Products (CAC/RCP 52-2003).S. 37.. Review: Assessing and managing risk due to consumption of seafood contaminated with microorganisms. Sivertsvik M. and natural toxins. Armstrong D. Spoilage and shelf life extension of fresh fish and shell-fish. Quality and quality changes in fresh fish. 7. Jehanno D. parasites. Food and Agriculture Organisation. 243-60. 20 .. Gram L. Dalgaard P. Davey K. 283-94.. 9. Kautter D. A review of modified atmosphere packaging of fish and fishery products – significance of microbial growth.J. 87-118. 14. Critical Reviews in Food Science and Nutrition.. Rome. References Huss H. 19. 1996. Smith J. Spoilage and shelf life of cod fillets packed in vacuum or modified atmospheres. 5-7.R.J. Gram L. activities and safety. 121-37. 1995.H.H..H.FISH AND SEAFOOD 1. 8. 291-304. 33. Food and Agriculture Organisation of the United Nations. 2003.A.. Reddy N. Huss H. The role of seafood in bacterial foodborne diseases. 2. Journal of Food Safety. 2002. Shelf life extension and safety concerns about fresh fishery products packaged under modified atmospheres: a review. Food Control. Codex Alimentarius Commission. Ed. 2000. International Journal of Food Microbiology. 3. 27. Food Control. Microbiological spoilage of fish and fish products.R. 2. 91-8.P.. 1993.E. 6. 87121. Incidence and growth of psychrotrophic Clostridium botulinum in foods. Jeksrud W. Italy. Rome. International Journal of Food Microbiology. Microbes and Infection. 12. Huss H..
Conway E. International Journal of Food Microbiology.M. 23. Flick G. Hendrie M. 419-29. AVI Publishing. 1987.. Letters in Applied Microbiology. Huis in't Veld J.C. Gram L. Martin R.E. Børresen T..J.N. Hebard C.A. 33.. Lipid oxidation in seafood. Sea Fish Industry Authority.J. 305-17. 29. in Fresh Fish. The micro-determination of ammonia. Pedrosa-Menabrito A. Gill T. Occurrence and significance of trimethylamine oxide and its derivatives in fish and shellfish. 1983. 21 16.... 1982. Washington D.. Flick G.E. 1959. 22-36. Ed. 1. 19. 42-54. 1995.E. Leblanc P. Edinburgh. National Advisory Committee on Microbiological Criteria for Foods.J. 117-27.J. Guidelines for the Handling of Fish Packed in a Controlled Atmosphere. Non-enzymatic oxidation of seafood lipids.G.E.H. Huss H. Journal of Agricultural and Food Chemistry. 1-18. 27.. Quality and Quality Changes. 245-67.edu/haccp/compendium/chapt08. Journal of Food Quality..J.. 1988. 36. Shelf life extension of fresh fish . Changes in purine nucleotide of red lateral muscle of rainbow trout.H. Biochemical Journal.M.E.. Microbiological spoilage of fish and fish products. March 20. 27.M. Surette M. An absorption apparatus for the micro-determination of certain volatile substances.CHILLED AND FROZEN RAW FISH 15. Quality and Quality Changes. 1985. 19-22. Huss H. 11. 21. 21. Nature. Reginstein J. Khayat A. Gill T. 28. Technomic. 1995. 25.J. Part I . 354-8.. 1415.E. 37. 121-37. Fate of Aeromonas and Yersinia on modified-atmospherepackaged (MAP) cod and trout.M.A.. 1995.J. Saito T.spoilage of fish.J. Arai K. 184. 2007.a review.. Eds. Bacteria active in the spoilage of certain seafoods. 34. Martin R. 130-2. 20. 24.H. Microbial and biochemical spoilage of foods: an overview. Schwall D. Journal of Applied Bacteriology. Yajima T. 1933. . Autolytic changes in fish post mortem. Hebard C. Westport. 26. Biochemical basis of post-mortem nucleotide catabolism in cod (Gadus morhua) and its relationship to spoilage. Herbert R. National Advisory Committee on Microbiological Criteria for Foods. Byrne A. 17. Dalgaard P. in Advances in Seafood Biochemistry. 30. 1992. 41-50. 18.ucdavis. Ratnayake W. Ed.htm Anon.H.. Eds.. 1971. Qualitative and quantitative characterisation of spoilage bacteria from packed fish. Martin R. Food Technology. 1992. in Chemistry and Biology of Marine Food Products. 33.S. in Fresh Fish. Lancaster. Adopted. Ackman R. http://seafood. Rome.R. Slade A. 26. 1995. Huss H. 22. Chemical composition. Flick G. Food and Agricultural Organisation of the United Nations. 1996. Shewan J. Gibson D.. International Journal of Food Microbiology. Vacuum or Modified Atmosphere Packaging for Refrigerated Raw Fishery Products. Rome. Food and Agricultural Organisation of the United Nations. International Journal of Food Microbiology. Davies A. 1996..
1973. II.J. 32. Dalgaard P. Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish.do?uri=CONSLEG:1996R2406:20050602:E N:PDF Sumner J. 334. Blackwell Scientific Publications. Journal of Bacteriology. Growth and activity of Shewanella putrefaciens isolated from spoiling fish. 33..H.. Rome. Nychas G.B. 38. Huss H. 1992.M. 174. Drosinos E.F.... 33. Miller A. 61-6. 1982... 323-9. Myers C. Do Tropical Fish Keep Longer in Ice Than Temperate Fish: The Circumstantial and Definitive Approaches. 1984. 109-36.. Dixon N. 13.L. 18-21. Brochothrix thermosphacta. 51-62. 34. Pseudomonas fluorescens.eu/LexUriServ/LexUriServ. Journal of Applied Bacteriology. Scanlan R. Sumner J.H. Do fish from tropical waters spoil less rapidly in ice than fish from temperate waters? Food Technology in Australia. Karnop G.E. 4... 2005.M. 62-70. 22 . 37. a dominant microorganism in Mediterranean fresh fish (Sparus aurata) stored under modified atmosphere.. 71. International Journal of Food Microbiology. 2002. Environmental Biotechnology. Brady P. The effect of storage temperature on shelf life. Cohen D.. Jørgensen B. Processes in Pathology and Microbiology. Fish spoilage bacteria – problems and solutions. 262-6. Nealson K. Italian Journal of Food Science. Food and Agricultural Organisation of the United Nations. Volatile compounds produced in sterile fish muscle (Sebastes melanops) by Pseudomonas putrefaciens. The inhibition by CO2 of the growth and metabolism of microorganisms. 6689-97. Libbey L. 72-80. 1989.R.. 311-43. Saffarini D. Magno-Orejana F. Archiv für Lebensmittelhygiene.J.A. and regulation. Venkateswaran K. in Fresh Fish. 1984. Applied Microbiology. 40. Taussig M.europa. Role of proteolytic bacteria in fish spoilage. 44.M. Gorczyca E. Applied and Environmental Microbiology. 9. 1985. Kell D. 39. Huss H. 41. physiology. FAO Fisheries Report.H. Localisation of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens.R. and an Achromobacter species.S. 36. Lee J. 67. Myers J. Satomi M. 36. 42. 43. 1996. Iron and manganese in anaerobic respiration: environmental significance. Gram L. 35. 1994. 48. Occurrence and importance of proteolytic bacteria as spoilage indicators. Ed. Oxford. Annual Review of Microbiology. 317. 3429-38. Dalgaard P.FISH AND SEAFOOD 31. 26. Gram L. 328-9. 1989.. Vogel B.. Quality and Quality Changes. COUNCIL REGULATION (EC) No 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products http://eurlex. Rome. 1995.H..
1973-1998. New York. International Journal of Food Microbiology.P. Thompson S.K.P. Loessner M.. 46. Ljungh A. Ray B. 57. 20. Myers M. 1-34.H. Histamine fish poisoning revisited. 1987. Fish and Fish Products (Microorganisms in Fish Products).. 23. Lindroth S. 85-95. Oxford.. Kluwer Academic/Plenum Publishers. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures.. 2006. 1991. 1986. Aeromonas and Plesiomonas as food.. 53. The International Commission on Microbiological Specifications for Foods. Garcia G.C... Journal of Food Protection.J. 47. Bishop R.. Wadström T. MacKinnon L.and waterborne pathogens.L. Applied and Environmental Microbiology. 2003. detection and growth of Listeria monocytogenes in seafoods: a review. Genigeorgis C. Microbial Ecology of Food Commodities.N. Olley J. Presence. 1991. Griffin P.L. Miller A. Sampling for Microbiological Analysis: Principles and Specific Application.. E and F in salmon fillets stored under modified atmospheres at low and abused temperatures.M. 2005. 1661-6. 181. Woodruff R. 51. Braden C. International Journal of Food Microbiology. 2000.M. 181-96. Surveillance for Foodborne-Disease Outbreaks . Majeed K.. Hammond R. Lynch M.. Whiting R. 181-8. Bean N.. 17-34. Huss H..K. 12... 50. Blackwell Scientific Publications. Food Technology. 23 . 50. Daniels N.M.. Modern Food Microbiology. 58. Springer. Jay J.A. Altekruse S.United States. Heat resistence of Listeria monocytogenes in vacuum packaged pasteurised fish fillets. 303-12. 56.. International Journal of Food Microbiology. 2000. 69. 55 (SS10). Painter J. PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico... 49. 52.H. 55. 2005. Golden D. 1997. Panicker G.W. 48.. Wilson S. Bej A. 2194200. Vibrio parahaemolyticus Infections in the United States. Journal of Infectious Diseases. Smith J. 12. Embarek B. Lehane L.A. USA. 1993. 54. Ed. Morbidity and Mortality Weekly Report. The International Commission on Microbiological Specifications for Foods. 1-37. 1998-2002. 100-3.C. International Journal of Food Microbiology.CHILLED AND FROZEN RAW FISH 45. International Journal of Food Microbiology. Use of risk assessment to reduce Listeriosis incidence. 330-6.K. Microorganisms in Foods 2... Embarek B.. 1994. Mac Rae I.J. Risk of growth and toxin production by Clostridium botulinum non-proteolytic types B. The International Commission on Microbiological Specifications for Foods. in Microorganisms in Foods 6. Slutsker L... 51 (4). 172-249.
. Handling and processing of scad.M. 1992.. 15. 1986. Olley J. Trolle G.. Mathematical Modelling Used for Evaluation and Prediction of Microbial Fish Spoilage. 1984.E. Journal of Food Protection. in Seafood Science and Technology.S.L.. Cann D. Villemure G. Cho Y. Huss H..H. 1989. Changes in microflora and chemical compounds in mackerel stored by partial freezing.. Food and Agriculture Organisation. 1984.. 127-34. 1997.. Eds. Microbiological quality and shelf life prediction of chilled fish. 741-50.G.A. International Journal of Food Microbiology..D.. Torry Research Station. in FAO Fisheries Report No. Jones Y. Sumner J.9. 55. Bulletin of the Faculty of Fisheries (Hokkaido University). Ed.D. 317 (Suppl. Huss H. Application of predictive microbiology to assure the quality and safety of fish and fish products. 317-21. 51.. Gibson D. Technomic Publishers.1983. 295-307. 1. 1989. 1961.1 Further Reading Processing Anon.P. Ravn Jørgensen B. 1985. Shinano H. 13-32. 73-89.H. Borgstrom G...H. International Journal of Food Microbiology. 1. Hobbs G. 35. 271-85. Ogden I. Akiba M. 1992. Studies on the microbiological ecology of mackerel stored by the method of partial freezing. Huss H.. 1.. Rome. Torry Research Station of the Ministry of Agriculture.. Bulk storage of cod fillets and gutted cod (Gadus morhua) under carbon dioxide atmosphere. Smith G. Ed. Gibson D. Fisheries and Food. Taoukis P. 1988.. Do Tropical Fish Keep Longer in Ice than Temperate Fish: the Circumstantial and Definitive Approaches. Journal of Food Science.M. Gram L. Picard G. Prediction of shelf life and safety of minimally processed CAP/MAP chilled foods: a review. International Journal of Food Microbiology.M. International Journal of Food Microbiology. Aberdeen. 65-72. 62-70. Magno-Orejana F. Torry Research Station of the Ministry of Agriculture. Further Studies on Marine Fish Stored Under Modified Atmosphere Packaging. 24 . Fisheries and Food. McMeekin T. in Fish as Food. Shahidi F.2 Spoilage Shewan J. The Microbiology of Seawater Fish. Dalgaard P.. Academic Press. Simard R.FISH AND SEAFOOD 1.9.). Food and Agriculture Organisation of the United Nations. Ross T. 6. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures.9 1.. in Torry Advisory Note No 93.C. Estimation of the bacteriological quality of fish by automated conductance measurements. Labuza T. Florida. Aberdeen.. Houston N. Lancaster. 4. Kitts D.
2197-203. Inhibitory effect against pathogenic and spoilage bacteria of Pseudomonas strains isolated from spoiled and fresh fish. Journal of Applied Bacteriology.. Amsterdam. Applied and Environmental Microbiology. Modified atmosphere packaging of fish and fish products. Otero A. 25 ..4 Standards and criteria Council Regulation (EC) Number 2406/96 of 26 November 1996 laying down common marketing standards for certain fishery products. 479-88. International Journal of Food Microbiology. Huss H.eu/LexUriServ/LexUriServ.. 137-46.9.. 322-6. 1992. Gram L. International Journal of Food Microbiology.9. 5. Elsevier Science. 4. 1.H. Garcia M. Effect of culture age.H.3 Pathogens Condon S. London. Gram L. Huss H.R.do?uri=CONSLEG:1996R2406: 20050602:EN:PDF Davies A. Detection of specific spoilage bacteria from fish stored at low (0 °C) and high (20 °C) temperatures. in Fish processing technology. Klausen N. 1997.H. Eds.CHILLED AND FROZEN RAW FISH Einarsson H. 1987... 1987. Jacobsen M.europa. pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila.. Trolle G. 1.. Sala F. Ed. Liston J. 1992. A rapid method for detection of histamine producing bacteria. in Quality Assurance in the Fish Industry. 20023.K. http://eur-lex. Blackie Academic & Professional. 65-72. Predicting the shelf life of cod (Gadus morhua) fillets stored in air and modified atmosphere at temperatures between -4 °C and +16 °C. Huss H. Hall G. 1993.J.L. 72.M. 59.
Sushimi tuna). 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. the exceptions being Sushimi (high quality raw fish).g. and crustacea such as shrimp and crabs. 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. although recent developments in improved supply chains. CHILLED AND FROZEN PREPARED FISH PRODUCTS Dr. seafood (with the exception of some crustaceans) remains uncooked until the point of consumption. Few seafood products are sold ‘ready-to-eat’.2. clarity of eyes. smell etc.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. This is no longer the case with many processed products now available in stores. but still acceptable and meeting all safety criteria. products of lesser quality. The majority of seafood products produced require a cooking stage before consumption.). it does not necessarily apply to the toxins that may have been produced. due to the early cessation of spoilage. are used in chilled and frozen prepared meals since many of the attributes on which quality is assessed are not present or noticeable (e. colour of gills. 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. smoked (cold) salmon. emphasising the high quality of the seafood and the gourmet nature of the prepared products. colourings etc. or could be disguised by sauces. smoked (hot) mackerel. 27 .g. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 2. Even though heating prior to consumption will eliminate pathogenic microbes. The same used to be said for the difference between fresh and frozen products.
Doubts have been raised over the microbiological status and handling practices of reef-caught fish. The market spans fresh fish counter sales and frozen recipe dishes with fish as the predominant source of protein. in particular. They can be with or without skin and may contain pin-bones although the major skeletal bones are removed during filleting. The expected shelf life attainable for this format is usually up to 8 days. or in MAP depends on catch conditions and treatment. Prepared natural fillets are fillets of fish without any additional coating. but these are now finding favour as products in their own right. often using the simplicity of a microwave oven. they are usually supplied filleted with head on. from both wild capture and aquaculture sources..2 Definitions The traditional focus of the chilled and frozen fish market has shifted over the last few years. Shelf life on ice. and pathogenic Vibrio spp. Seafood is the term used descriptively to include all categories of product. prawns. Wet or chilled/frozen whole fish are whole fresh fish that may or may not have been filleted. Microbiological hazards include Salmonella. which can be farmed. At the same time.. they are usually of a better standard than species such as tilapia. etc. It is sold from wet fish counters on ice or pre-packed in modified atmosphere packaging (MAP). They are typically flown into export markets. Frozen fillets may have a surface layer of 28 . but should be equivalent to cold-water products. 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. have shown the increasing importance of convenience foods. where the preparation has already been carried out. the processing industry has developed a wide range of value added products. Nevertheless. There is also a limited market for filleted material. This includes all categories of seafood including trout and other freshwater species. and this has produced a greater availability and diversity of products in the marketplace. as they tend to lose colour and sheen after defrosting. rather than frozen. Warm-water/reef-caught fish were initially supplied to add colour and variety to wet fish displays. shellfish and molluscs. Marketing-style descriptions and process definitions are presented below to acquaint the reader with the variety and forms of seafood available. Listeria spp. and these are increasingly being presented with an ever-expanding range of accompaniments. including freshwater fish. and are well known for their diet consisting of waste materials.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. molluscs. 2.
which may. but may be increased (up to 1 month) by packaging methods like MAP or VP. Hot-smoked fillets are prepared by brining fish fillets in a 26% salt solution with added natural colours to provide a range of product types. The result is fish with a typical residual salt content of 1. These products are packaged in separate high-care facilities to minimise contamination with microbes either from the workforce or from raw products. and salmon.CHILLED AND FROZEN PREPARED FISH PRODUCTS frozen water that forms a protective glaze. Hot-smoked fish are prepared from brined. or pre-packed in MAP or vacuum packaging (VP). and typically uses natural smoke passing over the fillets suspended on racks in a kiln (oak chippings. A core temperature of around 27 30 ºC for approximately 100 minutes is a typical processing condition. This process in effect cooks the product and reduces or eliminates microbial contaminants. The pâtés are sold from segregated ‘wet fish’ counters as a ready-to-eat product. the ingredients premixed. 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.2 days) longer than that of non-prepacked fish.. 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. Cold smoking is essentially a flavouring process. mackerel. mackerel.5 .g. resulting in a product that is ready-to-eat without any further cooking. salmon.5%. They are then kilnsmoked to a minimum core temperature of 66 ºC for 2 minutes. or pre-packed frozen with added glaze. be flavoured with herb dips or peppercorns. swordfish steaks). sawdust with other woods adjusted for flavour profile and burn duration).g. Again. and the product then shaped and heat-processed to cook the product before being chilled. natural fillets of fish such as herring. The expected shelf life of these products is up to 8 days (or longer for some chilled. Fish steaks can be cut from a natural fillet or loin (e. a protective glaze may be added to the frozen steak. They are sold from wet fish counters. Added colours can be included with the brine solution to meet specific requirements for products e. Fresh fillets are sold from wet fish counters. tuna steaks) or band-sawn from a frozen block (e. which is approximately 30% (or 1 . cod). Some products may be salted for flavour only (e. Cold-smoked natural fillets are prepared from a range of cold water fish including herring. A typical cold smoking process may pretreat the fillet by dipping in a 26% brine solution with a retention time of approximately 3 minutes. annatto with smoked haddock. in VP or through mail order. haddock. The shelf life for MAP fillets is typically up to 5 days.2.5% in the aqueous phase.g. 29 . VP products) because of the high salt content acting as a barrier to spoilage. etc. Pâtés are prepared with raw fish.g. in addition. with a minimum residual salt content of 3.
and formed into round cake shapes. ‘fingers’ before being coated in batter and/ or breadcrumbs. 30 . rice or pasta.g. and heat-fixed. e.) Raw material is typically fish fillet. mixed whitefish. tuna. Prawns. etc. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. which may include molluscs. Breaded fillets/steaks and battered fillets are either natural or shaped fillets. e. salmon) coated and sold as chilled products.. and seafood mixes. coated in batter or breadcrumbs. flavours.g.FISH AND SEAFOOD Seafood recipe meal centres (frozen or chilled) are places where these products are supplied without a source of carbohydrate. haddock. Surimi is the insoluble protein fraction of fish after washing and processing. long-life seafood mixes using sorbate and benzoate as preservatives. Other ingredients may be natural or reconstituted. and in the shell. binders. or cut from a preformed frozen fish blocks into shapes. Fish cakes are products produced by mixing fish with potato and herbs. The protein source can be fish fillet or blocks. This ‘catch-all’ category includes seafood salads. jellied eels. It is often used as a protein supplement.or a five-stage process. 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. frozen blocks or canned fish and may include molluscs or prawns that may or may not have undergone a minimal heat process. rice. pasta or potato. cephalopods. fish nuggets/bites with dips and seafood-based snack products. it can be used as a seafood analogue. taramasalata. The cakes are usually coated using a three. They are pasteurised products and treated as high risk raw materials. With further processing. and may also include additional ingredients such as fresh herbs and spices. Band-saw dust and fish trimmings from fishblock preparation materials may be used. Natural prawns from cold or warm waters can include those peeled.g. There is an increasing market for high value fresh or thawed fish (e. addition of ingredients such as colours. with an expected shelf life of around 7 days. Other. The majority are products made from various types of whitefish (cod. Scampi are tails of Dublin Bay prawn or langoustine.
The concern is that fish may be visibly acceptable but any subsequent washing process will have little effect on removing contaminants. which may become a focus of contamination deep within the muscle. The Food and Drug Administration (FDA) has produced guidelines for handling and processing of tuna. specifying internal temperature limits for fish and for limiting temperature abuse during handling and grading (1). skin. Dalgaard et al. 6). and ingredients added to the final product. and have emphasised the origin and sustainability of the fish as key marketing points. remain on the surface of the fish fillet except where “gaping” or separation of the muscle blocks occurs. Only after death. In summary. Grading is based on size of fish (>25 kg). and the potentially toxic by-products of histamine-producing bacteria (5.3 2.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2.1 Initial Microflora Seafood Other chapters have discussed in some detail the indigenous spoilage. on the whole. Tuna that does not meet the highest of grades based on colour. Frequent rebaiting and prompt removal of caught fish is essential to limit rapid spoilage. Recent public opinion has favoured the use of dolphin.e. 2). i. species (Bluefin>Bigeye>Yellowfin>Albacore). it may be used for other products or markets depending on the product specification. the microbial flora of seafood is associated with the gills. including seasonal variation. handling or preparation stage. is not used for sushi or other high risk (and high value) products. Contamination will. fat content. After capture and during any subsequent processing. and pathogenic flora associated with seafood. and processing conditions such as dehydration or poor filleting practices. see Table 2. product handlers. and intestines of the fish. or visible damage from gaff marks (4). A primary role in fish spoilage has been assigned to this organism. 31 . searing or bar marking). and an increased risk from a potentially unsafe product (especially those that receive a minimal heat process. other potentially pathogenic and spoilage bacterial contaminants may be introduced from a wide range of sources including the environment. and his co-workers have described a large celled microorganism found in spoiling fish and identified it as Photobacterium phosphoreum (3). For a summary of background information on the hazards associated with fisheries products and ingredients. as a result there is an increase in spoilage rate thereby reducing the product shelf life. whilst gaping is also dependent on a range of other factors that effect the muscle composition. sensory assessment of freshness etc. storage. Tuna caught and discarded as unsuitable for sushi or other high-risk uses is often diverted for canning.and turtlefriendly long lining to the less discriminatory netting. The edible tissue of some species of fish are more susceptible to gaping than others.I. will the saprophytic flora start to invade the surface flesh (1.3.
pseudomonads.3. Shigella. cereus Carbohydrates Potato flake and powder Rice Pasta Flour Dried herbs and spices Dairy products including butter. Pseudomonas. histamine Cryptosporidium. mushrooms. and. B.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. moulds Bacillus. Staphylococcus. staphylococcal toxin. protozoa and viruses B. coryneforms. by the HACCP team or by a hazard and risk assessment programme. enteric viruses 2.I provides an overview of common raw materials. Clostridium perfringens. intermediate processing. streptococci. and milk Cheese Water/Ice Bacillus. Bacillus subtilis B. Enterobacteriaceae. Clostridium Lactobacilli. Table 2. cereus Bacillus licheniformis. yeasts Pathogenic microflora/toxins Salmonella. In addition. Aspergillus. cream.FISH AND SEAFOOD 2. anaerobic spore-formers. Fusarium. Moulds Enterobacteriaceae Salmonella.4 2. cereus Staphylococcus aureus. Penicillium. moulds Bacillus Bacillus. and so are the potential microbial contaminants associated with them.4. Listeria. nuts and fruit Associated microflora Lactobacilli. toxin. TABLE 2. Vibrio cholerae. Escherichia coli. product assembly. shelf life determination.2 Non-seafood ingredients The range of ingredients used in the preparation of chilled and frozen prepared seafood products is very diverse. yeasts Lactobacilli. 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. raw materials. and associated spoilage and pathogenic microorganisms. moulds Bacillus. Aeromonas. however. moulds. coliforms.1 Processing and its Effects on the Microflora Selection of raw materials/ingredients It is important that the potential microbial contaminants of all ingredients. cereus B. spore-formers. Clostridium botulinum. Staph. be considered at an early stage during new product development. 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. for chilled products. Bacillus cereus. 32 .
All have the same intention. this may be an incidental effect. up to -5 °C so as to improve the cutting and handling properties during processing.4. For some components.3. 2.1 . Where the raw materials are frozen. 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. to evenly raise the temperature of the entire frozen product from -20 °C or lower. It is recommended that the principles of HACCP are applied to novel processes. Block-sawn dust that is required for use in fish cakes is swiftly transferred to a chiller to avoid water loss and spoilage from process contaminants.2.CHILLED AND FROZEN PREPARED FISH PRODUCTS Any preparation stage or process should be suited to the raw material. The use of water for rapid thawing of product is not recommended since it not only rapidly thaws the product surface. Intermediate processing and use of chill for storage of work-in-progress will help to optimise available life of perishable components. Effects on the microflora are minimal during tempering (7). as a heat process is often required for functional or organoleptic reasons. the core temperature. and potential opportunities for growth of foodpoisoning bacteria. as the full complexity of the processes used for specific products cannot be covered in a general text.4. but it also poses a risk of cross-contaminating other products undergoing tempering. but will also need to ensure that the optimum shelf life can be safely obtained. and contamination from equipment to clean fish muscle. Band-sawn materials or fish dice are normally promptly further processed. or the use of microwave technology. but it is assumed that any band sawing or dice process will introduce heat from blade friction. Generic examples have been used to illustrate only the major points. 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. Figures 2.1 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. Rapid controlled cooling of heat-processed components will help to minimise spoilage. line extensions and process reviews. defined holding time at a chill temperature (0 – 3 °C).5 illustrate basic principles in the preparation of chilled and frozen recipe dishes.4. 2. depending on the size of the fillet 33 .3 Preparation of fish component of chilled and frozen recipe dishes 2. there are a number of distinct methods used.
FISH AND SEAFOOD processed.2). Individual Quick Frozen (IQF) fish fillets or steaks may also be coated using a similar process. frozen in a nitrogen blast freezer and weighed into sacks for later processing. On arrival at the processing plant.4. The scampi meat (tails) is removed intact from the shell using potable water whilst still frozen. 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. quickfrozen. may not rise above 0 ºC during processing.1.Shaped Fillets. The “blown” meat is graded.3.4.1. Fig. are placed into plastic boxes and covered in ice to preserve their condition.1 (2.2.1 Whole-fillet / scampi Scampi (Nephrops norvegicus).1C .7 days. after landing. The processing of three examples types are described in Figure 2. 2.Fish Cakes). with the coating heat-fixed. 2. Any broken pieces or fragments of frozen tail meat are kept for use in the production of a reformed scampi product.Scampi. the scampi is washed in potable water to remove foreign bodies.1. 2. by mincing and mixing with a binding agent (as described in section 2.1B . It is then size-graded. chilled or sold chilled on defrost with an expected shelf life of 6 . Frozen cores are usually batter-enrobed.3. Raw material preparation stages for chilled or frozen coated fish products 34 . drained and then refrozen for storage until required for further processing.1A .
fish fingers). The mix is shaped and formed ready for packing or for enrobing.1. 2.1. as the physical structure of the material has been destroyed. because of its nature. iii) Fresh/fillet/minced fish block.. more seriously. pathogens such as Salmonella and Listeria. Fish raw material can be obtained from a number of sources i) By-product of fish shaping or cutting/dicing process . and water is added to rehydrate the components.4. It is for this reason that all by-products from primary processing. Where frozen blocks of fillets are used they will be “tempered” (temperature raised to around -7 °C) to facilitate efficient cutting with a band-saw to a specified thickness.off-cuts or belly flaps. such as aerobic spore formers and Staphylococcus spp. Therefore. The remaining dried ingredients are mixed with the fish.3. Waste fish generated as a by-product of another process will always be susceptible to spoilage and water loss. or those where the trimmings. additional spoilage organisms may be introduced.7 days.block-sawn dust. that are to be reused for other foods. 35 . material will need to be transferred swiftly to a chiller.2 Shaped products This type of product falls into two categories. those that use frozen blocks of whole fillets. but.4. with an expected shelf life of 6 . Spoilage of fillet pieces or trimmings will not be appreciably different from that of fillet. ii) Fish-filleting waste . Waste material produced from this type of processing is often reused in the third category (fish cakes). with the coating heat-fixed. Fish cake cores may be batter-enrobed. This is especially the case with block-sawn dust. Shapes may then be cut from the individual slabs using either hydraulic shape cutters or continued cutting with band-saw type blades (e. once the dry components have been rehydrated. This type of material will be susceptible to contamination by Enterobacteriaceae and possibly Staphylococcus spp. Herbs or seasonings used in the recipe may contribute additional spoilage organisms or.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. unless it is poorly handled. spoil more rapidly than the dry ingredients. quickfrozen. are handled and stored as any other food product. off-cuts and by-products from other processes (maybe minced) are reformatted give a final product that is still 100% fish. or promptly processed to avoid further quality loss. or sold as a chilled product after thawing.g. chilled.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. from potato flake. The fish component of the cake will.3.
g. and possibly Staphylococcus spp.or fivestage) frozen or chilled. If a crumb coating is required.2 illustrates the typical stages of enrobing or breading. spices. raising agents. starch. which is product-dependent and not always required. and to carry flavourings where required. and if required. a. or sold as a chilled product after thawing. Scampi tails are particularly susceptible to contamination with Enterobacteriaceae. A large variety of product formats and types can be batter-enrobed. i. egg. rusk or crumb. vibrios and other fishassociated spoilage organisms. Over time. flour. Natural fillets d. with the coating heat-fixed. The product then passes along a wire mesh belt through a ‘waterfall’ enrober that gives an even coating of batter to the entire product. lactobacilli from the milk powder and flour. Breaded or battered fish fillets are fully heat-processed before consumption. e.7 days. e. Fish products. because of the intensive handling required during processing. a characteristic flora will develop from the batter components.4 Battering/enrobing process Figure 2. The function of this is to improve the adhesion of subsequent coatings. with an expected shelf life of 6 . Bacillus spp. Products are preformed and presented to the process (either three. The first stage. chilled. from flour. Tempered band-sawn materials b..4. and Enterobacteriaceae from flour. which typically consists of flour. absorb any excess moisture on the surface of the fish portion.e. Batter contains water.g. IQF/pre-formed fillets c. some of this initial flora 36 . potentially. fish cakes Portions or fillets may be batter-enrobed. is the addition of a pre-dust. 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. any excess is removed with an air knife. quickfrozen. such as Listeria and Vibrio spp. milk powder and spices. 2. and may include milk powder. rapid automated processing with the minimum of delays will minimise product spoilage by pseudomonads. rapidly multiply if strict temperature control was not maintained. the crumb is poured onto the product followed by repeat enrobing and crumbing to complete the five-stage process. Control of temperature limits the potential for growth of spoilage organisms and pathogenic bacteria. which can occasionally be isolated from protein-handling processes. as well as ensuring that there is minimal contamination from human processors.FISH AND SEAFOOD In all of the above examples. Initially. Spore-forming anaerobes such as C. perfringens could.
Fig. 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. Staph. cereus can proliferate under certain conditions. but because batter is heat-set.CHILLED AND FROZEN PREPARED FISH PRODUCTS associated with the ingredients will be diluted out. and. Flow diagram outline of a typical three. but it is believed that poor hygiene regarding product and food handlers.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. dry batter mixes are all potential sources. this is not of great significance to the microbiological standard of the finished product. in the case of B. With inadequate temperature control of batter systems. aureus and B. cereus. The FDA has issued clear guidelines on acceptable 37 .2. contaminants such as Enterobacteriacae can proliferate and develop during the process. time-limitation or production run temperature control. It is advisable that some control mechanism is employed to prevent toxinforming organisms from multiplying to dangerous levels. replaced in part by process contaminants and bacterial flora from seafood. The origin of contamination for the two organisms is often unclear.2.
Flow diagram outline for recipe dish ingredients preparation 2.5 Recipe products These products involve the combining of seafood with other food ingredients as part of a recipe. and may include molluscs or prawns that may or may not have undergone a minimal heat process during their primary processing. pasta or potato. Vegetable quality is an important characteristic often overlooked in the selection of vegetables for frozen and chilled products. to produce a specific product.5. 2.FISH AND SEAFOOD temperature regimes and run times permitted for batter enrobers. There are two main categories.g. A temperature no greater than 10 ºC for 12 hours cumulatively or 21.1 ºC for no more than 3 hours is recommended (1).4. as this could introduce off-flavours and reduce shelf life. Well-managed breading and pre-mix systems that are segregated from batter enrobers present little hazard (7). Fig.2. For example. rice.4. 38 . The vegetables must withstand processing. but they are supplied without a source of carbohydrate e. Figure 2.3.1 Seafood recipe meal centres (frozen or chilled) These products can be defined as the central component of a meal. canned fish may be used in their preparation owing to it having wider availability and low wastage.3 illustrates vegetable processing and treatment for inclusion in chilled and frozen fish recipe dishes. The protein source can be fish fillet or block. and must not show evidence of disease.
rice or pasta. Although the processes are claimed to be superior to using unmodified hypochlorite for washing. Salmonella typhi.e. frozen block or canned fish. The presentation of vegetables has a significant effect on the product quality. loss of quality. all-year-round fresh vegetables are preferred. Washing with a biocide will at best achieve a 2 . chopping and mixing of material. Blanching of vegetables can be an effective means of reducing or removing surface contamination. will disperse the saprophytic flora of vegetables . and may include sauce accompaniments. and E. encourage more rapid decay and loss of structural integrity.5. coli O157. However. coryneforms. or breadcrumbs. For example. treated as a highrisk ingredient. and protected from any post-processing contamination. but also as components in recipe meals. for example. 2. Raw material is typically fish fillet. In an attempt to improve washing and reduce chlorine taint. 39 . Any assumptions regarding the application of good agricultural practice to produce should be made with caution. 8) will. and reduce spoilage organisms and potential pathogens. i. The opportunity for growth of L. The majority are products made from various types of whitefish (cod.CHILLED AND FROZEN PREPARED FISH PRODUCTS The products will often be presented with vegetable toppings. pastry. and may include molluscs or prawns that may or may not have undergone a minimal heat process.4. not just as accompaniments. they should be stored under temperature-controlled conditions. and mixed whitefish). the use of night soil or incorrectly applied manure or sewage sludge for ground preparation may contaminate produce with a wide range of pathogens. control is more onerous.3 log reduction in bacterial numbers on the surface of vegetables. Obvious signs of vegetable soft rot due to mechanical damage or infection by Erwinia spp. larger diced pieces allow for easier handling and a reduction in spoilage but are less attractive to the consumer. coliforms and pseudomonads throughout the mix. Products must be washed thoroughly to remove pests and soil.lactobacilli. haddock. spore-formers. (7. acidification of hypochlorite and chlorine dioxide have been proposed and recommended as an improved alternative. with the accompanying release of additional nutrients by processing. Once vegetables have been blanched. However. For premium products. monocytogenes should be taken into account when setting shelf life or holding times of vegetable components. even those with pH-modified dressings. A number of examples of direct faecal contamination of vegetables exist in the literature (8). with cases of foodborne disease traced to Listeria monocytogenes. colour and water will occur.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. This is a concern for chilled products that may have a shelf presence for up to 6 days including day of manufacture. if the process is not appropriately controlled. any processing stage. For example.
) to the product (12. and an increased risk of contamination from L. the preparation process must ensure the eradication of all vegetative pathogens from the ingredients. On hydration. 11). Placing such products in a standard chill store to cool down could result in condensation. Sauces or marinades are added to fish to broaden appeal or to accompany components. or use barrier or hurdle principles to preclude the growth of pathogenic microorganisms. 13). This is a particular issue especially where products are heat treated to eliminate vegetative cells. Typically. cereus). If the sauce contains modified starches for freeze/chill stability.1 Sauce manufacture The variety of ingredients and components used in sauce manufacture is extremely wide. Sauces can be multi-component preparations or more simple preparations. Chilled pasta is becoming more popular than dried pasta as a component in many product lines. monocytogenes and other bacteria that may be present on chilled surfaces such as ceilings. but temperature-abused pasta can also spoil rapidly by mould growth. B. a process equivalent to at least a 5D Listeria kill will be necessary. should be chilled to <5 °C as quickly as possible using either blast chillers or other equipment specifically designed for this purpose.FISH AND SEAFOOD Food-poisoning incidents traced to rice and caused by B. and then cooled slowly allowing germination and re-growth. 2. cereus are well known and are still a common issue in the UK (9). An incident involving Staph. if subjected to temperatures between 20 and 40°C. 40 . such as butter garnish with black pepper. For this reason all products. Alternatively. cereus. where composite meals are heat-treated. a heat process in excess of 73 ºC will be required to rehydrate this component fully. Products or dried ingredients contaminated with either spores of aerobic bacteria (e.4. Potato toppings made from flake or powder will normally have little associated spoilage flora. with the range always expanding as new tastes and cuisines are developed. spore-formers can proliferate if adequate control of temperature is not maintained.g. Spices and herbs added to form toppings along with breadcrumbs or other coating materials will similarly contribute a spore loading (primarily Bacillus spp. aureus toxin is well documented (10. rely on the use of microbiologically sound ingredients.4. Appropriate handling facilities or equipment will be required to protect the product.6. such as high-risk/low-risk barriers and dispensing and/or holding at above 63 ºC. can germinate and rapidly multiply. especially in those products that have a vegetable component or rice accompaniments.6 Preparation of non-fish components of chilled and frozen recipe dishes 2. Manufacturers of chilled products with an extended shelf life must recognise the potential for growth and spoilage of psychrotropic strains of B.
5 illustrate the recipe dish ingredients preparation and assembly and packaging. 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.herbs.2. Flow diagram outline for recipe dish ingredients preparation Time limitation may minimise the potential for spoilage by process contaminants or at the dispensing unit. flour products. Acidified or low-pH sauce formats using acetic acid or vinegar normally require a lesser heat process for the equivalent effect. The format uses flavour compounds . Marinade-type products are becoming more popular. Figures 2. 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. 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.4. etc. that of a dairy-based sauce.4 and 2. spices.CHILLED AND FROZEN PREPARED FISH PRODUCTS Fig. Without barriers. and handling of processed sauce can be less rigorous than. 41 . respectively. 13). for instance. suspended in an acidified sauce with an oil component.
The oxygen-free conditions will however allow other anaerobic bacteria such as C.1 Vacuum packing A range of fresh chilled products including salmon (smoked and fresh portions).5. the method replaces air in the sealed packaging with a defined mixture of gases.4. 2.FISH AND SEAFOOD Fig.4. as it dissolves in the surface moisture to form a weak acid. which may restrict either microbial growth or enzyme activity.7 Effects of packaging on chilled products Products with minimal additional ingredients may use packing techniques designed to minimise microbial growth. These include: 2. Increased levels of carbon dioxide will also result in a slight decrease in the pH of the surface tissues. so replacing the oxygen necessary for aerobic growth of spoilage microorganisms. botulinum to grow.7.2 Modified-atmosphere packaging Originally developed for the red meat industry.7. Flow diagram outline assembly and packaging of recipe dish 2. unless the temperature is strictly controlled to below 5 °C throughout the supply chain. The objective of this method is to deny the aerobic bacteria (both spoilage and pathogens) the oxygen that they need to grow. generally carbon dioxide and nitrogen. and tuna loins are packed by placing the product in a plastic pouch and removing all the air.2. 42 .4.
and Yersinia spp. but little on in-pack spoilage.5. Beyond this. by a loss of texture. 2. where the protein component is nearly always heat-processed. the slow development of ammonical and sulphuric ‘off’ odours and flavours(16). This is also true of chill storage and the distribution chain. (20). both organisms are capable of growth at chill temperatures.. The fish component of recipe dishes. and in influencing the final eating quality of the product. initially. However.10 ºC) Enterobacteriaceae are often found to proliferate and produce a characteristic type of spoilage that includes. have both been suggested as possible spoilage or pathogenic contaminants of chilled recipe meals (18). at slightly higher temperatures (10 ºC). advanced spoilage gives further changes in texture and the production of characteristic ‘off-odours’.5 Spoilage During processing. Aeromonas spp. It is not clear whether the additional contaminants from components or processing will accelerate this typical seafood-type spoilage. the microflora of interest will be mainly psychrophilic or cold-adapted (14) spoilage organisms such as Pseudomonas spp. but they will also be readily adapted to the nutrient source and conditions. but a conclusive case for pathogenicity has yet to be presented for Aeromonas spp. followed by moisture loss. phosphoreum. except for some crustacea.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. Not only will these organisms have a numerical advantage over other proposed spoilage organisms. Spoilage of this material is not expected to differ greatly from the processes described elsewhere. temperature control is important in preserving chilled integrity of ingredients. Vibrionaceae will succeed (17 ). 43 . Nonetheless. in numerical terms. The literature has good information on spoilage of individual components of recipe meals. as this organism is a common contaminant of freshwater fish species (19). This is considered to be the one major difference between fish and meat products. is typically uncooked or at best lightly processed to retain eating characteristics. The case for growth and contamination of products with Aeromonas is stronger. Spoilage will be most readily detected in proteinaceous components of ready meals. Shewanella putrefaciens and Ph. but is not limited to. At refrigeration temperatures (5 .1 Chilled storage For chilled products or chilled components of frozen products. a large number of nutritionally non-fastidious spoilage organisms are commonly isolated from seafood. and finally flavour loss. and therefore will have a greater opportunity to dominate other spoilage flora numerically over the course of product shelf life (15).
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). As a note of interest. Bacillus spp. this should also take into account shelf life of product components used in the assembly of the final product. which would warn the consumer of the potentially unsafe nature of the product.. For example.FISH AND SEAFOOD 2. By retaining the intrinsic spoilage flora of seafood. auto-oxidation of fats/oils). It is not unreasonable to expect that spoilage bacteria release many extracellular enzymes. eliminating nematodes in products that do not have a heat treatment step. the shelf life of products is limited by the natural spoilage process described earlier. However. freezing also has a protective role in improving the safety of raw materials by. 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. released by a pseudomonad with demonstrable activity to 0 ºC and a peak in production at 10 ºC (15). Spoilage can be either a function of bacterial growth or the microbial release of free enzymes into solution. where the variety of components and ingredients used in manufacture is varied and complex. for example. Products that are developed with an extended shelf life beyond the 6 . producing characteristic off-flavours. This also ensures that any post-process bacterial contamination or spore-forming bacteria in the product will not grow. and Aerobacter spp.g. a recent report examining fish gut bacteria from cold waters characterised a proteolytic enzyme. The type and nature of the enzymes and their activity will depend on the bacterial species. within dairy products are well known. 44 .3 Enzymic spoilage Post-harvest changes in the quality of fish due to enzymic activity and rigor mortis are well known and understood (10). Serratia spp. Still.5. These have been proved to impart off-flavours. spoilage should not be considered a purely microbial phenomenon as taints and off-flavours can develop by non-microbial means (e. however. This reinforces the need for prompt processing and temperature control of ingredients and finished products. The conditions and freezing process most favourable for good-quality products will.5.2 Frozen storage Maintenance of product temperature at <-18 ºC will not permit bacterial growth or enzymic spoilage. 2. botulinum.. Pseudomonas spp.7 days typical of fish products/recipe meals must consider the potential for growth of non-proteolytic C. Other examples of bacterial proteolytic and lipolytic enzymes produced by Alicaligenes spp. Characterisation of the enzymes has demonstrated residual activity even after a heat process that would destroy vegetative cells (24)..
45 . 2. typhi. spices and herbs and vegetable shoots have been reported (12. air. and some effects related to product type and time to toxin production have been noted.CHILLED AND FROZEN PREPARED FISH PRODUCTS 2. cereus. 13.. Inappropriate holding times and temperatures will. and many raw or processed foods (26).2 Clostridium botulinum There are reports of rapid spoilage of vegetables by C. Germination of spores requires favourable conditions. These are C. Hazards attributable to the non-seafood components are listed below and this information. low oxygen or anaerobic conditions. etc. any risk of C.1 Bacillus cereus This is a common contaminant of dust. 2. agricultural run-off or sewage can contaminate in-shore species. or a heat shock followed by holding at a temperature suitable for growth. perfringens contaminating seafood will depend on the source of raw material. Therefore. have been linked with food poisoning and should also be considered. Cases quoting contamination of seafood with organisms such as S. botulinum when held in favourable conditions for growth (28) i. with pathogenic microorganisms or viruses. along with previous discussion. are dealt with elsewhere in this book. and Campylobacter are recorded in filter feeding molluscs such as mussels. especially molluscan filter feeders. histamine intoxication. in some cases. Additional hazards from marine bio-toxins. Incidents in such diverse materials as cream. botulinum type E and Vibrio parahaemolyticus (25).6 Pathogens: Growth and Survival There are only two species of pathogenic bacteria that can truly be said to occur naturally in fish. A number of other Bacillus spp. soil.104/ g should be treated with some concern (26). perfringens in seafood noted a transient population of spores in the guts of fish feeding adjacent to rivers and sewage outfalls (29). Counts of between 103 .3 Clostridium perfringens A study examining C.6. permit rapid growth of B.6. 2. 27). Vibrio spp. Numerous quoted figures exist in the literature regarding cell counts required before toxin production occurs. In addition. This may occur where storage under gas such as carbon dioxide is used to reduce ripening and so prolong storage. may be of benefit within any risk assessment for chilled and frozen prepared fish products. and this issue may be of greater consequence for warmwater. farmed or free-living crustacea (30).6. water.e. potato.
Chilling may help Shigella to survive. L. All the above points should be considered within any risk assessment of the potential for growth and toxin production by Staph. An acceptance has now developed in the UK of the presence at low levels of L. and will therefore cause “food poisoning” even in a product that is cooked before consumption. These include inadequate refrigeration and preparing foods far in advance of planned service. aureus (35). 46 .4 Listeria monocytogenes Interest in this organism has been characterised by a number of very serious incidents associated with a variety of product types (27). However. aureus enterotoxins in foods.6. 33) due to its ability to grow at low temperatures and form “biofilms” on surfaces. 2. indigenous or other. infected food handlers. at the present time. and inadequate cooking or holding within the temperature growth range (27).6. when introduced or inoculated into sterile components of chilled prepared recipe dishes. Rates of contamination with smoked seafood are much higher than those with unsmoked products. and this is believed to be due to a tolerance to brine used in the smoking process (32). the restrictions on growth rate seen with seafood will not apply.6. monocytogenes as a contaminant in foods (31). but under the appropriate conditions. 2. Sources of contamination are many because of the widespread distribution of this organism in the environment. Shigella spp.6 Staphylococcus aureus There are a number of underlying reasons for the development of Staph. It is unclear. will colonise factory environments in a similar fashion except for possibly Vibrio spp. These conditions will allow the production of the toxin. if any other potentially pathogenic organisms. the possibility of a sanitiser-resistant biofilm colonising processing environments should be viewed with some concern (32). 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. growth may occur in foods (26). monocytogenes has been frequently isolated from fish-processing plants and is thought to be able to colonise this type of environment (32. monocytogenes tends to grow poorly at refrigeration temperatures in seafood (33). 2.FISH AND SEAFOOD Vegetable raw materials are potentially contaminated through soil conditioning with animal manure. which is not affected by any subsequent cooking process.5 Shigella spp. L.
pastry. etc. but provide the industry with an indication of the microbial level at which action is required to ensure product safety. unless the criteria is for a pathogenic organisms where only non detection is acceptable.A dairy-based sauce that includes a non-pasteurised cheese will contribute significant aerobic total counts. 2.7 Viruses Concerns over contamination of seafoods with viruses (due to species-specific nature of viruses) are generally limited to molluscan shellfish from waters or crops contaminated with sewage. are not mandatory.III based on IFST Guidelines (39)) provide example guidelines for seafood products. and their association with similar materials. Microbial criteria and the monitoring of microbial levels within a product are important for monitoring trends within production systems. the Institute of Food Science and Technology (IFST) (39) or the British Retail Consortium (BRC) (40). although the only seafood products specifically identified are raw “shelled and shucked products of cooked crustaceans and molluscan shellfish”.7 Published Microbiological Criteria Due to the complexity of many processed food products. with respect to the number and types of ingredients used. but these may be eliminated by a subsequent heat-treatment stage. cereus emetic toxin are often confused owing to similar symptoms. Microbial guidelines. such as batter mixes. For instance. The following tables (Table 2.CHILLED AND FROZEN PREPARED FISH PRODUCTS Illnesses caused by Staph. . The criteria themselves will have a range of levels that are dependant on the point in the processing or supply chain at which they apply. 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 not to final products.6. although due to long analysis times required. (36). aureus enterotoxins and B. 2. and as such the monitoring is for verification purposes rather than control.II and 2. Microbiological specifications do not necessarily relate to product safety. 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. pasta. the fresh product is often consumed before the results are obtained. 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. as published by for example. and are a recognised means of providing data on manufacturing standards. In the EU these are defined under European Commission (EC) Regulation 2073/2005 (38). It should be remembered that 47 .
TABLE 2. coli GMP* <104 <102 <10 Maximum Product-dependent 104 103 GMP* Not detected in 25 g Not detected in 25 g <102 <20 Not detected in 25 g <50 ppm (<5 mg/100 g) Maximum Not detected in 25 g 103 104 103 102 <50 ppm (<5 mg/100 g) *GMP = Values found immediately after production under good manufacturing conditions. cereus Staph. fish fingers. pasta. aureus V.III IFST Guidelines for processed foods . Maximum= Levels at end of stated shelf life 48 .II IFST Guidelines for part-cooked foods – e. monocytogenes B. coli GMP* <102 Maximum 104 TABLE 2. batter etc) should also be consulted when establishing the specifications for raw materials and final product.g. fish cakes Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Pathogens GMP* Maximum Criteria for absence not generally applicable Indicators and spoilage organisms Organism E.FISH AND SEAFOOD the microbial criteria for other products (rice. L. parahaemolyticus (warm-water fish) Histamine (scrombroid fish) Indicators and spoilage organisms Organism APC (heat treated) Enterobacteriaceae E. ready meals Storage: Frozen/chilled Use: Re-heated prior to consumption Pathogens and toxins Organism Salmonella spp.fish products. coated fish.
. Hackney C.. Aureli P. International outbreak of staphylococcal food poisoning caused by contaminated lasagne. Animals and Spoilage. International Commission on Microbiological Specifications for Foods. Journal of Food Science. 7 (24).G. Ltd. Journal of Hygiene. Collette R. Food Science and Technology Today. Kluwer Academic. 12. Journal of Milk Food Technology.R. 7. England and Wales Quarterly Report.M.L. Beuchat L. 59 (2).. 49 4. Letters in Applied Microbiology. Toxin Related Diseases: Poisons Originating from Plants. Brown T. 8. Food and Drug Administration. Taylor S. 70-2. 1.M.... Cambridge. 3. 2005. 215. 11. 9. Pathogenic microorganisms associated with fresh produce. 6. Van Nostrand Reinhold.T. Bacterial histamine production as a function of temperature and time of incubation.CHILLED AND FROZEN PREPARED FISH PRODUCTS An authoritative survey of specifications can be found in Shapton & Shapton (41). Martin R. Fish: microbiological spoilage and safety.C.C.. 1993. London. 1991. 166-73. 5. Fish and Fisheries Products: Hazards and Controls Guidance. Microbiology of processed spices. Outbreaks of foodborne illness in humans. Dalgaard P.. Pvt. Oxford. 1976. 47.K. Communicable Disease Report. Masuoka Y. 663-87. Huss H. Journal of Food Protection... Kumeta H... Hoshino T. in Fish Inspection Quality Control and HACCP: A Global Focus. Tu A. Washington D. Woolaway M. References US Food and Drug Administration. 373-8. Ohgiya S.J. 1997. Ishizaki K. 38 (11). Communicable Disease Report. Behling A. 10.J.8 1.. Incidence and levels of Bacillus cereus in processed spices. . Microbiology of Marine Food Products. 1991.W... 2001. Journal of Milk Food Technology. Lawyer R. 25 (1). Wienke A. 1311-7.L. 1997. Slavin J. 67-73. 15. Microorganisms in Foods 6: Microbial Ecology of Food Commodities. 14.R. 1976. Proper processing and handling of raw tuna. 1986. Bartlett C. Shewan J.R.. New York. 2. 204-16. 24 (5). 1962. Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. Latt T. Hobbs G. Recent Advances in Food Science. Eds. 96. Christiansen T.L.R.A.. 668 -70. 1982. 233-7. Mejholm O. Ward D. Sakamoto T. Importance of Photobacterium phosphoreum in relation to spoilage. Yumoto I. London. 1997. Letters in Applied Microbiology. The bacteriology of fresh and spoiling fish and some related chemical changes. 167-93. 2.R. Matsuyama H. Murrel H. IBH Publishing Co.. Taylor and Francis. Gilbert R. Bartram P. 39 (10).. 5 (3). Powers E.. 13.C..H.. 1997. Powers E.M.E.
Blackwell Scientific Publications. 2000. International Journal of Food Microbiology. Ray B. 24.. Leatherhead. SAB Symposium Series No. 11. 1988. 1977. International Commission on Microbiological Specifications for Foods. 1992. 25 (2). 309-16. 27.A. perfringens? International Journal of Food Microbiology.. PHLS Microbiology Digest. Mead G. 13 (1). 1996. Oxford. Effects of freezing and storage on microorganisms in frozen foods: a review. Fresh and processed fish and shellfish . 22.J. 59-72. Huss H.J. 21. Lund B. 1979.R. 28. 65. Blackie Academic and Professional. Modern Food Microbiology.E.L. 25.M. Leatherhead Food Publications.. Aspen Publishers. Halligan A. 655-60. 1974. Applied Microbiology. Journal of Food Safety.C. 1995. 32.H. Eds. Wood L.. 1996.H. 9. Leistner L. Speck M. Food Research International. Eds. 31. Occurrence of Listeria monocytogenes in fresh water fish farms and fish smoking plants. Keusch A.. 1993. Ronner U. Journal of Food Protection. 51-158 Saxby M. Curran D. 19. Huss H. in The Microbiological Safety and Quality of Food... Morgan D.. Jemmi T. Aeromonas and Verotoxigenic E. cereus and C. Clostridium perfringens in the environment. Granum P. London. Food preservation by combined methods. Report on Vacuum Packing and Associated Processes. Microorganisms in Foods 5: Microbiological Specifications of Food Pathogens. Food Microbiology. 274-86. Gram L.W. Microbiology in fishery science. Trolle G. 41-3. 29. Food Taints and Off-flavours..C. Glasgow. Food Control. 26. New York... Liston J. Anaerobes in relation to foods of plant origin.Yersinia. The Emerging pathogens . Microbiological guidelines for some ready-to-eat foods sampled at the point of sale: an expert opinion from the Public Health Laboratory Service.V. Gram L. coli (VTEC) . Andersson A. What problems does the food industry have with spore forming pathogens B. 4. Jay M. Detection of specific spoilage bacteria from fish stored at low (0 ºC) and high (20 ºC) temperatures. Gaithersburg. 333-6...K. Baird-Parker T. 1990. Brooks J.A Literature Survey.. 50 17. 28. Advisory Committee on the Microbiological Safety of Foods. Connel J. Ed. in Anaerobic Bacteria in Habitats Other than Man. 4 (1). Matches J. 1986. 20. 18. 1987. 40 (5). 28 (4). 472-506. Gould G. Loessner M.J. Blackie Academic Press. 1992. 155-9. 34-40. 351-72. 23. Liston J. Barnes E.J. 30. London. Golden D.C. 13..J. 1996. a foodborne pathogen? Review of the clinical data. 1994. Survey of the incidence of Aeromonas and Yersinia species in retail foods. Springer.R. 2005. Lund B. U. Walker S. Her Majesty’s Stationery Office. . in Advances in Fish Science and Technology.M. Is Aeromonas spp. Fishing News Books Ltd. Anon..FISH AND SEAFOOD 16.
2006.europa. London. Guidance on the Practical Implementation of EC Regulations on Microbiological Criteria for Foodstuffs. 50 (9). 39. Journal of Food Protection. http://eur-lex.G. 41. 372-6. McCarthy S. Gloucestershire. British Retail Consortium. Psychrotrophic Bacteria in Foods: Disease and Spoilage. 1987..F.L. Varnam A. Journal of Food Protection. 37. 35. 1991. 788-801.A.H. http://www.pdf Shapton D.eu/LexUriServ/ LexUriServ.do?uri=CELEX:02005R2073-20071227:EN:NOT Institute of Food Science and Technology. Development and Use of Microbiological Criteria for Foods. 1999. 1997. 38. Microbiological Criteria. Kraft A. Cambridge. Wolfe Publishing. Shapton N. Chilled Food Association. U.K. Phil Voysey.chilledfood.org/Resources/Chilled%20Food%20Association/Public%20 Resources/BRC_CFA_Micro_Criteria_Guidance_Ed_1. Smith J. 34. 36.A.uk/cday/posters2008/npd6. Incidence and survival of Listeria monocytogenes in ready-to-eat seafood products. 40. http://www. 51 .2. Foodborne Pathogens an Illustrated Text. CRC Press.CHILLED AND FROZEN PREPARED FISH PRODUCTS 33.pdf Commission Regulation (EC) No 2073/2005 of 15 November 2005 on Microbiological Criteria for Foodstuffs. Shigella as a foodborne pathogen. 1998.A. 1992.K. Chilled Food Association. Principles and Practices for the Safe Processing of Foods. U. 60 (4). London.. Institute of Food Science and Technology. Evans M.co.campden. Campden & Chorleywood Food Research Association. Woodhead Publishing Ltd.
3.S. In these forms. Bivalves feed by filtering large volumes of water across their gills to obtain oxygen and food. octopi. clams. Pelecypods have little need for locomotion. Pelecypoda.1. can hold the shells tightly shut against enemies (1). slugs. cockles and scallops. where it is eventually swallowed and digested. completely encloses the body. and Cephalopoda. the foot is compressed to form a muscular spade for digging and the head is greatly reduced. Jr. Food and Drug Administration 22201 23rd Dr. chitons. Gastropoda. Many microorganisms ingested by shellfish survive the digestive process (3. In most species. chemical contaminants and other impurities on the mucus of the gills. clams. which includes oysters. mussels. MOLLUSCAN SHELLFISH Carlos Abeyta. scallops.1 3. snails. oysters. mussels. squid. and scallops. Box 3012 Bothell WA 98041-4421 United States of America 3. Other species of shellfish such as crabs. they also trap bacteria. the shell. Strong muscles. cockles.E. The class having species of economic and public health importance is the Pelecypoda. and 2. For example. Pacific Regional Laboratory Northwest U. whelks. abalones. viruses. Scaphopoda. S. and tooth shells are a group of soft-bodied animals that usually secrete external protective shells.1 Definitions General features of the mollusc The phylum Mollusca. lobster. oysters and clams can process about 3. the adductors. and shrimp. while 53 .7 litres of water per hour. Special tracts of cilia move this mucus toward the mouth. The class Pelecypoda includes all of the bivalves such as clams. P. Plankton and algae form the bulk of their diet from seawater. composed of two valves hinged together dorsally. lying well within the mantle cavity. respectively. The term shellfish in this context is limited to oysters. In the process of filtering water. O. The phylum consists of five classes: Amphineura.6 litres. The digestion process requires less than 2 hours in actively feeding adult bivalves (2). 4). cockles. mussels.
South America Pacific – Japan Quahaug (also known as hard clam and hard shell clam) Coquina clam Hen clam Butter clam (also called the Washington clam) Mogal clam Little neck clam (also called rock Cockle Pacific littleneck) Atlantic – North American/Europe Atlantic – Europe/North America Japan Atlantic/PacificNorth America Donax spp. The habitat of oysters and mussels are sessile. The following are descriptions of the various bivalves and their forms of preparation for human consumption (6): TABLE 3. but maintain contact with the water by means of a siphon tube (5).North America Saxidomus nuttali Venus mortoni Protothaca thaca Meretrix spp. many species of bivalves are edible (Table 3. Whatever contaminants are in the water will eventually get into the shellfish.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 .1. Clams are benthic and live burrowed in the mud. and usually at higher concentrations than found in their habitat. 3. Mactra sachalinensis Saxidomus giganteus Japan Pacific – North American Saxidomus nuttali Protothaca staminea or Paphia staminea 54 .2 International commercial products Globally.I). are not as vulnerable to contamination through pollution of their beds as the bivalves. 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.FISH AND SEAFOOD commercially valuable.
Pacific – North America/ Australia/New Zealand Pacific – North America Crassostrea gigas Western oyster (also called Olympia oyster) Cockle Cockle Sydney rock oyster Rock oyster Dredged oyster Cockles Common cockle Ostrea lurida South America Japan Australia New Zealand New Zealand Ostrea chilensis Ostrea laperousei Crassostrea commercialis Crassostrea glomerta Ostrea lutaria North Atlantic – Europe/ North Africa Pacific – North America Cardium edule Cardium corbis Cardium aculeatum Cardium tuberculatum Spiny cockle Knotted cockle Mussels Blue mussel Atlantic/Mediterranean Atlantic/Mediterranean North Atlantic – Europe Pacific – New Zealand Pacific – North America Europe Atlantic/Mediterranean Mytilus edulis Common mussel Horse mussel Bearded horse mussel Mytilus californiaus Modiolus modiolus Modiolus barbatus 55 .MOLLUSCAN SHELLFISH Gulf clam Pismo clam Surf clam (also called bar clam) Freshwater clam Oysters Common oyster Portuguese oyster Blue point oyster (also called American oyster) Pacific oyster Europe Europe Atlantic – USA Ostrea edulis Crassostrea angulata Crassostrea virginica Gulf of Mexico Pacific – North America Atlantic – North America Titaria cordata Trivela stuttorum Spisula solidissima Japan Corbicula spp.
1. and two symmetrical shells (valves) held closed by two large muscles and joined by a dorsal hinge joint.FISH AND SEAFOOD Mussel Mussel Mussel Scallops Scallop Coquille St. geoduck. extended above the surface to maintain water currents needed for respiration and feeding. cherrystone. Buried clams leave tubes. hard-shell or soft-shell clam. littleneck. Most clams are only a few centimetres in their maximum dimensions. mostly marine bivalve molluscs. Jacques Iceland scallop Bay scallop Weathervane scallop (also called Alaska scallop) Atlantic North East Atlantic North Atlantic West Atlantic Pacific – North America Pacific – North America Pacific – Japan Common scallop Sea scallop (also called giant smooth) Japan Atlantic – North America Pecten varius Pecten maximus Chlamys islandica Argopecten irradians Pecten caurinus Pecten aequisulcatus Pecten laquaetus Pecten yessoensis Penten magellanicus or Placopecten magellanicus Pecten meridionalis Pecten novaezealandiae Pecten jacobaeus Aequipecten gibbus Chlamys opercularis Chlamys varius Amusium balloti Mediterranean – South East Europe New Zealand Australia Mytilus galloprovincialis Mytilus canaliculus Mytilus planulatus Commercial scallop Australia New Zealand Great scallop Calico scallop Queen scallop Variegated scallop Scallop saucer Atlantic/Mediterranean Atlantic – North American Atlantic Atlantic/Mediterranean Australia 3.1 Clam Clam is the common name of a large group of often edible. or siphons. Different species are variously called quahog.2. laterally compressed. A mantle tissue encloses the body of the clam. and other names. The muscular foot is used to burrow in mud or sand. but 56 .
boiled and then sun dried for 3 to 10 days before packing in boxes (Hong-Kong) Smoked: meats usually canned in edible oil.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. Dried: shelled meats. all in cans. having rough and irregularly shaped shells that are closed by a single adductor muscle. uncooked. washed. then heat-processed. Live: in shells. Soups: oyster stew. oyster soup. Semi-preserved: meats cooked and packed in spiced vinegar. or shelled meats. either whole or minced. the meats are smoked and packed in oil in cans or jars. 3.1. Live: in shell Fresh: in shell or shelled meats. Canned: meats removed from the shell by steaming. Dried: Hoshigai (Japan) shelled meat.2. uncooked (shucked). clam madrilène and clam liquor. Frozen: shelled meats. Smoked: meats. tidal waters or in brackish water in the mouths of rivers. They live free on the bottom or adhere to stones or other objects in shallow water along the seacoasts. Fresh: in shell. hermetically sealed. skewered with bamboo sticks. in cans with either brine or clam liquor. uncooked meats packed unprocessed. Oysters lack a foot or have a reduced foot. after precooking and brining. Canned soups include clam chowder. Frozen: in shell. trimmed and packed. These molluscs feed on minute plants and animals carried to them by the current. 57 . or shelled meats. and no siphon. sun-dried. oyster bisque. packed in weak brine. smoked meats are also canned. Canned: clams are steamed open and the meats removed from the shells.
in which the individual branches. Salted: cooked meats bottled in brine. scallops can swim or leap by a series of rapid closings and openings of the valves. vinegar solution. scallops possess a single large adductor muscle. the shell is often characterised with a lustrous nacreous or pearly lining inside their shells. Fresh: cooked meats. or packed with jelly. boiled. with the posterior ends of the shells (valves) broad and rounded. Unlike other sessile bivalve molluscs. Internally. Canned: cooked meats. In some species. composed of striated muscle fibres (used for fast swimming) and unstriated muscle fibres (used to hold together the two halves of the valves). mussel in butter sauce. The common scallop is about 5 cm long. Semi-preserves: cooked meats with vinegar-acidified brine and spices. Bait: used live extensively for baiting fish hooks for line fishing.FISH AND SEAFOOD 3. Smoked: meats may be canned in edible oil. gutted. and held apart. mussel paste. Fresh: shelled meats (often only the adductor muscle and the roe are eaten. sauces.) Dried: peeled.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. are united.4 Scallop Scallops are marine bivalve molluscs living in shallow waters of protected bays with sandy or muddy bottoms. The adductor muscle is the only edible part of the animal. The shells are fluted and round with a broad. brine.2. also with other ingredients. by interlocking tufts of hair-like cilia. They have a radially ribbed shell with the edge undulated and swim by opening and closing the valves. 3. 58 . flattened ear or wing at the hinge area.1. The posterior halves of the valves contain a large adductor muscle that holds the valves closed. and the anterior end smaller and pointed. in mussel liquor.1. smouldered and afterwards dried (Japan). They have filibranch gills. Live: whole with shells. cooked meats packed in dry salt for transport. or filaments. The familiar marine mussel is asymmetrical.2.
Live: in shell.1. depending on length of journey. and the environment. pH. salinity.2. nutrient concentration and pollutants of the water column. they accumulate pathogenic microorganisms from polluted waters. 3. Vinegar cured: bottled in malt vinegar after brining. and butter. also bottled in brine. Because molluscan shellfish are filter feeders. Fresh: meats removed from the shell by boiling. The microflora are dependent upon a number of factors such as season. The following pathogenic microorganisms causing illness in humans are characteristically associated with molluscan shellfish: Vibrio parahaemolyticus Vibrio cholerae O1 Vibrio fluvialis Vibrio hollisae Vibrio vulnificus Vibrio cholerae non-O1 Vibrio mimicus Vibrio furnissii 59 . Raw and partially cooked shellfish. clams and mussels. in sauce. Canned: in brine. mostly in oysters.2 Initial Microflora The microflora of molluscan shellfish directly reflects the environment from which the shellfish is harvested. Whatever contaminants are in the water will eventually get into the shellfish (7). 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.MOLLUSCAN SHELLFISH Frozen: shelled meats. The initial microflora of molluscan shellfish is comprised of the natural commensal microorganisms and the microorganisms accumulated from the water during feeding. Salted: meats either lightly or heavily dry salted. Primarily inhabits shallow waters near tidal zones where they bury themselves in the sandy or muddy bottoms. 3. Canned: in own juice.5 Cockle A marine bivalve mollusc having a shell with convex radially ribbed valves. Cockles range from about 1 to 15 cm in length and have two valves of equal size. which affects the temperature. are vectors of disease transmission (8).
shellstock (live) which is shellfish in the shell. Campylobacter coli Bacillus cereus Hepatitis non-A.1 Processing and its Effects on the Microflora Oysters. 15). The microflora of molluscan shellfish at harvest consists predominantly of Gramnegative rods while the Gram-positives constitute a minor portion of the flora. Flow diagrams of the major processing steps of molluscs are presented in Figures 3.FISH AND SEAFOOD Aeromonas hydrophila group Staphylococcus aureus Shigella spp.1 and 3. Bacillus spp. Enterococci spp. responding to the environment in which they are harvested. At harvest. 12.. clams. but have never been cultured (14.. and shucked shellfish that is shellfish. 3. (9.. Water temperatures affect the concentrations of microflora in the water. whole or in part. There are microorganisms. nutrients. In summary.3 3.. Campylobacter jejuni Escherichia coli Hepatitis A Norwalk virus Other viral agents A variety of commensal microflora are found in molluscan shellfish. The largest groups found are the Pseudomonads. Pseudomonas spp.1. For example. Other microorganisms found in lesser numbers are the Achromobacter spp. 11. Alcaligenes spp.. 3. 10. Moraxella spp.1.2. mechanical harvesters.3. Acinetobacter spp. non-B Snow Mountain virus Plesiomonas shigelloides Salmonella spp..000 /g. rakes. pollution point sources and hydrological events will greatly influence the initial microflora in bivalves. and yeast cells of Rhodotorula rubra and Trichosporon spp. spirochetes of the genus Cristispira and Saprospira. Micrococcus spp.1 Shellstock 3. They are termed viable non-culturable microorganisms. Temperature..1 Processing description The method of processing shellstock begins with the harvester collecting shellstock by the utilisation of tongs. and cockles The processing of bivalves takes two forms.3. from which one or both shells have been removed. and Aeromonads. salinity. or by 60 . molluscan shellfish have an Aerobic Plate Count (APC) of approximately 1000 to 100. the initial flora composition and concentration of bivalves are highly variable..1. have been observed microscopically in the digestive tract of Eastern and Pacific oysters. mussels. 13). that have never been cultured on laboratory media. Flavobacterium spp.3. Vibrios. observed microscopically.. dredges.
for transportation to the point of sale.MOLLUSCAN SHELLFISH hand (Figure 3. Thirdly. There are three primary reasons for wet storage: first. shellfish can be harvested from remote areas at convenient times and held live for brief periods at locations close to the point of sale. Shellstock are given a wash to reasonably free them of bottom sediments and detritus as soon as practicable. wet storage may be used to increase palatability by increasing salt content of shellfish harvested from low salinity waters. If by boat. Shellstock may be wet-stored at the processing facility in tanks of clean seawater.3. The shellstock is transported dry. 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. Fig. usually in open-mesh containers and/or burlap sacks. by truck or boat.1).1. after harvesting. Generic flow diagram of live shellfish from harvester to point of sale 61 . or immediately placed in refrigerated trucks. or in bay-approved shellfish-growing waters prior to sale or processing. in containers from which they can be retrieved easily. Shellstock are placed in drycooler storage containers in the facility. Water used for shellstock washing is from approved growing areas. or from other sources approved by the State Shellfish Control Agency (SSCA). usually the boat will be docked by the processing facility with shellstock piled on the deck of the boat or dock. Secondly.
and the sanitary practices applied during harvesting and shipping. involving 691 cases was attributed to oysters contaminated in a wet storage area (27). metabolic waste accumulates in the shell serving as nutrients for bacterial growth. 26). 22).3. there is evidence that in the absence of adequate refrigeration. without adequate refrigeration. Klebsiella. to avoid possible contamination of the shellstock. Over a period. Shellstock should be refrigerated at temperatures at or below 10 ºC (50 ºF). and Citrobacter species) and Vibrios. Several studies have established that pathogenic Vibrios and other bacteria may be present in marine sediments throughout the year (21. synthetic seawater for the purpose of de-sanding and salt uptake. 23. 24. Several studies have established the increase of bacterial counts in shellstock during post harvest period before the shucking process (18. 25). with the exception of A. The washing of sediment and detritus soon after harvesting will prevent further contamination. some pathogenic organisms will survive and multiply in shellfish (20.1. Washing shellstock helps prevent quantities of sediment and bacteria being mixed with. Shellfish in wet storage tanks are similarly subjected to pollution if the tank water is obtained from a polluted source. Oysters will remain alive in the shell for approximately two weeks if kept at the recommended temperatures. 21. however. At harvest. shellstock are not refrigerated on the harvest boat. Enterobacter. hydrophila (18). Generally. 16.FISH AND SEAFOOD 3.2 ºC (45 ºF) or below (19). At this temperature the multiplication of faecal coliforms (E. shellstock are placed in refrigerated trucks for transport or placed in wet storage. The safety of shellfish is predicated by the cleanliness of the growing area waters from which they are harvested. Furthermore. vulnificus. including V. Excessive sediment on the shells and dead shellfish may increase bacterial 62 . Food and Drug Administration (FDA) recommends that potentially hazardous food such as shellfish be held at 7. coli. Wet storage operations are highly variable and may range from temporary storage near-shore in approved areas to on-shore tanks using re-circulating. and thus contaminating the shucked meat at the point of sale. If long delays occur before reaching the processing facility and/or refrigeration. Harvest conditions are another factor that influences the growth of bacteria in shellstock. when it reaches the processing facility. are prevented. The most important factor in controlling bacterial growth in shellstock is temperature. for a considerable length of time.2 Effects of microflora during processing The initial microflora found on or in shellstock is influenced by sources of shellstock. Generally. 17).1. with prolonged storage the microbiological quality of the bivalve decreases. Shellfish may contain higher levels of pathogens than are found in the water in which they grow. An outbreak of infectious hepatitis in Sweden in 1956. bivalves tightly close their shells thus trapping water and microflora. Water used for shellstock washing should be of good sanitary quality. handling. the bacterial counts in the meats will increase. There is a positive relationship between sewage-polluted shellfish and enteric disease transmitted by shellfish (8. and temperature storage of shellstock to point of sale.
2.1. Sealed containers are placed in boxes and covered with crushed ice for chilling or refrigeration and transportation.2). In addition. thus contributing to high bacterial counts in the final product (28). clothing becomes very soiled. whereas cotton become wet and 63 . and packing shellfish into containers (Figure 3. Washing of meats with potable water is accomplished by placing the meats on a skimmer table (the stand-supported. 3. Under these conditions if the shuckers enter the packing room. If shellfish are not reasonably clean at the time of processing. Thus.3. the meats are in a tank of water and agitated by the introduction of air from the bottom of the tank.2. for the effects of microflora during the processing of shellstock. 3. Due to the nature of the shucking operation. Meats are collected in either perforated or non-perforated buckets for delivery to the washing station.2 Shucked shellfish 3. are applied to the shucking operation of shellfish. In the blower-washer. the knife is used to cut the remaining adductor thus freeing the shellfish (meats). 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. Shucking and washing of meats under sanitary conditions prevents the proliferation of bacterial populations.3. perforated tray in which shucked shellfish are spray washed and/or drained) or in a blower-washer (a tank-like device for immersion washing of shucked shellfish). To remove the shellfish attached to the remaining valve.1 Processing description The processing of shucked shellfish is limited to washing of shellstock. the tank is opened through a drain gate and chute. It presents a risk of cross-contamination from the non-molluscan animals to the bivalves. Shuckers routinely use either cotton or rubber gloves during the shucking operation. removal of shellfish (shucking). Studies have shown the use of rubber gloves reduces the contamination level. After washing of meats.1. washing and culling shellfish prior to storage is essential.3. or subjected to splash from foot traffic.1. through which the washed meats are discharged for rinsing and then packed in containers. washing. Containers are placed in packing boxes for shipping at refrigerated temperatures.2 Effects of microflora during processing The same considerations as previously discussed. and fish.MOLLUSCAN SHELLFISH loads in the tanks and lead to increased microbial levels in the shellfish during storage. lobsters. shellstock is subjected to contamination when stored on the floor in standing water. a considerable quantity of the adhering material will be mixed into the shellfish during the shucking process. the finished product may become contaminated. Another potential risk with the use of wet storage is the commingling of bivalve molluscs with other marine species such as crab.
2 ºC or less. and gloves help prevent the contamination of shellfish.FISH AND SEAFOOD retains bacteria during the shucking thus increasing the bacterial load in shellfish (29). There is a tendency to store such articles on the shucking benches or in packing rooms. and sanitised before each filling. cleaned and delivered to the packing room or prechilled and placed in temporary refrigeration (≤ 2 hours) at 7. 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. Fig 3. aprons. Shucking containers are rinsed clean with running water. Shucked meats are thoroughly drained. within one hour. Separate rooms or lockers for storage of clothing. Shucking buckets are completely emptied at the packing room and no overage is returned to the shucker.2. 2. 4. 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. 64 .
Refrigeration and shipping of shucked shellfish in wet ice is highly recommended. Pseudomonas. The spoilage process in shellfish may be divided into three stages: increase of acidity. All equipment coming in contact with the shellfish such as colanders. showed that organisms such as Achromobacter. whereas bacterium such as Pseudomonas. Proteus. where there is a breakdown of temperature control or prolonged storage of shellfish. As described previously. Studies have shown that bacterial growth is significantly reduced at storage temperatures of less than 7. these include lactobacilli. and Pseudomonas fluorescens are the responsible agents of spoilage in oysters held at various temperatures (10.4 Spoilage There is a lack of information in the literature regarding spoilage microorganisms in shellfish. Glycogen is hydrolysed by the initial flora found in shellfish. Achromobacter and Flavobacterium levels decrease during storage.8% (37). Shellfish. 3. The environment has an effect on the bacterial population found in shellfish. keeping shellfish refrigerated is the most effective method of retarding deterioration and spoilage. streptococci. and proteolysis (10). Micrococcus. Controls that should be applied include using clean packing equipment. glycogen levels range from 0. anywhere during the process of shellfish from harvesting to the point of sale. blowers and other utensils that have cracked. This process changes the bacterial profile to microorganisms favouring low pH. For example. Alcaligenes.MOLLUSCAN SHELLFISH Unacceptable practices in the packing step can affect the microflora of shellfish. In summary. 36). 33.2 ºC. shucking pails. Glycogen. Earlier studies. skimmers. rough. and packing the shellfish in a timely manner. are an excellent medium for growth of bacteria. clean gloves. at the turn of the century. Temperature abuse will result in high bacterial counts. Shucked shellfish are held and transported at temperatures of 7. 32). 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. Flavobacterium. 34. Lactobacilli seem to be the predominant microflora in shellfish. is stored in the bivalve as a source of energy. and yeasts. Shucked shellfish are an excellent medium for the growth of bacteria. a carbohydrate. The initial flora found in shellfish is the major factor of spoilage in bivalves. or inaccessible surfaces are apt to harbour accumulations of organic material in which bacteria may grow.2 ºC or less. spoilage will occur. These organisms may play a significant role in the spoilage processes. 65 . abundant gas production. particularly shucked meats. particularly oysters stored at 7 ºC.47 to 6. 35. in oysters. and that storage in wet ice is the most effective method for refrigeration of shucked meats (30. 31. Much of the work has been done with oysters and clams. thus producing acid and gas (38).
and very common in Japan. lot and retail outlet. of V. V. The ability of the bacterium to cause gastroenteritis is highly correlated with the production of a heat-stable haemolysin (42). Descriptions of outbreaks or reports of growth of the organism in shellfish are presented. parahaemolyticus illness is usually a result of eating raw or partial cooked shellfish (8. headache and fever.5. parahaemolyticus. parahaemolyticus multiplies rapidly at temperatures above 20 ºC. although non-hemolytic strains have been associated with illness (43. and harvesting of shellfish in the cooler months.FISH AND SEAFOOD 3. parahaemolyticus causes gastroenteritis lasting 24 . only a small percentage of strains in the marine environment are potentially pathogenic (45). where outbreaks occur regularly. 41).1 Vibrios Vibrios are Gram-negative facultative rods naturally occurring in the marine water (estuaries. Large numbers of bacteria must be present to reach potentially infective levels of >105 colony forming units per g (39). and estimates based from a hospital study show less than 0. during processing.000 population/year (46). parahaemolyticus from shellfish is low. ranging from epidemic cholera to sporadic cases of diarrhoea.e. 44). nausea. They can be introduced to the shellfish from the environment or added during processing. Fortunately. Diseases caused by Vibrios are enteric in nature.1.48 hours with abdominal pain.5 cases/100. from harvesting to finished product (i.5 Pathogens: Growth and Survival This section deals with bacterial pathogens frequently present in shellfish. shellstock and packing of shellfish in jars) (29). Sporadic cases are frequent along the coast of the United States. V. One of 250 cases reported involved shucked products with low levels. The incidence of V. The control of growth and survival of these organisms can be reduced by the use of good manufacturing practices including immediate and proper refrigeration. Products involved were from the same processor. diarrhoea. 3. widespread in both polluted and unpolluted waters. there were over 250 cases of illness on the West Coast of North American extending from California to Canada. Major outbreaks are associated with the warmer months. These organisms can be present in the shellfish from the point of harvesting. coastal areas). The majority of the illnesses were associated with the consumption of raw or partially cooked oysters and clams. parahaemolyticus possibly producing illnesses (40). 66 . Between May and September 1997.1 Vibrio parahaemolyticus V. approximately 23 cfu/g. there is a recent report of low levels of V. It is a common marine bacterium.5. 3. however. and to the point of sale. prevention of cross-contamination. This bacterium is present in all the major processing steps of a shellfish plant. primarily in brackish or saltwater.
cirrhosis. cholerae. however. abdominal cramps. Illnesses are usually associated with the consumption of raw oysters as described in Table 3. It is one of the most severe foodborne infectious diseases. the organism has been found in the Gulf of Mexico.II (8. The bacterium is widespread in estuarine waters particularly in warm waters of >20 °C. Oysters are the most hazardous food associated with V. vulnificus. Hazards from this bacterium can be controlled by thorough cooking of shellfish. The bacterium is an estuarine species that causes wound infections. like V. or immunosuppression. and preventing cross contamination once the product is cooked. The widespread distribution of this bacterium is comparable to other studies in the Gulf and Atlantic Coasts (52.48 hours of onset. Death has been reported to occur within 36 hours from onset. nausea. 53. vulnificus causing septicaemia and death has been associated with the consumption of oysters (8. They are sensitive to cold. vulnificus are fever. 55. Onset of symptoms occurs in 48 hours and can be severe. those who have liver disease. and nausea within 24 . V. with a fatality rate of 50% for individuals with septicaemia. diabetes. 46.0. 23 of 24 major USA-West Coast estuaries were positive for V. fever.MOLLUSCAN SHELLFISH Control of V. 57. All isolates are considered pathogenic.8 case/100.5. cholerae non-01 is ubiquitous in the estuarine environment. Healthy individuals are susceptible to gastroenteritis. The organism attaches to the intestine and release of a toxin is suspected.000 population/year (8. parahaemolyticus the estimated annual incidence rate is very low.5. leukaemia. High-risk individuals.2 Vibrio vulnificus The symptoms of V. Primarily.1. and vomiting. V. V. cholerae non-01 has been associated with septicaemia. 46.4 .1. 49). Control of V. parahaemolyticus growth in shellfish meats is temperature dependent. vulnificus is by rapid refrigeration during warm weather months. lasting 6 . chills. 54. these individuals are advised not to eat raw shellfish. Again. Inadequate refrigeration will accelerate the growth of V. and rapidly become inactive at low storage temperatures. It causes diarrhoea. cholerae non-01 is a common bacterial cause of molluscan shellfishassociated illness. in the range of 0.3 Vibrio cholerae non-01 Illness caused by V. which have been implicated in shellfish-related disease outbreaks (31).7 days. cholerae non-01 is generally a less severe gastroenteritis than that caused by V. 3. are particularly susceptible to primary septicaemia. Control of oyster-associated 67 . 56). gastroenteritis. primarily in immunocompromised individuals with a mortality rate exceeding 50% (50). cholerae non-01 (51). 48). or primary septicaemia. 58). 3. The occurrence of V. the organism has also been found in cooler temperatures from estuaries of Washington and Oregon (47). parahaemolyticus. In one study.
acidity and heat. Gulf coast environment. abdominal pain. these organisms cause gastroenteritis lasting 24 . 3. 61. fluvialis. particularly in oysters. nausea. In developing countries. widespread in both polluted and unpolluted waters. fever. cholerae 01. salinity. headache and fever. jejuni requires reduced levels of oxygen (3 .4 Vibrio cholerae 01 and 0139 Symptoms of epidemic cholera vary from mild watery diarrhoea to acute diarrhoea with characteristic rice water stools. and V. respectively. and nausea. and harvesting of oysters in colder months when Vibrio counts in water are the lowest (59). whereas excellent sanitation facilities in developed countries are responsible for the near eradication of epidemic cholera. 63). processors should know the product source. and illness lasts 7 . imported product from a country experiencing an epidemic. Onset of disease is 6 hours to 5 days. vulnificus.2 Campylobacter jejuni C. There are several other Vibrio spp. nausea. Illness can include abdominal cramps.1.5. e. including V. They are common marine bacteria. 3. associated with shellfish-borne illness outbreaks. Onset of illness is from 3 to 5 days after ingestion of contaminated food. 68 . V. diarrhoea. and southern Asia including India and Bangladesh. and the transmission of the disease by consumption of raw. parahaemolyticus and V.5 Other Vibrios spp. dehydration. Illnesses are associated with the warmer months. preventing crosscontamination of cooked seafood. V. Serotype Ogawa biotype El Tor and 0139 are responsible for epidemics in South American. V. drying. the pathogenic severity is lower than V. is the same as previously discussed for the non-01 strains. (8.5%) and 2 .5. cholerae 01 in molluscan shellfish. It causes diarrhoea. or cross-contaminated shellfish (57.48 hours with abdominal pain.FISH AND SEAFOOD V.g. Control of infection from these organisms is no different from the other Vibrios. low pH. It is considered a fragile and sensitive organism to environmental stresses such as oxygen. There is evidence of strains of V.1. hollisae. cholerae 01 establishing in the U. cholerae non-01 illnesses is by cooking seafood thoroughly. vomiting.S. In general.10 days. however. cholerae 01 is transmitted by faecal contamination of food or water. 60).10% carbon dioxide for optimal growth conditions.5. In addition. The control of V. 3. and shock. Death may occur after severe fluid and electrolyte loss. The presence of these strains appears to be in low numbers and non-virulent (59). undercooked. The frequency of infection of these organisms is no different from other Vibrios. Poor sanitation and contaminated water supplies will spread the disease. 62. mimicus.
V. Recently. jejuni survived for over 20 days in market-shucked oysters stored at 4 ºC.3 Other organisms of concern 3. however.500 cells/gram of Campylobacter may cause illness (64.MOLLUSCAN SHELLFISH TABLE 3. Although. V.a (8) Species Hard clams Oysters Soft Clams Mussels Scallops No. they survive well at refrigerated temperatures. They have been implicated in limited shellfishassociated gastroenteritis cases (8. the incidence of Campylobacter infection is low from the consumption of molluscan shellfish.4 to 114 cells. In one case. cholerae 01 and non-01.5. of Cases 4 362 0 1 0 a Includes V. have been isolated from shellfish beds throughout the world (67. Proper care should be taken if the finished product is to be packaged in modified-atmosphere packaging (MAP). 78) and P. Campylobacters do not survive well in the environment due to the organism's lack of resistance to environmental factors. 81. 76. 83. 85). Campylobacter appear to survive well in oysters during storage. 73). whereas at 10 ºC Campylobacter survive only for 5 days (71). parahaemolyticus. and have been implicated as the causative agent in illnesses from the consumption of raw oysters and clams (68. 72. jejuni were recovered. V. C. 68.1 Aeromonas hydrophila group and Plesiomonas shigelloides A. Campylobacter spp. infection occurred among participants of a firemen's banquet.3. shigelloides (79. 82. 84.5. The seasonality of 69 . 66). mimicus. jejuni begins in the harvest of shellstock from approved waters. jejuni does not multiply in shellstock at refrigerated temperatures. 3. 80) are ubiquitous in the marine environment. of Incidents 4 279 0 1 0 No. V. and their immediate proper refrigeration.II Molluscan shellfish associated outbreaks caused by Vibrio spp. and V. ranging from 0. from Pacific shellstock oysters purchased from a retail grocery store (74). This finding is significant in that C. Sixteen of 28 participants became ill with campylobacteriosis after ingesting raw clams. C. To reduce the risk of shellfish-borne campylobacteriosis. Human feeding studies suggest that about 400 . 70. although their role as a pathogen is highly controversial and circumstantial. hydrophila group (75. the organism is being continually inoculated into the environment by waterfowl and farm run-offs and thus eventually contaminating shellfish growing waters. vulnificus. 65. 71). control of C. however. fluviallis. 69. 77. hollisae Infective dose varies with host susceptibility and virulence of the strain.
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. vulnificus in molluscan shellfish. being isolated more often during the warmer month.000 MPN per gram1 negative for staphylococcal enterotoxin or when the viable MPN count is <10. cholerae V. 87) of the outgrowth of A.600. after 7 days storage of shucked oysters at a shellfish processors’ cooler kept a 5 ºC Aeromonas counts ranged from 500. vulnificus 1 Acceptable Levels in Molluscan Shellfish <500. Enforcement action is considered on a case-by-case basis taking into account all the factors associated with the specific situation. Pathogen monitoring of Selected High Risk Foods Domestic including: milk. FDA/ Administrative Guidelines. dried milk/whey drink powders. 3. FDA/Administrative Guidelines 3 Source (91).6 Published Microbiological Criteria The published microbiological criteria below are included as a guide. jejuni V. They survive well in shellstock or shucked oyster meats held at refrigeration temperatures.000 cfu/g1 <230/100 g2 <1000 per gram. FDA established action levels or levels of concern for certain microbial pathogens in molluscan shellfish. Caution should be exercised by individuals with haematological malignancies (88). They are easily destroyed with mild cooking temperatures.FISH AND SEAFOOD Aeromonas and Plesiomonas is similar to the Vibrios. suggesting that these individuals are at higher risk when consuming raw or partially cooked shellfish containing these bacteria.000/gram (29). Note: Although molluscan shellfish are not included in the selected high risk food. In one study. The Agency will consider enforcement action against the shipment of molluscan shellfish if the levels exceed as stated. non-dairy frozen desserts. coli Salmonella V. and where indicated are intended to be used for official control purposes. parahaemolyticus Staph. action is considered on a case-by-case basis taking into account all the factors associated with the specific situation.000 to 3. Control of these organisms in shellfish products is limited. Similar results were seen by other investigators (86. aureus C. soy products. Test Standard Plate Counts E. 4 FDA and ISSC recommending establishment of an educational program to alert at risk population of presence of V. 70 . hydrophila group stored at refrigeration temperatures. prepared sandwiches. coli Enterotoxigenic E. 2 Source (90).0001 negative for the presence3 Under review4 Source (89).
14. water and sediment from Galveston Bay. 544-8. 9. 13. in General Zoology. in Fishery Bulletin 1964 No. Applied Microbiology. 11.S.. 42. Applied and Environmental Microbiology. Colwell R. Effects of harvesting waters and storage conditions on yeast population in shellfish.E. 30. Eds. 17. 58. 1983. 7.. 12. Microbial flora and level of Vibrio parahaemolyticus of oysters (Crassostrea virginica). U. 6. 1960... Tubiash H.B. Ward D. 105-8. Fleet G. New York. 13. in Multilingual Dictionary of Fish and Fish Products. 15.K. 33643. 8. Microbiology of shellfish: bacteriological study of the natural flora of pacific oysters (Crassostrea gigas). 1940. A typhoid fever epidemic caused by carrier bootlegging oysters. 82-7. 1975.D. Applied Microbiology.R.C. Vanderzant C... 231-51. Clinical Microbiology Review.S. Ray S. Saunders Company.R.A.H. References The Phylum Mollusca.C.7 1. 36. 1968. 33-6. Old H. Fish and Wildlife Service... 755-76. Quantitative and qualitative commensal bacterial flora of Crassostrea virginica in Chesapeake Bay. Lovelace T.S. 1984.J.. Proceedings of the National Shellfisheries Association.. Galtsoff P. American Journal of Public Health.N. in Microbiology of Marine Food Products.. 29. 633-40. 44. 64. Veliger. 8. Bacterial flora of the market oyster. Biology of the oyster in relation to sanitation. Fate of enteric bacteria in estuarine sediments and oyster faeces. Liston J. 71 . 1995. Rowse A. Villee C. Scanning electron microscopy of Cristispira species in Chesapeake Bay oysters. Bernard F.F. Ed. Infectious diseases associated with molluscan shellfish consumption. Occurrence of the spirochaete genus Cristispira in western Canadian marine bivalves..A. Hood M. 1981. Philadelphia. 7. Pfeifer D.W. The American Oyster. 2. 19-34. American Journal of Hygiene.R.. Nauman R. 1970. Washington D. Eliot C. 1936. Eds. 6. W. Galtsoff P. 1994. Microbiology of bivalve molluscan shellfish. 447-52.R. Van Nostrand Reinhold. Thompson C. Smith F. 26. Jr. Journal of Milk and Food Technology. Applied and Environmental Microbiology. Journal of the Mississippi Academy of Sciences. Government Printing Office. Schmidt C.MOLLUSCAN SHELLFISH 3. 10.M.K. Gill S. American Journal of Public Health. 16. Organisation for Economic Co-operation and Development. Colwell R.E. 1991. Hackney C. 245-7. 1982. Wiley-Blackwell.L. 30.W. 3. 71-6. 1963. Cook D. 1973.A.R. Journal of Food Protection. 104-9. 1926.R. Rippey S. Tall B. Cook D.S. 46. 5. Walker W. 1964. Viability and release Salmonella charity and Escherichia coli from oyster faeces. Jr. 4. 419-25. Microbiological characteristics of Dungeness crab (Cancer magister). Lee J. 72-8. USA.
1989. 26. 5. Journal of Food Protection. Liston J. Establishment of critical control points in west coast oyster processing plants. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. 1214. 1992. oysters. Kelly C. Ruple A.C.J. 324..A. Beck W. Rodrick G.C. Presnell M.. FDA Gulf Coast Technical Services Unit..H.W. 1995. 994-1002. I. in International Symposium Vibrio parahaemolyticus. Special Report U. Abeyta C. 40..A. 1490-5. 31.E. and Manillal clams (Venerupis japonica).F. Cook D. 1954. 47 (5). Vaughn M. Journal of Food Science. Bacteriological changes in live shellfish: Pacific oysters (Crassostrea gigas).. Applied and Environmental Microbiology. U. Public Health Service. Part II Harvesting. Presnell M. 23. Saikon Publishing Co. Kelly C. Fujino T. Tokyo.. Trost P.S. U. The influence of time and temperature on the bacteriological quality of shell oysters during processing and shipping. Fraiser M. Food and Drug Administration... Applied and Environmental Microbiology. during depuration. Hepatitis epidemic conveyed by oysters. 1964.. crabs and mullet.. Bacteriological survey of an oyster bed in Tangier Sound. Ltd. Eds. U.W. Survival of enteric organisms in shellfish.H.M. Williams L. Crassostrea commercialis. Vasconcelos G. Incidence of Samonellae in clams. LaRock P. and storage. 22. Journal of Food Protection. Washington D. 50.A. 1221-8.D. 19.S.S. 27. 25. Colburn K. Sadaguchi G. 53. Bacteriological studies of commercial shellfish operations on the Gulf Coast. Stott R. Food and Drug Administration. 72 . Temporal occurrence of Vibrio species and Aeromonas hydrophila in estuarine sediments. 1973.B. 24. P-1. 4.. Koburger J. Fleet G.A.. 343-5. Food and Drug Administration. Crassostrea virginica and Mercenaria campechiensis. 30. Jones W. Effects of storage on microbial loads of two commercially important shellfish species. Timetemperature effects on the bacteriological quality of stored shellfish. Blake N.. 167-71. Native Littleneck clams (Protothaca staminea).A.. Applied and Environmental Microbiology. Sakazaki R.S. American Society for Microbiology. Behaviour of pathogenic bacteria in the oyster. 1971... 69.. 21. 52. 1983. Olympia oysters (Ostrea lurida). 343-9. Hood M. 1984.B. Public Health Service Technical Report. 28. 32.G. Handling and Shipping Shellfish. 1205-10. Jr. Chesapeake Science.B. Roos B. Hoff J. Takeda Y. Ed. 1985. Arcisz W.FISH AND SEAFOOD 18.W. 989-1003. F61-9. 20. 29.S. seafood-handling procedures on Vibrio parahaemolyticus. Kaysner C. Influence of U.. 1956. re-laying.. Public Health Report. Maryland..W.. Wekell M.J. 123-8. 45. Ness G. in National Shellfish Sanitation Program Manual of Operations.J.E. Ericksen T. 1961.W. Svenska Lakartodningen. Son N.A.. 1967.T. 1980.
41. Thompson C.M. Wekell M. Studies on pathogenic halophilic bacteria. 107-8.M.. 2891-3. West Coast States and British Columbia. Ishibashi M. Arcisz W.N.W. Parasitenkunde. Infectionskrankheiten und Hygien. Academic Press. Zentralblatt für Bakteriologie. 2.W.. 2820-2.. Libonati J. 73 .L. Kelly C. Geographic and monthly variation in composition of oysters. 1989. Bothell.. (2). Kaysner C. 1349-51. Joseph M. 40.P. 1989. Leitch J. 1. Bulletin of the Johns Hopkins Hospital.. 1961. 41. 42. Crassostrea virginica. Serological and hemolytic characteristics of Vibrio parahaemolyticus from marine sources... The bacterial content of market oysters. 25. Journal of Bacteriology.. Yokohama Medical Bulletin. 128-31. Vanderzant C. 39.B. 1916. 13-7. 133-47. 35.A. Public Health Reports.G. Morris J. Borgstrom G.. Obara Y. 41. Epidemiology and spectrum of Vibrio infections in a Chesapeake Bay community. Microbiology of shellfish decomposition.S. In vitro haemolytic characteristics of Vibrio parahaemolyticus: Its close correlation with human pathogenicity. 1997. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States west coast.. Novak A. Powers C. Journal of Infectious Disease.. Watsky D. 27.D. 1147-9. Applied Environmental Microbiology. in Fish as Food. Journal of Bacteriology. Journal of Infectious Disease. 985-93. 1954.. 204-5. Bacteriological examination of oysters stored at low temperatures. Journal of Food Science.. Hlady W. Akiyama S. Stott R.S. 46.F. Miyamoto Y. Fieger E. Geiger J. Bacteriological control of oysters during processing and marketing.A.A.. Kelly M. The bacterial flora of market oysters.C. Takikawa I. 1976. Peeler R. 43. 60.L. 353-7. Jacobson M.. Stroh E..D. Kato T. 313-22.. 1969. Journal of Food Protection. 1979. Janda J.MOLLUSCAN SHELLFISH 32. Shellfish Specialist. 44. U. Israel E. 33. Washington Report October 14. 1958.A. 273-80. 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. 100. 36. Grodner R.E. kanagawa-negative Vibrio parahaemolyticus from patients and the environment in the Pacific Northwest.. Sidwell V. 1937.. Kaysner C.. Volume 1. 53. Abbott S.. Food and Drug Administration. Vibrio parahaemolyticus illnesses.M.. Jr. Bacteriological findings in Baltimore oysters. Abeyta C.. Journal of Clinical Microbiology. Journal of Clinical Microbiology. Yamai S.G.. Joseph S. Urease-positive. New York. 69. Takeda Y. Sample T. Berry F. U.T. 1997. Vibrio infections associated with raw oyster consumption in Florida. 47. 38. DePaola A. Hoge C. Marine Fisheries Review. 27. 1926... 45. 1914. 1987.M.. 97. 37. 1989. Tanikawa E. 38. Ward W.M. 34. Takizawa K.. 160. Ed. Jr.. Loomis A..
Journal of the American Medical Association. 91-103. Riddle C. DePaola A.A.F. Vibrio vulnificus: Man and the sea.. Snyder J. 1475-8.. 41. Blake P. Blake P. Lee J. Roderick G.. 57. 312. 1982. 1. Wilson T.B. Zywno S. 45.. Jr...T. 53.O. Puhr N. sediment and waters of the U.B. Morris J. 1983. 1984.A. McPhearson R..R.T.N.J. Lancet. Jr. Microbial Ecology. Non-01 Vibrio cholerae in shellfish.. Francis D.M.M. Blake P... Duran W. Jr. 1987.D.. Colwell R. 49. Tacket C.O. R.. Feeley J. Journal of Food Protection... Jr. Kaysner C.G.. Hunt J. Incidence of Vibrio cholerae from estuaries of the United States West coast.I. Jr. Applied and Environmental Microbiology.R.M..K.M. Siebeling..G. Cholera . 37... Colwell R.G. Bradford H.. 1982.. 74 .W.A. Ecology. McFarland L.A..M....J. cholerae in Louisiana.R. Applied and Environmental Microbiology. 2050-3.G.O. Wathen H.M. Jr.... 656-8. Plaard R. 1988.T. Vibrios in the Louisiana Gulf Coast environment. epidemiologic.. 1344-8. 305.M. McCay C.D. Hollis D. 58... Adams M. 10. Colwell R. Hickman F. 55. 558-61.. Non-01 Vibrio cholerae bacteraemia: Case report and review. An epidemiological study of V.L.M. Becker R. 54.. 50. Barrett T.. Small P.B. Hug A. 61. Wachsmuth I. Roberts N.A. 305-9.S.A. Motes M. 52. Wilson R. Illness caused by Vibrio damsela and Vibrio hollisae. Blake P.G. Kaper J..W. Bradford H.. Jacobs R. Craig J. 1294-7. Wazenski T. Leitch J. 59. Twedt R. 1012-7. 149.F. Safrin S..A. Characterisation of Vibrios cholerae isolated from oysters.. West P. Hug M. Annals of Internal Medicine.. Twedt R. 60. Tacket C. 5972... 1985.W. John Wiley & Sons Inc.. McFarland L... Jr..R. 1981. 343-50.FISH AND SEAFOOD 48... serology. Review of Infectious Disease. New England Journal of Medicine. 1980.. Johnston J.G. Cholera and other vibrioses in the United States. Maden J. Allegra D. New England Journal of Medicine. Caraway C. 53.. Morris J.E.F. Hebert S. 94. Brenner F.. 1983. 1981. 1985.. 802-6..A. 275-83. 46.J.G.. Morris J. 51.P.T. Black R..E.G. DePaola A. Kaper J. 1984. Davis B. Stott R. 1979. Presnell M.. and enterotoxin production of Vibrio cholerae in Chesapeake Bay. Pons V. Morris J.C.A. in Vibrios in the Environment.E. 56.... Clinical features and an epidemiological study of Vibrio vulnificus infections.M. Wekell M. Non-O group 1 Vibrio cholerae gastroenteritis in the United States: Clinical. Spite G. Ed. Rahman R. 8.A possible endemic focus in the United States.V. Conte J.C. and laboratory characteristics of sporadic cases. Weaver R. Miller H.E.. 299-312.. New York. Abeyta C. Ecological relationship between Vibrio cholerae and planktonic crustacean copepods. Journal of Infectious Disease..W.. 253. Joseph S..T... Applied and Environmental Microbiology. Feldman R.. Lockman H. Becker S. Peeler J. Gulf Coast. Applied and Environmental Microbiology.
Jr. in Campylobacter II: Proceedings of the Second International Workshop on Campylobacter Infections. Plenum Press.G.F. 71. Abeyta C. British Medical Journal. Davis J. Moore J.E. 1995. 1983. in Campylobacters. 256. 11. Levine M.E... Jr.G.S. Blake P. Newell D.I.. Abeyta C. Infective dose of Campylobacter jejuni in milk. 1991.G.. Wareing D. Wilson T. EF-6. Blaser M.. Wekell M. Public Health Laboratory Service..M.G. Griffin M.. Helicobacters and Related Organisms.J..A. Wilson I..MOLLUSCAN SHELLFISH 62.A. 169-71. 1983.... New York.. 67. 69.S. Alam A. 99. 1980. C. Helicobacters and Related Organisms. 51-64. Kaysner. Journal of Infectious Disease. a newly recognised Vibrio species. in Campylobacters. 72.A. 1993. A method for isolation of Campylobacter spp.. Wekell M.R.X. infections in man. Johnston J. 472-9.E. Skirrows M.A.P. Microbial Ecology in Health and Disease.A.. Matsuda M. S41. Clements M. 157. 65. Ketley J.. Eds.P. 13. Experimental Campylobacter jejuni infection in humans.. 1987. Plenum Press.M... 80. Shandera W.R.. Isolation of vibrio-like group. Austin S.R. 1996.. Pearson A. Ketley J. Brussels.E. Brenner D. 64. 101-4.G. Journal of the Medical Society of New Jersey..R. 68.H. Murphy P.. London. Jones D. The occurrence of Campylobacter jejuni and Campylobacter coli in Sidney rock oyster.B.. Morris G.K. coli and lari) from shellfish and the marine environment.K.G. International Journal of Food Microbiology. 257. (jejuni.M. Annals of Internal Medicine. Journal of Clinical Microbiology. Journal of Food Protection. 75 .. Black R.M...L. Black R. Wilson T. Wilson I. Disease from infection with Vibrio mimicus.. Campylobacter gastroenteritis associated with raw clams. Abeyta C. Phenotypic and genotypic characterisation of urease-positive thermophilic Campylobacter (uptc) isolated in N. 4.R. Eds. Hunt J. Arumugaswamy R. 323-5. Kaysner C... Dalley E..M.. 56.M.A. Proudford R. 1996. 621-4. Hughes T.D. Campylobacter jejuni in a Washington State shellfish growing bed associated with illness.A. Clements M..W. Fitzpatrick M.J. Eds.. Jr. Ireland. 66.. Huges T.J... 4.A. 73.D. Journal of Rapid Methods and Automation in Microbiology. 1988. Newell D.M. 63.M. Studies of Campylobacter jejuni infection in volunteers. Murphy P. Deeter F. Feldman R. Blaser M. 282.M. Part 1 Abstracts: Shellfish as a possible source of thermophilic Campylobacter spp.K.A. Feldman R.J. Kaysner C. Stott R.. 4. New York.L.. 1980. Incidence and survival of thermophilic campylobacters from shellfish growing water: media evaluation. 1981. from patients with diarrhoea. 1584. 70. Wareing D. Huq M.G. Levine M.. Davis B.. 6-9 September 1983. 607-9. Robinson D. Moore J.. Rowe B.
Applied and Environmental Microbiology. 1985. Effects of processing and storing oyster meats on concentrations of indicator bacteria.F. 1990.. 690-4. Kaper J. Trost P.A. Incidence of motile aeromonads from United States west coast shellfish growing estuaries. 1987.D. Plesiomonas shigelloides: An opportunistic food and waterborne pathogen.. 80.. Seidler R. Frequent isolation of Edwardsiella tarda and Plesiomonas shigelloides from healthy Zairese freshwater fish: A possible source of sporadic diarrhoea in the tropics... Bark D.J.. 598-601. Journal of Food Protection. Aeromonas hydrophila: ecology and toxigenicity of isolates from an estuary. Kain K... Kaysner C.C.. Wekell M.. Report (1/13/84). Robinson J. 1988. 78..R.W. 1984. 55... Journal of Applied Bacteriology. Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species. Applied and Environmental Microbiology.. 5.W. Annals of Internal Medicine. Vandepitte J. Hunt J. 77. 998-1001. Kelly M. 36.M. Clinical features. Herrington T.. In vitro and in vivo pathogenicity of Plesiomonas shigelloides. Infection and Immunity.M. Esch G. Department of Health and Human Services/Public Health Service/Food and Drug Administration. Gracey M. Sullivan J. 849-55.. Hood M. Cook D. 1146-9. 1997. Tzipori S. Koburger J.A. 53. Partridge K.L.. Blake P.A. The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish. Miller M. Fla. Cooper M..M. 39. 81. Levine M. Colwell R. Holmerg S.. Stott R. Burke V. Hickman-Brenner F. Fliermans C. Applied and Environmental Microbiology. Abeyta C. 47. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness.H. 475-9.M. Wekell M. Wekell M... 359-77. Kaysner C. Stelma G. 76..M.D.J.. 1981.D..M.. 1986. 50.A. Hazan T..T. Journal of Clinical Microbiology. Jr. 87. Journal of Food Protection. 84. 1989.. Petersburg. GA. Beaman J..A.. Journal of Rapid Methods and Automation in Microbiology.. Robins-Browne R. 48. Wachsmuth I.P. Ruple A. Herrington D. 1980. 86.. Abeyta C. 75. 1989. Van Damme L. 27..L. Oyster borne disease outbreaks in St.J. vibrios and Aeromonas hydrophila. Journal of Food Protection. 52. Farmer J. 85. III.K.M. Journal of Food Protection. and treatment of Plesiomonas shigelloides diarrhoea.N. 223-47... Jr. 979-85. Tall B. 343-9. 83.. Hirsch R. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. Plesiomonas enteric infections in the United States. 47.B.L.W..A.R. 731-8. 76 .A. Journal of Food Protection. Kaysner C. 105. Baker R. Tenge B.. epidemiology. 79. Prevalence and distribution of Aeromonas hydrophila in the United States.. 7-12.FISH AND SEAFOOD 74. 1978. Lockman H. 52. Jr. Atlanta.C. 449-57. Singelton F.A.J. 82.. Peterson D. 1984.. Abeyta C.
Quality Control. 1995.. Food and Drug Administration Compliance Program Guidance Manual.L.1987. in U. U. Clams.. 3. Chapter 5: Foods.C. Slavin J.. Washington D.S.. Technomic Publishing Co.12). Smith J. 1991.C. Food and Drug Administration. 1987. U. U.E. Food and Drug Administration. Collette R. Government Printing Office .600 (CPG 7119.H..C. Martin R.S. Food and Drug Administration. U. Ed. Ed.MOLLUSCAN SHELLFISH 88. Inc. 77 . National Academy Press. mussels. Program 7303.S. 2000. Food and Drug Administration.. fresh or frozenadulteration by bacteriological contamination Sec..S.S.Pathogen monitoring of selected high risk foods (FY 88/89). 90. Baird-Parker T. 91. 63-8. in U. Food and Drug Administration.S..A. Food and Drug Administration. Sherlock C. Food and Drug Administration. Does Aeromonas hydrophila preferentially colonise the bowels of patients with hematologic malignancies? Diagnostic Microbiology and Infectious Disease. Gaithersburg.C. Seafood Safety. in National Shellfish Sanitation Program Manual of Operations. 560. The Microbiological Safety and Quality of Food.W. Ed. U.030. Food and Drug Administration Compliance Policy Guides.8 Further Reading Ahmed F. Gould G. 1995.. Lund B. Washington D. 7. and HACCP: A Global Focus. oysters. U.S. Part I Sanitation of Shellfish Growing Areas. 1997.S. Lancaster.M.R. Washington D. Pittsurgh. Burdge D. 89. Fish Inspection. Food and Drug Administration.E. Aspen Publishers Inc.W.
(deep-water shrimp).5 and 1.1 Definitions Crustaceans are invertebrate animals and represent a class of the phylum Arthropoda. Pandalus borealis (deep-water shrimp). crayfish and shrimp (1). and frozen in 1 and 2. Pandalus montagui and Crangon crangon (inshore shrimp).0 kg polythene bags.. with additional water to form a glaze. lobsters and crabs have ten legs. Shrimp can be purchased in the raw or cooked state in a variety of forms: Entire shrimp. They possess a segmented body. peeled product. M.4. e. CRUSTACEAN SHELLFISH Linda Nicolaides. lobsters. Crayfish.0 kg polythene bags. Peeled ready-to-cook shrimp are raw.5 and 1.g. crabs. with shells and heads on or off are usually packed. packed and frozen in 0. Breaded shrimp and scampi are battered and coated in breadcrumbs and sold frozen to the consumer for cooking prior to consumption. Parapeneopsis spp.. Species eaten from tropical waters include Penaeus spp. jointed limbs and a chitinous exoskeleton.5 kilo pack (boxes or bags).. Peeled and cooked shrimp are ready-to-eat after controlled thawing. FRSPH Ethical Trade and Food Management Group Natural Resources Institute University of Greenwich Central Avenue Chatham Maritime Kent ME4 4TB United Kingdom 4. and Trachypenaeus spp. prawns and scampi are temperate species commonly eaten by man and include members of the families Nephrops norvegicus (scampi). Such products are usually only partially cooked before freezing.Ph. and are usually aquatic. the front pair ends in claws. Shrimps. and available in 0. 79 .
for crabs and lobsters. packed into polythene bags. Microbiological risks associated with crustacean shellfish increase in proportion to the degree of handling that the product undergoes. lobster and crayfish are available for the consumer owing to the growing demand for ready-to-eat products. tropical waters.g. their feeding and living habits. this presents more favourable conditions for growth by both spoilage and pathogenic bacteria. lobsters and crayfish are traditionally kept alive after capture and then cooked to kill immediately before consumption.g. For example. Pasteurised crabmeat is edible meat removed from the exoskeleton by hand. e. However.2 Initial Microflora As with finfish. spoilage bacteria are of little importance as. during the peeling of cooked shrimp. Spoilage bacteria associated with temperate crustaceans include Pseudomonas spp. traditionally. Poor hygienic procedures will increase the probability of contamination with bacteria of public health significance. the season. The structure of the circulatory system of crabs is not closed. Following capture or harvest. stored and transported as frozen product. boiled to kill the crustaceans and cook their meat prior to consumption. the geography of the area they are captured from or are grown in. storage above the recommended storage temperature. Control measures to prevent these potential hazards occurring include training and supervision of operatives as well as Good Hygienic Practices (GHP). If these products are subjected to temperature abuse. particularly after it has been cooked. there is a growing market for fresh lobster tails or cooked crabmeat in crab shells. shrimp die immediately after capture. hence. which means that the haemolymph can be a reservoir of bacteria. so that spoilage may occur during landing and transportation to the processing plant. with the increasing demand for ready-to-eat/cook crustacean products. Shewanella putrefaciens and 80 .FISH AND SEAFOOD Crabs. the level and type of the initial microflora of crustacean shellfish will reflect a combination of factors. particularly for members of the genus Vibrio (2). only live carcasses are processed. and the temperature and quality of the waters in which they exist. which include the environment from which they have been captured or harvested. the potential exists for crosscontamination from both the processor and from the environment. 4. Spoilage begins following death once the immuno-response system of the crustacean shellfish fails to function. e. The microbial flora of crustaceans from temperate waters differs from that of crustaceans from warmer. lobster tails and cooked crabmeat presented decoratively in their shells.. As with finfish. Cooked and chilled. In comparison. or frozen crab. the flora will change depending upon the methods of handling and/or the environmental conditions to which they are exposed. it is important that shrimp are handled and iced correctly in order to keep them in as fresh and uncontaminated a state as possible.
Hence. so they are either sold on the local market or are processed in local processing plants. corynebacteria. Shrimp might be further processed with the shell on. glazed and frozen (see Figure 4. uncooked prawns and shrimp is presented in Figure 4. The rate of spoilage will be a function of time and temperature. Micrococcus spp. lobsters and crayfish are harvested from cages or pots containing bait. 4. chilled sea water. After capture and transport to the processing plant. 4. peeled and cooked. or cooked. In contrast. the genera associated with tropical species include Enterobacteriaceae. Shrimps can be processed whole. As the muscles of crustacean shellfish decompose rapidly after death dead animals are rejected. 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. The claws are usually bound together to prevent the animals fighting and causing damage to each other. bacterial contamination and spoilage are inevitable.3. and will be dependent upon whether the crustaceans were harvested from cool or warmer waters. When the shrimp arrive at the processing plant they are washed and then sorted into size. and Achromobacter spp. Pseudomonas spp. up to 16 days for tropical species compared with 8 . thus. They are then taken to market alive in either holding tanks. the rate of deterioration of the crustacean shellfish by these organisms is reduced.CRUSTACEAN SHELLFISH members of the Acinetobacter-Moraxella group. graded according to size.10 days for temperate water species (3).1 Prawns and shrimps An example of the process used for preparing frozen. from the time of capture and processing. they are washed.. Shrimp will then be glazed and frozen in a raw state. In comparison shrimps die quickly after capture.. or with the shell and central vein removed. especially if they are stored in ice or iced water. chilled 81 . Scrupulous hygiene is important at this stage to prevent the cross-contamination of the shrimp tails. These animals can survive out of water if the surrounding environment is kept cool and damp. whether GHP are in place. there is evidence that the shelf life of tropical crustaceans is longer than that of temperate-water species stored correctly in ice . however..3 Processing and its Effects on the Microflora Crabs. with “head on” or “head off” whereby the edible tail is removed from the head. A large proportion of shrimps are produced by aquaculture. The latter process can expose the shrimp muscle to incidents of cross-contamination if GHP are not followed. Alcaligenes. or transported directly to processing plants. Vibrio spp. by removing the head and thorax a source of enzymes is eliminated that will prevent the occurrence of black spots.1).1. As with finfish. gills and thorax. or are found in brackish water around the shores.for example.
Breaded products introduce an added risk of spore forming bacteria. should be followed to ensure that the product is not re-contaminated with microorganisms present in the processing environment or from the workers. Fig. and packed to meet the customer’s specification. Good Manufacturing Practice (GMP). including GHP. 4. After cooking. Flow diagram .FISH AND SEAFOOD frozen. 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. particularly the pathogens Bacillus cereus and Clostridium perfringens originating in the flour. spices and cooking batter.1.frozen uncooked prawns 82 .
wet and ‘soapy’ or ‘jammy’ to the touch. If hygienic procedures are not followed.3. whilst boiled crustaceans have a pleasant slightly sweet smell.2 Crabs and lobsters Crabs and lobsters should be alive when boiled or sold.4. there is no associated spoilage problem. 4. contaminating microflora may include enteric bacteria. the joint between the tail and carapace is open and the limbs hang loosely. dull and sticky.1 Prawns and shrimps Freshness indicators for prawns and shrimps are for the shells to be crisp and dry and feel sharp and cool to the touch. putrefaciens and Pseudomonas spp. there is a new hazard of contamination of the cooked meat by workers during its physical removal from the exoskeleton. putrefaciens. spoiling prawns and shrimps have shells that are soft. the meat is either repacked and sold in the shells. However. with the growing demand for ready-to-eat products from crab and lobster meat. for crabs.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. immediately prior to boiling. slightly iodine smell. since crabs and lobsters are traditionally consumed immediately after cooking. An offensive odour is also produced. indicative of protein degradation and the growth of the principal spoilage bacterium Sh.2 Cooked crabs and lobsters As indicated earlier. Heat is produced if the shrimps decompose in a confined space. with visible decomposition or green discoloration seen through the membrane exposed by the dropping tail. bacteria of public health significance. and a strong ammoniacal odour is produced. In comparison.4. The shell will be soft. 83 . there is no indication of life. Processed crab and lobster meat can also be bought as packages of meat that have been subjected to a pasteurisation step. or packaged and processed. With spoiled crabs and lobsters. or in hermatically sealed cans that have undergone an appropriate heat treatment. 4.CRUSTACEAN SHELLFISH 4. If boiled. If the tail drops. the tightness of limbs will indicate whether they were killed by immersion in boiling water or. and have a sweetish. Once removed and cleaned. 4. bright and dry appearance. viruses or parasites. The shells have a crisp. they include Sh.
2 Listeria spp.5 4. 4. and usually for both. and to increase testing of many other domestically produced seafoods under the General Pathogen Surveillance Programme (14. in 24. although efficient cooking will quickly eliminate it (6). However. Results of a survey carried out in England and Wales between 1987 and 1989 demonstrated that L. the first in a series of Class I recalls was issued in May 1987 to retrieve nearly 4 tonnes of tainted crabmeat (11). often contains Salmonella and other enteric pathogens (8) and has been demonstrated to contaminate crustaceans growing in these environments. even if GMP is followed. animal manure. shrimps and cockles sold (17). Following this. in compliance with the Food and Drug Admistration’s (FDA) ‘zero tolerance policy’ for L. They are mesophilic microorganisms usually residing in the intestinal tracts of human beings and warm-blooded animals.5. monocytogenes was absent from 40 samples of cooked prawns. The pathogen was later identified in frozen raw shrimp (13) and lobster tails (14) . which required automatic detention of all Mexican frozen crabmeat that tested positive for Listeria and Escherichia coli (12). monocytogenes in contaminated crustacea was recognised in the United States of America (USA) when the pathogen was isolated from Mexican frozen cooked crabmeat (10). shrimp and shellfish (11). Hence.5% of frozen raw shrimps from different countries (13). monocytogenes in cooked ready-to-eat crustaceans is recognised as a potential risk to selected groups of individuals: the very young and 84 . 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. 5. Further surveys identified Listeria spp. monocytogenes in cooked ready-to-eat foods. The above results prompted the FDA to develop a compliance programme for the pathogen.1 Pathogens: Growth and Survival Salmonella spp. as well as for Salmonella. Although foodborne illness had not been associated with the product. 7). cooked and smoked fish. Moreover. in domestic/imported shrimps (15). One of the earliest accounts of the presence of Listeria monocytogenes in crustaceans was by Russian workers in 1959 (9). a common pond fertiliser used in aquaculture. fish fingers.5. nearly 30 years elapsed before the potential hazard of L. Aquatic birds and the hands of workers have also been identified as sources of the pathogen. The genus Salmonella contains a wide variety of ‘species’ pathogenic for man or animals. later surveys in 1990 recovered the pathogen from retail raw. Salmonella cannot be completely removed from raw shrimp during processing. 16). An import alert was put in place in June 1987. 6. The presence of L.FISH AND SEAFOOD 4.cooked prior to consumption.
the causative agent of cholera. 24. Once the competing microflora has been removed by a heat treatment. combined with the degree of handling of these types of product. cholerae non-O1. V. The main source of contamination is the hands of processing personnel. 25). which was eventually attributed to a ship discharging its bilge into the coastal waters of Peru. botulinum type E at 10 °C but not at 4 °C (20).3 Staphylococcus aureus Staph.70% of healthy individuals carry toxin-producing strains of Staph. although more recently the non-O1 type has also been demonstrated to cause foodborne illness in crustaceans. therefore.g. 4. cholerae is divided into two groups. GMP should be followed in the processing plant to ensure that the final product is held at 3 °C. especially in products that have been vacuum packaged. 4. although contamination by types A and B may occur from the environment. 22) as well as from undercooked crustacea when harvested from contaminated waters (23. was associated with seafood consumption. Vibrio parahaemolyticus and Vibrio vulnificus are species that have been reported to be associated with raw crustaceans harvested from estuarine waters (21. it is important that the cooking stage is controlled to eliminate the pathogen from the product and that procedures are in place to prevent any cross-contamination of the cooked product. aureus because of the ubiquity of this organism in the processing environment. is widely distributed in aquatic environments. cholerae. as part of the natural bacterial flora. about 50 . aureus is not a member of the intrinsic flora associated with crustacean shellfish. The pathogen has resistant spores. 4. pregnant women and the immuno-compromised.5. so is able to survive heat treatments that eliminate vegetative cells.CRUSTACEAN SHELLFISH old. particularly crabmeat (18. botulinum type E is commonly associated with fish and fishery products. whilst boils and cuts are also common sources of the pathogen.4 Clostridium botulinum C. 85 . which can become colonised by V. e. An epidemic that began in South America in 1991. Crabs are covered in a chitin exoskeleton. the pathogen can grow at temperatures as low as 3 °C. To date.5. Control should be achieved by promoting GHP and staff training programmes. 19). cholerae O1 and V. Vibrio cholerae. aureus on their hands. However. monocytogenes has been attributed to crustaceans.5 Vibrio spp. V.5. The former. pasteurisation and cooking. Crustaceans have been linked with food intoxications attributed to Staph. no incidence of foodborne illness caused by L. Researchers have demonstrated that inoculated shrimp supported toxin production by C.
However. Caliciviruses. although A.FISH AND SEAFOOD and caused major economic losses to South American countries that export shrimp to the United States. hydrophila is able to grow well at refrigeration temperatures (0 . 4. hydrophila and A.g. non-B Hepatitis viruses (31). Hepatitis A virus presents the most serious hazard. to date. sobria are known to be fish pathogens and are commonly isolated from ponds used for aquaculture. been implicated in a case of foodborne infection associated with crustaceans. Parasitic infections are controlled by either cooking infected material prior to consumption.5. 31. vulnificus is an invasive and lethal pathogen associated with wound infections. V. However. occurs in crustaceans from warm environments. whilst 5. parahaemolyticus. jejuni has not.7 Aeromonas hydrophila and Aeromonas sobria A. however.30 MPN/g) (27). the potential hazard exists as C. It is more commonly implicated in countries where crustacean shellfish are eaten raw. Astroviruses and non-A.9 Viruses Crustaceans grown in waters contaminated with sewage may become carriers of viruses such as Hepatitis A.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. V.8% of samples contained C. South America. A. Norovirus. coli. a marine halophile. jejuni was isolated from 36 (15%) of 240 samples of freshly hand-picked blue crab (Callinectes sapidus) meat. 4. hydrophila has been isolated from prawns (29). they have not been confirmed to be causal agents of foodborne illness associated with crustaceans (28). 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. Of these.5. but has the potential to cause fatal foodborne illness (26).5. or freezing at a temperature of -23 °C for 7 days (30. 32). in a survey carried out in the United States between 1977 and 86 . 4.6 Campylobacter jejuni and Campylobacter coli C. The organism is widespread in estuarine waters and is a common isolate from harbour water in warm climates.8 Parasites The use of animal excreta in aquaculture has also been associated with foodborne parasitic infections of crustaceans.5. e. 4. particularly trematodes (6). Japan. Quantitative levels were below limits of detection in all cases (<0.
CRUSTACEAN SHELLFISH 1981. A summary of microbiological criteria for crustacean shellfish is listed below. Both Hepatitis A virus and Norovirus are stable pathogens and are relatively resistant to inactivation (34). 4. produced or imported into the European Union (EU). The risk of such hazards is far higher with fin fish and molluscan shellfish.10 Allergens Allergens have not been reported to be a significant hazard with crustacean shellfish. Therefore any microbiological criteria need to be part of this HACCP-based system to verify that the system is under control. 38. Provisions laid down concerning the use of additives would also apply to these types of products. although it is one of the European Union (EU) legislated allergens. consistent with that in the stomach.6. 4. They can survive low pH levels. which results in a persistent and apparently permanent loss of short-term memory (36). 2073/2005.1 EU Legislation Microbiological standards for cooked crustaceans and shellfish.6 Published Microbiological Criteria The global legislative requirement for controlling the safety (and quality) of crustacean shellfish is to apply a preventive. Norwalk viruses can also withstand heating to 60 °C. 4. 39). or chlorine concentrations similar to those found in drinking water.5. only 0. 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. No foodborne incidents have been attributed to this source to date. 4. The toxin causes amnesic shellfish poisoning (ASP) when consumed. from farm to fork (37. 87 . which could be transferred to the meat during unskilled removal from the shell (35). Selected parts from this regulation is presented below.47% of outbreaks attributed to Hepatitis A virus were attributed to a foodborne origin (33).5.11 Chemical pollutants and biotoxins Researchers have demonstrated that crabs feeding in waters contaminated with algae do have potentially high levels of algal toxin domoic acid (DA) in their viscera. are included in the Commission Regulation (EC) No.
FISH AND SEAFOOD
22.214.171.124 Microbiological criteria applicable to the production of cooked crustaceans (Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs) 126.96.36.199.1 Chapter 1 - Food safety criteria
L. monocytogenes Prepared crustacean products fall into the category described in the regulation as “Ready-to-eat foods available to support the growth of L. monocytogenes other than those intended for infants and for special medical purposes”. During capture and preparation there is a zero tolerance for the presence of L. monocytogenes on the product. However, to take into account the issues of natural habitats being contaminated with low levels of this pathogen there is a slight loosening of the regulation at the point of sale and for the duration of the products shelf life (refer to Table 9.I, Food Category 1.2). If the level of L. monocytogenes should be above this level then the consignment is considered to be unsatisfactory. Salmonella All cooked crustaceans and molluscan shellfish should be free from Salmonella at the point of sale and for the duration of their shelf life (refer to Table 9.I, Food Category 1.16). Should any sample test positive for Salmonella then the product is declared to be unsatisfactory. 188.8.131.52.2 Chapter 2 - Process hygiene criteria
Process hygiene criteria for E. coli and coagulase-positive staphylococci in shellfish and shucked products of cooked crustaceans and molluscan shellfish are laid down (refer to Table 9.II, Food Category 2.4.1). 4.6.2 International requirements
International requirements are based upon having a risk-based, preventive management system in place at all stages of the supply chain. The requirements for this preventive approach is described as follows (37): • Fish and fishery products should be prepared in plants certified by the local competent authority. All certified plants should comply with the GHP (39); • The fisheries industry should take responsibility, implement and maintain safety management systems based upon HACCP; • The national competent authority is responsible for the certification of fish processing and manufacturing plants, verification of effective systems, and
issuing of certificates of compliance for export products. This includes auditing and inspection programmes; • National surveillance and monitoring programmes should be in place to demonstrate that all identified hazards are under control, e.g. biotoxins, and to identify potentially emerging hazards; Within this requirement is the flexibility for countries and trading blocks to set specific standards required to deliver an equivalent requirement with those at national level. Such standards need to be science-based, especially if they are more stringent that those recommended by the Codex Alimentarius Commission. Verification that GHP have been followed during the handling and preparation of crustacean shellfish for consumption is usually done by assessing the level of E. coli and/or coagulase-positive staphylococci in the products. Legal requirements generally require that these groups of indicator organisms are present in levels <100 cells/gram of product. Pathogenic bacteria such as Salmonella spp. should be absence is 25 grams multiplied by the number of subsamples tested. 184.108.40.206 United States The FDA brought into force a Federal Mandate Seafood Rule in 1995 which forms the foundation for good practices required for the handling and processing of imported fish and fishery products into the U.S. The best practices include GHP and HACCP (37). All fish and fishery products produced in and imported into the U.S. after 17 December 1997 are required to demonstrate that HACCP principles were applied to their handling and processing. The USFDA has a zero tolerance for Salmonella in fish and fishery products; this includes farmed shrimp. The US FDA considers that the presence of pathogenic microorganisms in seafood is an indicator of adulteration (32). 4.7
Encarta 1997. Encarta 97 Encyclopedia World English Edition. Microsoft. The International Commission for Microbiological Specifications for Food. Fish and Fish Products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. The International Commission for Microbiological Specifications for Food. New York, Kluwer Academic/Plenum Publishers. 2005, 174-249. Cann D.C. Bacteriology of shellfish with reference to international trade, in Handling, Processing and Marketing of Tropical Fish. Ed. Tropical Products Institute. London, Ministry of Overseas Development. 1976, 377-94. D’Aoust J.Y., Gelinas R., Maishment C. Presence of indicator organisms and the recovery of Salmonella in fish and shellfish. Journal of Food Protection, 1980, 43, 679-82.
FISH AND SEAFOOD 5. 6. 7. 8. 9. 10. 11. Fraiser M.B., Koburger J.A. Incidence of salmonellae in clams, oysters, crabs and mullet. Journal of Food Protection, 47, 343-5. Reilly P.J.A., Twiddy D.R., Fuchs R.S. Review of the occurrence of Salmonella in tropical shrimp. FAO Fisheries Circular No. 851, 1992, (FIIU/C851). Reilly P.J.A. Salmonella and Vibrio cholerae in brackishwater cultured tropical prawns. International Journal of Food Microbiology, 1992, 16, 293-301. Twiddy D.R. Antibiotic-resistant human pathogens in integrated fish farms. ASEAN Food Journal, 1995, 10, 22-9. Olsufjew N.G., Petrow V.G. Detection of Erysipelothrix and Listeria in stream water. Zhurnal Mikrobiologii Epidemiologii i Immunobiologii, 1959, 30, 89-94. Anon. First Listeria finding in crabmeat confirmed by FDA. Food Chemistry News, 1987, 29 (14), 38. Ryser E.T., Marth E.H. Incidence and behaviour of Listeria monocytogenes in fish and seafood, in Listeria, Listeriosis and Food Safety. Eds. Ryser E.T., Marth E.H. New York, CRC Press. 2007, 617-54. Anon. Mexican crabmeat, Greek pasta automatically detained. Food Chemistry News, 29 (19), 43. Anon. FDA checking imported domestic shrimp, crabmeat for Listeria. Food Chemistry News, 1987, 29 (24) 15-7. Archer D.L. Review of the latest FDA information on the presence of Listeria in foods. WHO Working Group on Foodborne Listeriosis, Geneva, Switzerland. 1988, 15-9. Anon. FDA to sample shrimp for Salmonella, Listeria. Food Chemistry News, 1988, 30 (36), 9-10. Anon. Monitoring of changes to listeriosis problem urged. Food Chemistry News, 1989, 31 (20), 13-7. Richmond M. Report of the Committee on the Microbiological Safety of Food. London, HMSO. 1990, 133-7. Swartzentruber A., Schwab A.H., Duran A.P., Wentz B.A., Read R.B. Jr. Microbiological quality of frozen shrimp and lobster tail in the retail market. Journal of Food Protection, 1980, 40, 765-9. Wentz B.A., Duran A.P., Swaartzentruber A., Schwab A.H., McClure F.D., Archer D., Read R.B. Jr. Microbiological quality of crabmeat during processing. Journal of Food Protection, 1985, 48, 44-9. Garren D.M., Harrison M.A., Huang Y-W. Clostridium botulinum type E outgrowth and toxin production in vacuum-skin packaged shrimp. Food Microbiology, 1994, 11, 467-72. Molitoris E., Joseph S.E., Krichevsky M.I., Sindhuhardja W., Colwell R.R. Characterisation and distribution of Vibrio alginolyticus and Vibrio parahaemolyticus isolated from Indonesia. Applied and Environmental Microbiology, 1985, 50, 1388-94.
12. 13. 14.
15. 16. 17. 18.
CRUSTACEAN SHELLFISH 22. Varma P.R.G., Iver T.S.G., Joseph M.A., Zacharia S. Studies on the incidence of Vibrio cholera in fishery products. Journal of Food Science and Technology, 1989, 26, 341-2. Madden J.M., McCardell B.A., Morris J.G. Jr. Vibrio cholerae, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 525-42. Twedt R.M. Vibrio parahaemolyticus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 543-68. Oliver J.D. Vibrio vulnificus, in Foodborne Bacterial Pathogens. Ed. Doyle M.P. New York, Marcel Dekker. 1989, 569-600. Pearson A.M., Dutson T.R. Quality Attributes and Their Measurement in Meat, Poultry and Fish Products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994. Reinhard R.G., McAdam T.J., Flick G.J., Croonenberghs R.E., Wittman R.F., Diallo A.A., Fernandes C. Analysis of Campylobacter jejuni, Campylobacter coli, Salmonella, Klebsiella pneumoniae and Escherichia coli 0157:H7 in fresh handpicked blue crab (Callinectes sapidus) meat. Journal of Food Protection, 1996, 59 (8), 803-7. Buchanan R.L., Palumbo S.A. Aeromonas hydrophila and Aeromonas sobria as potential food poisoning species: a review. Journal of Food Safety, 1985, 7, 15-29. Palumbo S.A., Maxino F. Williams A.C., Buchanan R.L., Thayer D.W. Starch ampicillin agar for the quantitive detection of Aeromonas hydrophila in foods. Applied and Environmental Microbiology, 1985, 50, 1027-30. Wekell M.M., Manger R., Colburn K., Adams A., Hill W. Microbiological quality of seafoods: viruses, bacteria and parasites, in Seafoods: Chemistry, Processing Technology and Quality. Eds. Shahidi F., Botta J.R. Glasgow, Blackie Academic and Professional. 1994, 196-219. Schantz J., McKerrow J.H. The dangers of eating raw fish. New England Journal of Medicine, 1989, 320, 1143-5. Ahmed F.E. Seafood Safety. Seafood Safety. Committee on Evaluation of the Safety of Fisheries Products, Food and Nutrition Board, Institute of Medicine. Washington D.C., National Academy Press. 1991. Blackwell J.H., Cliver D.O., Callis J.J., Heidelbaugh N.D., Larkin E.P., McKercher P.D., Tayer D.W. Foodborne viruses: their importance and need for research. Journal of Food Protection, 1985, 48, 717-23. Kapikian A.Z., Chanock R.M. Norwalk group of viruses, in Virology. Eds. Fields D.M., Knipe D.M. New York, Raven Press. 1990, 359-77. Horner R.A., Postel J.R. Toxic diatoms in western Eashington waters (US west coast). Hydrobiologia, 1993, 269/270, 197-205. Perl T.M., Bedard M.S.M., Kosatsky T., Hockin J.C., Todd E.C.D., Remis R.S. An outbreak of toxic encephalopathy caused by eating mussels contaminated with domoic acid. The New England Medical Journal, 1990, 322, 1775-80.
23. 24. 25. 26.
34. 35. 36.
FISH AND SEAFOOD 37. Ababouch L., Gandini G., Ryder J. Causes in Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. Australian Government. Australian Quarantine and Inspection Service Hazard Analysis and Critical Control Point (HACCP). A Guide to Compliance with Export Control (Fish and Fish Products) Orders 2005. FAO/WHO. General Hygiene Requirements. Rome, Food and Agriculture Organisation of the United Nations. 2003.
Further Reading Spoilage and pathogens
Gram L., Huss H.H. Fresh and Processed Fish and Shellfish, in Microbiological Safety and Quality of Food Volume 1. Eds. Lund B.M., Baird-Parker A.C., Gould G.W. Maryland, Aspen Publishers Inc. 2000, 472-506. Ababouch L., Gandini G., Ryder J. Causes of Detentions and Rejections in International Fish Trade. FAO Fisheries Technical Paper 473. Rome, Food and Agriculture Organisation of the United Nations. 2005. International Commission on Microbiological Specifications for Foods. Fish and fish products, in Microorganisms in Foods 6. Microbial Ecology of Food Commodities. Ed. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1998, 130-89. Sutcliffe P., Disney J. Handling, Processing and Marketing of Tropical Fish. London, Tropical Products Institute. 1977. Connell J.J. Control of Fish Quality. Oxford, Fishing News Books. 1995 Ward D.R., Hackney C. Microbiology of Marine Food Products. New York, Van Nostrand Reinhold AVI. 1991 Pearson A.M., Dutson T.R. Quality Attributes and their Measurement in Meat, Poultry and Fish products. Advances in Meat Research Series. Volume 9. London, Chapman & Hall. 1994.
4.8.2 Pathogens - general
International Commission on Microbiological Specifications for Foods. Microorganisms in Foods 5. Characteristics of Microbial Pathogens. Eds. International Commission on Microbiological Specifications for Foods. London, Blackie Academic and Professional Press. 1996. Varnam A.H, Evans M.G. Foodborne Pathogens - An Illustrated Text. London, Wolfe Publishing Ltd. 1991. Doyle M.P. Foodborne Bacterial Pathogens. New York, Marcel Dekker. 1989.
and marinating processes. and salting is done by stacking the fish in layers. rather than for preservation itself. Simon Derrick The Grimsby Institute Humber Seafood Institute Europarc Grimsby DN37 9TZ United Kingdom 5. alternating fish layers with a layer of salt. SMOKED AND DRIED FISH Dr.5. CURED. Wet salting or brining. fish is immersed for a few hours in a relatively low concentration brine. Usually. and a preliminary operation to some smoking. dependent on the country or the type of product. to assure a salt concentration in the fish flesh that is appropriate for flavouring purposes prior to the use of other preservation techniques. Fish can be salted in three different ways or their combination: Dry salting. This can be called ‘kench’ curing and is mainly used for non-fatty white fish. The immersion in concentrated brines for long periods is usually used for fatty species and for longer-term preservation. drying. The fish is immersed in a brine solution in which different salt concentrations can be used according to the process. split and opened out flat. drying and smoking processes. 93 . both to preserve and to create a variety of new products. and the extracted moisture is allowed to drain away. The fish is headed. The processes take anywhere from a few hours to a few days or even months. where the salt replaces moisture in the fish flesh. Fish curing is practised worldwide. Salting is both a method of preserving fish. It includes the salting. and before significant spoilage takes place.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. their combinations and/or variations. and is sometimes known as ‘pickling’.
the ice sublimes and the vapour is removed from the fish by vacuum pumps. the preservative effect is due to the low pH used in the process. Smoking is an ancient process for preserving and flavouring food. and the fish can be immersed in such brines for long periods. The principle of preservation of the first method is the inactivation of enzymes and killing the non-spore-forming organisms. 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 . thus delaying spoilage for a few days. without other preservation methods applied after boiling. is used as a preservative instead of salt. There are some marinating processes where salt is used for the first cure and a firmer product is obtained. Included in this group are some lightly preserved products where no acid is added to the marinade. such as ‘gravad’ fish. Heat is transferred to fish from heated air or a heated surface. or boiling it with dry salt and water. This is a mixture of a dry and a wet salting process. evaporated water is removed by vacuum pump. The products are usually consumed raw. usually vinegar. Drying is the preservation of fish by removing the moisture content of the fish flesh by evaporation. Longer shelf life is achieved in the products when salt is added. this ‘brine’ can be collected and placed again over the fish several times. After the freezedrying process. Food products can be dried in three different ways: Air or contact drying. are added to the marinade. where an organic acid.FISH AND SEAFOOD Dry / wet salting. The product is dried by contact with heated surfaces or by radiation. It has found limited application with fish. Usually. and the air movement removes the vapour. Salt. When the process is running in a closed system. this was done naturally by exposure of the fish to the air. the product rehydrates quickly but does not retain reabsorbed moisture on cooking. Vacuum drying. as concentrated brines are formed. Salted-boiled fish are the products obtained on either boiling fish in a brine. Marinades are acidified brines. it can also be named ‘pickling’ or ‘blood pickling’. thus reducing the water activity of the product. thus making the product unsuitable for microbial growth. as well as other flavourings (sugar. As the water is drawn out of the flesh by the salt a brine is formed. Thus. The fish is mixed with dry salt but in a watertight container. Marinating or pickling. Freeze drying. This is the common method of drying fish. Fish in contact with refrigerated plates freeze. depending on the type of product. spices). Traditionally. Salt-boiling. under normal pressure. prior to or during the boiling process. It is not commonly used for fish products.
The smoke itself may contain spores of moulds. After the salting step. 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. which impart the flavour of smoke to fish. smoking is now used mainly to give the fish a characteristic flavour.2 Initial Microflora The initial microflora are those of the raw fish from which the products are made. Usually. Cold-smoking. 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. fish can be smoked by means other than the direct action of smoke. sporeforming bacteria or osmophilic moulds. 95 . or by means of smoke-flavouring processes with liquid smoke. such as injection or dipping in a solution of liquid smoke. Usually. process temperatures are over 60 °C. leaving only elevated levels of carbon monoxide. Smoked-flavoured fish. It can be smoked either traditionally. Also. the sugar used in some products may contain yeasts and moulds as well as spore-forming bacteria. Hot-smoking. the fish is subjected to heat for a period that does not coagulate protein. 5. It can be injected into the product. Liquid smoke is a natural aqueous condensate of wood smoke that has been aged. Clear smoke. by exposure to the wood or sawdust smoke. With the exception of some Asian countries. In this process. or by means of smoke flavouring processes. including pathogens. which irreversibly combine with the haemoglobin in the tissue to maintain the red colour. It is commonly used in place of traditional smoke. A dry or wet salting step is always used in the process before exposure to the wood or sawdust smoke. or added to the brine with other flavourings. Smoking can be either traditional. the salt used in the processes may contain halophilic bacteria. However. In this process. the fish is subjected to heat for a sufficient period of time to coagulate protein throughout the fish. Smoked fish is prepared by subjecting the fish to the direct action of smoke from wood or wood sawdust burning. temperatures below 30 °C are used. The process was designed so that the smoke is filtered to remove all smoke particles. and filtered to remove tars and particulate matter. The different spices used in the immersion brines are also a potential source of microorganisms. SMOKED AND DRIED FISH of smoke.CURED. or liquid smoke. and the preservative effect is only slight.
FISH AND SEAFOOD 5.4 184. all using salting steps.2 105. sometimes the salt crystallises onto the surface of the fish.0 .80º brine (184.4% salt (Table 5.3 Processing and its Effects on the Microflora Owing to the wide variety of products.0 158.2 g of salt per litre of brine). Different brine strengths are required according to the curing processes and/or the type of product. smoking or marinating processes. it is impossible to establish standardised processing stages in the curing of fish. Salt in the water phase is the amount of salt compared with the total amount of water and salt in the fish.6 264. When a milder brine. a 0º brine saturation means pure water.2 237. or salt in the water phase.8 211.I). and salt content. For smoking salmon. it is important to state the temperature. about 50º (132.8 79. This should be distinguished from the salt content.6 g of salt or more per litre of brine) may cause salt burn in this way. When working with salting. etc. a 90º or above brine (237. % 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. Salt concentration means the percentage of salt in the water contained in the fish.6 132. Thus.0 g of salt per litre of brine).4 52. Therefore.I Brine strength at 18 °C (after Bannerman (2)) Brineometer degrees 10 20 30 40 50 60 70 80 90 100 96 Weight of salt (g/litre of brine) 26.8 211. the fish is usually salted with a 70 . The dissolution of salt in water is dependent on the temperature. it causes swelling because the fish absorbs water from the salt solution (1). When a 100º brine is used. it is important to understand the concept of salt concentration. The strength of the solution is described as degrees of saturation (brineometer degrees). TABLE 5. Salt in the water phase links the two conditions of moisture and saltiness. A 100º brine solution means that the solution is saturated and no more salt can be dissolved. At 18 °C a 100º brine is produced when the solution contains 26. processes and producers’ private recipes. which is the percentage of salt in the whole weight of fish that is water plus protein plus fat. only the most common processes can be described here. is employed.
and salt can speed this oxidation process. are recommended to help make the product whiter. Usually it is followed by other preservation techniques such as drying and/or smoking.15 . When pure salt is used. This condition is known as ‘putty fish’ (4). and for a heavy salting a ratio of 1:3 salt:fish is used (1). which include calcium and magnesium. the fish is mixed with one part of salt per eight parts fish.15% calcium and magnesium. ‘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. low in calcium and magnesium. For a light salting. can lead to the decomposition of the centre of thick muscles before it reaches the level of salt concentration sufficient to prevent spoilage. and the salt has penetrated throughout the fish (3). calcium and magnesium ions of impure salts can form a barrier to the passage of sodium ions from the surface to the bulk of the fish.1 ppm and 10 ppm. it tends to lead to a slight yellowing and softening of the product. when the air humidity is high. However. lean white fish is used for this purpose. 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.3. mainly in tropical conditions. addition of levels of 0. Salting can be done in a number of different ways.1 Dry salting / Kench curing Usually. It is common for the dried salted cod in the Mediterranean countries to be made using solar salts. The fish is then stacked in piles of about 2 metres and the water is allowed to drain away. caught in North Atlantic waters. This is because fatty fish turn rancid quickly. The most common fish used in the northern hemisphere is cod (Gadus morhua). 5. The salt used should be of food-grade quality. 97 . for which there is a high demand in Mediterranean countries. however. can be dried further to produce ‘klipfish’ or ‘bacalão’. respectively.0.1. On the other hand.1 Salting Salting of fish for preservation purposes has been used since pre-history. if very pure sodium chloride is used for curing purposes.3. It takes about 3 weeks until proper salt penetration and moisture loss are achieved. and a ‘putty’ fish product is also obtained (1).1.35% and 0. and such delays. and essentially free from iron and copper (3). which consequently inhibits the diffusion of the water from the bulk of the product to the surface. respectively (1). are likely to cause hydroscopicity of the product and. The calcium and magnesium impurities. the fish is restacked with fresh salt after about a week. outgrowth of halophilic bacteria may occur. See Figure 5. when rock salt is used the ‘dun’ patches caused by moulds are prevalent (1). having a short section at the end of the tail. ‘Green cured’ split cod with about 60% of the backbone removed. SMOKED AND DRIED FISH 5. producing characteristic pink patches. Also.CURED.
For large fish it is advisable to split it before salting. and the uptake of salt is dependent on the type of fish (fatty or lean) as well as its presentation (whole. the process is mainly for flavouring purposes prior to the use of other preservation processes.90 are rarely achieved. Fig.1. Where fish are salted in their whole form.1. Only small fish can be salted whole. The greatest barriers to salt penetration are the skin and scales. such as 98 .3.FISH AND SEAFOOD Dry salting can also be used for small species without the need for opening the fish flat.5. such as Staphylococcus aureus could survive and grow at such water activities in the product. eviscerated and/or split). Food-poisoning organisms. although toxin production would be unlikely (4). it appears that water activities much less than 0. when a light brine is used. Processing of dried salted cod 5.2 Wet salting or brining As referred to above. so that the risk of spoilage is high.
TABLE 5.03 4. SMOKED AND DRIED FISH smoking.85 3. the product obtained is called gaspé-cured (5).II. 5.91 2.64 4.41 5.CURED.70 6.49 4.33 5.98 5. a saturated brine is used.84 1.90 (1). the fish enzymes play an important role by maturing or ripening the product.78 2.76 4. The main objective of the brining method of salting fish is to limit air penetration during processing. if split fish is dry-salted.25 5.19 3.04 5. For example.23 8.76 4.76 99 . sardines. The fish are kept immersed in their own brine near saturation point for several days. and can be collected and replaced again over the fish.85 3.23 3.06 4.96 1.60 2.5 4.44 2.26 3.1.78 6.41 6. This is why it is the preferred method for fatty fish.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) 2.23 2.0 2. with pressure being applied to the top of the container to help the brine to drain away. Histamineforming bacteria do not generally grow at water activities below 0.09 8.45 3.61 3.3. It is then taken out and dried in the sun or in kilns until about 35% moisture in the product is obtained. See Table 5. limiting the amount of rancidity that can develop.41 3.86 2.25 5.30 2.78) (1). Here.65 2.0 4. and a red burgundy colour will develop in the product.73 1.76 4.35 4.05 7.26 2.10 1.98 6.26 1.5 3.11 9. can be cured in a saturated brine solution for 6 days and.88 5. after being drained and pressed at 465 kg/m2 for 15 hours. thus. they can be stored at 28 °C. Histamine formation is of importance when using scombroid and some clupeidean species. Anchovies are the main fish used for these purposes.26 4.11 4. herrings and other fatty fish can be used.41 4. the brine is then replaced over the fish.74 9.47 6. but as it is placed in containers.3 Dry / wet salting In this process. the liquid that comes out of the fish forms a brine. with a shelf life of about 4 weeks (water activity (aw) about 0.17 5.17 4. headed and gutted. The brining step for smoking purposes also helps the product to develop a desirable glossy appearance.51 5.80 5.16 7. the fish is dry-salted.23 2.00 3.26 7.98 6. There are several variations of this process.76 4.22 6.95 3.99 2.46 4.09 8.70 3. For preservation purposes.61 3.66 5.66 5.51 3.5 10. Fresh sardines.03 2. the water that comes out of the fish forms a brine in which the fish stays immersed for 2 .5 3.56 6.44 2.3 days.0 3. but mackerel.54 5.76 4.11 3.
and 6% salt.2 months. The cooked portions are poured into bowls containing gelatin dissolved in water (10% solution) and sometimes a small amount of vinegar. 5. In a second step. it can be repacked in milder brine (1 . because of the salt content of the product. only the nonspore-forming organisms are eliminated in the process and. flavouring (sugar. Usually. herring or other fatty species are used. Other preservation techniques can be used for extending the shelf life of these products. outgrowth of bacterial spores after processing may occur (4). or can be boiled by being placed in a basket.6 days without refrigeration (1). The first process can give a longer shelf life. and packed (3). until the flesh is soft.FISH AND SEAFOOD 5. and lasts for 3 .7%) for as long as 3 weeks. Another variation uses sour cream instead of the second brine step. salt. Traditional jellied eels are a product where pieces of fish are cooked in boiling water. up to 1 . containing 1 kg of salt for every 25 kg of fish. Afterwards. Herring can also be cured for several days in an 80 to 90º brine with 2. which is dipped into a boiling brine for 10 . but it has further smoking.30 minutes. even if the product is hermetically sealed. is made of different varieties of fish. and even death.3. then packed in barrels with a 70º brine. and spices) is added to the brine. The fish can be soaked in a high concentration of vinegar (about 4 . Ceviche. There is epidemiological evidence that these products have been the cause of some 100 . They will have a salt content >6% (w/w) in the water phase. the occasional implication of pindang in cases of foodborne illness. they are packed for consumption within 2 weeks at chill temperature (6). or a pH <5 and they are stabilised for a limited period. The Indonesian ‘pindang’ is made by boiling the fish in a saturated salt solution and it is sold as is. a Peruvian product. As referred to above.2 Salt-boiling Fish can be boiled after being dry-salted in a watertight container and heated for at least 1 hour. All these products are ‘semi-preserves’. they can be cut and cured for 3 days in a solution with 3% white distilled vinegar. usually requiring refrigeration below 10 °C for longer-term storage. The product is usually consumed raw with the high strength acid solution softening the bones enough for them to be eaten. giving a cooked appearance to the product. However.5% 120grain distilled vinegar. After this. Once cooled. has a shelf life of several years. The lime juice plays the role of the vinegar. whereas the latter method preserves the fish for only a few days. ‘ikan kaju’.3. underlines the need for careful control (4). usually flavoured with onions and hot chillies. drying and salt-boiling steps until a hard translucent block is obtained (4). Another salt-boiled Indonesian product.2% acetic acid and 2 .3 Marinating / pickling Fish can be brined in solutions based on organic acid instead of salt. A much firmer product is obtained when the fish is salted prior to acidification.4% of salt) for distribution and sale (1). placed in a marinade of lime juice.
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). sugar and spices. 5. Foodborne pathogens. are commonly dried in Asian countries.5. Stockfish products have a golden brown colour and are very hard. some fish species such as moray eel and varieties of dog fish.) where no acid is added to the process. 101 .0) (7). Fresh dill is placed between the layers of fish. In some coastal areas of Portugal. bacterial toxins and parasites (7). Italy and Spain (5). 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. such as squid. gravad fish. are likely to survive in these mild-processed products. used as an appetiser.3.CURED. Dried shrimps are widely used in Asia. the temperature and the amount of pressure applied (5). SMOKED AND DRIED FISH foodborne diseases related to the presence of biotoxins. The most common method used for fish products is the air-drying process. Matjes are made of fat young herring. At pH levels below 4. and the flesh will break down. jellyfish.4 Air drying This is probably the oldest method of fish preservation. Some bacteria and enzymes are still active in the presence of acid and salt. the fish is dried naturally in a relatively cold environment. mixed with salt. are generally air-dried at sea. and the stacks are placed in the refrigerator. They are not cooked prior to consumption and they have a limited shelf life even at refrigeration temperatures. marinated products will keep for several months at a temperature of 4 °C. such as Listeria monocytogenes and Clostridium botulinum. then mild-cured with special seasonings and packed in barrels with 80 º brine (5). Shark pectoral fins. still made without prior salting on a large scale. split and dried gadoid species) are traditionally made in cold climates. Stockfish is extensively used in Portugal. It takes about 2 days to marinate completely. under a light pressure. In Scandinavian countries. while in the latter case there is a provision for fans or other means of mechanically moving the air (1). which are placed in a mixture of salt. are sundried and. Other products. gutted. sea cucumbers and seaweeds. In the first case. it will last for many years (5). It is important to assure the quality of the raw material. Gravad fish is a Scandinavian product made of salmon fillets. Provided the product is kept in dry and fairly cool conditions. after frying. Dried octopus is also a very common dish in the interior of the country. but sometimes it is dried without prior salting. and a fine product is obtained. South America and the Caribbean. Usually. depending on the thickness. Stockfish (dressed. by using either solar heaters or mechanical dryers.There are several products named marinades (matjes. etc. it being preferable to use frozen salmon or to freeze the product at the end of the process to avoid parasite survival. the fish is dried after salting or brining. They are an important dried product. air flows naturally. a very fine delicacy for Oriental food lovers.
there is an increasing demand from people that have emigrated to industrialised nations for traditional products. typically in stews and soups. In many such artisanal fisheries the drying is simply conducted by spreading the fish on the beach. that may have contaminated the product during the drying process. To prevent this. Whilst the low aw of the final product will inhibit microbial growth. Spain. more commonly nowadays. C. Thus. and increasing amounts of such products are making their way into the market place. Although controls are in place to ensure that products processed in such relatively uncontrolled conditions do not enter the supply chain. France. Whilst the production of dried fish products is tightly controlled in industrialised countries.83 (9)).the lower the humidity. or wooden drying rack to dry in the sun. if the rate of water removal is too high in comparison with the diffusion rate. making a water barrier and consequently. In conclusion. It is important to balance the drying rates and quality/safety issues. the better the moisture transfer. Italy and South America. and the water must be changed several times before cooking.g. the skin will form a hard case (case-hardening). aureus) or spores (e. some of which are listed in Table 5. botulinum). toxic secondary metabolites of the moulds Aspergillus flavus and Aspergillus parasiticus are not produced at aw <0. Owing to restriction of fishing areas for Spanish and Portuguese vessels since the 1970s. However. The amount of moisture transfer from the fish to the air is dependent on the relative humidity . the diffusion of the bulk water to the surface of the fish is blocked. by warm air circulating in a drying chamber. the ‘green cured’ cod is rinsed or washed with water and then hung to dry either by the sun and breeze or. Portuguese and Spanish fisheries cannot satisfy their own countries’ demand for cod. The product is dried to a final water content of about 25 to 38% (3). but fast enough to avoid spoilage caused by bacterial and enzymic activities (1). Traditional processing conditions provide ample opportunity for product contamination from a wide range of hazards and sources. which leads to mould growth (aflatoxins. and has the primary objective of creating a product with specific taste and textural characteristics.III. the product can be held for months at room temperature. When no more water can be removed from the fish. 102 .75 (1). the quality is more difficult to control. and it is exported to Portugal. They must be soaked in water. nor subsequent cooking operations will affect the presence of toxins (e. the fish must be dried slowly enough to avoid case-hardening. The fish must be stored in reasonably dry conditions to avoid moisture absorption.g. neither it. Iceland and Canada. tarpaulin. A highly demanded dried fish product in Mediterranean countries is dried salted cod.FISH AND SEAFOOD When the fish is sun-dried in tropical conditions. spoilage and possibly C. Staph. 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. the protein must be set or denatured with low-temperature drying before applying higher temperatures. dried salted cod (bacalão) is now mainly produced in Norway. skin up. After dry salting. Dried salted products must have a water activity below 0. botulinum growth might occur in the bulk of the product.
swordfish. In the mechanical kiln. There are many fish species that are used for smoking purposes . aureus and Saccharomyces cerevisiae (10). Staph. trout. is mainly a process for flavouring fish rather than a preservation technique. such as Escherichia coli. Blends of various woods can be formulated to produce sawdust with specific flavours. herring. mussels and many others from tropical countries.CURED. as referred to above. It has been reported that the smoke from mahogany. Aroma and flavour are a blend of smoke components (3). clams. Maintaining the temperatures of pyrolysis between 425 and 200 °C.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. making full description impossible here. The production of smoke by wood or sawdust burning can be done in a mechanical kiln or in traditional chimney kilns. haddock. However. Proper cleaning and preparation prior to smoking and washing of the fresh or frozen fish by water spray or a continuous 103 . sprats. botulinum (spores) Pesticides / Chemicals Physical Glass / sand etc. are some of the ways of reducing the polycyclic aromatic hydrocarbon (PAH) compounds (10).salmon. white mangrove and abura has an inhibitory effect on some microorganisms. aureus (toxin) C. temperature and humidity can be automatically controlled and the process is faster because the directed air movement draws the smoke through the fish.3. tuna. The smoking procedures vary widely. It can be used for cold or hot smoking. softwood colours the product quickly. Preventing the occurrence of carcinogens during smoking has been an issue for the past several years. eel. using electrostatic filtration of the smoke. oysters. and the use of smoke generated by superheated steam or liquid smoke. Hardwoods are considered better because of the milder flavour they impart to the food.5 Smoking Smoking today. SMOKED AND DRIED FISH TABLE 5. Top-quality fish must always be used. 5. cod. mackerel. cod roes. The quality of the smoke is related to the type of wood used.
or packed in retortable foil pouches. wooden boxes or master cartons (10). or quick-frozen and stored at -18 °C (2). after being smoked need to be chilled immediately to 4 °C before being packed. as well as many other products prepared in the same way. it will reduce the salty flavour. Cod roes.and hot-smoking is the temperature of the process. but it will increase towards the end of the process (10). canned or bottled. whether cold. are not cooked prior to consumption. Depending on the type of product. and the cold smoke is denser. spices. This difference is crucial for safety issues because the flesh is not cooked. However. thus allowing the deposit of smoke on the fish surface.FISH AND SEAFOOD water flow system with chlorinated water (25 . but a ‘safety’ (i. but this concentration will again depend on the type of product and the length of the brining stage.or hot-smoked. tuna and swordfish. usually dry-salted and cold-smoked. The fish can be dry-salted using a weight of salt 5 . giving the product the desired appearance and flavour (4). the excess salt is removed with water. the highly desirable cold-smoked salmon. All the smoked products.e. are boiled before smoking (12). Sugar can be added together with the salt.10% of the weight of the fish. filleted. temperatures and times used in processing coldsmoked fish are very favourable for the proliferation of food-spoilage and foodpoisoning microorganisms. The maintenance of temperatures below 30 °C avoids protein coagulation. 104 . such as smoked salmon-trout. Sugar. Wet salting or brining is preferable to dry-salting when smoking fillets because. however. covering the fish surface completely (10). permitted colourings or smoke flavours can also be added to the product in the brining step. skin off or on. it will take up salt twice as fast as fresh fish (10). Whether the product is prepared headed. smoked cod fillets and finnan haddock are usually cooked before consumption.5. the following step is salting. Thus. shrink packaged. Also. 70 – 80 º brine is commonly used.1 Cold-smoking The major difference between cold. low temperatures will slow the uptake of salt. owing to the elution of soluble proteins. it will double the drying time in the kiln to get the same total weight loss as for dry-salted fillets (11). If frozen fish is used. The length of the smoking process for such products is much greater than for the hotsmoked fish. all cold-smoked products should be cooked before consumption. Thus. an attractive glossy pellicle on the fish fillet surface is achieved after draining. Drying should be minimal at the start.50 ppm) should always be done just before process initiation (3). 5. In cold-smoking. pasteurisation) temperature is not achieved in any stage of the process. which is important for the appearance of these products (4). The salting period is dependent on the size of the fish and its presentation (whole. they can be vacuum packaged. When frozen/thawed fish is used. Some herring products. skin on or off). Brining at 10 15 °C is recommended (10). as it retains more moisture. temperatures must be kept under 30 °C throughout the process. gutted or filleted. thawing must be done slowly for 12 . If wet salting is chosen.3.18 hours at 4 °C.
but they should be cooled to a temperature of 10 °C within 3 hours. Smoked products can be also packed under controlled-atmosphere packaging. 5. If the refrigeration chain is maintained. and the recommended gas mixtures are 60% carbon dioxide and 40% nitrogen (15). as. Smoked salmon fillets are usually vacuum packed whole or after slicing. are of concern when smoking fish from tropical waters (13). The protein must be set or denaturated with lower temperatures before the application of higher temperatures to avoid case-hardening. Hot-smoked products require a longer cooling stage. which underlines the danger of storage at abuse temperatures of >10 °C (13). the products are cooled. Neither the smoking temperature nor the salt content is enough to kill the organism. further cooled to 3 °C within 12 hours and then packed. and their sensitivity to salt (13). Vacuum packing extends the shelf life of the lightly smoked products by suppressing the growth of other bacteria and moulds. secondly. 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. typically mesophilic. botulinum type E present an even longer time than before to grow and produce toxin (14). can grow with salt levels of 10% in the water phase. Length of the smoking process at each stage will depend on the species and the type of product. a partial cooking period at 50 °C. such as Yersinia enterocolitica. On the other hand.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.3. Furthermore. 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 present. The exception is vacuum packed fish. refrigerated storage and vacuum packaging still allow its growth (12). a drying period at 30 °C.CURED. a final cooking period at 80 °C (12). in order to produce toxin. can be isolated from seafood. there is no point in the process that can fully assure the absence of L. if a 105 . thirdly. an initial humidity of 70% seems to be preferable because it is a good compromise between maximum smoke adsorption and minimum casehardening (1). stored at 4 °C and packed. A typical temperature set is as follows: firstly. if they are taken from a chill room at 3 °C and packed. SMOKED AND DRIED FISH During cold-smoking. See Figure 5. monocytogenes. Vacuum packing is recommended only for coldsmoked salmon. thus allowing any psychrotrophic C. gravad salmon (also called gravad lax or gravlax). These products should be vacuum packed at room temperature.2. they are more sensitive to sodium chloride and unlikely to grow in smoked products (13). condensation will occur inside the pack. and hotsmoked salmon (kippered) (3). Although other human pathogens that can grow at refrigeration temperature. This temperature should be maintained during storage and distribution (3). Afterwards. but the proteolytic types A and B. During smoking. Pathogenic vibrios.
FISH AND SEAFOOD product is packed too warm without being cooled to room temperature. and moulds will grow on the product (12). Process of cold-smoked sliced salmon 106 . Fig. it will have a reduced shelf life. 5.2.
including ‘umami’ (16.5. 5. apart from the improved appearance. synthetic smoke flavours. then hot smoking and finally sun drying it. Aspergillus ostianus.CURED. 17. hot-smoked fish can be classified as a ‘heat-treated or pasteurised’ product (7). 22. When refrigerated at 3 °C. The composition of commercial liquid smoke can vary widely depending on the method of manufacture. Growth of these moulds on the tuna results in the extraction of moisture. to have no further advantage over other packing methods for other fish (12). including Eurotium repens and Eurotium rubrum. such as yeast derivatives. are usually 80º brined for 5 . and substances unconnected with smoke but with a smoky flavour or smell. Boiled or steamed clams. They will keep well frozen. chilled or freeze-thawed. Mackerel. (Penicillum cyclopium and Penicillium puterellii) (19). are used. The product is removed from the drying cycle. white fish about 8 days. The classical liquid smoke is 107 . fatty fish will remain in good condition for 6 days. Hot-smoked products made from white fish will usually keep better than those from fatty fish. 21. rewetted and remoulded to give the desired shape and consistency. a very expensive and fashionable smoked product in the Netherlands. are usually hot-smoked after being immersed in 80º brine or dry-salted (6). Masmeen is usually prepared from bonito tuna by eviscerating and filleting the fish. Katsuobushi. Natural smoke extracts. moulding it into long thick sections. but mechanical drying is used instead of sun drying. cooking in brine. The production method is similar to masmeen.6 hours and smoked for 3 . similar to masmeen.3½ hours using the typical temperature set for hot-smoking referred to above (12). Eels. Aspergillus tamarii) and several Penicillium spp. and many closely related Aspergilli (Aspergillus ochraceus. is a boiled smoked fish product that is used in Japanese cooking to impart flavour. A type of dried hot smoked tuna called masmeen or masmin is produced in the Lakshadweep islands of the Indian Ocean.3 Liquid smoke Smoke-flavoured fish represents a small percentage of the total smoked fish industry output. Vacuum packaging retards the onset of rancidity when fatty fish is used. In terms of safety. oysters and mussels are usually wet-salted in 50º brine for 5 minutes and hot-smoked at 80 °C for less than 1 hour (12). They are generally not cooked prior to consumption.3. resulting in a characteristic flavour (20. they can be stored for 6 months at -30 °C and for even longer when frozen in vacuum packs (12). SMOKED AND DRIED FISH There are several species that are usually hot-smoked and the process can vary markedly. break down of fats and the moderation of smoke flavour. 23). Aspergillus oryzae. which means that any bacterial contamination and growth after the heat treatment is a serious hazard that must be controlled by Good Manufacturing Practices (GMP) and factory hygiene/sanitation after the heat treatment (7). The product is often allowed to go mouldy by incubating it in a chamber containing a natural fungal flora. Hon kare bushi is a form of Katsuobushi from which shavings are cut to make aromatic stocks for use in Japanese dishes (24). but seems. 18).
4 Spoilage Brining. Whilst neither the CO or the carboxymyoglobin complex are harmful in themselves. so as to impart an unnatural red colouration to the flesh.3.8% and still provide protection for 7 days (25). both of which are considered potential carcinogens. it has been reported that the liquid smoke-treated product has less pronounced aroma and flavour (25). smoking. The European Scientific Commitee for Food. the salt level could effectively be reduced from 4. There are two groups of chemicals of concern in smoke: polycyclic aromatic hydrocarbons (PAHs) and nitrosamines. 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.03 μg/kg of benzopyrene. 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). and less than 0. drying.5.FISH AND SEAFOOD smoke condensate that is dissolved in water or oil. injecting it into the fish flesh. consistency of flavour and colour. or smoke extracts in organic solvents. pickling and marinating fish were originally used to lengthen the short shelf life of fresh fish. botulinum types A and E spores in several fish species that were stored at 25 °C for 7 and 14 days (25). 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. nutritional neutrality and antioxidant activity (27). It has been shown that liquid smoke can reduce the formation of PAHs (3).6 to 2. When liquid smoke was used. or applying it as an atomised spray within a modern automatic smoke kiln. Liquid smoke in combination with sodium chloride in a hot-process smoke flavoured fish was found to be effective in preventing growth and toxin production associated with the growth of C. According to some recent research. applying it as a dip after brining.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. established in its guidelines that smoke flavourings added to the food product must assure a level lower than 0. 5. Liquid smoke was considered to provide advantages of safety. Fresh fish spoilage may be measured by 108 . spraying is preferable because it is easiest to control flavour and acceptable with this method (3). In experiments comparing traditionally smoked fish with liquid smoke. Smoke-flavoured fish processed using liquid smoke can be prepared by including liquid smoke in the brine. 5.06 μg/kg of benzoanthracene in the final foodstuff (28).
the fish flora are transferred to the brine mix. Once these changes are defined. a very dry product is required to produce 'shelf-stability'. a process similar in microbiological effect to that of saturated brining. otherwise. Usually. contact with the bacteria 109 . Stronger and longer brining has an effect like that of pickling. When acid marinating or smoking. a product may become undesirable for consumption as a result of changes in its sensory characteristics. but. spoilage is due to endogenous microorganisms and enzymes of the fish. mainly comprising corynebacteria and Gram-negative bacteria from the fish and water (29). When heavy brining or salting is important for product shelf life. as the fish are added. Micrococcus spp. This is not usually the case for smoked fish as the residual salt concentration is not high enough to inhibit all the non-halophiles. contaminant bacteria from other materials used to produce the product. Some brines are re-used over a period of days and a load of 10. For some products. the shelf life may be estimated and investigation made into the causes of the loss of sensory quality. Brining.. however. which are found in the salt. and of smoke to add chemical preservatives. both of these will affect the spoilage patterns. marination or drying). for some. and.CURED. pickling and dry-salting all introduce bacteria to the fish. or extended refrigerated storage.3%). Spoilage needs to be carefully defined for each product. and coryneform bacteria. the consequent reduction in the pH will help to inhibit spoilage organisms. the fish are first cleaned (known as ‘rousing’) by a salt rub to remove any outer slime. and high concentrations are usually required to inhibit their growth fully. The use of drying and/or salt to lower the water activity. Salts used for brining have been found to carry a microbial load of between 10 and 1000 organisms per gram of wet salt. also. the fish are cut and splayed to allow greater surface contact with the salt. solar salts investigated carried the highest load and included halophilic organisms as a result of their production method (open-air evaporation of sea water). chemical oxidation. The microflora attached to the fish is relatively unaffected by light brines used before smoking. the production process will modify the microbial flora present on the product. Endogenous marine fish bacteria prefer to grow in the presence of salt (marine concentration approximately 2. Most marinated and smoked fish products are first brined or salt-pickled. or their products require refrigerated storage. limits the types of microorganism capable of growing. therefore.000 cfu/ml of brine may build up. Even saturated brines allow some halophilic organisms to grow. In salts.. the bulk of the microflora comprises Bacillus spp. these bacteria tend to continue to dominate the flora in the liquid. which may also be produced to describe the quality attributes and their loss over time for processed fish. for others. Once placed into brines. Within the limits of safety. SMOKED AND DRIED FISH grading schemes or quality indices. Upon salt packing. these halophilic organisms may grow and cause spoilage. rock and manufactured salts did not usually contain halophilic bacteria (29). Often.g. the pickle and dry-salting preservation processes require further processing for significant shelf life increases (e. Pickling and/or the anchovy process uses solid salt extraction of fish fluids to form a brine solution (cure).
lactic acid bacteria (LAB) and moulds. A. although it may occur in other salted fish. ‘Pink’ describes a condition associated with surface growth of the pigmented halophilic bacteria Halococcus spp. Aspergillus sydowii. powdery growth.. spoilage of these pickled and dried fish in refrigerated storage tends to be due to chemical changes (textural changes due to proteolysis. or after drying if the fish is stored in excessive humidity. Unlike ‘pink’. The metabolism of the organisms will produce off-odours. 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). The microbiology of heavily salted products is dominated by Gram-positive. may also be found in the salt used in production. 33). as the water activity of saturated brine solutions is low enough to inhibit most bacteria. The bacteria are osmosensitive and will lyse in water. ‘Sliming’ or ‘putty’ fish is a condition that occurs in poorly or insufficiently salted cod (e. Therefore. most notably Penicillium citrinum and Penicillium thomii. and Aspergillus penicilloides. though only the latter two species were considered likely to have grown on the fish (32). repens and E. halophilica was isolated 110 . which. The condition describes the growth of the halophilic moulds (Wallemia sebi. pisce was the main fungus isolated. rancidity due to lipolysis. yeasts. yeasts and fungi. (syn Oospora halophila) and Polypaecilum pisce are two white halophilic moulds that are associated with spoilage of salted fish (31. together with a range of Aspergilli. Eurotium amstelodami. ‘pink’ and ‘dun’. Sporendonema sebi)). flavus. Bacteria from these two genera also grow sufficiently in brine and even salt to cause pink discolouration. like the ‘pink’ organisms. Studies conducted on dried and salted fish from Indonesia (31. Basipetospora halophila. including hydrogen sulphide (26). if not too contaminated. it was found in about 50% of samples examined. B. At the pre-drying stage. including A. sebi is considered to be the main spoilage agent of dried fish produced in temperate regions (30). sometimes covering the whole of the fish with a whitish. and still produce a reasonable dried product. it is possible to wash off the infected parts. ‘dun’ is not characterised by the production of offensive odours. the best way of controlling the organisms is to refrigerate the fish. Insufficient salt penetration into the flesh leaves areas where Gram-negative rod-shaped bacteria and micrococci grow and cause a putrefactive spoilage.FISH AND SEAFOOD in the salt is made and the flora will develop accordingly in the blood pickle. gaspé). (syn. protein breakdown. W. rubrum are also common isolates from dried fish. 32. Several Penicillium spp. halotolerant or halophilic micrococci. Their growth produces noticeable spoilage of the fish appearance but is also controlled by refrigeration. A number of spoilage organisms can grow and produce conditions such as ‘sliming’. and Halobacterium spp. 34) have shown that P. spore-forming bacteria. or fat breakdown) caused by enzymes rather than by microbial growth.g. Aspergillus wentii. This condition is often associated with case-hardening. niger. were also isolated. E. Only halophilic microbes may grow. The bacteria form coloured colonies on the surface of salted fish prior to drying. as little if any growth will occur below 5 °C on salted fish (29).
Cold-smoked fish will contain microbial flora representative of that of the raw fish. Reports for vacuum packed cold-smoked salmon have identified more than one organism associated with spoilage.747. Vibrionaceae and Enterobacteriaceae may all play some part in shelf life limitation. so the most probable spoilage organisms will be spore-formers or post-smoking contaminants. with a lower aw limit for growth of 0. The salt in the water phase must be high enough to inhibit the growth of C. Aspergillus spp. The preservative effect of the smoke will inhibit some spoilage microorganisms. 33. Refrigeration is necessary. The brining to 3 . most marine bacteria are moderately halophilic and are not effectively inhibited by these low salt concentrations. For more information on spoilage see Table 5. SMOKED AND DRIED FISH infrequently at low levels. will have a different residual post-smoking flora from 'hot-smoked' fish. Technical studies demonstrate the ability of many of the xerophiles above to grow at typical water activities seen in dried salted fish (32. The heat during hot-smoking will kill vegetative microorganisms but not all spores. Aspergillus clavatus and Aspergillus niger. flavus. including A. In Vietnam. It has been reported that the type of salting (dry. Refrigeration of hot-smoked fish will greatly reduce the number of organisms able to grow and will lengthen the shelf life.or wet-salting) in the cold-smoked salmon process leads to different microflora growth at the onset of spoilage. has been isolated from dried fish (35). it was.CURED. able to grow on dried salted fish. which does not create the salty conditions required for spoilage by those halophilic microorganisms. where a high internal temperature is reached. Marine vibrios were the dominant flora in smoked salmon that was drysalted. or any smoking process where temperatures in the interior of the fish do not reach >50 °C for a significant period of time. botulinum type E. not the growth of spoilage organisms. Fish processed by cold-smoking. however.IV. so the bacteria spoiling these products will be psychrotrophic or psychrophilic.V gives details of pH ranges and aw minima for the growth of some important spoilage microorganisms and pathogens (38). LAB. around the water activity of saturated sodium chloride. Table 5.. and microbial spoilage is more likely. but this will not prevent the growth of spoilage bacteria. whereas a mixture of Enterobacteriaceae and LAB were prevalent in brineinjected salmon (37). as already stated. Smoked fish products tend to have a light brine treatment.5% sodium chloride in the water phase will inhibit some bacteria. 111 . Pseudomonadaceae. Vacuum packing limits chemical oxidation. but often colour changes due to chemical oxidation of the smoked product will limit the shelf life. but. 36).
Dun. boiled. halophila Halophilic spoilage organisms Halophilic spoilage organisms. sebi and B. + to +++ strength of preservative effect 4 Post-process contaminants 112 .FISH AND SEAFOOD TABLE 5. .IV Spoilage of salted. Enterobacteria and PPC Spore formers and PPC Variable Salt marinades Salted then boiled Brined then boiled Salted then dried Canned Fish Gravad. 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.W. marinated and smoked fish Spoilage controlled by pH Heat Ant1 Ref2 Process Product Aw Probable spoilage organisms None Fresh fish spoilage organisms Halophilic spore formers and PPC4 Halophilic spore formers and PPC Halophilic organisms: Pink-Halococcus spp.not used/unchanged. Histamine producers Halophilic spoilage organisms Spore formers and PPC None Acid-tolerant spoilage organisms Xerophilic moulds Fresh fish spoilage organisms. and Halobacterium spp. Matjes v3 + ++ v - +++ ++ ++ - v +/- ++ ++ ++/++/- Pindang Bacalao + +++ Gaspé cure Anchovie ++ Anchovies ++ - - - ++ ++ Saturated brined fish Salted then boiled Salted. LAB.variable.
Before.5 5. aureus Yeasts Moulds Lactic acid bacteria pH 5. Examination of 165 outbreaks of botulism caused by fish products showed that smoked. smoked and dried. under optimal conditions.5-10.V pH ranges and aw minima for growth of microorganisms (38) Organisms Most Pseudomonadaceae C. pressure on the traditional methods of curing.5% of salt in the water phase and great care must be taken to avoid any temperatures >10 °C (13). It has been shown that. botulinum type E and production of toxin.97 0. and as botulism is a serious hazard in foods. In the United States. the fermented products being the most dangerous group (7). but the presence of the mesophilic types A and B is of concern in fish from warmer waters (13).5-10 4. Since neither the smoking nor the drying parts of this curing process are particularly severe. However. 3% sodium chloride in the water phase is enough to inhibit growth of type E for at least 30 days (39). the non-proteolytic type E strains can grow in up to 5% sodium chloride.5-8.75 0. although approximately 10% salt in the water phase is necessary to fully inhibit the mesophilic types A and B strains (14). the products were heavily salted.5 5.5-8. but the concentration of salt in smoked salmon or trout (usually varying from 1 4% salt) is not high enough to prevent growth. The presence of salt in cured products has a great effect on the growth of the bacteria. SMOKED AND DRIED FISH TABLE 5. although the combination of the two factors.8 1. popular demand for healthy foods has led to a change in the tastes preferred by the public and. botulinum type E. The concentration required to prevent growth at room temperature can vary from as low as 3. types A and B are not affected by 3 . could not grow and produce toxin. respectively.61 Organisms Most Pseudomonadaceae C. botulinum Staph. with this change.5 4.8 0. A number of surveys have shown that fish from temperate waters is mainly contaminated with the psychrotrophic C. the importance of cold-chain integrity must be emphasised. sodium 113 .5% to 5%. botulinum.CURED. temperature ≤5 °C and salt ≥3% in the water phase is sufficient to prevent any growth of type E.5. salted and pickled products accounted for 10.9 0. aureus Most spoilage bacteria Most spoilage moulds Halophilic bacteria Xerophilic moulds Aw 0. botulinum type E Staph. Sodium nitrite in conjunction with sodium chloride is known for its ability to inhibit C.86 0. 9 and 8 of the outbreaks. and C.1 Pathogens: Growth and Survival Clostridium botulinum In the last few decades.5 0-11 3.97 0. when present in the raw material.0-9. but if the fish products are stored at temperatures <10 °C.
The FDA established that hot. coli all occur on fish products as a result of contamination from animal/human reservoirs. as they require more handling. its occurrence in a final product is due to contamination during processing by an infected asymptomatic carrier with poor personal hygiene. they can survive lightly cured processes when the sodium chloride content is less than 5% and water activities are higher than 0.5. It is a halotolerant bacterium able to grow at water activities as low as 0.94 (7). gills and guts as well as in several lightly preserved fish 114 .0% salt in the water phase (30). As they are poor competitors.30% of the population positive for enterotoxin-producing strains (41).4 Listeria monocytogenes The psychrotrophic L. growth and toxin formation may occur. but. Thus. are particularly susceptible to contamination and. Shigella and E. 5. they do not grow well in the raw material. However. Infection by Shigella sonnei has increased in the UK. they will grow rapidly (7). Sliced smoked fish products.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.86 and 10% sodium chloride. if storage temperatures are over 10 °C.2 Staphylococcus aureus This is a ubiquitous organism but its main reservoir and habitat is animals and. aureus growth. if it contains sodium nitrite ≥100 ppm. the human nose. 5. related to the consumption of a raw smoked fish meal. but requires higher temperatures for toxin production (>10 °C). it should contain not less than 3.5% salt in the water phase. In foodstuffs.5. when the bacterial load of the product is lower. and the minimum pH for growth is 4. if the person handling the product is a carrier of Staph. monocytogenes is a ubiquitous organism that has been detected in fresh fish skin. Usually.4. The organism is mesophilic with a minimum growth temperature of 7 °C.3 Enterobacteriaceae The pathogens Salmonella. the organism can survive most of the light curing processes. 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. Once the product is contaminated. An outbreak of Shigella in an Italian cruise ship. but. throat and skin. The human carrier rate can be as high as 60% of healthy individuals with an average of 25 . has been reported (42). The degree of concern in coldsmoked salmon is low unless temperature abuse >10 °C occurs (13). As with other organisms. the organism may contaminate the product. 5.or cold-smoked fish should have at least 3. aureus. more importantly.5. the risk of infection can be eliminated by proper cooking. after processing. if recontamination occurs and storage conditions are favourable.5.0 .
most of them are of little concern to public health. are considered unsafe.the round worm Anisakis spp.95 (45). monocytogenes’ in 25 g of the finished product. Some LAB. the CT 3162-FAIR program.5. the only concern to public health is when eating raw or uncooked fish products. Thus. has also been detected in cod (7). cold-smoked fish. Pseudoterranova dicipiens. marinades (ceviche) (43) and gravad fish (44). Two outbreaks in the United States have been reported to be linked to salmon (10). a number of fish products. As they are heat-sensitive. They have complicated life cycles. As freezing to -20 °C and maintaining this temperature for at least 24 h will kill all the nematodes. favours growth of psychrotrophic LAB. gravad fish. This includes matjes herring. widely used as starter cultures in food processes. lightly salted caviar. which. and other local traditional products (7). The round worms.CURED. and in CT95-1207FAIR). Marine LAB is part of the natural flora of chill-stored vacuum packed salmon and cod. 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. SMOKED AND DRIED FISH products such as cold-smoked fish. do not survive 1 min at 60 °C.0. Unlike some European countries.6 and water activities below 0. or nematodes. from seawater fish.9.8 . when an outbreak of nine cases of listeriosis was suspected to have been caused by gravad or cold-smoked trout (47). such as ‘lightly preserved’ fish products.45 °C. salt levels of 8 . As its presence has been reported in lightly preserved fish products whose processing chain cannot fully assure the absence of the organism.2. from freshwater fish. the USA established ‘zero tolerance’ for the organism. a short period of freezing either of the raw material or of 115 . monocytogenes in levels at or above 102 per gram (46). Vacuum packaging (VP) of sliced smoked fish creates a microaerophilic atmosphere. both being detected in cold-smoked salmon.514. 5. which means that the process must conform to a level of ‘no detectable L. there were no reported outbreaks related to cured fish. researchers are looking for other ways of preventing the pathogen’s occurrence or growth. The use of LAB in fish preservation is a topic that has been studied in projects (UP.5 Parasites Despite the presence of parasites in fish being very common. pH 4. Its limiting conditions for growth are 0 . monocytogenes pathogenic for only specific segments at risk. in combination with curing agents and refrigerated storage. which means that cooking a fillet 3 cm thick for 10 min at 60 °C will kill any worms present (48). which considered L. have been shown to have an anti-listerial activity in many food products. Of microbiological concern in the northern hemisphere are two parasites . Until 1997. a round worm. and the tape worm Diphyllobothrium spp. which include intermediate hosts. in the EU Fisheries R&D-FLAIR program.12%.92 .
<107 ≥107 N/A <100 <20 100 .<100 20 .FISH AND SEAFOOD the final product should therefore be included in the processing as a means of controlling parasites (7). texture.<103 N/A 100 . ‡ On occasions some strains may be pathogenic. These foods fall into category 5. salami. monocytogenes) (cfu/g unless stated) <106 106 . perfringens B.<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. cholerae Vibrio parahaemolyticus L. E.<104 100 . † Guidelines for aerobic colony counts may not apply to certain fermented foods. aureus C.<100 200 . monocytogenes Staph. and unpasteurised yoghurt.<104 104 . the organism should be identified # N/A denotes not applicable 116 . If the Bacillus counts exceed 104/g. Acceptability is based on appearance. 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. 5.<100 10 .100 >100 N/A Pathogens Salmonella spp.<100 20 .VI Guidelines for the microbiological quality of ready-to-eat smoked fish.<104 20 .6 Published Microbiological Criteria TABLE 5. coli O157 & other VTEC Vibrio. cereus and Bacillus subtilis# <20 <20** <20 <10 20 . soft cheese. smell.<104 present in 25 g ≥103 >100 ≥104 ≥104 <103 103 . 48h ± 2h) Indicator organisms‡ Enterobacteriaceae E. coli (total) Listeria spp. for example. (not L.
in Fish Processing Technology.M. Wareing P. 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. 1994. Moody M.8 ppm (saxitoxin equivalent) 20 ppm domoic acid. Regenstein C. Marinades. New York. Horner W. monocytogenes Salmonella spp. Eds. 1982. 1984. salting and smoking). 334. where 30 ppm is permitted Paralytic shellfish poisoning* Amnesic shellfish poisoning * *These criteria are applied to all fish 5. 1997. 82 (revised). New York. 4.F. Torry Advisory Note No. Eds. 3. botulinum * Limit of tolerance 1x 103 ETEC/g. 117 . Catching. Fernandes R. Van Nostrand Reinhold.7 1. 6. 1979. 9. coli (ETEC) L.H.A. in The Seafood Industry. 7. Leatherhead. 120-38. 1990. Fernandes R. London. Wareing P. 254-82 5. 381-406. parahaemolyticus Vibrio vulnificus C. Urner Barry Publications Inc. Hall G. SMOKED AND DRIED FISH TABLE 5. 8. 56. USA.E.CURED. Flick G. 2. Torry Advisory Note No. Van Nostrand Reinhold. 1993. in Micro-Facts: The Working Companion for Food Microbiologists. FAO Fisheries Technical Paper. Horne J.. Chapman & Hall. Preservation of fish by curing (drying. 3273. Bannerman A.M. Handling and Processing Eels. Martin R. 2007. Introduction to Fish Technology. Doré I. Birnie K.F. Ed. cured and dried fish. Hot Smoking of Fish. Leatherhead Publishing. Food spoilage fungi. Torry Advisory Note No. except in the viscera of dungeness crab. 1991.. or presence of toxin 0. Assurance of Seafood Quality. Smoked. Huss H..VII US Food and Drug Administration (FDA) guidelines related to ready-to-eat fishery products (45) Product Ready-to-eat fishery products (minimal cooking by consumer) Organisms and/or toxins Enterotoxigenic E.. McLay R. The Smoked and Cured Seafood Guide. 37(revised).W.. * Staph. aureus * Vibrio cholerae V.. Flick G. References Regenstein J.
... Kinoshita Y..P. Botulism and Fishery Products. Aoshima H. 56 (6).FISH AND SEAFOOD 10. 21. 18. 13. Lancaster. 6-dimethoxy-4-methylphenol by Aspergillus glaucus and their possible contribution to Katsuobushi flavour. 1989.C. Dillon R. Bioscience. 335-40.. Shuto Y. 1990. Kinoshita Y. Yamauchi H. Torry Advisory Note No.. Ninomiya M. 1386-7. 51-81. Flavour constituents of dried bonito (Katsuobushi).. Technomic Publishers.C. Food Product Design. Martin A. 17.M. 53 (4).W. 840-6. Ogiwara Y. 1981. 843. 553-72. Yokayama S.R. Hayashi K. Biotechnological Applications. 23. 5 (revised). Biotechnology and Biochemistry. Oxidation of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavour. and Biochemistry.. Martin A. London. Sakakibara H.. Bannerman A. 1992. Shiemori Y.. Shuto Y. Doi M. Bioscience. 2008. Kanayama T. Torry Advisory Note No.. 6 (4).E. 12. 1995. Shuto Y. 22. 2002. Degradation and omethylation of phenols among volatile flavour components of dried bonito (Katsuobushi) by Aspergillus species. Teeny F. 22 (revised). Biotechnology and Biochemistry. Processing Cod Roes.A.. 25. Doe P. Chapman & Hall. The discovery of umami. Packing Fish in a Modified Atmosphere. Food Chemistry. 1982. Recommendations for the Preparation of Smoked Salmon. Pelroy G. Peterson M. Ed. Agricultural and Biological Chemistry. Matsui M.. 2324-5. 19. Huss H. Mitou M. What’s in a name? Are MSG and umami the same? Chemical Senses. Journal of Food Protection. Cann D. 20. 27 (9).K. Eklund M. Doi M. Control of biological hazards in cold smoked salmon production. Torry Advisory Note No.. Food Control. Matsui M.. 88. Druschitz A. 15. Hosokawa M... 14. O-methylation of 2.. 6 (6).. Lindemann B. 508. Cann D. Patel T. Microbiological control for fish smoking operations..B. 1980. 17 (9).E. Horne J. Inhibition of Clostridium botulinum types A and E toxin production by liquid smoke and NaCl in hot-process smoke-flavoured fish.. Jeppesen V. 59 (12). 108 (3).F. 1995. 1977. 16. Effect of dried bonito (Katauobushi) and some of its components on GABA-A receptors. 958-60. 2007. Food Reviews International.H. Perusing Asian soup options. Doi M. 118 . 1998. Ninomiya Y. Halpern B. 11. Doi M. Embarek P. 61 (8).. 1051-5. 1984. 1994.. Asymmetric reduction of acetophenone by Aspergillus species and their possible contribution to Katsuobushi flavor. Yajima I.M.. Biotechnology. Bannerman A.M.. 1997. 45 (10). 935-41... Fish Drying and Smoking: Production and Quality. 24. Paranjpye R. 18. Torry Advisory Note No. Bioscience. in Fisheries Processing..
1941. 123-9. 2005..E.A. 43.. Smoked and smoke-flavoured fishery products. Journal of Applied Bacteriology. 22. Diyaolu S. 35. Maga J. V.. Hocking A.B. 88-90.T. Press Inc. Antimicrobial properties of smoke. 1985. Food Research International. Nahrung.C.. 5. Frank M.R. 38. Incidence of Listeria monocytogenes in an acidified product. US Food and Drug Administration.A. Borgstrøm G. Studies on salt fish. 1979. CRC. Lucas F. 31. Sirvas S.. CEC. London. 197-208. 39. Journal of the Fisheries Research Board of Canada. Journal of Tropical Medicine and Hygiene. Smoke flavourings. Blackie. National Academic Press. Fuchs R. Huss H. Mycotaxon. Anggawati A.K. 37. Ed. Hocking A. 1991.F.Y. Food and Drug Administration. 1996. 1971.. Washington DC.D. 40.. 28. Importance of autolysis and microbiological activity on quality of cold-smoked salmon. Institute of Medicine. 1995. Pszczola D. 38 (3). 98100. 119 . Ed. Scientific Committee for Food. Lancet. 41. 1996. Gill T. Studies on Sporendonema epizoum from ‘dun’ salt fish. Food and Drug Administration.. Academic Press. 1961. Fungal flora of South Vietnamese fish and rice..M.V. Springer Science. 42. Ed. The control of botulism in hot-smoked trout and mackerel. in Reports of the Scientific Committee for Food (34th Series). Taylor L. 74 (2).I.J. Cox J.CURED. Loessner M. Effects of sodium chloride and relative humidity on growth and sporulation of moulds isolated from cured fish. in Fish and Fishery Products Hazards and Controls Guide. 29 (2). Townsend J. Rockville.. 1986. An outbreak of Shigella. Luxembourg. The microbiology of sea-water fish. 70-2. in Fish as Food. Pitt J..A. 1994.A.. Boca Raton. 164-9. Seafood Safety. 30. Wheeler K. Hess E. Hansen L. 34. 27686. Pitt J. Shewan J.1988. Sprouse R. Cann D. Hocking A. 348..D. 27. 1995.E. Ahmed F. 1991.D. 29. Pitt J. Oxford.F. Letters in Applied Microbiology. Food Microbiology. in Smoke in Food Processing. 3 (4).O. Jay J.. ceviche...I. 74. 1994. Tour highlights production and uses of smoke-based flavours.M. Wheeler K.H. 311-7.I. 33. Journal of Food Technology.. Fungi and Food Spoilage. 12. New species of fungi from Indonesian dried fish. Ed. Simini B. 181-8. 74. 218-25.. 32. Adebajo L. SMOKED AND DRIED FISH 26. Interactions among xerophilic fungi associated with dried salted fish. 36. Food Technology. 1993. 351-7. Golden D. Fungi associated with Indonesian dried fish. 49 (1).S..D. 14. Røntved S. 742. Modern Food Microbiology. Commission of the European Communities. 1997.M. 107-11. Hocking A. New York. Committee on Evaluation of the Safety of Fishery Products.
W.D. Journal of Food Protection. Elsevier Science Publishers B. Mentzing L-O.. 48. Liston J. Loncarevic S. Washington D. Association of Food and Drug Officials. US FDA Centre for Food Safety & Applied Nutrition. Susuki E. Appendix 5: FDA and EPA Guidance Levels. 1989. salted. Persson I.N. 1992. Amsterdam. Loncarevic S.. Gibbs P. Ribeiro C. 55 (4).. 47.. Inhibition of Clostridium botulinum types A and E toxin formation by sodium nitrite and sodium chloride in hot-process (smoked) salmon. 1976.W. 5-7.. Tham W. An outbreak of listeriosis suspected to have been caused by rainbow trout.. Canadian Journal of Microbiology.fda. Danielsson-Tham M-L. 49-59.. 1980. 1982.R.A. Pelaez C. 1996. Food and Drug Administration. 188 (3). Richards J. US FDA Centre for Food Safety & Applied Nutrition..M. 2000.. Fletcher R. Eklund M.. 5.L. 863-6. 3.gov/~comm/haccp4x5. Danielsson-Tham M. 5 (1). 22.. de Louvois J. Pathogenic microorganisms of importance in seafood. Donovan T.S. and smoked fish establishments good manufacturing practices. Ed. Huang Y. Moral A. Peterson M. in smoked and "gravad" fish. 833-41. Round Worms in Fish. Wooten R.. 135-85. Huss H..J.. 45.8 Further Reading Gibson D.. Clostridium botulinum and its importance in fishery products.. Bolton F. 35 (11) 2904-7.. The effect of NaCl on the upper temperature limit for growth and enterotoxin synthesis by Staphylococcus aureus.. Unnerstad H. Tham W. Pelroy G. Journal of the Association of Food and Drug Officials..C. 1997. Ericsson H. Little C. Growth and production of toxin of Clostridium botulinum type E in rainbow trout under various storage conditions. Journal of Clinical Microbiology.html Gilbert R.. Nye K.. Keeping quality of vacuum packed smoked sardine fillets: microbiological aspects. Communicable Disease and Public Health. Jacobsen M. Harrison M. http://www.. 1995.. Hobbs G. 163-7. Prevalence of Listeria monocytogenes and other Listeria spp. 13-8..J. Advances in Food Research.. Hurst A. Cured. 197-209. 1991. Torry Advisory Note No. Roberts D. Eds. 1980.E. in Fish and Fishery Products Hazards and Controls Guide. 26 (4). in Quality Assurance in the Fish Industry. 80. 232-6. 507-10. Garren D..V.C.. Hughes A. 120 . 2001. Acta Veterinaria Scandinavia. 58 (8).cfsan. Zeitschrift für Lebensmittel-Untersuchung und –Forschung. 1994.. Eklow A. Paranjpye R.A. Beltran A. Davies A.M. Cann D.A. Food Control. Incidence and growth of psychrotrophic Clostridium botulinum in food. 46.FISH AND SEAFOOD 44. 37 (1). Journal of Food Protection. 45 (9)..H.. Microbiological guidelines for some ready-to-eat foods sampled at the point of sale.
Histamine production by psychrophilic flora.. Contamination pattern of Listeria monocytogenes and other Listeria spp. Incidence of Listeria in hot and cold-smoked fish. International Journal of Food Microbiology. 1995. 1994. detection and growth of Listeria monocytogenes in seafoods: a review. 1075-80.. and temperature on the growth of Vibrio parahaemolyticus. Combined effects of water activity. 1974.. Fuchs R. Paranjpye R. Hitchins A. Incidence and sources of Listeria monocytogenes in cold-smoked fishery products and processing plants.CURED.T. Pelroy G.. 11. 1992.. Slade A. 94-8. Nicolaides L. Journal of Food Protection. pH and sodium chloride. Lashbrook L. 1734. 1994. 25 (5). 27 (6). Yndestad M.. 394-6. Interacting effects of pH. 866-70. Keusch A. 61 (12).R.J. Davies A. Poindexter C. 1994.R. Presence.A. 309-16. 25. Liston J... Food Microbiology. 1988. Bayliss A. and salt concentration on growth and survival of Vibrio parahaemolyticus. Ratnam S. 1987.. Occurrence of Listeria monocytogenes in freshwater farms and fishsmoking plants. 4389-95. 19-27. Roberts T. 502-8. 42.. SMOKED AND DRIED FISH Sutherland J. 1992. Predictive modelling of growth of Staphylococcus aureus: the effects of temperature. 14. 1995. 844-6. temperature.K. International Journal of Food Microbiology. 21 (3). Applied Microbiology. 1973.R. Elsevier. Competition of food bacteria with Listeria monocytogenes during enrichment culture.. Journal of Food Protection. Ben Embarek P. in Seafood Quality Determination.. Kramer D.E. Food Technology.. 1995. van Spreckens K. Beuchat L. Jemmi T.. 23 (1). 293301. Dillon R. Fate of Aeromonas and Yersinia on modified-atmosphere packaged (MAP) cod and trout. Letters in Applied Microbiology. International Journal of Food Microbiology.A. 217-36. 1991. Journal of Food Safety. Caugant D. Letters in Applied Microbiology. Beuchat L. New York. 58 (5). Poysky F. Rørvik L. in a salmon slaughterhouse and smoked salmon processing plant. Taylor S. Efficacy of high sodium chloride concentrations for the destruction of Listeria monocytogenes. 19.N. Eklund M.S.. Peterson M. Marine toxins of microbial origin..A.C. Eds. 1994. 121 . Tran T. Hudson J. 309-18. solute.P.. Patel T. 55 (11). Prevalence of Listeria in smoked fish. Letters in Applied Microbiology. 11.. Applied Microbiology.178-80. Dallas H.W.
the best-known of the fish sauces (gyoshoyu) of Japan is shottsuru. typically in the range 15 .25% and are used mainly as a condiment. 6. Martin Adams Faculty of Health and Medical Sciences University of Surrey Guildford Surrey GU2 7XH United Kingdom 6. Those to which only salt is added tend to be mainly autolytic.1. lactic fermentation can play a significant role. For example. The fish sauces are produced on the largest scale. 123 . The principal fish sauces and pastes of south-east Asia and their local names are listed in Table 6. It is produced from the sandfish Arctoscopus japonicus and has the rice-based enzyme preparation. they can be divided into two categories: fish/salt products and fish/salt/carbohydrate products. Sugar can also be added to the Indonesian fish sauce bakasang and the Thai product nam-budu. though some are produced elsewhere. Fermented fish products are largely confined to east and south-east Asia. the most important of all fermented fish products. FERMENTED FISH Prof. As with most traditional products that are still produced principally on a cottage industry or domestic scale. The relative importance of these two activities depends on the formulation of the product. in economic terms. Essentially. whereas in those where a carbohydrate source such as rice or sugar is also added.6.1 Fish/salt products such as the fish pastes and sauces tend to contain relatively high levels of salt. there are numerous variants of some common themes and a host of local names used to describe them. koji. producing the fish sauces and pastes. they are exported to European. the term ‘fermented’ describes a spectrum of processes that ranges from the largely autolytic degradation of fish protein. added during preparation.1 Definitions As applied to fish products. Carbohydrates are added to some fish sauces during their preparation.I. to those processes in which the activity of lactic acid bacteria (LAB) plays an important part. North American and other markets and are.
6. made from fish livers in Greece. a variety of products can be produced within a single country. to products analogous to other lactic fermented foods such as salami.I Fish sauces and pastes of south-east Asia Country Sauce Amber/brown liquid. and pissala.II. 124 . who had adopted them from the Greeks. These have long since fallen from use. may be direct descendants. although garos.FISH AND SEAFOOD TABLE 6.1. In Europe.III. where the bacterial production of lactic acid is a major feature. and the more widely known Worcestershire sauce a very distant relative. fish sauces known as liquamen or garum were ubiquitous condiments for the Romans. This is well exemplified by some of the fish/salt products of Thailand described in Table 6. Fermented fish products are of very minor importance outside Asia.2 Fish/salt/carbohydrate products range from those that resemble the fish sauces and pastes in the sense that extensive autolysis has occurred. 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. salty taste. A selection of Asian products is presented in Table 6. 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. made from whole fish in France.
8 months Main dish TABLE 6.FERMENTED FISH TABLE 6. mam-seeing sikhae som-kay-pa-eun.g.g. funa-suchi.II Fermented fish/salt products common in Thailand Name Type of fish Form Approx. mam-chao. pa-khem. cencalok burong-isda. burong-dalag.4 months Condiment and main dish Whole 1:3 18 months Condiment Whole 1:3 After 3 . pa-chao. the mixture is then boiled and bottled - Condiment Nam-khoei Shrimps.12 months 10% raw sugar added. pla-chao. e. som-fak/ som-fug 125 Malaysia Philippines Thailand . crustaceans Mackerel eviscerated fish Variety of fish Whole 1:4 Condiment Pla-thu-khem Whole 1:3 2 .III Fish/salt/carbohydrate products of south-east and east Asia Country Japan Kampuchea Korea Laos Products I-sushi. som-pa. crustaceans Variety of fish Variety of brackish or marine fish 1:4 3 . shrimps. som-pa-keng pekasam. burong-ayungi. e. burong-bangus pla-ra. salt/fish ratio Production period Type of use Hoi-dong Molluscs Whole or without shells Whole - Main dish Kapi (fish or shrimp paste) Nam-pla (fish sauce) Nam-budu Small fish. tai-suchi phaak.3 months Main dish Tai-pla Bowels 1:3 6 . pla-som. mam-pa-kor. ayu-sushi.
in Sweden. 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. it was found that several different sources were used for the fish . malt or flour (1).IV) and in the way in which they are handled prior to processing means that the initial microbial levels are far from uniform. rakefisk or surfisk. which employ lower levels of salt than Asian products. is a traditional. and counts ranging from below 104 /g to in excess of 107 have been reported. These are often described as fish/salt products. and the initial microflora is likely to be considerably larger in these cases. for example. fermented trout. the most popular varieties of which are rakørret. freshwater fish are the usual raw material. For example. Norwegian gravlaks. and old descriptions of their production often refer to the addition of whey. or buried salmon. made from herring. 126 . Here. paddy ponds beside rice fields used for collecting fish when the field has dried up.IV Fish species used in lactic fermented fish products of the Philippines Product names Burong ayungin Burong bangus Burong dala Burong gurami Burong hito Burong kanduli Burong tilapya Balao balao Burong hipon Local Ayungin Bangus Dalag Gurami Hito Kanduli Tilapya Tagunton Suwahe English Silver perch Milkfish Mudfish Goramy Catfish Sea catfish Tilapia Shrimp Shrimp Scientific name Therapon plumbeus Chanos chanos Ophicephalus striatus Osphronemus goramy Clarias batrachus Arias manillensis Tilapia nilotica Macrobrachium spp. however. in Norway and surstrømming. in a survey of fermented fish products in north-east Thailand. 6. The diversity of fish used in fermented products (see.the flooded rice fields. and a large local freshwater reservoir (2). Table 6. relatively mild-tasting product. Penaeus indicua Many of the fish used in fermentation are uneviscerated. 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. This could lead to quite marked variations since the same fish can be obtained from a variety of sources. especially in fish/salt products. A modern account of their production. TABLE 6. and their microflora tends to reflect their local environment more than that of marine species. some fermented fish products remain popular. There are also more heavily fermented products. which has been increasing in popularity over the last 50 years.FISH AND SEAFOOD In Northern Europe.
either directly or as the partially saccharified rice they produce. however. Mucor. 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. they can contribute to the initial microflora. Hansenula. traditional starter culture preparations such as koji (Japan).1 – 6. sufficient to saturate the water phase present and produce an aw of 0. were found to be the predominant isolates. angkak (Philippines and Thailand) and look-pang (Thailand) are used. Bacillus spp.FERMENTED FISH Where other ingredients are used. 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.organisms that play an important part in the production process but can also contribute to the ultimate spoilage of the product. where the total counts dropped from above 107 cfu/g to below 103 cfu/g after 14 days and below 102 after 40 days. mean that microorganisms have been generally held to play little or no role in the process that follows. 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. This. after one month. 6. In another microbiological study of four fish sauces. 4). or boiled or roasted rice. Endomycopsis and Saccharomyces . suggesting that halophilic bacteria do play an important role in the production of fish sauce (8). in numbers reaching 108 /ml after 3 weeks of fermentation but declining thereafter. however.3 6.1 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. Similar results were seen with patis. principally species of the genera Rhizopus. probably reflecting their ability as spore-formers to survive rather than any capacity to multiply under the prevailing conditions (5). in many cases. More recently. Similarly microbial inactivation by irradiation has not been found to adversely affect the properties of Korean fermented squid (7). 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.74 or below. These preparations contain a mixed microflora of moulds and yeasts.3. In some cases. its contribution is likely to be insignificant compared with that of the fish microflora in terms of both spoilage potential and public health concerns. and studies 127 .4. the Philippine fish sauce. Generally. Other halophilic species have been identified from fish/salt products. (3. When this is a simple carbohydrate source such as sugar. and the anaerobic conditions that prevail in the bulk of the material. Studies have shown that the number of bacteria declines rapidly during the production of the Thai fish sauce nam pla and that. which included nam-pla and patis.
Patis (Condiment) 128 . 6.FISH AND SEAFOOD have demonstrated the production of halophilic proteinases from fish sauce isolates (9 .2. Belacan (Condiment) Fig. Fig.1. 6.12).
6.4. This is more rapid when whole fish are used since the head and viscera contain higher concentrations of proteolytic enzymes than other tissues. Ika-Shiokara (side dish) Fig. and muscle cathepsins have been shown to be inhibited at relatively low salt concentrations (13. the 129 . 14). 6. eviscerated fish are sometimes used to slow the autolytic process. thought to be largely the result of autolytic breakdown of the fish tissues. In fish sauce production.FERMENTED FISH Fig. however. To produce semi-solid fish pastes as opposed to fish sauces. Myulchijeot (Staple food) The extensive proteolysis and liquefaction that occur during production are.3.
8. Their main feature is that the provision of a source of carbohydrate allows a lactic acid bacterial fermentation to occur. 6. filtered and bottled. Fig. particularly glutamate. Figure 6.5. 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. and the low pH produced contributes a second protective barrier or hurdle against the growth/survival of the normal fish-spoilage organisms and pathogens.3.6 – 6. snack) 130 .2 Fish/salt/carbohydrate products of Asia This category encompasses an even greater diversity of products than the fish sauces and pastes. for example. volatile fatty acids and nucleotides. 6.5). In contrast to free-swimming fish.V and Figures 6. Some examples of products and their formulation are presented in Table 6.FISH AND SEAFOOD high salt level used also assists in the osmotic extraction of a supernatant rich in amino acids. Hoi-Malangpu-Dong (side dish.
25-1 Fish:salt:roasted rice: garlic 10:1:3:1 Fish (minced):salt:rice: garlic 10:0. N. C. the prefix 'pla' is replaced by 'kung'. north-east. N. Kung-Chom (staple food.6.FERMENTED FISH TABLE 6. NE 10 . Ang-kak: red mould rice obtained by fermenting rice with Monascus purpureus. C NE. N S 5 . C NE. south of Thailand. NE Formulation Fish (whole or pieces): salt: roasted rice. 3:1:0.5:2-3:1 Fish:salt:boiled rice: ang-kak 3:1:3:0. side dish) 131 .5-1. central.2-4 Fish (pieces): salt:khaomark (fermented rice). north.4 are also produced using shrimps.7 days 5 .12 days 3 .20 days 3 Pla-som 4 Pla-jom 5 Som-fak 6 Pla-paengdaeng NE.6. NE.10 days 5 days Main dish Main dish Main dish Condiment and main dish Note: Products 1 . N. S.03 Preparation time 6 .3 years Condiment and main dish 2-3 months 3 weeks 2 weeks 2 weeks 6 .12 months Main dish 2 Pla-jao C. Fig.V Fish/salt carbohydrate products of Thailand Name 1 Pla-ra Region C. 3:1:1-3 Fish:salt:boiled rice: garlic 10:2:1:0. In these cases.12 months Shelf life Use 1 .
7. The LAB isolated from these products are those with more marked salt tolerance but.6. typically in the range 2 ..3. while they are able to grow at high salt levels.10%. then this will take longer to develop than if lower salt levels are used.VI). This is readily explained by the susceptibility of LAB to elevated salt concentrations.1 The use of higher salt levels results in a longer production phase but a better keeping quality product. This can be clearly seen from the data on reported shelf lives of a number of fish salt carbohydrate products and their respective pH ranges and salt contents (Table 6.2. their growth and metabolism are nonetheless slowed. Phillipines Journal of Food Science and Technology. (from Arroyo et al. Red burong isda 132 . the salt is of far greater importance in determining the product’s long-term stability. but there is considerable variation in this. giving a reduced rate of acid generation. It also indicates that. 106) Fig. Generally. of the two factors contributing to shelf life . 2. these products are salted at lower levels than the fish sauces and pastes. If acidity is a desired sensory property of the product. A survey of fish/salt/carbohydrate products in Thailand (15) noted that there were two empirical rules governing formulation:6.FISH AND SEAFOOD The extent to which lactic fermentation occurs depends largely on the salt content.salt and lactic acid.
1 .0 .9 4.2 4.9 . 106) Fig.5 .0 .9. Philippines Journal of Food Science and Technology.3 5. White balao-balao TABLE 6.8. 133 .9.5 .3 .8 .6.VI Shelf lives reported for fish salt carbohydrate products Product Pla-som Pla-paeng-daeng Pla-ra Pla-chao Pla-chom Som-fak Salt 2.4 .8 .12 months 1 .5.2 years 2 weeks 2 weeks Source: A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries (16).5.3 years 1 . 2.0 .6.2 4.8 220.127.116.11 3.8 pH 4.2 7.6 18.104.22.168.4.0 Shelf life 3 weeks 6 ..7 .FERMENTED FISH (from Arroyo et al.1 4.4.9 4.
efficient lactic acid production will depend upon the presence of amylolytic LAB or saccharification by other microorganisms or enzymes that may be present. TABLE 6. alternatively. Garlic. being similar (18. When added as boiled or roasted rice alone. they increase the amount of soluble sugars produced and thus the range of LAB that can grow. This results in a more active lactic fermentation and a lower pH. In several products.FISH AND SEAFOOD 6. 19). the rice may be added in a partially saccharified form such as kao-mark (Thailand). a number of amylolytic species have been described and an amylolytic strain of Lactobacillus plantarum has been isolated from burong bangus in the Philippines (17).3. inulin. Rice is the most common carbohydrate source used. Although the ability to ferment starch is not widespread in the LAB.9) compared with the white product (pH 4.0 . The effect of this will be to decrease the product’s final pH and increase the amount of total acidity.VII). of course. For example. also lower the unit cost of the product and this is probably a significant factor in many cases. Addition of carbohydrate will. 21).1 . also serves as a valuable source of the more readily fermented carbohydrate. a survey of burong-isda in the Philippines reported consistently lower pH values for red burong-isda where ang-kak was used (pH 3. a saccharifying agent such as koji (Japan) or ang-kak (the Philippines) may be added separately (Table 6.VII Fish/salt/carbohydate products in which saccharified rice or a saccharifying starter culture is used Product red burong-isda i-sushi. Where they are used. 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 . such as salt content. In som-fak 9% of LAB isolated were able to ferment rice whereas 19% could ferment garlic and omission of garlic from the formulation resulted in lower acid production during fermentation (20.3. although the final population of LAB may be similar. a common ingredient in these products.5) despite other factors.2 Inclusion of more carbohydrate gives a faster fermentation and a stronger acid taste Increasing the proportion of carbohydrate in the formulation decreases the product’s buffering capacity and provides the LAB with more substrate on which to act.2.4.
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. such as wrapping in banana leaves. This suggests that use of oxygen-impermeable wrapping films could extend the shelf life of such products.7 to below 4. and there is a marked regional variation in the preferred type. red pepper powder and garlic in the ratio fish:millet:pepper:garlic of 80:7. which ranged from 7. 6. lead to an increase in pH. 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). Cleaned pieces of eviscerated fish flesh are salted with 6% salt overnight before mixing with cooked millet. but they are likely to behave similarly to lactic fermented products produced from other commodities. presumably correlating with the salt content. Generally.2.4 Spoilage The ASCA handbook on fermented foods in the region ascribes shelf lives to various fermented fish products ranging from 2 weeks in the case of some of the low-salt lactic fermented products to 5 years for some of the high salt sauces and pastes (16). It seems that a lactic fermentation takes place in many types of pla-ra but is not significant in others.5 in the first 2 . the level of salt employed is on the high side for fish/salt/carbohydrate products.17. Any loss of acceptability must arise from chemical or physical changes occurring over prolonged storage. the salt levels in the latter are sufficient to rule out microbiological spoilage in all but the most exceptional of cases. around 12%. The growth of yeasts and moulds could. the pH drops from 6.5. There is little published information on the factors that limit the shelf life of the low-salt products.2 at the end of fermentation. The mixture is allowed to ferment for 2 . Traditional packing techniques used with lactic fermented fish products.8 . will not reliably exclude air. Results of a product survey were consistent with this. 135 . The Thai product pla-ra exemplifies these basic rules as well as the difficulties associated with attempts to classify such products. One study found that any effective measure to exclude air from a fermenting fish/carbohydrate mix prevented yeast growth (23). A combination of irradiation and chill storage has been shown to prevent both over-acidification and fungal growth in som-fak. allowing other spoilage organisms to grow. is something of an exception since millet is used along with garlic as the carbohydrate source. in turn. but in most cases it is probable that yeast and mould growth would be an important cause of spoilage. and surface mould growth has been shown to be a cause of rejection in the Thai product som-fak (24).7 up to 6. which is sufficient to prevent lactic fermentation by all but the most halotolerant LAB. reporting the product’s pH to range from 4. the Korean fermented fish product. Clearly. thereafter declining to about 4. markedly extending the shelf life (25).3 weeks at around 20 °C. Excessive growth and acid production by the LAB could render products too sour for acceptability.FERMENTED FISH Sikhae.5:9:3.3 days.9%.
outbreaks of botulism are the only well documented instances of illness associated with fermented fish products. the high salt concentrations preclude the growth of all food-poisoning bacteria and those initially present would not survive well the prolonged exposure to high salt during production. typically 30 °C. In the fish sauces and pastes. The epidemiological evidence points to greater awareness of the importance of these factors since outbreaks have declined over recent years (27). however. The combinations of pH and salt in many fermented products are likely to preclude growth of the organism completely and. In Japan. The fish/salt/carbohydrate products depend on the combination of salt and pH/lactic acid for their safety.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. There is little in the way of documented information associating fermented fish products with foodborne illness.5 Pathogens: Growth and Survival Many fermented fish products are stored at ambient temperatures. outbreaks of 136 . fish products are the most common source of botulism. V. parahaemolyticus has been reported growing down to a pH of 4. 6. are Vibrio parahaemolyticus and Clostridium botulinum. fish can potentially be contaminated with almost any foodborne pathogen as a result of improper handling and storage. the pH decreases below 5 and safe processing is recognised to depend on proper salt levels and temperature control until the pH drops. The Scandinavian fermented fish product rakefisk was responsible for 16 (84%) of the outbreaks of botulism recorded in Norway in the period 1961 90. 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. Improper storage of fish prior to processing could result in the production of toxins capable of persisting through to the product. but we are unaware of any reports of this happening. although these limits apply when all other factors are optimal (26). As in Norway. Because of the severity of the illness it causes. and failure to achieve a sufficient combination of inhibitory factors could lead to their growth or survival. viable numbers will probably decline quite rapidly. The bacterial pathogens most commonly associated with the living environment of fish. under conditions inimical to growth and high ambient storage temperatures. After capture. Their safety therefore is critically dependent on the quality of the raw materials used and the inhibitory effects of the fermentation process. which accounted for 88% of outbreaks in the period 1952 .8 and at salt levels as high 10%.87. and consumed without cooking. During fermentation. Several measures have been suggested for reducing the risk of botulism. and the most frequently implicated product is the fish-salt-carbohydrate fermented product Izushi.
1990. Institute of Nutrition. Lutefisk. 6. viverrini were a strong risk indicator and that consumption of fermented fish products was an independent risk factor in the contraction of cholangiocarcinoma (31). botulinum. giving an alkaline fermentation.FERMENTED FISH botulism have become less frequent in recent years. Infection with the trematode Opisthorchis viverrini is associated with cholangiocarcinoma. They are packed into containers such as buckets or pits in the ground and left at ambient temperature to putrefy. Salting levels are often insufficiently low to prevent growth of C. Kasemsarn B. particularly in south-east Asia and the western Pacific regions (30). Raw materials subject to this process include seal flipper. Microbiology and chemistry of fermented fish. Rakefisk and Herring in the Norwegian Tradition. 137 . 2. whale meat. 1989. 3. 31. Liston J. they could act as an important vehicle of transmission.. One study with the trematode Haplorchis taichui. Baseline Study (1987-1988). Fermented Fish Products Study (Thailand) Phase 1. though 27% survival was still recorded (33). Saisithi P. salt content was found to have the greatest impact on the viability of larvae. Dhanamitta S. botulinum and the production of toxin. Botulism has also been associated with a number of traditional foods of the peoples of northern Canada.. 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 these are often used in the production of fermented fish products. Many of these products are described as fermented. A case-control study conducted in northeast Thailand showed that elevated levels of antibodies against O.6 1.. Greenland and Alaska. With this particular organism. and salmon heads and eggs. Yhoung-Aree J. 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. Novus Press. References Riddervold A.M. Thailand. affecting an estimated 40 million people. walrus nose. showed that inclusion of fermented fish in the Thai dish lab-pla improved inactivation of the metacercariae. although the absence of any source of fermentable carbohydrate means that the process is putrefactive.. The more widespread current use of non-traditional containers such as glass or plastic jars is thought to be one contributing factor (28). Foodborne trematode infections are emerging as a major public health problem. 1966. Puwastien P. while increasing the acetic acid content of the formulation had little or no effect (32). Mahidol University. 105-10.. Journal of Food Science. Oslo. An investigation of survival of the fish parasitic nematode Anisakis in marinated herring has given some indication of what these studies might find. which is generally consumed raw (29). and warm weather can result in rapid growth of C.O. beaver tail and paw. which will persist into the product. Where these products are consumed without final cooking. Dollar A.
185-92. SK33 isolated from fish sauce fermentation. Nov. Journal of Fermentation and Bioengineering. Crisan E. 11. 2008. Microflora of four fermented fish sauces. Lee S.. Takada K. 353-63.R. 138 . 1395-401. 63. 8. 1985.W. microbiological and chemical properties of salted and fermented squid. Olympia M. 141-2.K. 12. Saono S.M. Dymsza H. Aslam Z.FISH AND SEAFOOD 4... Sinsuwan S. 2000.. 61-73. 1972. Catheptic activity of fish muscle. Sands A. 15.. Itoh T.. Hiraga K.. A Concise Handbook of Indigenous Fermented Foods in the ASCA Countries. 7. 1976.K. Tanasupawat S. Journal of Food Technology. Phil. Bioscience. Orejana F.. Applied Microbiology.. International Journal of Systematic and Evolutionary Microbiology.. Takano M. Tsunemi M. SR5-3 isolated from fish sauce: purification and characterisation. Effects of gamma radiation on sensory qualities. 11. from fermented shrimp paste in Thailand. International Journal of Systematic and Evolutionary Microbiology. 3649. 37. 1957. Pakdeeto A. Rattagool P. Halophiles in Thai fish sauce (Nam pla). Journal of Food Science. Dhamcharee B.S. Fermented fish products of south east Asia. 6. Protein hydrolysate from fish.J. 70. 2007.. 643-7. 381-8. Fukuda H. 2006. Journal of Food Science and Technology. Yasir M. Kyong-Haeng L.. Journal of Scientific and Industrial Research (India). New York. Some chemical changes in fish silage. 17.. Thawai C. Backhoff P. Lim J.. Salinococcus jeotgali spp. Tropical Science. 3. 57... Kudo T. Lentibacillus kapialis spp. Yongsawatdigul J. Constantinides S..H. 934-9. 25. 1979.. 124-30. a traditional Korean fermented seafood. Liston J.. Thongthai C.V. Production and characterisation of NaCl-activated proteinases from Virgibacillus spp. Canberra. Byun M. Tanasupawat S.. Plenum Press... 10. Dong-Ho K. Nov..H. Rodriguez-Valera F. 17-30. 16. 106-8.. 1975.T. Ono H.. Process Biochemistry. Government of Australia. isolated from jeotgal. Im W. 6338. Reddi P.. Chung Y. 43. Jae-Hun K. in General and Applied Aspects of Halophilic Microorganisms. Ed. 80. 13. Cooke R. Ahn H. Hull R. Namwong S.A.. 29..R... 5. 9. Rodtong S. 14. 2007. 1986....M.T. 57. Oda K. 1991. Velankar N.. A halophilic serine proteinase from Halobacillus spp.. Characterisation of starch-hydrolysing bacteria isolated from a fermented fish and rice food “burong isda” and its amylolytic enzyme. 16A. Agents of proteolysis and its inhibition in patis (fish sauce).D.R.. Adams M.. 1995. Moonmangmee S. Kaneko Y. Biotechnology and Biochemistry. Suntinanalert P. Yook H. Journal of Food Protection.
2002. Marcel Dekker. Epidemiology of human foodborne botulism. Blackie. 2002. Erikson U. 23. 103.. Journal of Agricultural Science (Tokyo).. Volume 5: Microbiological Specifications of Food Pathogens. Benjakul S. Ed.. Tanaka M.. 27. Food Chemistry. 139 . Paludan-Müller C. 38. 28.H. New York. burong-isda.A. Canada Communicable Disease Report..H. Gram L. Moody L. 274-86. Characterisation of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation. 19. Gram L. Paludan-Müller C. Martin J. 1987.. Journal of Agricultural Science (Tokyo). Valyasevi R. 92. 1992. 25. 138-44. Dodds K.. 22. 30. Fermentation and spoilage of som fak. Visessanguan W. 28 (6).B. Yamprayoon J. Eds. in Microorganisms in Foods. Avurhi J. 28. 1998. Effect of irradiation on properties and storage stability of Som-fug produced from bigeye snapper. Hauschild A. Hauschild A.. 1995. Huss H. The influence of oxygen accessibility on the growth of yeast in fish/rice fermentation.. a Thai low-salt fish product. 20. in the Philippines.L. Riebroy S. Wainwright R. 426-35.M.L.A. Isaac-Renton J. London.W. 1-4.. in Clostridium botulinum: Ecology and Control in Foods. Gram L. 219-29. 3. 105-12.. 1983. Patrick D. 1996. 305-22.K. Journal of Applied Microbiology. 26. Dawar M. Sakai H. Control of foodborne trematode infections... Ostergaard A.H. a Thai low-salt fermented fish product. International Commission on Microbiological Specifications for Foods. New York. 69-104.H. Kozaki M. The fermented fish food. Rustad T. Hazards from northern native foods. Dodds K... Huss H... Tropical Science. Sakai H. 1815.. Vibrio parahaemolyticus. Two outbreaks of botulism associated with fermented salmon roe – British Columbia. WHO Study Group.. in Clostridium botulinum: Ecology and Control in Foods. 1990. Hauschild A. 24.D. 29.. Caldo G.. Yeast-flora in red burong-isda a fermented fish food from the Philippines.H. Huss H. Genotypic and phenotypic characterisation of garlic-fermenting lactic acid bacteria isolated from som-fak. Fung C.W.. Journal of Food Science and Technology. 1992. Wedell-Neergaard C. International Commission on Microbiological Specifications for Foods.W. Ben Embarek P. Traditional fermented fish products with special reference to Thai products. ASEAN Food Journal..D... 46. 1999. 307-14. 28.. Caldo G. 1983. 104-6.B. 21. Eds. 27. 2007.H. Marcel Dekker.. Kozaki M. World Health Organisation Technical Report Series.FERMENTED FISH 18. International Journal of Food Microbiology. Saisithi P. 3-10. Owens J.
Genetic and environmental determinants of risk for cholangiosarcoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom. Thai traditional food preparation. Bloemsma B. Survival of Anisakis larvae in marinated herring fillets.. Horst K. Wongsawad C. Hanai S.. Ruamsuk A. 110-1.. Honjo S. northeast Thailand. Survival of Haplorchis taichui metacercariae in lab-pla. 2005. 2001.. 29. Fermented flesh foods. Sripa B.. Southeast Asian Journal of Tropical Medicine and Public Health. Huss H.R. 854-60.J. 32. 36. 6. 117.R.. Aspen Publishers Inc.7 Further Reading Adams M. Nithikathkul C... International Journal of Cancer. 23. 2005.FISH AND SEAFOOD 31. Progress in Industrial Microbiology. Todoroki T.. Chuboon S.R. Roepstorff A..H... Uchida K. Gaithersburg.. Fermentation and Food Safety. Kikukawa H. Kittiwatanachot P.. Adams M. 15998. International Journal of Food Science & Technology.. Deerasamee S.. 1986. Miwa M. Sriplung H. Jedpiyawongse A. 140 ... 661-70. Srivatanakul P. 33. 1995. Nout M.
7. Another book dealing with the clinical effects of a variety of poisons and toxins.M. Ph. Increasingly sophisticated instrumentation (e. most notably the azaspiracids and the cyclic imines as well as some additional polyethers. a number of new toxins and toxin families have been identified. Scientific and Cultural Organisation) has been updated (7). known and unknown. Wright.. An emerging technology is the use of immunosensors (1 . several scientific reports have described their efficacy and potential. It remains to be seen if this approach gains momentum in the future. While some of these toxins only occur in seafood from a restricted geographical region.g.D.C. FISH AND SHELLFISH TOXINS Jeffrey L. physiology and detection of marine and freshwater toxins has also appeared (8). Although most of these antibody-based test kits have yet to receive international regulatory approval. Since the original chapter in this series was published (6). produced by the Intergovernmental Oceanographic Commission (IOC) of UNESCO (United Nations Educational. The manual on harmful marine algae and their products.. FCIC UNC Wilmington Center for Marine Science Marvin Moss Lane Wilmington NC 28409 United States of America 7. and is often necessary in light of the complex toxin profiles that can occur in shellfish tissue. Nevertheless. including the well known marine toxins 141 . the export of exotic seafood that may contain such toxins can lead to unexpected public health issues. more traditional methods such as the mouse bioassay continue to be used in many countries to warn of the presence of toxins.1 Introduction Shellfish and finfish toxins continue to be a threat to public health and local fisheries around the world. C. Another important development is the introduction of sensitive and selective antibody-based detection methods for a number of toxins. Liquid Chromatography/ Mass Spectrometry/ Mass Spectrometry -LC/MS/MS) is employed to identify and confirm the presence of known toxins. In the last decade seafood monitoring has intensified and detection methods have continued to develop in selectivity and sensitivity. and a second updated edition dealing with the pharmacology. 5).3) and some practical applications for detecting shellfish toxins have been reported (4.
Consequently. can become contaminated with these toxins. PSPs are watersoluble and heat stable so cooking has little or no effect on PSP toxicity. and a tropical species Pyrodinium bahamense (16).FISH AND SEAFOOD has been produced (9). even crustaceans such as lobsters. Many of these species are extremely common and are found throughout the world in a variety of cold.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. Not all the toxins found in shellfish extracts are microalgal products. An account of this meeting detailing recommendations and scientific advice on marine biotoxins has been published (10). dealing with the detection methods and regulatory requirements for many marine toxins has been published (12). 7. a great variety of shellfish from all latitudes. and sub-tropical waters and habitats (15).2. with intoxications occurring in many countries even today (15). all of which contain a tetrahydropurine skeleton (Figure 7. All PSPs degrade rapidly in alkaline conditions. The Ncarbamoyl sulfates are readily hydrolysed to STX. A brief report on the status of rapid detection methods for phycotoxins in the European Union (EU) has appeared (11).1). In addition. Gymnodinium catenatum. and a review on the risk assessment of marine toxins has also been produced (14). and another.g. 7. Around twenty PSP toxins are known. Saxitoxin Group instead of the Paralytic Shellfish Poisons). neoSTX and Gonyautoxins (GTX) 1-4. The remarkable frequency of neurotoxins from marine dinoflagellates has been noted (13). 142 . some are the result of biotransformation processes by the filter-feeding host. 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. temperate.1 Origins and distribution The PSP toxins are produced by several dinoflagellate species belonging to the genera Alexandrium. 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. Saxitoxin (STX) and neosaxitoxin (neoSTX) are the parent members of the group and are the most toxic.
and fingers within 5 . this is never sufficient to render the seafood safe to consume.2. the less potent C toxins are readily hydrolysed by acid to the more potent GTX toxins. tongue. Cooking does not destroy these compounds and although some of the toxins may be extracted into the cooking water.2 Toxicology PSP toxins affect the central nervous system of mammals.FISH AND SHELLFISH TOXINS STX GTX2 GTX3 GTX5 (B1) C1 C2 NEO GTX1 GTX4 GTX6 (B2) C3 C4 R1 H H H H H H OH OH OH OH OH OH R2 H H OSO3 H H OSO3 H H OSO3 H H OSO3 - R3 H OSO3 H H OSO3 H H OSO3 H H OSO3 H R4 H H H SO3 SO3 SO3 H H H SO3 SO3 SO3 - Fig. 143 . However.1. Structure of saxitoxin and neosaxitoxin and their sulphated derivatives 7. 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).30 minutes of consumption. leading to paralysis within a few hours and in extreme cases death by asphyxiation due to respiratory paralysis. 7. 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. and humans seem to be particularly susceptible. Human symptoms begin with tingling and numbness of the lips.
Early post-column methods were somewhat time-consuming and required three separate HPLC runs to complete the analysis. An antibody-based kit that can detect STX in shellfish is commercially available (RIDASCREEN®). and more standards of individual PSP toxins are required to validate the complete profile to toxins that could occur in contaminated shellfish tissue. as well as other parts.3 Detection methods The safe limit set by most countries for PSP toxins is 800 µg/kg (0.3. 23. including Japan. and while simple and relatively inexpensive to operate. a criticism has been that such methods are time consuming.3 Tetrodotoxin Tetrodotoxin (TTX) possesses a unique chemical structure and was the first marine toxin to be chemically characterised.2). The mouse bioassay has been in place for decades and the method has been standardised. Two approaches have been taken here. 17). However it remains as the monitoring method of choice in many countries. China. and has also been evaluated by a number of EU laboratories. Most intoxication events have occurred in South East Asia. Since its discovery. TTX most commonly occurs in many species of puffer fish. or conversion before HPLC separation (pre-column derivatisation) and these have recently been thoroughly reviewed (12. namely oxidative conversion to fluorescent derivatives after High Performance Liquid Chromatography (HPLC) analysis (post-column derivatisation). Taiwan. and a second rapid dipstick kit (MIST Alert™) is available for the qualitative detection of positive and negative samples (19). other derivatives have been reported.1 Origins and distribution Unlike many other marine toxins there is no temporal distribution of TTX episodes. One approach has been to take advantage of the ready conversion of these toxins to fluorescent derivatives. 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. Thailand. and over ten analogs are known (22.8 µg/g) of shellfish or seafood tissue. While the chemical detection methods are most promising for the future. This kit was found effective in all the samples tested in Europe (20) and was applied recently in a survey of Scottish shellfish (21). After some development and various trials. specifically 144 . a pre-column method has received Association of Analytical Communities (AOAC) approval (18). 7. Alternative HPLC methods have subsequently reduced this to a single HPLC run. 24) (Figure 7. There continues to be a significant effort applied to the development of alternative chemical and biochemical detection methods for these toxins. 15.FISH AND SEAFOOD 7.2. The latter test is accepted by the United States Food and Drug Administration (US FDA) for use in the United States National Shellfish Sanitation Program. and islands of the South Pacific. 7.
respiratory failure and coma. and in mild cases results in distal muscle weakness. In Japan. 25. All of the observed toxicity is secondary to the action potential blockade.3. hypotension. Fig. However TTX has been found to occur in a remarkable variety of genetically unrelated species including gastropods. gonads and roe (24). and blocks diffusion of sodium through the sodium channel. 26). 27). 7. this has expanded to include bacteria within the Alteromonas and Pseudomonas genera. 7.2 Toxicology Like the PSP toxins. Since then. and even newts. horseshoe crabs. and once again the mouse bioassay has been used to detect the presence of the toxin. where puffer fish of the genus Fugu are a delicacy. raising speculation that TTX is produced by a bacterial source. Structure of tetrodotoxin 7.2 mg of purified toxin can be lethal. preventing depolarisation and propagation of action potentials in nerve cells. and ingestion of larger amounts leads to cardiovascular effects including bradycardia.3. 23. Typically this has been accomplished using HPLC followed by post 145 . the blue-ringed octopus.3 Detection methods TTX can be extracted from suspect tissue using dilute acid conditions. A review of TTX detection methods has been published (12. 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. nausea and vomiting. Severe poisoning causes paralysis and respiratory failure. TTX acts on the central nervous system by binding extracellularly to site 1 of voltage-gated sodium channels. The onset of symptoms usually occurs rapidly.FISH AND SHELLFISH TOXINS in the liver.2. This was indeed confirmed and a number of marine Vibrios have been shown to be capable of TTX production (24. it is served in speciality restaurants with qualified chefs to properly prepare the fish. An oral dose of 1 . and more recently by an actinomycete Norcardiopsis dassonvillei (26). molluscs.
but all of them are considerably less toxic than DA itself (29).FISH AND SEAFOOD column oxidation to the fluorescent derivatives.1 Origins and distribution The cosmopolitan diatom Pseudonitzschia multiseries was identified as the culprit organism in the 1987 poisoning event (31). Fig. but 146 . Currently 12 isomers of DA have been reported (see Figure 7.4 Amnesic Shellfish Poisoning (ASP) Toxins The parent compound of this group is domoic acid (DA). 7. Immunoassay methods have also been investigated. a polar glutamate agonist.3. 7. Surveys by a number of groups have identified over a dozen species of Pseudonitzschia capable of producing DA. However this method has been criticised for being complex. These isomers arise as a result of isomerisation and location of the double bonds in the side chain of the molecule. Structure of domoic acid and three naturally occurring photoisomers 7. and while promising. need further development (24). but it is known that such congeners are less toxic than DA itself (30).3 for examples). Since the original episode of human poisoning following consumption of DA-contaminated shellfish in Canada in 1987 (28).4. both the toxin and the diatoms capable of producing it have been found to be distributed worldwide. 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).
2 Toxicology DA is an acidic amino acid and potent neurotoxin unaffected by cooking or steaming. another diatom species Nitzschia navisvaringica.10% of the administered dose. Levels of DA in the other strains are generally <1 pg/cell. 33). Recently. herbivorous fish become toxic after ingesting strains of Pseudonitzschia spp. cormorants and pelicans (34). scallops. and Pseudonitzschia seriata) are considered to be high producers of DA (>10 pg/cell). seizures. The effects of DA may be augmented by the excitotoxic effects of glutamate and aspartate in the sample. multiseries. as does impaired renal function which increases serum concentration and residence time of the toxin. and sea animals such as sea lions and even whales (35). and in locations around New Zealand (32.. To a great extent. 7. Victims display a variety of symptoms including headache. Another factor is the movement of the toxin through the food chain. Four deaths were attributed to the Canadian incident and years later a few seriously affected individuals still display chronic neurological deficits. Okinawa and more northerly parts of Japan. including persistent short term memory deficits (36 . The original 1987 event was followed some years later by a more widespread episode along the western coast of the US and today toxin-producing strains of Pseudonitzschia have been found in the Gulf coast of the US and along the western shores of the US and Canada.FISH AND SHELLFISH TOXINS only three strains (P. disorientation and coma. Penetration of the blood brain barrier is poor in healthy subjects. The oral absorption of DA is around 5 . the Mediterranean. has been reported as a medium-level producer of DA (33). 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. For example. which has been well documented in studies along the west coast of the US. and is mainly distributed in the blood. nausea. Indeed. Pseudonitzschia australis. found in the Pacific around Vietnam. and review of the toxicologic pathology of DA has been published (36). 147 . 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. southern and western portions of Ireland. the Philippines. 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. and this has led to ASP toxicity in birds. and clams. 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.39). but any impairment of the blood brain barrier results in additional risk.
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. and this is typically the method of choice.30 ng/g) with a rapid clean-up step using a strong anion exchange resin. However. are the more potent source of toxins – often very low cells/ml of seawater can result in human poisoning. are amenable to laboratory culture and consequently are used in many experiments involving studies of DSP toxins. the heterotrophic dinoflagellates Dinophysis spp. 49). Since then.. The probes have been employed in trial studies in New Zealand (44) and Scotland (43) to detect P.4. shellfish tissue can be efficiently extracted with aqueous methanol (1:1). and an immunosensor method has also been reported (4). A rapid immunodiagnostic kit is now available to detect DA in phytoplankton and is reported to work satisfactorily in this application in the field (43). dinophysistoxin-1 (DTX-1). and dinophysistoxin-2 (DTX-2) (Figure 7. and consequently were often overlooked as the real source of the toxins. produce a variety of other 148 . though this method is most commonly used to analyse plankton and seawater (42). In the context of this chapter. DTX-2 has been found to be extremely widespread throughout many European maritime countries and is often the major DSP toxin present. Provided the concentrations in shellfish are high enough (>50 µg/g shellfish tissue). but just over 15 years ago a third toxin. australis. DTX-2 was discovered in contaminated Irish shellfish (46).FISH AND SEAFOOD 7.. 48. Of additional interest is the use of molecular probes originally developed to detect the presence of cells of potentially toxic Pseudonitzschia spp. The benthic Prorocentrum spp.3 Detection methods DA is water soluble and can be extracted from shellfish tissue by the acidic PSP extraction method although this may result in some decomposition. DSP toxins are produced by dinoflagellates of the genera Prorocentrum and Dinophysis (47.4). An EnzymeLinked ImmunoSorbent Assay (ELISA) kit (Biosense) for DA has been approved by the AOAC. Dinophysis spp. 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. Alternatively. Detection levels can be further lowered by conversion of DA to the fluorescent fluorenylmethoxycarbonyl (FMOC) derivative. in Monterey Bay.5 Diarrhetic Shellfish Poisoning (DSP) Toxins The active agents responsible for DSP incidents are a group of polyethers that include okadaic acid (OA). To further complicate matters. Pseudonitzschia fraudulenta and Pseudonitzschia delicatissima were found to be less effective. the term “DSP toxins” will only be used to refer to these three polyether compounds and any derivative based on them. and these methods have been reviewed (40. 41). 7. California. The sensitivity of the LC/UV (ultra violet) method (detection limit ~ 1 µg/g) can be improved (20 . Since their first characterisation it was believed that the only causative agents of this group were OA and DTX-1 (45). although in the latter study the probes for Pseudonitzschia pungens. as can LC/MS.
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. In comparison. Dinophysis acuta. Dinophysis fortii. and include Dinophysis acuminata.4. Norway and Japan were likely due to DSP toxins produced by a Dinophysis spp. are also recognised as a threat to human health. are recognised as DSP toxin producers. Today over ten Dinophysis spp.5. and Prorocentrum belizeanum. which include the pectenotoxins and the yessotoxins. Structure of okadaic acid and the two related DSP toxins 7. and DTX-6) all of which are rapidly converted to OA when the plankton cells are ruptured (48). DTX-5. 149 . Lately these compounds.. such as Prorocentrum lima. This was probably not the first DSP episode. The parent acids are the only DSP congeners to be found in shellfish tissue. In this case Dinophysis was suspected as the culprit organism. 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. Dinophysis caudata. and Dinophysis norvegica among others (47. 49). DSP toxins produced by benthic Prorocentrum spp. but do not have the same mechanism of action as DSP toxins nor induce the same symptoms. Fig. While these esters are not toxic in vitro. they can be converted to the parent toxins by nonspecific esterases or lipases either in the producing organism or in the host shellfish. 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.FISH AND SHELLFISH TOXINS lipophilic compounds that are toxic in the mouse bioassay. 7. analysis of Prorocentrum cultures have uncovered an additional series of diol esters of OA and sulphated diesters (called DTX-4. Prorocentrum maculosum. and anecdotal evidence now suggests that several seafood poisoning episodes during the 1960s and 1970s in the Netherlands.
Human symptoms include diarrhoea. Immunoassays have been developed for OA detection but an inherent problem of these methods has been the low cross reactivity with DTX-1. 7. The onset of symptoms can occur within 30 minutes of consuming contaminated shellfish and generally last about 2 . even with the minimal of clean up steps (48). 7. 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). and DTX-2. and a detection limit of 100 µg/kg has been reported (12). DSP-Check is a commercially 150 . 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. DSP episodes have been reported worldwide and are included in many monitoring programs (47). In recent years. which can lead to an underestimation of the total DSP content in a sample. but the method is complex and matrix effects as well as interferences with other lipophilic compounds such as the pectenotoxins compromise the method. At the first Meeting of the EU National Reference Laboratories on Marine Biotoxins and Analytical Methods and Toxicity Criteria in 1996.160 µg OA equivalents/kg of whole shellfish tissue has been implemented in some countries. some careful clean-up steps are required to avoid such matrix interferences and to simplify data interpretation (48). New Zealand and Chile.3 Detection methods Once again the mouse or rat bioassay is relied upon to detect DSP toxins in contaminated seafood. 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. and abdominal pain. A regulatory level of 80 . Preparation of a fluorescent 9-anthryldiazomethane (ADAM) derivative followed by HPLC offers reasonable sensitivity. various European countries.2 Toxicology The DSP toxins are potent inhibitors of the serine/threonine phosphatases PP1 and PP2A.FISH AND SEAFOOD Perhaps as a consequence of these multiple sources of DSP toxins. Since the late 1970s. 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). DSP episodes have been reported from Japan. Due to the lipophilic nature of the toxins. This regulatory level includes the possibility of LSPs in the DSP fraction being tested. All the DSP toxins have around a thousandfold greater affinity for PP2A than PP1. 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.5. vomiting.3 days. nausea.5. 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. it does not provide information on the DSP toxin profile. and while this provides critical seafood safety information.
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). Indeed. As a result these compounds are now being referred to as lipophilic shellfish toxins (LSTs) (49). fluorometric protein phosphatase inhibition assays have been found to perform better than colorimetric assays.6.5 and 7. for many years these compounds were included in the DSP toxin complex. Some representative examples of both groups of toxins are shown in Figures 7. 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. with a claimed detection limit of 20 µg/kg. and another rapid antibodybased test kit for DSP toxins has also been reported (53). and the latter group are usually found to be responsible for any reported shellfish toxicity (59). 7.FISH AND SHELLFISH TOXINS available ELISA test kit that can detect both OA and DTX-1 with comparable sensitivity. Structure of the principal pectenotoxins and one of the seco-acid derivatives 151 . with good agreement with the mouse bioassay and LC–based methods (54. Fig. over 20 PTXs are known (56) and more than 30 YTXs have been reported (57. Within this group. 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. 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. However. they are toxic in the mouse bioassay and can result in false positives and hence are included here for completeness.5. In yet another approach. 55). 58). 7.
6. 7. many parts of Europe and the Mediterranean. 57. 58). The sources of YTXs have now been identified as the dinoflagellates Protoceratium reticulatum and Lingulodinium polyedrum. Structure of the two parent yessotoxins. but nevertheless the toxins are extremely widespread and have been found in various shellfish in Norway and Scandinavia. New Zealand and South America (49. YTX and homo-YTX 7.6.FISH AND SEAFOOD Fig. eastern Canada. 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). which may co-occur in blooms with other organisms. 7. Oxidation of the methyl group at C-43 to a carboxyl function as in PTX-6 significantly reduces toxicity (500 µg/kg body weight) as does hydrolysis to PTX-2 seco acid (>5000 µg/kg body weight) a common metabolite found in shellfish extracts.6. As is typical for many algal species.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). While 152 . the toxin content and profile may vary among different strains. Japan.2 Toxicology PTXs show varied toxicity in the mouse bioassay (59).
Although there is no evidence that yessotoxins are responsible for human illness. Their toxicology and mechanism of action remain uncertain. some of which are found as glycosides and sulfates. a regulatory level of 1 mg/kg shellfish tissue has been set. The structures and chemical data including the molecular weight of each member have been collated (56). Acetone extracts of shellfish tissue are dried and re-suspended in 60% aqueous methanol before partitioning against dichloromethane. The EU has recommended an extraction and solvent partition process to separate the polar YTX compounds from the more lipophilic PTX and DSP compounds.3 Detection methods Typically these toxins are co-extracted with DSP toxins following acetone or aqueous methanol extraction of the shellfish tissue. chemical analysis is best carried out by LC/MS and several approaches have been described (57. There are no clear cases of human illness resulting from ingestion of PTXs (59). Many YTX toxins contain a conjugated diene system. Because of the number and complexity of the YTX group. However. 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). While LC/UV detection is possible for PTXs. but various congeners are acutely toxic in the mouse bioassay following IP injection. the PTXs show considerably less toxicity when administered orally. 58). it may be compromised by other biological materials in the matrix. For many years it was believed that 153 . 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. There are two skeletal types known as PbTx-1 (or brevetoxin A) and PbTx-2 (brevetoxin B) (Figure 7.7). which is further compounded by differences in the experimental design. but a fluorescent derivative has been prepared for others. The YTX group has not been reported to cause human illness. The crude extract may be used directly in the mouse bioassay.6. which aids detection by LC/UV. Estimates of acute toxicity of the parent compound following IP injection vary considerably. The YTXs are retained in the aqueous methanol fraction while the PTX and DSP toxins are concentrated in a dichloromethane fraction. 64).FISH AND SHELLFISH TOXINS demonstrating considerable toxicity following IP injection. though its cell effects may be linked to modulation of calcium channels (62).7 Neurotoxin Shellfish Poisoning (NSP) Toxins NSP episodes are caused by a family of fused polyether compounds known as the brevetoxins (63. Some of the PTXs are potent cytotoxins in vitro (60). and this is believed to be due to hydrolysis of PTX-2 to the seco acid derivative. 7. 7.
Structure of the two parent brevetoxins. These are biotransformation products of principally PbTx-2 by addition of certain amino acids to the conjugated aldehyde function (66). They have also been associated with fish kills along the eastern mid-Atlantic coast of the US. 66). In the Gulf of Mexico. whelks. which are found in Japanese waters (66). and one of the brevetoxin metabolites BTX-B2 7. oysters. Fibrocapsa japonica. 7. and Heretosigma akashiwo. verruculosa off the mid Atlantic coast of the U. Many of these species are associated with massive fish kills. was chemically characterised as PbTx-2 (67). 154 .S.7.FISH AND SEAFOOD NSP toxins only occurred in the Gulf of Mexico and along the western coast of Florida.1 Origins and distribution The brevetoxins are produced by the dinoflagellate Karenia brevis (formerly known as Gymondinium breve and Pytochodiscus brevis) a dinoflagellate found in the Gulf of Mexico. A complicating factor both in terms of establishing the profile and toxicity of brevetoxin-contaminated shellfish has been the discovery of the so-called “brevetoxin metabolites”. and recently the major toxin associated with an ichthyotoxic unialgal bloom of Chatonella cf. and off the coast of New Zealand (65). 66). Fig. and cockles (65. namely Chatonella marina. but in 1992/1993 they were identified as the culprits in a human poisoning event in New Zealand (65. brevetoxins are found in mussels. Interestingly.7. the same toxins have been reported to be produced by three genera of Raphidophytes. PbTx-1 and PbTx-2.
In this application. and it has also been used in studies to quantify occupational and recreational exposure to aerosolised brevetoxin exposure (73). While the mouse bioassay continues to be used. 7.3 Detection methods Three basic approaches for the detection of NSP toxins have been recently reviewed (71). 7. and an action level has been set at 800 µg/kg (or 80 µg PbTx-2 equivalents/100 g or 20 MU/100 g or 4 µg/mouse) of shellfish tissue to trigger regulatory action. The brevetoxin metabolites are reported to be less toxic than the parent compound (70). Ingestion of brevetoxins results in a variety of symptoms including parathesia. and in severe cases seizure. pre-treatment or clean-up of the shellfish sample is essential in order to obtain reproducible results. nausea.8 Ciguatoxins Known for centuries and originally referred to as ciguatera by Spanish explorers in the Indo-Pacific ocean and Caribbean. with hot spots in the coral reef areas of the Caribbean and French Polynesia. 66. vertigo. Ciguatera toxins (CTXs) are extremely potent and occur in finfish rather than shellfish. beginning with herbivores that feed on benthic dinoflagellates associated with reefs and which are 155 . considerable progress has been made in the development of LC-MS methods to routinely monitor for brevetoxins in shellfish. which can result in a cascade of associated events in a cell (66). and diarrhoea. and mammalian body fluids without pretreatment or dilution (72).FISH AND SHELLFISH TOXINS although recently it has been shown that herbivorous fish can store brevetoxins in their flesh (68).7. The mouse bioassay is used as the primary detection method. 7. The human health implications following exposure to NSPs have been reviewed (69). with a mean duration of symptoms of around 1 day. the competitive ELISA method correlated most closely with the mouse bioassay results. A second generation competitive ELISA assay has been developed that has successfully quantified brevetoxins in a variety of biological and environmental matrices including seawater. malaise.2 Toxicology Brevetoxins bind to site 5 of voltage-gated sodium channels.7. Outbreaks of ciguatera poisoning are confined to tropical and sub-tropical regions of the ocean. shellfish homogenates. In a comprehensive multi laboratory evaluation of five different detection methods as possible replacements for the mouse bioassay. but this method is now used routinely in New Zealand and the US to monitor for the presence of brevetoxins (68). this family of toxins has proven to be a challenge both to identify and monitor. Exposure to brevetoxins can also occur through inhalation of brevetoxin aerosols resulting in irritation of the upper respiratory tract (65. 69). The mean time to onset of symptoms is around 3 hours.
and only a few structures have been identified. but more are suspected to exist. The diversity of benthic phytoplankton that exist on reefs further complicates the picture. Such microalgae contribute to the primary production of tropical and sub-tropical reefs and live in epiphytic association with bushy red. Structure of the Pacific variant of ciguatera toxin (P-CTX-1) and the Caribbean variant (C-CTX-1) 7. 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. 7. although the dinoflagellate Gambierdiscus toxicus is considered to be the major source of these toxins. Indeed even the chemistry of CTXs from the Pacific differs subtly from those found in the Caribbean (Figure 7. green and brown seaweeds and also occur in sediments and coral debris.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. lipid soluble polyethers. and other genera of neighbouring reef dinoflagellates can further contribute to an extremely complex profile of toxins in finfish.8).FISH AND SEAFOOD transferred through the food chain to the larger carnivores. Culprit species producing CTXs may simultaneously produce unrelated toxins such as maitotoxin.8. Fig. complex. a group of closely related polyethers that are considered the source of ciguatoxins. CTXs are large. Distribution through the food chain begins with 156 .
Differences in potency among the CTXs correlate directly with their binding affinity for the sodium channel. which in turn are consumed by carnivores such as snappers. A good knowledge and careful observation of the symptoms in the bioassay (Minimal 157 . only trace amounts of CTXs can cause illness following consumption of contaminated fish. although not all CTXs are equipotent.76). Even when suspect samples are available. and in fact CTX poisoning is often only identified after consumption of contaminated fish. Globally. pruritus. sharks.3 Detection methods As a consequence of their potency. 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. though the mouse bioassay continues to be relied upon as the most common method of detecting CTXs. and the sensation of loose teeth (77). The onset of symptoms can occur rapidly within 24 hours.8. groupers.FISH AND SHELLFISH TOXINS grazing by herbivorous fish including the sturgeon fish and parrotfish. which can return following the consumption of alcohol and certain foods such as oily fish (74.8.1 µg of ciguatoxin can result in human illness (74. Another symptom that can develop over time is extreme fatigue. 75. they bind to voltage-dependant sodium channels in the cell membrane causing them to open. 78). 75). Local island populations attempt to avoid poisoning by catching only smaller fish.2 Toxicology Ciguatoxins (and maitotoxin which is often co-produced with CTXs) are extremely poisonous. but fish toxicity can vary widely depending on where the fish are caught and how long they have been feeding in that area. and it is estimated that ingestion of as little as 0. weeks or even months (74 . and inducing membrane depolarisation resulting in prolonged symptoms indicative of damage to nerve tissue. and temperature sensation reversal. myalgia. Nevertheless CTX poisoning continues to be a threat to local and visiting populations. and moray eels. and there are suggestions that such incidents are increasing and spreading worldwide (74). arthralgia. barracudas. the absence of a strong chromophore means that simple detection methods based on UV or fluorescence-based detection methods are ineffective. 7. there are neither standards nor an official testing program. and although the gastronomical problems may dissipate within a day or so. all of which can be linked to the effect of CTX on excitable membranes. Patients have reported symptoms lasting weeks and even months. Like the brevetoxins. The pharmacological effects following CTX exposure are numerous and include severe gastrointestinal distress and neurological symptoms. 7. where cold objects feel hot and vice versa. These include paresthesias (numbness and tingling of the extremities). which can be confused with chronic fatigue syndrome. the neurological symptoms can persist for days. Other symptoms include a metallic taste.
7.30%) and AZA3 (5 . Acetone extracts of fish tissue are subjected to solvent partitioning. high-throughput screen (74). almost 20 additional derivatives have been reported (83 . only a few laboratories are able to undertake such analyses.5) have been fully characterised by Nuclear Magnetic Resonance (NMR) and MS analysis. Recently. 84). although specificity and sensitivity remain major hurdles (74) and the presence of DSP toxins affects the test (80). Consequently.FISH AND SEAFOOD Lethal Dose (MLD) 0.9).9). and only a few structures (AZA 1 .g.20%) present in lesser amounts (Figure 7.25 µg/kg) can provide useful clues as to the toxins present but more specific and informative assays are required. and the corrected structure was published in 2004 (86) (Figure 7. 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). following consumption of contaminated mussels Mytelius edulis (82). with AZA2 (10 . and another dealing with the chemistry and detection methods has been published (85). 158 . specific for CTX activity. Usually the additional AZA derivatives found in shellfish tissue are hydroxylated derivatives (e. Usually the major AZA derivatives found in shellfish are AZA1 (50 . and following synthesis of the molecule some time later. Immunoassays may be an alternative detection method (79). minor revisions of a few stereocenters were made. This is then subjected to LCMS/MS analysis. and it is likely that most of these compounds are biotransformation products formed in the shellfish tissue. A sodium channel-specific assay using mouse neuroblastoma cells has been described although this approach has still to be developed in a routine. but this only has application in a clinical setting (81).9 Azaspiracids Azaspiracid (AZA1) was originally identified in 1995 as the causative agent for a poisoning episode in Ireland. followed by clean-up on silica and aminopropyl cartridges. which features characteristic fragment ions corresponding to sequential losses of three molecules of water.85). Since the original discovery and identification of the toxin.80%). which are based on an LC-MS/MS approach. another in vitro sodium channel-based assay. Recent reviews of this new group of toxins have appeared (83. and the separation of these polyethers from the lipid-soluble fraction of fish provides an additional challenge. The majority of these have been proposed based on LC/MS analytical data. at C-3 and C-23). Chemical detection methods are severely compromised by the lack of standards. has been developed.
88.FISH AND SHELLFISH TOXINS Fig. Ensis siliqua. oysters (Crassostrea gigas. and variable field results have led to suggestions that the actual producing organisms are Dinophysis spp. 90). 7. There has been considerable debate over the source of azaspiracids. AZA toxins do 159 . as in other cases of shellfish toxins.1 Origins and distribution The azaspiracids were first isolated from contaminated shellfish. and some time after the original episode in Ireland. Mytilus galloprovincialis).9. These symptoms are not unlike those experienced following the consumption of DSP toxins.3 days. Ostrea edulis). clams (Tapes philipinarum.9. cassipes produces AZAs in culture.9. 84. which can persist for 2 . it was reported that the heterotrophic dinoflagellate Protoperidinium cassipes was the planktonic source of the toxins (87). which are ingested by P. AZA occurrence has been reported by several European countries including France. Donax spp. and off the coast of Morocco (83. 89). The bivalves affected include mussels (Mytilus edulis. Portugal. While this result is not necessarily in debate. Following the original episode. and to date no long-term effects have been recorded (83. Scandinavia. cassipes (83). 7.2 Toxicology Consumption of AZA contaminated shellfish can result in acute symptoms that include nausea. and to date this has been the only source from which these toxins have been found at toxic levels. but current data indicates that unlike the DSP compounds. scallops (Pecten maximus). vomiting. these molluscs were never considered to be the biogenetic source. it has been impossible to establish that P. Structure of the three principal azaspiracids found in shellfish 7. and most recently in a crustacean (Cancer pagurus). cockles (Cerastoderma edule).). diarrhoea and stomach cramps. However. In fact the routes of trophic transfer of AZAs through the food chain ending in contaminated shellfish remain unknown.
but all contain a spirocyclic or cyclic imine function within their structure. 7. and closely related ptierotoxins (Figures 7. which are particularly amenable to detection by positive ion electrospray mass spectrometry. 91. The limited amounts of AZA toxins available continue to hinder a complete understanding of their mechanism of action. 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. abnormalities in the thymus and spleen.12). Currently LC/MS and LC-MS/MS analysis is the preferred chemical analytical detection method for AZAs. The mouse bioassay has been used to monitor shellfish extracts. 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).10 – 7. 85). and ASP.10 Cyclic imines These toxins are represented by several distinct structural types. and necrosis of epithelial cells on the microvilli (91). 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). Using the same approach. though the possibility exists that they may inhibit other phosphatases. However. but unlike DSP monitoring methods which use only the hepatopancreas of the shellfish for extraction. it is believed that the hydroxylated derivatives AZA4 (470 mg/kg) and AZA5 (<1000 mg/kg) are considerably less toxic. and erosion and bleeding in the stomach. the pinnatoxins. 92). Key fragment ions are useful in identifying the various AZA compounds that can occur in shellfish tissue (83. Injection of a lethal dose of AZA1 in mice (>150 µg/kg) caused vacuole formation and fatty acid accumulation in hepatocytes. This common cyclic imine feature is believed to be a common pharmacophore of this group of toxins which comprises the gymnodimines. Due to their relatively recent discovery and limited knowledge of their toxicological properties and impact on human 160 .9. the spirolides. although autopsies conducted after 4 and 8 hours revealed various gastrointestinal (GI) abnormalities including accumulation of fluid in the ileum. PSP. 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.3 Detection methods The regulatory level for AZAs in shellfish has been set by the EU at 160 µg AZA equivalents/kg shellfish tissue. Once again. 7. 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. AZAs also display cytotoxicity towards various mammalian cell lines (83. parenchymal cell pyknosis in the pancreas. Importantly these pathological outcomes were reported as being different from those caused by DSP.FISH AND SEAFOOD not inhibit the phosphatases PP1 and PP2.
FISH AND SHELLFISH TOXINS health. surf clam and New Zealand abalone. led to the identification of a new marine toxin with an unprecedented structure named gymnodimine (96) (Figure 7. scallops. it was proposed that a dinoflagellate identified as Gymnodinium spp. 7. 95). but they are included here for completeness.10.1 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.10. they are sometimes referred to as emerging toxins of uncertain human health concern. selliformis (97). 161 . Pacific oyster. Subsequent taxonomic studies revised this again to a new dinoflagellate K. 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 . At the time of the initial observation of gymnodimine.1. two additional congeners..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.520 Da (94. The structure is novel in that it contains a spirocylic imine group and a single tetrahydrofuran ring embedded within a macrocyclic structure. 7. cockles. 95). Structure of the cyclic imine gymnodimine 7. gymnodimine B and C were isolated from the dinoflagellate Karenia selliformis (formerly Gymnodinium selliformis). Some time later. In only two cases (gymnodimines and spirolides) has a planktonic source been identified. Fig.10). 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.10. Reviews of the current status of the cyclic imine group were published by Molgo and Cembella (94.
95) (Figure 7. and so spectroscopic detection by UV methods is not favoured.1.10. and lack of responsiveness to stimuli. A later study using purified toxin established an LD50 dose (Lethal Dose. They were originally discovered in extracts of mussels (M. 7. and short-acting cholinesterase inhibitors such as neostigmine or physostigmine protected mice against the toxic effects of gymnodimine.1. all or nothing” mouse bioassay response to gymnodimines and cyclic imines in general continues to be a useful monitoring approach.to10-fold reduction of toxicity (LD50 600 . Fractions are then dried and re-suspended in methanol or acetonitrile in preparation for LC/MS analysis (94. Most cyclic imines lack an extended chromophore.2 Toxicology A review of the toxicology of all cyclic imines has been published (98). 95).10. Since their original discovery which resulted in the characterisation of spirolides A-F (99. together with a lack of evidence of human intoxication. finishing with 100% methanol.10. Based on this diminished oral toxicity. However. This led to the suggestion that gymnodimine acts by blocking nicotinic acetylcholine receptors at the neuromuscular junction.2 Spirolides The spirolides comprise the largest group of cyclic imine compounds.100). 7. abdominal breathing.11).FISH AND SEAFOOD 7. The original observation of toxicity in New Zealand followed from IP injection of lipophilic extracts in the mouse bioassay.900 µg/kg). Oral administration of the toxin to mice by gavage resulted in about an 8. cyclic imines are readily observed by electrospray mass spectrometry in the positive ion mode. and this is the detection method of choice.3 Detection methods The characteristic “fast-acting. The time lapse between ingestion and death is between 5 and 15 minutes. In particular. paralysis of the hind quarters and eventually complete immobility. The structure of the spirolides bears a strong resemblance to both gymnodimine and 162 . 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. Reduction of the imine group eradicated the toxicity. There is a rapid onset of symptoms including respiratory distress. Whether isolated from dinoflagellate cells directly or from shellfish tissue. and there was a further ten-fold reduction in toxicity when the toxin was supplied with food. 50%) of 96 µg/kg. 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. edulis) and scallops (Placopectin magellanicus) from eastern Canada that displayed abnormal symptoms in the mouse bioassay. it has been stated that regulation against gymnodimine is not required (98).
707 µg/kg. R1 = CH3) of the cyclic imine ring are the most toxic of the group (98). While oral toxicity is considerably less than by IP injection.11.1 Origins and distribution At the time of the original discovery of the spirolides in shellfish from the coastal waters of eastern Nova Scotia. Structure of four spirolide toxins A-D 7. 7.11. these are the toxins that are also most resistant to acid hydrolysis of the imine function 163 . Since then the spirolides have been found to be globally distributed and are found in shellfish or plankton fractions along eastern Canada. the Bay of Biscay and most recently from Mediterranean shellfish and plankton in the Adriatic (102).2 Toxicology The spirolides are the most studied group of the cyclic imines. fasted mice). fasted mice). 7. They have also been reported in the southern hemisphere in Chilean mussels and in New Zealand shellfish and plankton (94.2. All data reported here are for desmethyl spirolide C. 95).8 µg/kg).10.FISH AND SHELLFISH TOXINS the pinnatoxins (vide infra).2. Fig. 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. southern Ireland. Interestingly. the gulf of Maine. This proved to be the case and eventually the dinoflagellate Alexandrium ostenfeldii was identified as the culprit organism (101). those spirolides possessing the vicinal dimethyl grouping (see Figure 7. by gavage (87 .10. their localisation in the digestive gland of mussels and scallops led to the suspicion that plankton were the real source of the toxins. the western coast of Scotland. The LD50 values vary considerably depending upon whether the compound is administered by IP injection (5 . or by oral ingestion (381 .166 µg/kg. in fjords in Denmark and Norway.
Once again this relies on the sensitivity of these cyclic imines to detection by positive ion electrospray mass spectrometry. 95. To date. The pteriatoxins were isolated from a different mollusc but bear a strong structural resemblance to the pinnatoxins (Figure 7.2. A number of methods following this procedure have been described (94. 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. While the mechanism of action remains to be clearly defined. However.FISH AND SEAFOOD (100). Three members of this group have been reported.10. The compounds are zwitterionic and therefore somewhat more polar than the spirolides.12.3 Pinnatoxins and pteriatoxins Pinnatoxins A .10. Structure of the pinnatoxin group and the closely related pteriatoxins 164 . 7. and in fact these compounds share a number of other structural features. Fig. 7. the mouse bioassay can be used to detect these fast acting toxins.3 Detection methods As discussed for gymnodimine.12). which includes two diastereomers. 7. consistent with modulation of acetyl cholinergic receptors (103).B contain a spiro imine moiety similar to that found in the spirolides. 102). experiments have shown upregulation of muscarinic and nicotinic receptors in mice following dosing with spirolide C. four members of the group have been identified – two of them being the C-34 diastereomers.
The LD99 (Lethal Dose.100 µg/kg). new toxin groups such as the azaspiracids and cyclic imines have also been reported.11 Conclusion There has been considerable progress in the detection of known marine toxins.3 Detection methods No formal detection methods for these compounds have been reported. to date action or 165 .10.2 Toxicology There is little information describing the toxicology of these compounds (98). 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. there is no compelling evidence to suggest that they have resulted in human toxicity (98). With increasing knowledge of the chemistry of toxins. Only the acute toxicity in mice following IP injection has been reported. but the mouse bioassay data and LC-MS can be used to monitor suspect seafood for these compounds.180 µg/kg).3. while pinnatoxin D was essentially inactive. arise from an algal source . The LD99 dose for the pteriatoxins is reported to be in the range (8 . 7. Despite this progress. 7. has come the realisation that a “toxin profile” can be a complex mixture of many different classes of toxins. There is strong circumstantial evidence to suspect that these toxins. This prompted a chemical investigation of Pinna muricata that ultimately led to the identification of the pinnatoxins in 1995 (104). have long been suspected of causing human illness after ingestion. like the gymnodimines and spirolides. and in some cases it is known that one toxin can potentiate or augment the toxicity of the other.1 Origins and distribution Shellfish of the genus Pinna. Coupled with this. but this remains to be conclusively established (106). This is generally a result of increased monitoring and continually improving detection technologies and methods. and more advanced means of detection. and later the closely related pteriatoxins were isolated from the digestive glands of another Okinawan bivalve Pteria penguin (105). However. research and certified standards of the key toxins are often unavailable or in short supply and in the future considerable logistical effort will be required to address this issue.3. 99%) values for pinnatoxins A-C are in the range (22 .10.most likely a dinoflagellate. which occupy tropical waters around Okinawa and the south east coast of China.FISH AND SHELLFISH TOXINS 7. which in many cases are now found to be more widespread than previously believed.10. 7.
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Boca Raton. Bourdelais A.M. Schrank K. Lenzi A. 71. Prockop L. Manzo L. Buonocore M.. Fernandez M..... Baden D..S. in FAO Food and Nutrition Paper 80. 2002. Pierce R. 2008. Ed.. 551-8. Flewelling L.P. Bottein M-Y.E. 2004. Dickey R. Bourdelais A... Environmental Health Perspectives. Boca Raton. Gannon D. Botana L.. CRC Press. Competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater. 2004. Abraham W. Whitney P.E.G.. Neurologic Shellfish Poisoning (NSP). Wang Z. Naar J. are potent ichthyotoxic neurotoxins that accumulate in fish. 50.S. CRC Press.. Tomas C. 1598-603. like ciguatoxins. Jacocks H.. Baden D. Poli M.. New fish-killing alga in coastal Delaware produces neurotoxins..W. Ed. 2008. Naar J... Weisman R. Pierce R... Kubanek J. Dickey R. Lancaster J.FISH AND SHELLFISH TOXINS 65. Henry M. 117-71.G.E. Locatelli C. 187-94.G. 131.. Wolny J.. Pierce R.N. Tomas C. Marine Drugs. Muscle Nerve. 707-23. Watkins S..A. 110. 2007.... in Seafood and Freshwater Toxins. Dechraoui M-Y.. Butera R..G. 110. Botana L...J.. Ayyar R. Zaias J. Chest..D. West Indian Medical Journal...H. Gandini C... in Seafood and Freshwater Toxins. Baden D.C. Kirkpatrick B. Abraham W. Food and Agriculture Organisation.P. Flewelling L. 479-516.. CRC Press.M. Brevetoxins. Naar J.D. Bean J. Wanner A... 2008. Granholm A. Ed.... Reich A. Backer L.E. Fleming L.. Toxicon. Kubanek J. in Seafood and Freshwater Toxins.E..C.A.S. Bienfang P.. 74. Fleming L. 456-79. 76..S. 2004. Poon-King T. Backer L. Ciguatera toxins: Chemistry. Botana L.. 2000. Literature review of Florida red tide: implications for human health. Rome. Baden D. 171 . Abbott J.M. Toxicon. shellfish and mammalian body fluid. 2007. 68.M. Marine Biotoxins.. Ramsdell J. Benson J. Ciguatera fish poisoning in industrial ship crew members: a retrospective study in a seaport general practice in Trinidad and Tobago. Granade R. 67. Detection of brevetoxins in the twenty-first century. Kirkpatrick B. Mild ciguatera poisoning: case reports with neurophysiological evaluations. 73. 69. Harmful Algae.. Zaias J. Food and Agriculture Organisation of the United Nations. Boca Raton. Backer L. 179-85. 75.G..P.R.A.J.. Reich A. 220-6. 23. Fleming L. Naar J. 6... 77. Baden D.. 2008. toxicology. Intrinsic potency for synthetically prepared brevetoxin cysteine metabolites BTX-B2 and desoxy-BTX-B2.A. 53. 3. Landsberg J...B. Ed. Freidman M. 70. Steidinger K. Environmental Health Perspectives. Cheng Y. 465-70.. Aerosolised red tide toxins (Brevetoxins) and asthma. Poon-King C. Bossart G. 99-115. 72... Clark R. Johnson D.. and detection. 2002.. The molecular and integrative basis to brevetoxin toxicity..M.. Ramsdell J. 66.. 50. 519-50.. Squicciarini D. 825-34.. Ciguatera fish poisoning: treatment prevention and management.. Food and Agriculture Organisation.. 2007. Chen A. 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.
3. 2. procedure.3 Stages of a HACCP Study The HACCP system consists of the following seven basic principles: 1. Establish procedures for verification to confirm that the HACCP system is working effectively. 4. hygiene managers. food microbiology. Each stage is discussed below in detail. Determine the CCPs.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Step . Establish documentation concerning all procedures and records appropriate to these principles and their application.A point. food chemistry or engineering. 6. from primary production to final consumption. Establish the critical limit(s). procedures. Establish a system to monitor control of the CCP. An effective HACCP plan is best carried out as a multidisciplinary team exercise to ensure that the appropriate product-specific expertise is available. operation or stage in the food chain including raw materials.1 is a flow diagram illustrating this 12-stage logic sequence. Establish the corrective action to be taken when monitoring shows that a CCP is not under control.3.The application of methods. 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. 7. Figure 8. Verification . tests and other evaluations. Conduct a hazard analysis. The team should also include 177 . 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.Obtaining evidence that the elements of the HACCP plan are effective. 5.1 Assemble HACCP team HACCP requires management commitment of resources to the process. in addition to monitoring to determine compliance with the HACCP plan. ingredient and packaging buyers. quality assurance or control. 8. Validation . 8.
1. Fig. biological. 8.FISH AND SEAFOOD personnel who are involved with the variability and limitations of the operations.g. 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. it may be obtained from external sources. chemical and/or physical). Logic sequence for application of HACCP 178 . If expert advice is not available on-site.
HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE 8.3. is a cooking process required?). marine toxins associated with particular fish and shellfish species. may need to be considered specifically.g. 8. physical and chemical structure (including water activity (aw). 8. it must be noted here that certain toxins (algal and shellfish) are not destroyed by heat treatments. fermentation. However. naturally present on fish from warm waters (> 15 °C). shelf life. there may be species-specific hazards related to certain fish and shellfish.1.1 Fish and seafood The HACCP team should ideally have access to expertise on the appropriate fisheries.g. storage and distribution conditions and instructions for use. including practices onboard fishing vessels.2 Describe the product It is important to have a complete understanding of the product. or in aquaculture operations. It is also important to identify the consumer target groups.3.3. such as children or the elderly. In addition. 8.). 8.4 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. aquaculture drug residues and potentially pathogenic Vibrio spp.3. processing conditions (e. etc. It should provide an accurate representation of all steps in the production process from raw materials to the end179 . heat treatment. pH. contaminated fish and shellfish should not be used. such as enteric viruses in shellfish. challenge testing and inoculation studies for evaluation of safety aspects are therefore essential for an effective HACCP team. If the processing does not include a kill step. Expertise on such inherent hazards. composition. for example contamination by foodborne pathogens as a result of time/temperature abuse or unsatisfactory handling practice. the only CCP that can render the product safe from pathogenic organisms is adequate heat treatment during preparation. Vulnerable groups of the population.).3.1 Fish and seafood Certain products may be contaminated or carry pathogenic organisms as a part of the natural flora. etc. These factors are likely to have a significant effect on some potential hazards.3 Identify intended use The intended use should be based on the expected uses of the product by the enduser or consumer (e. packaging.3. The description should include information such as the product name. which should be described in detail. freezing.
3. It is also important to include observation of production outside normal working hours such as shift patterns and weekend working. once determined. it should also include details of CCP steps.6 List all potential hazards associated with each step. Many fish-processing operations have relatively few steps and the flow diagrams appear simple. The operation should be observed at all stages. time/temperature data. It is essential that the diagram is accurate. 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. 8. For example.3. or circumstances of any reclaim or rework activity. features of equipment design. cleaning and hygiene procedures and storage conditions. because the hazard analysis and decisions regarding CCPs are based on these data. 8. conduct a hazard analysis.4. the qualitative and/or quantitative evaluation of the presence of hazards. Ideally. it is essential that the details of each step are fully appreciated and recorded. then any pre-drawn HACCP plans must be reviewed once the lines/products are finalised.1 Fish and seafood Examples of flow diagrams for specific fish products may be found in the appropriate product chapters. It may include details of the factory and equipment layout. However.FISH AND SEAFOOD product.3. 180 ● ● . 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. survival or multiplication of pathogenic microorganisms. ingredient specifications. and any discrepancies between the flow diagram and normal practice must be recorded and the diagram amended accordingly. 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. The analysis is likely to include consideration of: ● the likely occurrence of hazards and the severity of their adverse health effects. and therefore a detailed description of the procedure applied must be available. If HACCP studies are applied to proposed new process lines or products.
and there are an equally varied range of potential hazards associated with them. the hurdle effect.3. any food safety objectives or manufacturer’s food safety requirements.1 Fish and seafood The term 'fish and seafood' includes an extremely varied group of products. and the potential growth of Listeria monocytogenes on smoked fish. Therefore it is not possible. Vibrio spp. or desirable. The HACCP team should then determine what control measures exist that can be applied for each hazard. Pathogens may be part of the resident microflora of the living animal (e. the number of consumers potentially exposed and their vulnerability.). Many of the microbiological hazards associated with fish and seafood products are derived from the raw materials. 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. The incidence of psychrotrophic. to generalise about expected hazards and the reader is once again referred to the appropriate product chapter for additional advice on specific hazards. 181 . botulinum. non-proteolytic Clostridium botulinum on fresh fish is also sufficiently high that its presence may be assumed. modified-atmosphere packaging (MAP) of chilled raw fish may provide conditions suitable for the growth of psychrotrophic C. Note: it is important at this stage that no attempt is made to identify CCPs. For example. or poor control of batter mixes in frozen battered fish portions may allow growth of Staphylococcus aureus and production of enterotoxin.6. Salmonella spp.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE ● ● ● ● production or persistence of toxins. Hazards introduced during processing of fish products depend very much on the characteristics of the process. or may originate from polluted water or from post-capture contamination (e. there are particular hazards associated with contamination of shellfish by microorganisms from human sewage. One control measure may be relevant to several process steps. and viruses). For example. where a hazard is repeated. 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.g. since this may interfere with the analysis. 8. Hazards specific to certain types of product are detailed in the appropriate chapters of this manual.
8 Careful selection of sources for raw materials Adequate temperature control Effective sanitation Food handler hygiene Prevention of cross-contamination Establish critical limits for each CCP Critical limits separate acceptable from unacceptable products. it is necessary to specify both time and temperature for a thermal process.1 Fish and seafood Again.7. For example. However.3. critical limits should be specified and validated for each CCP. given the enormous variety of fish products and processes in use.3. it is not possible to generalise on likely CCPs. structured approach to decision making.1 Fish and seafood Specific product chapters provide information on criteria that may be used to set critical limits. Criteria used to set critical limits must be measurable and may include physical. application of the decision tree should be flexible. More than one critical limit may be defined for a single step. 8. then the process must be modified to include a control measure.7 Determine CCPs The determination of CCPs in the HACCP system is facilitated by a decision tree (Figure 8. It is prudent to set stricter limits (often-called target or process limits/levels) to ensure that any trend towards a loss of control is noted before the critical limit is exceeded. it can be said that effective control measures are likely to include the following: • • • • • 8.3. However.3.8. It is also essential that the HACCP team has access to sufficient technical data to determine the CCPs effectively. and its use may not always be appropriate. biological or sensory parameters.2) to provide a logical. Where possible. and the reader is referred to the appropriate product chapter. 8. Some examples relevant to fish products are: • • • Time and temperature limits for pasteurised and cooked products Temperature limits for chilled fish Brine concentration in cured fish 182 . If a significant hazard has been identified at a step where control is required for safety. chemical. but for which no control exists at that step or any other.FISH AND SEAFOOD 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.
H.HACCP IN FISH AND SEAFOOD PRODUCT MANUFACTURE Huss H. 14-9. Blackwell Scientific Publications. Bryan F. Volume 4: Application of the Hazard Analysis Critical Control Point (HACCP) System to Ensure Microbiological Safety and Quality. 33-44. 1992. 15 (1/2). International Journal of Food Microbiology. 188 . Development and use of the HACCP concept in fish processing. Oxford. 1992. Geneva. Mayes T. 1992. International Commission on Microbiological Specifications for Foods. 3 (1). WHO. Hazard Analysis Critical Control Point Evaluations: A Guide to Identifying Hazards and Assessing Risks Associated with Food Preparation and Storage. Food Control. Simple users' guide to the hazard analysis critical control point concept for the control of food microbiological safety.L.. World Health Organisation. Microorganisms in Foods. 1988.
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. eliminate or acceptably control pathogen contamination of food. and one that plays an important role in most countries’ food legislation. and conditions of raw materials. Microbiological standards have a useful role and help establish requirements for the microbiological safety and quality of food and raw materials. to personnel requirements. from structural conditions in the factory or process facility. including microbiological criteria. Hygiene is a general concept that covers a wide subject area. to sale or supply to the final consumer. A number of standards are provided in food legislation. A new package of European Commission (EC) hygiene measures became applicable on 1 January 2006 to update and consolidate the earlier 17 hygiene directives. 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. Procedures under the new legislation are based on risk assessment and management and follow a 'farm to fork' approach to food 189 . EC FOOD HYGIENE LEGISLATION Eugenia Choi and Dr.1 Introduction Hygiene is an important aspect of ensuring food safety. The new legislation was designed to establish conditions. have been replaced by several linked measures on food safety rules and associated animal health controls. Jenny Pfleger Leatherhead Food International Randalls Road Leatherhead Surrey KT22 7RY United Kingdom 9. transport and delivery vehicles requirements. with the intention of introducing consistency and clarity throughout the food production chain from primary production. final product specifications.9. however. under which food is produced to optimise public health and to prevent. the existence of microbiological standards alone cannot protect consumer health.
and sets out what those enforcing the provisions have to do. as last amended by Regulation (EC) No 806/2003. 2073/2005 on microbiological criteria for foodstuffs • The general hygiene requirements for all food business operators are laid down in Regulation 852/2004. European Communities L 268.FISH AND SEAFOOD safety with the inclusion of primary production in food hygiene legislation. storing. p. manufacturing. as last amended by Regulation (EC) No 806/2003. 190 • • . J. Regulation 854/2004 relates to the organisation of official controls on products of animal origin. 9. the following European Union (EU) hygiene regulations have applied: • • • Regulation (EC) No.B.1992. handling and offering for sale or supply to the consumer. Regulation 853/2004 supplements Regulation 852/2004 in that it lays down specific requirements for food businesses dealing with foods of animal origin. Council Directive 92/48/EEC of 16 June 1992 laying down the minimum hygiene rules applicable to fishery products caught on board certain vessels in accordance with Article 3(1)(a)(i) of Directive 91/493/EEC (OJ L 187. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin Regulation (EC) No. 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs Regulation (EC) No. Subsequently. 24.1991. N.9.7. packaging.9. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption Regulation (EC) No. 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. 41).2 Legislative Structure From 1 January 2006. processing. p.1991. transportation. existing hygiene Directives including those below were repealed: • Council Directive 91/492/EEC of 15 July 1991 laying down the health conditions for the production and the placing on the market of live bivalve molluscs (Off. 7. p.1). distribution.15). A number of more detailed implementing and transitional measures have been adopted at EC level. 24. Prescribed are detailed measures to ensure the safety and wholesomeness of food during preparation.
There are some exemptions. Wales and Northern Ireland). The national legislation in all four UK countries also applied the provisions of the EU Microbiological Criteria Regulation No. 9. for example. 852/2004 on the General Hygiene of Foodstuffs Food business operators must ensure that all stages of production.7.g.LEGISLATION • Council Directive 93/43/EEC of 14 June 1993 on the hygiene of foodstuffs (OJ L 175. and of hunting and fishing. 852/2004.1993.) in England. 19. national legislation in the form of a Statutory Instrument (S. etc. p. right through to the implementation of procedures based on HACCP principles. As regulations. is required to give effect to the EU regulations. e. The regulation lays down general hygiene provisions for which food business operators carrying out primary production must comply with as laid down in Part A of Annex 1.I. revocation of existing implementing legislation.14. Likewise Regulation 852/2004 will not apply to collection centres and tanneries meeting the definition of food business because they handle raw material for the production of gelatine or collagen. and also if the producer supplies small amounts of primary product to the final consumer or local retail establishments supplying the final consumer. 1 . with primary production. 2006 No. Additionally the requirements of EC Regulation 853/2004 must be complied with. Wales and Northern Ireland. food storage that is for private or domestic consumption. 2073/2005. 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. Although the regulations have the force of law. setting offences. This Regulation lays down general rules for food business operators on the hygiene of foodstuffs. Primary products are defined as products of primary production including products of the soil.I.11) The EU hygiene regulations apply to all stages of food production including primary production. penalties and powers of entry.3 Regulation (EC) No. the legislation is directly applicable law. The Food Hygiene (England) Regulations 2006 (S. of stock farming. and binding in its entirety on all member states from the date of entry into force. as amended) came into force on 11 January 2006 (separate but similar national legislation also came into force on that day in Scotland. domestic preparation or handling. which will be covered later in this chapter. and equivalent legislation in Scotland. processing and distribution of food under their control satisfy the relevant hygiene requirements laid down in Regulation (EC) No. particularly taking into account a number of factors ranging from ensuring food safety throughout the food chain to begin with primary production. 191 .
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Annex I – Primary production
Annex I (Part A) relates to general hygiene provisions for primary production and associated operations covering: (a) the transport, storage and handling of primary products at the place of production, (b) the transport of live animals, (c) for fishery products - transport operations to deliver primary products (which have not been substantially altered) from the place of production to an establishment Food business operators have the responsibility to ensure primary products are protected against contamination. Any community and national legislation relating to the control of hazards in primary production such as measures to control contamination resulting from surroundings e.g. air, soil, water etc., and measures relating to animal health and welfare, and plant health that may impact on human health should be complied with. Requirements for record keeping are also laid down. This relates to animal feed (nature and origin), veterinary medicines administered to animals (date given and withdrawal periods), any diseases, analysis of samples from other animals that might impact on human health as well as reports on animal checks performed. Part B of Annex I contains recommendations for guides to good hygiene practice. 9.3.2 Annex II – Stages other than primary production
Annex II of the regulation lays down additional general hygiene requirements that must be met by food business operators carrying out production, processing and distribution of food at all stages following primary production. A summary of Chapters I to IV of Annex II is provided as follows: 22.214.171.124 Chapter I Chapter I applies to all food premises, except premises to which Chapter III applies. Food premises must be kept clean and maintained in good repair and condition. Their layout should allow for this. The environment should allow good hygiene practices and provide temperature-controlled handling and storage conditions where necessary ,and enable foods to be kept at correct temperatures and be monitored.
Additionally, there are requirements for provision of adequate lavatories, basins, ventilation, lighting and drainage.
126.96.36.199 Chapter II Chapter II applies to all rooms where food is prepared, treated or processed, except dining areas and premises to which Chapter III applies. The design and layout of rooms should allow for good hygiene practices between and during operations. Therefore floor and wall surfaces, ceilings and windows should be constructed to prevent dirt accumulating. Surfaces where food is handled must be well-maintained and allow easy cleaning and disinfection, preferably using smooth, washable corrosionresistant and non-toxic materials. There should be facilities for cleaning or disinfecting, and for storing working utensils or equipment. Clean potable water and adequate provision for washing food is needed.
188.8.131.52 Chapter III Chapter III applies to temporary premises (e.g. marquees, market stalls, and mobile sales vehicles), premises used primarily as a private dwelling-house but where foods are regularly prepared for placing on the market, and vending machines. Premises and vending machines should be practically sited, designed, constructed and kept clean and maintained in good repair and condition so as to avoid the risk of contamination, in particular by animals and pests. Facilities should allow adequate personal hygiene, and surfaces in contact with food should be easy to clean. Enough potable water and storage arrangements for hazardous or inedible substances are required as well as adherence to food safety requirements.
184.108.40.206 Chapter IV Chapter IV applies to all transportation. Conveyances and/or containers used for transporting foodstuffs are to be kept clean and maintained in good repair and condition to protect foodstuffs from contamination and are, where necessary, to be designed and constructed to permit adequate cleaning and/or disinfection. Food should be maintained at appropriate temperatures.
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220.127.116.11 Chapter V Chapter V refers to equipment requirements. Adequate cleaning and disinfection is to be carried out frequently for articles, fittings and equipment in contact with food where contamination needs to be avoided. Equipment should be installed to allow adequate cleaning, and be fitted with the required control device.
18.104.22.168 Chapter VI Chapter VI refers to food waste. Food waste, non-edible by-products and other refuse is to be removed from rooms where food is present as quickly as possible to avoid accumulation. Such waste is to be deposited in closable containers, to allow easy cleaning. Refuse stores should allow easy cleaning and be free of pests. Waste must be eliminated hygienically in accordance with European Community legislation.
22.214.171.124 Chapter VII Chapter VII refers to the water supply. There must be an adequate supply of potable water, which should be used whenever necessary to ensure foodstuffs are not contaminated. There are also requirements for recycled water, ice in contact with food, use of steam and for the water used in the cooling process for heat-treated foods in hermetically sealed containers. Specific requirements are that clean water may be used with whole fishery products. Clean seawater, may be used for live bivalve molluscs, echinoderms, tunicates and marine gastropods; and for external washing. If it is used, there must be adequate facilities to ensure the supply does not create a source of contamination for the foodstuff. Recycled water used in processing or as an ingredient must not present a contamination risk. It must be of the same standard as potable water, unless the competent authority is satisfied that the quality of water cannot affect the wholesomeness of food in the finished form.
Additionally, ice that comes into contact with food or which could possibly contaminate it needs to be made from potable water or, in the case of chilling whole fishery products using clean water. Ice must be made, handled and stored under conditions that protect it from contamination.
126.96.36.199 Chapter VIII Chapter VIII sets out personal hygiene requirements for those working in a food handling area including clean protective clothing and the requirement that those carrying or suffering from a disease are not permitted to handle food. 188.8.131.52 Chapter IX Chapter IX covers provisions applicable to foodstuffs. A food business operator should not accept raw materials or ingredients, other than live animals, or any other material used in processing products, if they are known to be contaminated with parasites, pathogenic microorganisms or foreign substances, to be toxic or decomposed to such an extent that, even after the business operator applied normal hygienic processing , the product would be inedible. Raw materials must be kept under appropriate conditions throughout production, processing and distribution. In particular, temperature control (i.e. cold chain and food thawing) requirements are laid down.
184.108.40.206 Chapter X Chapter X lays down provisions applicable to the wrapping and packaging of foodstuffs to avoid contamination of any form. 220.127.116.11 Chapter XI Chapter XI lays down heat treatment requirements for food that is placed on the market in hermetically sealed containers. The main relevant process parameters (particularly temperature, pressure, sealing and microbiology) must be checked by the food business operator to ensure that the required heat treatment has been achieved. Also, the process used should comply with an internationally recognised standard (e.g. pasteurisation, ultra high temperature or sterilisation). 18.104.22.168 Chapter XII Chapter XII states training requirements for food business operators to ensure that food handlers are trained in food hygiene matters and in the application of HACCP principles.
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The Regulation requires that food business operators must notify the relevant competent authority of each establishment under their control that carries out any of the stages of production, processing and distribution of food, with a view to the registration of each establishment. Food business operators must also ensure that the competent authority always has up-to-date information on establishments, including the notification of significant changes in activity, and closure of an existing establishment. Food business operators must ensure that establishments are approved by the competent authority, following at least one on-site visit, when approval is required by the national law of the Member State in which the establishment is located, or under Regulation (EC) No. 853/2004, or by a separate decision adopted. Separate rules apply for businesses producing products of animal origin. 9.3.4 HACCP
Food business operators, other than at the level of primary production, and associated operations must put in place, implement and maintain a permanent procedure or procedures based on principles of the system of hazard analysis and critical control points (HACCP). Emphasis is placed on risk-related control, with responsibility placed on the proprietor of the food business to ensure that potential hazards are identified and systems are developed to control them. Under HACCP, food business operators must amongst other considerations, identify hazards to be prevented or eliminated or reduced to acceptable levels, identify critical control points (CCP) at which control is essential to prevent, eliminate or reduce hazards and establish critical limits at these points, implement effective monitoring procedures, and establish corrective actions in the case where a CCP is out of control. Procedures must be followed to confirm the above is in place and up-todate, as well as providing the competent authority with documents and records as evidence, when required. 9.4 Regulation (EC) No. 853/2004 Laying Down Specific Hygiene Rules for Food of Animal Origin
Regulation (EC) No. 853/2004 lays down hygiene rules for products of animal origin that apply in addition to the general hygiene rules of Regulation (EC) No. 852/2004. 9.4.1 Definitions
The following definitions apply for fishery products and live bivalve molluscs:
shucking. filleting. heading. gutting and removal of fins and. where appropriate. 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. to remove mud. where necessary. Factory vessel means any vessel on board which fishery products undergo one or more of the following operations followed by wrapping or packaging and. skinning. Freezer vessel means any vessel on board in which freezing of fishery products is carried out. including products packaged under vacuum or in a modifiedatmosphere that have not undergone any treatment to ensure preservation other than chilling. after preparatory work such as bleeding. Fresh fishery products means unprocessed fishery products. slicing. Bivalve molluscs means filter-feeding lamellibranch molluscs Marine biotoxins means poisonous substances accumulated by bivalve molluscs. or in natural sites. if necessary: chilling or freezing: filleting. reptiles and frogs) whether wild or farmed and including all edible forms. purification centres or dispatch centres in tanks or any other installation containing clean seawater. 197 . parts and products of such animals. shelling. slime or sand. to preserve or to improve organoleptic qualities and to ensure they are in a good state of vitality before wrapping or packing. such as gutting. and chopping. mincing or processing. followed by wrapping or packaging. live echinoderms. 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. in particular as a result of feeding on plankton containing toxins. whether whole or prepared.LEGISLATION Fishery products means all seawater or freshwater animals (except for live bivalve molluscs. and all mammals. heading. Conditioning means the storage of live bivalve molluscs coming from class A production areas. slicing. Prepared fishery products means unprocessed fishery products that have undergone an operation affecting their anatomical wholeness. live tunicates and live marine gastropods.
the code would be UK. EO. for other codes.2. EG.2 Fishery product requirements 9. EK.4. (ii) The mark must indicate the name of the country in which the establishment is located and written out in full or shown as a two-letter code in accordance with the relevant ISO standard. EY. ES. EC.2. a new mark must be applied to the product. EÜ.1 Application of the identification mark (i) The identification mark must be applied before the product leaves the establishment of production. the new mark must show the approval number of the establishment where this occurred.1. EF.4.4. 198 .2.2 Form of the identification mark (i) The mark must be legible and indelible. Extracts of the requirements of Annex II. In this case. 9. (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.1. 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.FISH AND SEAFOOD 9.1 Identification marking An identification mark needs to be applied following the requirements of Annex II. Section I of Regulation (EC) 853/2004 are given as follows: 9. (ii) However. EB or WE. (iv) If the identification mark is applied in an establishment within the Community. when a product's packaging and/or wrapping is removed or is further processed in another establishment. For the UK. the mark must be oval shaped and include the abbreviation CE. If an establishment manufactures both food to which Regulation 853/2004 applies and food to which it does not. the food business operator may apply the same identification mark to both types of food. please refer to Regulation (EC) 853/2004.4. the mark must show the approval number of the establishment. EK. and the characters easily decipherable and be clearly displayed. Section I of Regulation (EC) 853/2004. (iii) Additionally.
wrapping or packaging in another establishment. the approval number of the establishment and the abbreviation for the community where appropriate. (iii) In the case of liquid. 9.2. Part A and Chapter V apply to retail.2. reference should be made to the actual Regulation.3 Method of marking (i) The mark is to be applied directly to the product. Chapter IV. they would need to be obtained following Section VII on live bivalve molluscs. C and D.2 Hygiene requirements for fishery products The regulation details specific hygiene requirements for fishery products. an identification mark is not necessary if accompanying documentation contains the name of the country or its two-letter code. including vessels.4. tunicates and marine gastropods when placed on the market live. It may also be a tag made of a resistant material affixed in such a way that it cannot be removed. its wrapping or the packaging. it applies to those animals when not placed on the market live. the requirements accompany those of Annex II of Regulation (EC) 852/2004 199 . With the exception of Chapters I and II. echinoderms.1. the colourings used must be EC authorised colours. i. Parts A. processing. the mark may be applied to the outer surface of the container or packaging. for full requirements.e. 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 wrapping or the packaging. The requirements of this section accompany those in Regulation (EC) 852/2004. as amended. and are intended for further handling.LEGISLATION 9. or be printed on a label affixed to the product. This section does not apply to bivalve molluscs. it is sufficient to apply the mark to the exterior of that package only. Chapter III. Extracts of the requirements of Annex III. and fishery products carried in bulk. specifically relating to fishery products are given below.4. a) requirements for establishments. granulate and powdered products of animal origin carried in bulk. (iv) If products of animal origin are placed in a package destined for direct supply to the final consumer. Therefore. Section VIII of Regulation 853/2004. (ii) For products of animal origin that are placed in transport containers or large packages. (v) When the mark is applied directly to products of animal origin.
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. clean seawater may be used for the handling and washing of fishery products. i. they also include (i) the transport and storage of fishery products that have not been substantially altered (including live fishery products. heading. Structural and equipment requirements refer to the following points 1.. the production of ice used to chill fishery products and the rapid cooling of crustaceans and molluscs after their cooking. it must be placed somewhere that will not contaminate the water supply. or to handle or process them after harvesting. within fish farms on land) (ii) the transport of fishery products that have not been substantially altered (including live fishery products. this supplements the requirements of Annex II. The definition of fishery products covers primary production as well as its associated operations. if a vessel has water coming in that is used with fishery products.) 9.4. fishing and collection of live fishery products with a view to their being placed on the market. There is a derogation from point a. in that record keeping requirements stated by point 7 in Part A of Annex I of Regulation 852/2004 do not apply to operators involved in small-scale coastal fishing (as defined through Regulation (EC) 1198/2006) if the activity is under 24 hours. 2. Chapter VII of Regulation 852/2004. A. (a) primary production covers the farming. Requirements for all vessels Vessels should be made in a way that does not cause contamination with substances such as fuel and oil etc. and (b) related operations include any of the following operations if done on board fishing vessels: slaughter.1 Chapter I . surfaces that contact fishery products need to be made from cleanable corrosion-resistant material.e. Requirements for vessels designed and equipped to preserve fresh fishery products for more than 24 hours 200 .2.2. from the place of production to the first establishment of destination.FISH AND SEAFOOD c) for the water supply. gutting. removing fins. bleeding. refrigeration and wrapping.
(ii) Refrigeration equipment needs to keep fishery products in storage holds at not more than -18 °C. where necessary. tanks or containers for the storage of fishery products at the temperatures laid down in Chapter VII. (iii) In vessels equipped for chilling fishery products in cooled clean seawater. Holds and containers used for storing fishery products must allow its preservation under adequate hygiene conditions and. Requirements for factory vessels a) Factory vessels (i) These must have at a receiving area reserved for taking fishery products on board so each catch can be separated (ii) There must be a hygienic system for conveying fishery products from the receiving area to the work area (iii) Large work areas are needed to allow fishery products to be prepared and processed hygienically. and allow the monitoring and. Also. Requirements for freezer vessels (i) These need to contain freezing equipment in order to achieve a core temperature of no more than -18 °C. and not more than 0 °C after 16 hours. (v) Special equipment for disposing of waste/ inedible fishery products into the sea or a watertight tank and allocation of separate areas. (ii) Holds must be separated from the engine compartments and from the crew quarters by partitioning that prevent any contamination of the stored fishery products. Storage areas for the finished products also need to be sufficiently large and cleanable. Such devices must achieve a chilling rate that ensures that the mix of fish and clean seawater does not exceed 3 °C six hours after loading. to ensure that melt water does not stay in contact with the products. any waste processing unit needs to have a separate hold to store the waste. 201 .LEGISLATION (i) Such vessels need to have holds. tanks must incorporate devices that allow a uniform temperature throughout the tanks. where necessary. (iv) There needs to be somewhere for storing packaging materials and this is to be separate from product preparation and processing areas. (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. recording of temperatures. A temperature recording device must be present and placed where the temperature in the hold is the highest. 4.
must be chilled or. Livers and roes for human consumption must be preserved under ice (temperature to approach that of melting ice) or be frozen. b) Fishery products are to be handled and stored in a way that prevents the flesh bruising. c) Fishery products. 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. and products washed thoroughly. 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. Handlers may use spiked instruments to move large fish or fish that might injure them. b) Factory vessels on board which crustaceans and molluscs are cooked. where fishery products are displayed for sale.2 (i) Unloading and landing equipment that comes into contact with fishery products needs to be made of material that is easily cleanable. if this is not possible. landed as soon as possible.Requirements during and after landing 9.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.2.2. Chapter II . c) Factory vessels that freeze fishery products must have equipment meeting the requirements for freezer vessels as stated above. Parts that may present a danger to public health are to be removed quickly and kept away from products for human consumption. (ii) In the case of food business operators responsible for auction and wholesale markets etc. d) Any gutting or heading of fish on board is to be done hygienically and quickly after capture. the product must be kept at no more than -9 °C.4. provided no damage occurs to the flesh. e) In cases where whole fish intended for canning is frozen in brine. chilled and wrapped. do not have to meet the above requirements if no other form of handling or processing takes place on board such vessels. The brine must not be a source of contamination. other than those kept alive. B. 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 .
2. B. points 1 and 2. there are requirements that such premises should be well lit and vehicles emitting fumes should not be able to access the premises. and where necessary. C.LEGISLATION consumption. Requirements for frozen products An establishment on land that freezes fishery products needs to have equipment that meets that for freezer vessels as stated in Section VIII. Packaged fresh fishery products must be chilled to a temperature approaching that of melting ice. 9. (iii) Whole and gutted fresh fishery products can be transported and stored in cooled water on board vessels. part I. They should be wrapped. (ii) Heading and gutting must be carried out hygienically.3 Chapter III . Chapter I.2. Additionally. prepared or processed immediately after arrival at an establishment on land. They may also continue to be transported in cooled water after landing and be transported from aquaculture establishments until they reach the first establishment on land that carries out an activity other than transport or sorting. (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. Fillets and slices must then be removed from work tables as soon as possible after preparation. dispatched. handling fishery products A. C. 854/2004.Requirements for establishments. packaged and chilled as quickly as possible after preparation. Food business operators need to co-operate with the relevant competent authorities so as to allow them to carry out official controls as laid down in Regulation (EC) No. Requirements for mechanically separated fishery products (i) Raw materials used must satisfy the following 203 . (iv) Filleting and cutting is to be carried out in a way that avoids contamination or spoilage of fillets or slices.4. Gutting should ideally be done quickly once products have been caught or landed and afterwards be washed thoroughly. Requirements for fresh fishery products (i) Where chilled or unpackaged products are not distributed. including vessels. they must be stored under ice in appropriate facilities.
4.2. D. and (c) marinated and/or salted fishery products. they must be cooled rapidly. (iii) A document from the manufacturer. and whole fish must be gutted and washed beforehand. (iv) (wild) Atlantic and Pacific salmon. (ii) Compliance with the above is not necessary if epidemiological data indicates that the fishing grounds of origin do not present a health hazard in relation to the presence of parasites and this has been authorised by the competent authority. except when supplied to the final consumer. mechanically separated fishery products must be frozen as quickly as possible or incorporated into a product intended for freezing. 9.20 °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. (b) fishery products from the following species. stating the type of process they have undergone. 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. Requirements for cooking of crustaceans and molluscs After crustaceans and molluscs are cooked. must accompany fishery products referred to above when placed on the market. or a stabilising treatment. raw materials must have no guts.4 Chapter IV .Requirements for certain processed fishery products A. Requirements concerning parasites (i) The fishery products below must be frozen at no more than . (ii) The manufacturing process must satisfy the following Mechanical separation is to be carried out immediately after filleting. If no other method of preservation is used. cooling must continue until a temperature 204 . if the processing is insufficient to destroy nematode larvae. (iii) sprat. Once produced. (ii) mackerel.FISH AND SEAFOOD Only whole fish and bones after filleting may be used to produce mechanically separated fishery products.2.
the following conditions need to be met: A. 2074/2005. 205 . processing. cooked products are to be frozen or chilled to the temperatures stated in Chapter VII. 2074/2005). 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). Requirements for fish oil intended for human consumption (i) In relation to the raw materials used in their preparation and their transport. raw materials need to be chilled as soon as possible and should meet the temperature requirements 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. 2073/2005. separation. 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.LEGISLATION approaching that of melting ice is reached. C. However.2. 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. pressing. and after this. B.2. (ii) The production process for fish oil must ensure that all raw material intended for the production of crude fish oil is subject to a treatment including (where relevant) heating. centrifugation. refining and purification steps before being placed on the market for the final consumer. 2073/2005).Health standards for fishery products In addition to compliance with microbiological criteria laid down in Regulation (EC) No. and the raw material is processed within 36 hours after loading. 9.4. Shelling/shucking is to be done hygienically. Histamine Histamine limits must not be exceeded (limits are set in Regulation (EC) No. B.5 Chapter V .
Fresh. The requirements for histamine and parasites do not apply to whole fishery products being used directly for preparing fish oil intended for human consumption. Diodontidae and Canthigasteridae. may only be placed on the market in wrapped/packaged form and must be appropriately labeled to provide information to the consumer on preparation/cooking methods and on the risk related to the presence of substances with adverse gastrointestinal effects. 206 .Storage of fishery products (i) Fresh fishery products. 9. 9. In the case of whole frozen fish in brine intended for the manufacture of canned food. Parasites Fishery products are to be visually examined so that visible parasites are identified before being placed on the market.FISH AND SEAFOOD D.4.6 Chapter VI . it should be kept at no more than -9 °C. Toxins harmful to human health Fishery products derived from poisonous fish of the following families must not be placed on the market: Tetraodontidae. The scientific name must accompany the common name on the label.2. prepared and processed fishery products belonging to the family Gempylidae. (ii) Frozen blocks prepared onboard vessels must be adequately wrapped before landing. (ii) Frozen fishery products must be kept at a temperature of not more than -18 °C in all parts of the product. thawed unprocessed fishery products.2. and cooked and chilled products from crustaceans and molluscs are to be kept at a temperature close to that of melting ice. Molidae.4. and when fishery products are wrapped on board fishing vessels. Fishery products contaminated with parasites may not be placed on the market for human consumption.7 Chapter VII . 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. in particular Ruvettus pretiosus and Lepidocybium flavobrunneum. E.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.2.2.
2.).4. melt water must not remain in contact with the products. species belonging to the Gadidae family. provided that the journey is short and the competent authority allows this.LEGISLATION (iii) Fishery products kept alive are to be kept at a temperature which will not adversely affect food safety or its viability. species belonging to the Merlucciidae family.3 Other information: Rules relating to visual inspections for the detection of parasites in fishery products are stated in Annex II (obligations on the competent authorities) of Regulation (EC) No. The following in relation to TVB-N limits is extracted from the regulation: Unprocessed fishery products shall be regarded as unfit for human consumption where organoleptic assessment has raised doubts as to their freshness and chemical checks reveal that the following TVB-N limits are exceeded: (a) 25 mg of nitrogen/100 g of flesh for the species: Sebastes spp. 2074/2005. This covers Total volatile basic nitrogen (TVBN) limits and how to determine them.2. 9. 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.Transport of fishery products The above points for Chapter VII apply in relation to transport. Helicolenus dactylopterus. The temperature of frozen fishery products must not fluctuate above -15°C. 9. (c) 35 mg of nitrogen/100 g of flesh for the species: Salmo salar. (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 .8 Chapter VIII . (ii) If fishery products are kept under ice. The required temperature for whole frozen fish in brine intended for the manufacture of canned food during transport is not stated. (b) 30 mg of nitrogen/100 g of flesh for the species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp. Sebastichthys capensis.4.2.
Chapter IX applies to pectinidae harvested outside those areas. please refer directly to the regulation. 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. VIII and IX. for full requirements. Extracts of the requirements of Annex III. 9. A food business operator can only accept batch of such bivalve molluscs provided correct documentary requirements are presented. and point 3 of Chapter VII applies to retail.4. reference should be made to the actual regulation. shellfish species etc. Apart from the provisions on purification. this Section also applies to live echinoderms. they supplement the requirements of Annex II to that Regulation.3 The regulation details specific hygiene requirements for bivalve molluscs. 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.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. where the raw material complies with points (a). (b) In the case of other operations. as amended. tunicates and marine gastropods. taken from at least three different points and mixed together by grinding. however.FISH AND SEAFOOD Regulation (EC) No 853/2004. The copy of 208 . specifically relating to bivalve molluscs are given below. VI. Chapters V. The information required in the registration document is given in point 4 and includes the health status of the production area. Section VII of Regulation 853/2004. 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. Live bivalve mollusc requirements - 9. they supplement the requirements of Annex I to that Regulation. (b) and (c) of Part B(1) of that Chapter.3.4. The sample must consist of about 100 g of flesh.1 Chapter I . Member States may set limits at a higher level for certain species pending the establishment of specific Community legislation.
Requirements for production areas (i) The competent authority classes (classed A.3. C.2 Chapter II . 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. B. 209 . collection from: .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. processing or relaying. 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.LEGISLATION document must be kept for at least 12 months post dispatch or receipt or longer as stated by the competent authority. 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.4. B or C) production areas with fixed locations and boundaries for where live bivalve molluscs may be harvested and this must be complied with. 9.Class A means they can be placed directly on the market . poles or other fixed means are to identify the boundaries of the sites.Hygiene requirements for the production and harvesting of live bivalve molluscs A. Other requirements such as conditions for relaying in order to allow optimal conditions for purification and for permanent record keeping are also specified. (ii) Provided the requirements of chapter V are met.Class B means treatment in a purification centre or after relaying is done before it can be placed on the market . Transport needs to allow sufficient drainage for best survival conditions.
FISH AND SEAFOOD 9. (iv) yessotoxins.4. 800 micrograms per kilogram. containers and a label to identify purification. dinophysistoxins and pectenotoxins together. 1 milligram of yessotoxin equivalent per kilogram. requirements if there are many batches of live bivalve molluscs.3. (ii) amnesic shellfish poison (ASP).4.4. Before dispatch. the levels of marine biotoxins should not exceed the following limits.Health standards for live bivalve molluscs In addition to microbiological criteria and organoleptic characteristic requirements. 20 milligrams of domoic acid per kilogram (iii) okadaic acid. Requirements for dispatch centres Handling should not result in contamination to any degree. 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 9.5 Chapter V .3 Chapter III .3.3. 210 .Structural requirements for dispatch and purification centres Premises on land should not be subject to flooding by tides etc.4 Chapter IV . for: (i) paralytic shellfish poison (PSP). B.Hygiene requirements for purification and dispatch centres A. 9. 160 micrograms of okadaic acid equivalents per kilogram. Requirements for purification centres There are requirements for live bivalve molluscs to be washed. the purification system to allow them to resume /maintain filter-feeding activity. and there are requirements for tanks and water storage containers.
4.7 Chapter VII .4. The requirements for production areas as stated earlier apply to pectinidae and these can only be placed on the market through a fish auction. 9. including the identification mark. the date of minimum durability may be replaced by the words ‘these animals must be alive when sold’.4.2.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. dispatch centre or processing establishment.Wrapping and packaging of live bivalve molluscs (i) Oysters must be wrapped or packaged with the concave shell downwards.1).8 Chapter VIII .3. by way of derogation from directive 2000/13/EC. 211 . (ii) Once the live bivalve molluscs have been packed for retail sale and have left the dispatch centre. 160 micrograms of azaspiracid equivalents per kilogram.Identification marking and labelling In addition to general identification marking (contained in 9. must be waterproof.9 Chapter IX . Also.3. Finally. the species of bivalve mollusc and date of packaging (at least the date and month) are required. (ii) Individual consumer-size packages of live bivalve molluscs must be closed and remain closed after leaving the dispatch centre and until presented for sale to the final consumer.4.6 Chapter VI .3.4. they cannot be re-immersed in or sprayed with water. 9. Requirements of Chapters III and IV are to be complied with.3.LEGISLATION (v) azaspiracids. Registration document requirements also apply. the label.Specific requirements for pectinidae harvested outside classified production areas Requirements for harvesting and handling following harvesting and health standards for live bivalve molluscs must be met. 9. (Note: testing methods for detecting marine biotoxins are detailed in Annex III of Regulation (EC) 2074/2005) 9.
Fishery products in particular must comply with the requirements of Annex III of the regulation. they derive from: (i) poisonous fish. or - - 212 . In this regulation. (ii) fishery products not complying with the requirement concerning biotoxins. The official controls include audits of good hygiene practices and hazard analysis and critical control point (HACCP)-based procedures.FISH AND SEAFOOD 9. Section VIII. physical or microbiological checks or checks for parasites have shown that they are not in compliance with the relevant Community legislation. The requirements refer to hygienic conditions and regular inspections both on land and at port to ensure compliance of temperature and other requirements. This refers to official controls or production and placing on the market of fishery products. chemical.5. It is a requirement that food business operators give assistance to ensure that official controls carried out by the competent authority can be done properly. 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. or (iii) bivalve molluscs. general principles for official controls in respect of all products of animal origin falling within the scope of the regulation are given.1 Regulation (EC) 854/2004 Annex III Chapter III states that fishery products are to be declared unfit for human consumption if: organoleptic. 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. Therefore the health standards for fishery products as previously stated in Annex III. tunicates or marine gastropods containing marine biotoxins in total quantities exceeding the limits referred to in Regulation (EC) No 853/2004.5 Regulation (EC) No. Chapter V of Regulation 853/2004 apply. 9. echinoderms.
Also. In the remaining 10 % of samples. section VII. that microbiological criteria and chemical contaminants amongst other requirements are met.5.Official controls concerning live bivalve molluscs from classified production areas A. 213 . Live bivalve molluscs from Class B areas must not exceed.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. - - - - B. 4. coli per 100 g of flesh and intravalvular liquid. in 90% of the samples. live bivalve molluscs must not exceed 46.5. three dilutions MPN test specified in ISO 16649-3. Annex II 9. three dilutions Most Probable Number (MPN) test specified in ISO 16649-3. chapter II on hygiene requirements for the production and harvesting of live bivalve molluscs in Regulation (EC) 853/2004. Classification of production and relaying areas: This section reinforces that specified in Annex III. Requirements for the competent authority are stated in relation to the classification or a production or relaying area is stated.1 Chapter II .600 E.2. The reference method for this analysis is the five-tube. 000 E. live tunicates and live marine gastropods. as previously stated. Sampling plans and frequency are required. Regulation (EC) 854/2004. live echinoderms. Monitoring of classified relaying and production areas This is to ensure there is no malpractice. coli per 100 g of flesh and intravalvular liquid. A summary is given as follows: 9. The reference method for this analysis is the five-tube.000 E. coli per 100 g of flesh and intravalvular liquid. chapter V is reinforced. 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 46.2 This states official controls for the production and placing on the market of live bivalve molluscs.
opening or closing a production area.5. Additional monitoring requirements This concerns the competent authority in relation to monitoring of classified production areas where bivalve mollusc harvesting is forbidden and of relaying and production zones to ensure there is no malpractice etc.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. dispatch centres and processing establishments in order to confirm that health standards and other specific requirements are met. 9. boundaries or class of a production area or its closure. 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. F.FISH AND SEAFOOD C. establishes microbiological criteria for a range of foods. 2073/2005 on Microbiological Criteria for Foodstuffs Regulation (EC) No. 9. 9. 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. 214 .5. 2073/2005.Official controls concerning pectinidae harvested outside classified production areas These are to be carried out in fish auctions.2 Chapter III .6 Regulation (EC) No. Procedures for re-opening a closed production area are stated D. E. which has applied since 1 January 2006.3 Imports By way of derogation from Article 11(1) of Regulation (EC) No 854/2004. Food business operators' own checks Controls that food business operators have done are considered by the competent authority when classifying.
4 referring to fishery products. volume.II.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. area or batch. presence or number of microorganisms. with chapter 2.6. and/or on the quantity of their toxins or metabolites. ensuring that foods being placed on the market do not pose a risk to human health. The definition of ‘microbiological criterion’ means a criterion defining the acceptability of a product.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. based on the absence.I. 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.2 Process hygiene criteria Chapter 2 focuses on process hygiene criteria. 215 . The relevant criteria are outlined in Table 9.6. per unit(s) of mass. 9. Two kinds of criteria have been established: food safety criteria. a batch of foodstuffs or a process. 9.
monocytogenes 1.2 Ready-to-eat foods able to support the growth of L. other than those intended for infants and for special medical purposes(4) (8) L. monocytogenes 5 0 Absence in 25 g (7) FISH AND SEAFOOD 216 5 0 100 cfu/g 5 0 Absence in 25 g 1. monocytogenes.1 Ready-to eat foods intended for infants and ready-to-eat foods for special medical purposes (4) 5 0 100 cfu/g (5) EN/ISO 11290-2 (6) L.I Food safety criteria Sampling plan(1) n 10 5 Absence in 25 g EN/ISO 11290-1 c m M Products placed on the market during their shelf life Limit(2) Analytical reference method(3) Stage where the criterion applies Food category Microorganisms 1. 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.3 Ready-to-eat foods unable to support the growth of L. monocytogenes L.16 Cooked crustaceans and molluscan shellfish Salmonella EN/ISO 6579 Products placed on the market during their shelf life .TABLE 9. who has produced it EN/ISO 11290-2 (6) Products placed on the market during their shelf life 1.
17 Live bivalve molluscs and live echinoderms. coli (15) 1.26 Fishery products from fish species associated with a high amount of histidine (17) 9 2 200 mg/kg Histamine LEGISLATION 217 1.27 Fishery products which have undergone enzyme maturation treatment in brine. tunicates and gastropods 9 (18) 2 E.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 1.25 Live bivalve molluscs and live echinoderms. manufactured from fish species associated with a high amount of histidine (17) Histamine .
excluding sprouted seeds.1. 5 This criterion shall apply if the manufacturer is able to demonstrate.25. — — bottled or packed waters. biscuits and similar products. Other categories of products can also belong to this category. Dervin C. subject to scientific justification. — fresh. monocytogenes. 79. according to which the whole batch is to be deemed unsafe. E. including cocoa and chocolate products. The most recent edition of the standard shall be used. Assay of biogenic amines involved in fish decomposition. 2. wine. that the product will not exceed the limit 100 cfu/g throughout the shelf life. Particularly fish species of the families: Scombridae. 1 ml of inoculum is plated on a Petri dish of 140 mm diameter or on three Petri dishes of 90 mm diameter. m = M. In such a case the presumption laid down in Article 14(6) of Regulation (EC) No 178/2002. uncut and unprocessed vegetables and fruits. Clupeidae. that the product will not exceed the limit of 100 cfu/g throughout the shelf life. Scombresosidae. Coryfenidae.. to the satisfaction of the competent authority. Malle P.. Duflos G.4 or aw ≤ 0.0 and aw ≤ 0. sugar. Bouquelet S. 1996. coli is used here as an indicator of faecal contamination. when he is not able to demonstrate.92....94. 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. products with pH ≤ 5. products heat treated in their final package). when recontamination is not possible after this treatment (for example. AOAC Internat. to the satisfaction of the competent authority. Valle M. — bread. beer.FISH AND SEAFOOD 1 2 3 4 n = number of units comprising the sample. Engraulidae. honey and confectionery. soft drinks. Pomatomidae. A pooled sample comprising a minimum of 10 individual animals. live bivalve molluscs. c = number of sample units giving values between m and M For point 1. Malle P. This criterion shall apply to products before they have left the immediate control of the producing food business operator. Single samples may be taken at retail level. spirits and similar products. shall not apply. Relevance of matrix effect in determination of biogenic amines in 6 7 8 15 16 17 18 19 218 . Bouquelet S. 43-9. products with a shelf-life of less than five days shall be automatically considered to belong to this category. Products with pH ≤ 4. J. References: 1.1 . cider. 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.
Histamine in fishery products from fish species associated with a high amount of histidine: — 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. the mean value observed is ≤ m 2. Salmonella in different food categories: — satisfactory. a maximum of c/n values observed are between m and M 3. if the presence of the bacterium is detected in any of the sample units. if all the values observed indicate the absence of the bacterium. L. monocytogenes in ready-to-eat foods able to support the growth of L. tunicates and gastropods in relation to testing E. L. where the limit refers to a pooled sample. — unsatisfactory. The test results demonstrate the microbiological quality of the batch tested(1). AOAC Internat. if all the values observed indicate the absence of the bacterium. if the presence of the bacterium is detected in any of the sample units. if the following requirements are fulfilled: 1. — unsatisfactory. (1) 219 . monocytogenes in ready-to-eat foods intended for infants and for special medical purposes: — satisfactory. 1097-101.LEGISLATION plaice (Pleuronectes platessa) and whiting (Merlangus merlangus.I The limits given refer to each sample unit tested. excluding live bivalve molluscs and live echinoderms. J. — unsatisfactory. if the presence of the bacterium is detected in any of the sample units. if any of the values are > the limit. 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. no values observed exceed the limit of M. coli. if all the values observed are ≤ the limit. Interpretation of the test results relating to Table 9. — unsatisfactory. L. if all the values observed indicate the absence of the bacterium. 1999. 82. monocytogenes in other ready-to-eat foods: — satisfactory. — unsatisfactory. 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.
II Process hygiene criteria Limit(2) Analytical reference method(3) M 10 cfu/g ISO TS 16649-3 End of the manufacturing process End of the manufacturing process Improvements in production hygiene Stage where the criterion applies Action in case of unsatisfactory results Food category Microorganisms c 2 1 cfu/g m Sampling plan(1) n 2.4.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.
LEGISLATION 1 n = number of units comprising the sample. c = number of sample units giving values between m and M The most recent edition of the standard shall be used. if all the values observed are ≤ m. if all the values observed are ≤ m.K) Temperature control requirements 9. For example. — unsatisfactory. with chill holding tolerance periods. and particularly that perishable foodstuffs must not be kept at above the maximum recommended storage temperature. — unsatisfactory. coli in shelled and shucked products of cooked crustaceans and molluscan shellfish: — satisfactory. — acceptable. 9. if one or more of the values observed are > M or more than c/n values are between m and M. tolerance periods for chill-holding of foods and hot-holding variations. had not been 221 . and the rest of the values observed are ≤ m.7 Food Hygiene (England) Regulations 2006. 14 (Hygiene requirements specific to the U. 2006 No. 2006 No. which overrides the 8 °C requirement. E. S. 14 details temperature control requirements for foods in general. Hot-held foods (food having been cooked or reheated and is for service or on display for sale) must not be kept below 63 °C.7.1 In the U. The test results demonstrate the microbiological quality of the process tested. The defendant may be required to produce well-founded scientific proof to support his claims. The regulations provide for defences in relation to upward variations of the 8 °C temperature. The regulations prescribe a chilled food holding temperature of 8 °C or less. S. — acceptable. the defendant will need to prove that the food was on service or display. Coagulase-positive staphylococci in shelled and cooked crustaceans and molluscan shellfish: — satisfactory. if a maximum of c/n values are between m and M.I.I.K. Schedule 4 of the Food Hygiene (England) Regulations 2006. if one or more of the values observed are > M or more than c/n values are between m and M. if a maximum of c/n values are between m and M. but there is also a general requirement that foods must not be kept at temperatures that would result in a risk to health. 2 Interpretation of the test results relating to Table 9. and the rest of the values observed are ≤ m.II The limits given refer to each sample unit tested.
S. It should be noted that the temperature control requirements as detailed in Schedule 4 of the Food Hygiene (England) Regulations 2006 (S. such as that below. 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. 2006 No.I. preparing or processing the food recommending that it be kept at or below a specified temperature between 8 ºC and ambient temperatures. 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. for a limited period consistent with food safety. 14) do not apply to any food covered by EU Regulation 853/2004 on hygiene of products of animal origin or any food business operation carried out on a ship or aircraft. the 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. or at a temperature above 63 °C and to reheat food to a temperature of at least 82 °C (The Food Hygiene (Scotland) Regulations 2006. Schedule 4 of the Food Hygiene (England) Regulations 2006 contains several definitions.uk/foodindustry/guidancenotes/hygguid/fhlguidance/ 222 . For other food. and was kept at above 8 °C or the recommended temperature. 3). The permitted reasons are given below: to facilitate handling during and after processing or preparation the defrosting of equipment. Alternatively.S. 9. Recommended temperature: a specified temperature that has been recommended in accordance with a food business responsible for manufacturing. to or from premises (including vehicles) at which the food was to be kept at or below 8 °C or the recommended temperature.K.gov.food. 2006 No. the period for which it can be expected to remain fit for sale when kept in a manner consistent with food safety. was kept at above 8 °C or the recommended temperature for an unavoidable reason. or.FISH AND SEAFOOD previously put on display at more than 8 °C and had been kept there for less than four hours.I.8 Guidance In the U. 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.
2076/2005. 206). L226. 22. L139. 1791/2006 and 1020/2008 and as read with EC Directive 2004/41. 2076/2005. The revised text of Regulation (EC) No. L226. 2074/2005. 1688/2005. and Regulation (EEC) 1536/92 laying down common marketing standards for preserved tuna and bonito.6. 1). 22) as amended by Regulation (EC) Nos. 25. Regulation (EC) No. L226. Regulation (EC) No. 30. 1.LEGISLATION 9. Regulation(EC) No. 25. L139. 2073/2005 on microbiological criteria for foodstuffs (OJ No.2004.4.10 Further Reading Regulation (EC) No.4. 30. The revised text of Regulation (EC) No.10. Commission Regulation (EC) No. 853/2004 is now set out in a Corrigendum (OJ No. as read with the corrigenda at OJ No. 853/2004 of the European Parliament and of the Council laying down specific hygiene rules for food of animal origin (OJ No. 55). The revised text of Regulation (EC) No. 2076/2005. 9. as amended. 83) as amended by Regulation Nos 882/2004.6. Regulation (EEC) 2136/89. L338. 1441/2007. Regulation (EC) No. Regulation (EC) No. 854/2004 of the European Parliament and of the Council laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption (OJ No. 3) as amended by Regulation (EC) No. 1663/2006.2005. 2076/2005.2004. L139. Regulation (EC) No. laying down common marketing standards for preserved sardines.2006.2004.2004.6. 2074/2005 and Regulation (EC) No. 2074/2005.12. 2075/2005 and Regulation (EC) No. 30. 223 . 852/2004 of the European Parliament and of the Council on the hygiene of foodstuffs (OJ No. 1791/2006 and 1021/2008 and as read with EC Directive 2004/41. these are not be discussed here.4. 852/2004 is now set out in a Corrigendum (OJ No.2004. 854/2004 is now set out in a Corrigendum (OJ No. 14. L283. 2074/2005. 1662/2006. 1019/2008 and as read with Regulation 2073/2005. 62) as amended by Regulation (EC) No.2004. 25. namely.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.
22. as amended by Commission Regulation (EC) No 1022/2008 OJ L 338.12. 27 Commission Regulation (EC) No 2076/2005 laying down transitional arrangements for the implementation of Regulations (EC) No 853/2004. 21.2005. 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 278.10.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. 6) 224 . 83) Regulation (EC) No 104/2000 on the common organisation of the markets in fishery and aquaculture products (OJ L 17.1. 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 854/2004.2005. (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004 ( OJ L 338.12. 23.2000.2001.
1 Aeromonas spp.1.1.5 minutes.5 pH Under otherwise optimal conditions.3 . 10.000-fold increase in number is observed after one week in food systems (1).3 minutes. 10. The minimum pH value for growth is thought to be pH 5. a 10 to 1. The optimum growth temperature is 28 °C (2).10.5 µm 10.0 .1 10. At 5 °C.1.5.4 Heat resistance Aeromonads are sensitive to moderately high temperatures. PATHOGEN PROFILES 10.2 Oxygen requirements Facultative anaerobe. no growth is observed (1). under optimal incubation temperatures. Morphology Gram-negative rods.0 x 1.10 (3).1. and 2. 10.45 °C (1). and D-values at 45 and 51 °C have been reported as 29.1.5. respectively (1). 0.3. Some death will occur at this pH if the temperature is 4 °C. Aeromonas has the potential to grow over a pH range of ca 4 . 225 . At pH 4.1.3 Temperature Aeromonas grows at temperatures of 2 .
1 Morphology Gram-positive. All type A strains are proteolytic and type E strains are usually non-proteolytic. 6). 226 . many foods that are not obviously ‘anaerobic’ can provide adequate conditions for growth. Types A. to a lesser extent. 10. F are the types that are responsible for most cases of human botulism (5. botulinum grow reasonably well in the temperature range of 20 45 ºC.4 Heat resistance The vegetative cells of C.3 Temperature All strains of C.even as low as 3 ºC (7).2.2 Oxygen requirements Although C. botulinum types produce heat labile toxins. but the spores of this organism are more so. but the minimum temperatures required to inhibit Groups I (proteolytic group) and II (non-proteolytic group) are different.30 minutes.22. and groups I and II are those responsible for the botulism cases.2. but the interior of the food may do so.0 µm. types B and F can be either. spore-forming rods. B. botulinum is an obligate anaerobe.1. E and. 10. botulinum are not particularly heat resistant.2. botulinum are known. and is sensitive to > 4. 10.5% NaCl (4). A to G. but Group II strains are psychrotrophic.2 Clostridium botulinum Seven different types of C.FISH AND SEAFOOD 10. It is also important to note that the inclusion of oxygen as a packaging gas cannot ensure that growth of C. or at 90 ºC for a few seconds. at 85 ºC for 5 minutes. All C. 0. Group I will not grow at temperatures of 10 ºC or less. There are four main groupings of the organism. being capable of slow growth and toxin production at low temperatures .2.2. which may be inactivated by heating at 80 ºC for 20 .5 . Thus an aerobically packed product may not support the growth of the organism on the surface. 10.6 Aw/Sodium chloride Growth of Aeromonas is optimal in the presence of 1 .2% sodium chloride (NaCl) (1). 10.4 x 1. botulinum is prevented. forming at least seven different toxins.7 .
2. they may survive mild heat treatments (70 . 9).0 and aw > 0. 0.2. perfringens . vacuum packed. botulinum depends on solute. These concentrations correspond to limiting aw of 0.0. pH and temperature. grow on the surface of foods unless they are vacuum or gas-packed. It will not.1. botulinum are produced by type A and proteolytic B strains (D121ºC=ca 0.5 pH The minimum pH for the growth of proteolytic and non-proteolytic strains is pH 4. However. Note also that if humectants other than salt are used.6 and pH 5. The spores of Group II (non-proteolytic/psychotrophic) strains are less heat resistant than Group I strains. where oxygen levels are reduced. the minimum aw for growth may be lower. botulinum spores The most heat resistant spores of Group I C. parcooked meals with pH value > 5.85 ºC) and their ability to grow at refrigeration temperatures necessitates their control in foods capable of supporting their growth (e.is an obligate anaerobe.1 Clostridium perfringens Morphology Gram-positive spore-forming rods. These values have been established under carefully controlled laboratory conditions. 227 . therefore.g.3.21 minutes) (9). 10. 10.PATHOGEN PROFILES 10. 10.like other clostridia .10.3 .94 for Group I and 0. respectively (8. and 5% (w/w) NaCl is necessary to prevent growth of Group II organisms. 11)).2 Oxygen requirements C.9 x 2.0 µm. particularly by cooking. The organism will grow well in the centre of meat or poultry dishes.3.7 Characteristics of C. but under optimum growth conditions 10% (w/w) NaCl is required to prevent growth of Group I. safety margins must be introduced to allow for process variability.97) (10.97 for Group II (5).0 .3 10.2.6 Aw/ Sodium chloride The minimum water activity (aw) for growth of C. In commercial situations. 10.
depending on strain and other conditions. At cold temperatures. Salt concentrations of 6 . death of the vegetative cells of this organism usually occurs rapidly above this temperature (12.7 Characteristics of C. however. perfringens has the potential ability to grow within the temperature range 15 . The lowest aw recorded to support the growth of C. type of solute. 10. 15. will survive greater extremes of pH (and aw) (12.8% inhibit growth of most C. Spores. perfringens strains. vegetative cells die rapidly (12).5 pH C. (It is unusual for spores to be formed in foods after the growth of this organism (14)). perfringens is not a tolerant organism with respect to pH. perfringens in relation to food safety is the organism’s ability to grow extremely rapidly at high temperatures.5 minutes (17). Its optimum temperature for growth is 43 .3.it is destroyed by heating at 60 ºC for 10 minutes (14. the limits for aw are affected by temperature. pH. It grows best at pH values between 6 and 7 (the same pH as most meats).3. although prior growth at high temperatures or the presence of fat in a food will result in increased heat resistance. and their heat resistance is also affected by the heating substrate.4 Heat resistance Exposure to a temperature of 60 ºC or more will result in the death of vegetative cells of C. lower concentrations may be effective in combination with other factors. Under otherwise ideal conditions.93 and 0. While some growth can occur at 50 ºC. the enterotoxin is not heat resistant .FISH AND SEAFOOD 10. 10.45 ºC.3. In addition.97 (glycerol and sucrose respectively) depending on the solute used to control aw (13. 16).10 ºC.3. although C. perfringens appears to be 0. perfringens is not tolerant of low water activities.50 ºC. As in the case of other factors limiting the growth/survival of this organism. 10.6 Aw/Sodium chloride C. 13).3 Temperature The most significant characteristic of C. perfringens. The D-value for C. very limited growth may occur at pH values over the range pH ≤ 5 and ≥ 8. etc. 10. perfringens in roast beef at 60 ºC is 14. Some studies indicate that the presence of 3% NaCl delays growth of C. 18). perfringens spores The spores of C. 13). perfringens in vacuum packed beef (18).3. 0 . perfringens can vary quite considerably in their heat resistance. Recorded heat 228 .3.
monocytogenes is reported to be 45 ºC (20).5 x 0.2. so can be destroyed by pasteurisation. monocytogenes in crawfish tail meat at 55.2 has been suggested as the lowest working limit (22).98 and 0. 0.4 . pH < 5.4. monocytogenes is not a particularly heat resistant organism. However.0. -0. Morphology Gram-positive short rods. The upper temperature limit for the growth of L.64.19 minutes.5 . 10.albeit slowly .4.4. 229 . D-values for L. being potentially capable of growing . and temperature. and where acetic acid is used as an acidulant. growth at low temperatures can be very slow. the lowest limit for growth is likely to be considerably higher .4.PATHOGEN PROFILES resistance values at 95 ºC (D-values) range from 17. is between 30 and 37 ºC.at refrigeration temperatures down to.3 .2.3 Temperature Listeria monocytogenes is unusual amongst foodborne pathogens in that it is psychrotrophic. 10.4 10. In foods.0 µm.0 minutes for heat resistant spores to 1. 10. it is not a sporeformer.6 . requiring days or weeks to reach maximum numbers.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.2 Oxygen requirements Aerobe or microaerophilic.23. using hydrochloric acid as an acidulant). respectively (21).3 is the lowest value where growth has been recorded. Its optimum growth temperature. the organism is able to grow at pH values well below pH 5 (pH 4. It has been reported to have slightly greater heat resistance than certain other foodborne pathogens.8 minutes for heat sensitive spores (12). however. 10.4 ºC is probably a more likely minimum for growth in foods (19). 10. 1. Under ideal conditions.especially at refrigeration temperatures. 60 and 65 ºC were reported as 10.4 Heat resistance L.1 Listeria spp.4. however. or even below 0 ºC.
25).12% NaCl or more).FISH AND SEAFOOD 10.2 Oxygen requirements Facultative anaerobe.5. the optimum being 30 ºC (23).6 Aw/Sodium chloride L.6 Aw/Sodium chloride Plesiomonas can grow well at NaCl concentrations up to 4% (24.1 Plesiomonas Morphology Gram-negative.3 Temperature Plesiomonas can grow in the temperature range of 8 .3. It is likely to survive.5.5. 10. 10. 10.5. 10. at salt levels found in foods (10 .5 10. peritrichous flagella.97. 10.8.6 10.5 .5 (24.3 µm. monocytogenes is quite tolerant of high NaCl/low water activities.45 ºC. The bacterium may survive for long periods at aw values as low as 0. 25).90. It grows best at aw of ≥ 0.0 .5 .8 .5 pH Plesiomonas has a pH range for growth of pH 4.6. but has been shown to be able to grow at an aw level of 0.4 Heat resistance Pasteurisation at 60 ºC for 30 minutes has been reported as being effective in killing Plesiomonas shigelloides (24).83 (20). 0.0. 230 .0 µm. Morphology Gram-negative short rods. 0. 10.7 x 1.1 Salmonella spp.0 x 1 .4. 10. or even grow.1.5.5.
20 times more heat resistant than the average strain of Salmonella at high aw (27). and with an appropriate acid.0. D60 ºC values normally range from about 1 to 10 minutes.6. The growth of Salmonella is generally inhibited by the presence of 3 . the optimum being 35 . Some death will occur at pH values of less than about 4. Growth is slow at temperatures below about 10 ºC. or possibly 0. Salmonellae are quite resistant to drying. under otherwise ideal conditions.5. with a z-value of 4 .4% NaCl. However. substrate. depending on the type of acid and temperature.4 Heat resistance Salmonella is not a spore-forming organism. pasteurisation and equivalent treatments will destroy the organism under normal circumstances. some strains are able to grow at temperatures as low as 4 °C (26).9. therefore.6.5.5 pH Salmonella has a pH range for growth of pH 3. temperature and pH. 10. 10.37 ºC.48 ºC.3 Temperature Salmonellae can grow in the temperature range of 7 . and appropriate heat treatments must be determined experimentally for low-aw foods. strains vary in their ability to withstand heating. The optimal pH for Salmonella growth is between 6.945. high fat or low moisture levels (low aw) will reduce the effectiveness of heat treatments. 10.6. However.PATHOGEN PROFILES 10.6. Salmonella senftenberg 775W is about 10 .6.5 . 10. a heat resistant organism. although salt tolerance increases with increasing temperature (28). It is not. 231 .8 .5 ºC. Salmonella has the potential to grow at aw levels as low as 0. Furthermore.2 Oxygen requirements Facultative anaerobe.93 (as reported in dried meat and dehydrated soup).7. Salmonellae are quite resistant to freezing. depending on serotype.6 Aw/Sodium chloride Where all other conditions are favourable.
10.7.5 ºC. NB: The growth of Staph. the D60 ºC value is 2 .7 10.0 . however.1 Staphylococcus aureus Morphology Gram-positive cocci. 232 .).37 ºC (29). 10.7. stressed cells can also be less tolerant of heating.9 µm diameter. 0. As is the case with other bacteria.7 . Freezing and thawing have little effect on Staph.3 (optimum 7.7.7. aureus can grow within the temperature range 7 – 48. It can survive well at low temperatures. Staph. depending on aw.7. 10. Under most circumstances. fat content.2 Oxygen requirements Facultative anaerobe.5). aw and temperature (sensitivity to acid increases with temperature). under otherwise ideal conditions (29.0. Most strains of staphylococci can grow within the pH range 4. aw. aureus than citric acid).FISH AND SEAFOOD 10.2 to 9.4 Heat resistance Heat resistance depends very much on the food type in which the organism is being heated (conditions relating to pH.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. with an optimum of 35 . but may cause some cell damage (30).3 Temperature Under otherwise ideal conditions. 31). In meat macerate. The following relate to limits for growth only.7. aureus is more limited under anaerobic than aerobic conditions. 10. etc.20 minutes. the organism is heat sensitive and will be destroyed by pasteurisation. The limits for toxin production are also narrower than for growth. aureus viability.
2. An effect of increasing salt concentration is to raise the minimum pH of growth.8.6 Aw/Sodium chloride Staph. 34).0 (optimum 7. In general. 31) between 0. aureus can grow in 7 . an aw of 0. However.10% NaCl. 10. with a temperature range for growth of 10 . In general.8 10. and solute.8. atmosphere.8.7. Staphylococci are more resistant to salt present in foods than other organisms.8 x 1. especially vacuum packed foods (33) Temperature: pH: *Aw: Atmosphere: Heat Resistance: Enterotoxins are quite heat resistant. heating at 100 °C for at least 30 minutes may be required to destroy unpurified toxin (31.0 .83 . aureus is unusual amongst food-poisoning organisms in its ability to tolerate low water activities.4 .7. pH.2 Oxygen requirements Facultative anaerobe. Staph.43 ºC (35).7 Limits permitting toxin production 10 . * dependent on temperature.2 .3 Temperature The optimum temperature for growth has been reported as 30 ºC. but certain strains can grow in 20% NaCl.5 . 10.>0.99 aerobically under otherwise optimal conditions.87 and > 0. It can grow over the aw range 0.0. strain.6 µm 10.99 little or no toxin production in anaerobically packed foods. 233 .1 Vibrio cholerae Morphology Gram-negative curved rods. 0.46 ºC (optimum between 35 and 40 ºC) (very little toxin is produced at the upper and lower extremes) (31) 5. 10.PATHOGEN PROFILES 10.5) (29.7.86 is the generally recognised minimum in foods (32).9.
cholerae may persist on frozen fish for >180 days (36).9. 10. but not acid conditions.8. but only moderately sensitive to freezing (40). V. The D-value for V.4 Heat resistance V.8.0% NaCl (35).6. cholerae is sensitive to heat at temperatures of >45 ºC.9 10. 10. it has a pH range for growth of pH 5 . cholerae in crab meat homogenate at 60 ºC is 2. 10. respectively. 10.9.5 ºC).3 Temperature The minimum and maximum temperatures reported for growth in laboratory media.2 Oxygen requirements Facultative anaerobe.9.97 .1 Vibrio parahaemolyticus Morphology Gram-negative curved rods. with optimum at 7.8.984 (35). It can grow in the presence of 0. 234 . 37.4. 10. and it is sensitive to refrigeration temperatures (0 . parahaemolyticus is easily destroyed by drying.998. under otherwise favourable conditions. with an optimum of 0. declining in number during storage. 38).9.6 (35.5 pH This organism is tolerant of high pH values.FISH AND SEAFOOD Also. with an optimum of 37 ºC (39).0.1 . 10.6 Aw/Sodium chloride V.65 minutes (35). there is evidence that V. cholerae can grow over the aw range 0. are 5 and 43 ºC.
5 pH The pH range for growth is 4.9. parahaemolyticus will grow at aw levels as low as 0.11.9.5 minutes.5% NaCl.6 Aw/Sodium chloride V. 235 .8 .4 Heat resistance V. depending on solute. The D-values at 55 ºC for clam and crab homogenate are 0. it has been reported that cooking oysters for 10 minutes at 50 ºC should ensure destruction of the organism (43). 10.922.10.10. 10. parahaemolyticus is unable to grow in the absence of salt. with an optimum pH of 7. and 2.10.02 . V. parahaemolyticus are not heat resistant. and are readily destroyed by cooking. The organism can survive for long periods at freezer temperatures (42). 10. 10. 41).0. 10. but the optimum concentration is 2 .10. 10. 41).8.98 (35. its upper and lower limits for growth are 8 (although some reports suggest 10) and 0.3 Temperature The optimum temperature for growth has been reported at 37 ºC.4 Heat resistance V. respectively (39).4% (39.1 Morphology Gram-negative curved rods. respectively.2 Oxygen requirements Facultative anaerobe. The optimum concentration of NaCl equates to aw of approximately 0. with a temperature range for growth of 8 .6 (41).6 .10 Vibrio vulnificus 10.PATHOGEN PROFILES 10. vulnificus is heat sensitive.43 ºC (42).9.29 minutes.
5 . Beuchat L. in Foodborne Bacterial Pathogens. 10. Clostridium botulinum. Potential hazards associated with REPFEDS. Marcel Dekker Inc. Marcel Dekker. Great Britain.. London..L. Cliver D. 1994.E.L. Clostridium botulinum.5. Microbiology and Molecular Biology. New York. Bier J. Hui Y. 97-131. 1-19.L..D. 2001.5 pH The pH range for growth is 5 .N.H. in International Handbook of Foodborne Pathogens.P. New York. 2005. Stelma G. in Food Microbiology: Fundamentals and Frontiers. 329-49. Dodds K. Stelma G. Eds.. 6.H. Doyle M. 2003..W. in Foodborne Disease Handbook. 11. 236 . Clostridium botulinum. Notermans S. International Commission on Microbiological Specifications for Foods.0% NaCl. Washington D.K.W..R. 57-63. 1993. 1996. Austin J. 111-89. Eds.... 4.D.11 1.D. Peck M. Juneja V. 6 (1). Murrell K. Doyle M. 383-408.8 (42)..H.. 5. Dodds K. Kovacek K. Foegeding P. Lund B. Marcel Dekker. Clostridium botulinum and Clostridium perfringens. New York. Aeromonas hydrophila.. 357-67. Gorman J. Palumbo S.H. Aeromonas species.M. 121-76. New York. Eds. Food Control. in Foodborne Bacterial Pathogens. Blackie. Hauschild A. New York. Principles of control. 1-27.L.W.10. 5-19.R.. with an optimum pH of 7..W. New York. in Foodborne Disease Handbook. Cliver D.W. Abeyta C. Growth and heat resistance of psychrotrophic Clostridium botulinum in relation to ‘sous vide’.H. Miliotis M.O.6 Aw/Sodium chloride V.P. Gorham J. Smith J. Fratamico P. Aeromonas hydrophila group. Aeromonas. Montville T.. in Foodborne Pathogens. 2. Dodds K. 3. Eds. Gaze J. 1989. vulnificus grows over the range 0.M.O. Hauschild A.H.. Ed. 10. Faris A. ASM Press. Hauschild A..10. in Microorganisms in Foods.R. Diseases Caused by Bacteria.N. 279-303. Ed. Volume 1: Diseases Caused by Bacteria. 1995. Betts G.10. References International Commission on Microbiological Specifications for Foods. Eds.P. Kim J. Eds. Doyle M. Marcel Dekker. Volume 5: Microbiological Specifications of Food Pathogens. 9..D.. Eds..FISH AND SEAFOOD 10. in Clostridium botulinum: Ecology and Control in Foods. Ed. 7. Hui Y. 10. Caister Academic Press. Volume 1. Johnson E. New York.A.. 1989. Bhunia A.. 1993. Marcel Dekker.J. in Clostridium botulinum: Ecology and Control in Foods. Marcel Dekker. with an optimum of 2. 1994.M.C. Marcel Dekker.. Murrell K.5% (42). 8. Novak J.
International Commission on Microbiological Specifications for Foods.. 735-42. Listeriosis and Food Safety. 25.P.G. Banks J.An Introduction. 336 (8721).A. Marcel Dekker..T. London.P. Washington D. 229-40. Kim C.. 1991. Archer P. International Commission on Microbiological Specifications for Foods. 208-13. 13. 201-8. Volume 5: Microbiological Specifications of Food Pathogens. Juneja V. Diseases Caused by Bacteria.K. 1990. Gorham J. Journal of Applied Bacteriology. 15. 133-67. Volume 5: Microbiological Specifications of Food Pathogens.R. Lancet. Ed. in Microorganisms in Foods. Doyle M. Eds. in Microorganisms in Foods.R. Washington D. 33. in Foodborne Pathogens: An Illustrated Text. International Commission on Microbiological Specifications for Foods. Johnson E. New York.. 1990. Murrell K. 1989. 1994. Varnam A. 112-25. 68 (2). McClane B. in Foodborne Bacterial Pathogens. 982-6. 1996. Montville T. Volume 1. Eds. Marcel Dekker.H. Doyle M. 1989. ASM Press. 137-64. Evans M. 24. Academic Press. London. Blackie. in Foodborne Disease Handbook. Ed. in Food Microbiology: Fundamentals and Frontiers. 19. Eds. 20. Walker S. Emery D.O. in Microorganisms in Foods.G. New York. International Commission on Microbiological Specifications for Foods. Varnam A.M. 2006. 1989. in Food Microbiology .. Rinke H.J. in Foodborne Infection and Intoxication. Matthews K. 311-25. 26. Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs.D. 21. London.PATHOGEN PROFILES 12. Eds.A.. 1996.O. Ed.. Ryser E. Clostridium perfringens... International Commission on Microbiological Specifications for Foods. Lund B. Cliver D. 351-82. Nagaraja K. 22. Doyle M.. Growth of Listeria monocytogenes at refrigeration temperatures.A. Beuchat L. ASM Press. Foodborne disease due to Bacillus and Clostridium species.J. London.H. Ed.. Ed. Ed. Marth E. Cliver D. Plesiomonas shigelloides. Evans M. New York.. Marcel Dekker.G.M. 2007.C. London. 2005. Koburger J. 129-38. Riemann H. London. 17. Hui Y.. Labbe R.. Volume 5: Microbiological Specifications of Food Pathogens. Wrigley D. Listeria monocytogenes.H. 141-82. 14. Clostridium perfringens gastroenteritis. 23.J. Clostridium perfringens food poisoning.. Blackie. Elsevier. 2001.C.V. Clostridium perfringens.A.O. Marcel Dekker. Cliver D. 1990.J.. Plesiomonas shigelloides.R. Wolfe Publishing Ltd. International Commission on Microbiological Specifications for Foods. Clostridium perfringens. 16.P.. 237 ..P. New York.A. 157-62. 18.H. Eds. Avian Disease. Listeria. 191-243. Halvorson D. in Foodborne Diseases. Listeria monocytogenes. Blackie. 1996. Labbe R. Plesiomonas. in Foodborne Bacterial Pathogens. Clostridium perfringens. Montville T.
2005. London.. 4 (13). 1993. Vibrio cholerae. Eds. Ed. Ed. Applied and Environmental Microbiology.A.P.. Griffiths M. 1996. Japanese Journal of Experimental Medicine. Ed. 38.C. Jay J. Stress Response and Resistance. 1996. 39. 1983.A. in Microorganisms in Foods.A. Academic Press..P. Bhunia A. Reed G. 37. Staphylococcal intoxications.S.P. 327-52. 2005. 31. CDR Review. Twedt R. R15764. International Commission on Microbiological Specifications for Foods. 331-57.H. Vibrio parahaemolyticus. in Foodborne Pathogens. Wymondham. Caister Academic Press. 2005. Staphylococcal Food Poisoning. Microbiological Specifications of Food Pathogens. Volume 5. Eds. Volume 5.. Bergdoll M. London. Wilkinson B. 35. 13 (11). Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. Blackie.. Fratamico P. Marcel Dekker. Bergdoll M. in Foodborne Bacterial Pathogens.C. in Microorganisms in Foods. Cliver D. Cliver D. 543-68. Volume 5: Microbiological Specifications of Food Pathogens. Blackie. 8. Staphylococcus aureus as a food pathogen: staphylococcal enterotoxins and stress response systems. London. On the resisting power of Vibrio cholerae and typhoid Bacillus against low temperatures. Doyle M.J. Springer Science. 85-106.D. Blackie. Dairy.M. 1989. 1994. Cambridge. Tohyama Y. 28. in Microorganisms in Foods.M.S. Salmonellae. in Foodborne Pathogens: Microbiology and Molecular Biology. 217-64. Eds. 327-445. New York.S. International Commission on Microbiological Specifications for Foods. in Modern Food Microbiology.L. Golden D. in Foodborne Infections and Intoxications. Loessner M.O. London. 985-98. Marcel Dekker. D’Aoust J-Y.J. Woodhead Publishing Ltd. 1990. Ed. Experimental investigations concerning the prevention of epidemic diseases due to aquatic products (Fourth Report). 426-35. Golden D. 33. Vibrio parahaemolyticus. International Commission on Microbiological Specifications for Foods. 40. Ed. in Foodborne Disease. 34.FISH AND SEAFOOD 27. Foodborne illness (Part 1): Staphylococcal (“Staph”) food poisoning.K.R. 1996. Riemann H. 1989. International Commission on Microbiological Specifications for Foods. Smith J. Stewart G. 32.62. New York. 36. Cleland D. Ed. International Commission on Microbiological Specifications for Foods. London. Doyle M... 545-66. Loessner M. Microbiological Specifications of Food Pathogens. 273-84. Salmonella. 29. New York. International Commission on Microbiological Specifications for Foods. Warner R. Crowcroft N.M. Lee-Wong A.O. 2005.M. 1930. 523. Staphylococcal gastroenteritis.. in Understanding Pathogen Behaviour Virulence. 414-25. Ed. Staphylococcus aureus. 238 . Jay J. Academic Press. Food and Environmental Sanitation. Oliver J. 642. Gustafson J. 45 (3). 30.. Cholera: current epidemiology..
42.. Ruple A.PATHOGEN PROFILES 41.. Ed. Jay J. 436-9. Springer Science.J. International Commission on Microbiological Specifications for Foods. Volume 5. Vibrio vulnificus.. Golden D. Loessner M. International Commission on Microbiological Specifications for Foods. Cook D. New York. in Modern Food Microbiology. Cold storage and mild heat treatment as processing aids to reduce the numbers of Vibrio vulnificus in raw oysters. in Microorganisms in Foods. Yersinia.J. Journal of Food Protection..M. 657-78. and Campylobacter species.. Microbiological Specifications of Food Pathogens. 55 (12). Foodborne gastroenteritis caused by Vibrio. Jay J. London.M.W. 239 .D. Loessner M. Blackie. Golden D. 1996.A. 43.A. 1992. 2005. 985-9.
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org.gsi.CONTACTS Addresses of Trade Associations and Professional Bodies Association of British Salted Fish Curers and Exporters c/o Cawoods (Fish Curers) Ltd Estate Rd No 6 South Humberside Industrial Estate Grimsby North East Lincolnshire DN31 2TG United Kingdom Tel: + 44 (0) 1472 342248 Fax: + 44 (0) 1472 353100 Email: email@example.com 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: firstname.lastname@example.org@ifm.gsi.uk or v.gov.gov.ifm.seafish.uk Web site: www.defra.uk/ Department for Environment Food and Rural Affairs (Defra) Nobel House 17 Smith Square London SW1P 3JR United Kingdom Tel: + 44 (0) 207 2386000 Fax: + 44 (0) 207 2382188 241 Email: email@example.com Web site: www.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: firstname.lastname@example.org/ .org.gov.co.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.
dk Web site: www.uk Web site: www.htm Irish Fish Processors and Exporters Association 25 Kincora Avenue Clontarf Dublin 3 Eire Ireland Tel: + 353 1 833 7882/ 02770249 Fax: + 353 1 833 7882 Email: tormg.no/ Matís ohf (Icelandic Fisheries Laboratories) Skúlagata 4 101 Reykjavík Iceland Tel: + 354 422 5000 Fax: + 354 422 5001 Email: email@example.com. 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 Web site: www.FISH AND SEAFOOD Department of Agriculture and Rural Development.ie/templates/homepage.13 Postbox 6122 N-9291 Tromsø Norway Tel: + 47 77 629000 Fax: + 47 77 629100 Email: email@example.com/ Danish Institute for Fisheries Technology and Aquaculture (DIFTA) illemoesvej The North Sea Centre DK-9850 Hirtshals Denmark Tel: + 45 98 944300 Fax: + 45 98 942226 Email: firstname.lastname@example.org/s Norwegian Institute of Fisheries and Aquaculture Ltd Muninbakken 9 . Andersens Boulevard 37. Dublin Eire Ireland Tel: + 353 1 2144100 Web site: www.com Web site: www.difta.gov.gov.net Irish Sea Fisheries Board PO Box 12 Crofton Road Dun Laoghaire Co.uk/index/fisheriesfarming-and-food.dk Danmarks Fiskehandlere H.suite.matis.ifpea@eircom. Baelskaai 25 8400 Oostende Belgium Tel: + 32 59321876 Fax: + 32 59322840 Email: email@example.com 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.is Web site: www.fiskehandlerne.eapo.org/fishery/ European Association of Fish Producers’ Organisations H.org Web site: www. 1 1553 København V Denmark Tel: + 45 35 372023 Fax: + 45 35 372033 Email: Info@fiskehandlerne.is/ 242 .dk Web site: www.C.
fdf.org.co.uk Institute of Food Science and Technology (IFST) 5 Cambridge Court 210 Shepherds Bush Road London W6 7NL United Kingdom TeI: + 44 (0) 207 6036316 Fax: + 44 (0) 207 6029936 Email: firstname.lastname@example.org.CONTACTS Fishing Industry Research Institute 15 Lower Hope Street Rosebank 7700 Republic of South Africa National Fisheries Institute (USA) 7918 Jones Branch Drive Suite 700 McLean VA 22102 United States of America Tel: + 1 703 7528880 Email: email@example.com Web site: www.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 Web site: www.uk Department of Health Richmond House 79 Whitehall London SW1A 2NS United Kingdom Tel: + 44 (0) 207 2104850 Email: email@example.com CSIRO Marine Laboratories Castray Esplanade GPO Box 1538 Hobart Tasmania 7001 Australia Tel: + 61 3 62325222 Fax: + 61 3 62325000 Email: firstname.lastname@example.org Web site: www.org 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 New Zealand Seafood Industry Council Private Bag 24-901 Wellington 6142 New Zealand Tel: + 64 4 3854005 Fax: + 64 4 3852727 Email: firstname.lastname@example.org/ 243 .org.ac.ifr.uk Web site: www.seafood.uk/en/index.nz/sc-home Health Protection Agency (HPA) 61 Colindale Avenue London NW9 5EQ United Kingdom TeI: + 44 (0) 208 2004400 Fax: + 44 (0) 208 2007868 Web site: www.dh.au Web site: www.nz Web site: www.uk Institute of Food Research (IFR) Norwich Research Park Colney lane Norwich NR4 7UA United Kingdom Tel: + 44 (0) 160 3255000 Fax: + 44 (0) 160 3507723 Web site: www.ifst.csiro.cmar.
euro.who.int Web site: http://www.co. jsp Food Safety 10 .uk/scieh. Web site: www.asp Codex Alimentarius Commission (CAC) Viale delle Terme di Caracalla 00153 Rome Italy Tel: + 39 (06) 57051 Fax: + 39 (06) 57054593 Email: Codex@fao.uk/ Health Protection Scotland Clifton House Clifton Place Glasgow G3 7LN United Kingdom Tel: + 44 (0)141 3001100 Fax: + 44 (0)141 3001170 Email: email@example.com/web/indexen.se Web site: www.uk Web site: www.int/ SIK .who.00187 Rome Italy Tel: + 39 06 487751 Fax: + 39 06 4877599 Email: firstname.lastname@example.org Society of Food Hygiene and Technology The Granary Middleton House Farm Tamworth Road Middleton Staffs B78 2BD United Kingdom Tel: + 44 (0) 1827 872500 Fax: + 44 (0) 1827 875800 Email: admin@sofht.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 Web site: www.FISH AND SEAFOOD Society for Applied Microbiology Bedford Heights Brickhill Drive Bedford MK41 7PH United Kingdom Tel: + 44 (0) 1234 326661 Fax: + 44 (0) 1234 326678 Web site: www.who.co.1211 Geneva 27 Switzerland Tel: + 41 22 7912111 Fax: + 41 22 7913111 Email: firstname.lastname@example.org Web site: www.sfam.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 244 .uk Web site: www.int/foodsafety World Health Organization (WHO) Headquarters CH .sik.se/ FAO/WHO Collaborating Centre for Research and Training in Food Hygiene and Zoonoses Robert von Ostertag Institute Postfach 330013 D-14191 Berlin Germany Tel: + 49 (0) 188 84122155/ 2157 Fax: + 49 (0) 188 84122957 Email: firstname.lastname@example.org/index.who.int.nhs.who.org.euro.scot.nhs.
org Web site: http://www.ift.fda.fsai.org/ Institute of Food Technologists http://www.who.cfsan.org/cms/ Useful Web Sites Gateway to Government food safety information (US) http://www.uk/ 245 .foodscience.org/ The Association of Food.au/ International Association for Food Protection http://www.cdc.sofht.gov/ World Health Organization (WHO): Food safety programmes and projects http://www.uk/ Society of Food Hygiene and Technology (UK) http://www.gov/ Institute of Food Technologists 525 W. Van Buren Suite 1000 Chicago IL 60607 United States of America Tel: + 1 3127828424 or + 1 8004383663 Fax: + 1 3127828348 Email: email@example.com/pol/food/index_en.org/ Royal Institute of Public Health (UK) http://www.htm Centre for Disease Control and Prevention (CDC) (US) http://www.org.ie/ Food Science Australia http://www.gov/foodsafety/ Food Safety Authority of Ireland http://www. Beverage and Consumer Products Companies http://www.int/foodsafety/en/ European Commission: Activities of the European Union food safety http://europa.foodprotection.Center for Food Safety and Applied Nutrition 5100 Paint Branch Parkway College Park MD 20740-3835 United States of America Tel: + 1 888 SAFEFOOD (+ 1 888 723 3366) Web site: www.CONTACTS US FDA .co.fpa-food.riph.FoodSafety.ift.
FISH AND SEAFOOD 246 .
growth and survival in crustacean shellfish 86 Aeromonas sobria. growth and survival in chilled and frozen raw fish 19 of concern in preserved fish 115 Aspergillus spp. as spoilage organism in oysters 65 as spoilage organism in tropical crustacean shellfish 81 in molluscan shellfish 60 Alexandrium. 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 . 69-70 Aeromonas hydrophila. to test microbiological criteria 19 Aeromonadaceae. growth and survival in crustacean shellfish 86 Air drying 101 Alcaligenes. as autochthonous bacterial flora of fish 3 Aeromonas spp. as autochthonous bacterial flora of fish 3 in fish sauce 127 Bacillus cereus. source of PSP toxins 142 Alexandrium ostenfeldii.INDEX Achromobacter spp. 111 Astroviruses. 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. planktonic source of spirolides in shellfish 163 Allergens. causing spoilage of dried fish 110. breakdown of – as indicator of fish freshness 10-1 Azaspiracids 158-9 detection methods 160 origins and distribution 159 toxicology 159-60 Bacalao 97. in crustacean shellfish 86-7 ATP. preserved by dry/wet salting 99 Anisakis spp. as spoilage organisms in chilled recipe meals 43 growth and survival in chilled and frozen raw fish 17 pathogen profile 225-6 pathogenic microorganisms in molluscan shellfish 59-60. in crustacean shellfish 87 Alteromonas spp. as TTX producers 145 Amnesic shellfish poisoning (ASP) toxins 146-8 caused by domoic acid in crustacean shellfish 87 Anchovies. 102 Bacillus spp.
importance in lactic acid fermentation in fish products 130. as cause of spoilage of MAP fish 15 Calciviruses. 136. 113. of recipe fish meals 43 Cholera. cause of neurotoxin shellfish poisoning 153-5 origins and distribution 1545 toxicology 155 importance in wet salting of fish 96 Brining 93. in crustacean shellfish 86-7 Campylobacter coli. associated with consumption of fish/fish products 16.FISH AND SEAFOOD components of seafood ready meals 40 Bacterial pathogens. as autochthonous bacterial flora of fish 3 Clostridium botulinum. preserved by marinating 100 Chemical detection methods. 68-9 Capture and handling. affected by azaspiracids 159 affected by cyclic imines 161 Botulism. 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 . caused by Vibrio cholerae O1 17 Ciguatoxins 155-8 detection methods 157-8 origins and distribution 1567 toxicology 157 Clams. growth and survival in crustacean shellfish 86 growth and survival in molluscan shellfish 59-60. 226 Breading process 36-8 Brevetoxins. of fish – effects on microflora 5-6 Carbohydrate. description and uses 56-7 effects of processing on microflora 60-4 hot-smoking 107 Clear smoke treatments 108 Clostridium. of fresh fish spoilage 11 Chemical pollutants. 137. mould causing spoilage of dried fish 110 Battering process 36-8 Bivalves. fish sauce 123 Basipetospora halophila. for PSP toxins 144 Chemical indicators. 131 Carbon dioxide. 98-9 Brochothrix thermosphacta. as gas in modifiedatmosphere packaging 6 CCP Decision Tree (in HACCP plan) (Fig) 185 Ceviche. indigenous – in fish (Table) 4 non-indigenous – in fish (Table) 4 Bakasang. 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. growth and survival in crustacean shellfish 86 pathogenic microorganisms in molluscan shellfish 59-60 Campylobacter jejuni. in crustacean shellfish 87 Chilled prepared fish products 27-51 Chilled raw fish 1-25 Chilled storage.
smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 249 . for detection of DA 148 ELISA kit (DSP-Check). 152 Diphyllobothrium. of prepared fish products 44 Escherichia coli. of fish – analysis of 10 Diarrhetic shellfish poisoning (DSP) toxins 148-51 detection methods 150-1 origins and distribution 14950 toxicology 150 Dinophysis spp.INDEX pathogen profile 226-7 Clostridium perfringens. to determine fresh fish spoilage 11 Corynebacteria. smoked and dried fish 114 Enzymic spoilage. growth and survival in chilled and frozen raw fish 19 of concern in cold-smoked salmon 115 Domoic acid (DA). as risk in breaded shrimp and prawn products 82 growth and survival in chilled and frozen prepared fish products 45-6 pathogen profile 227-9 Coated fish products. hot-smoking 107 ELISA assay. growth and survival in crabmeat 84 growth and survival in cured. 106 Contacts 241-5 Conway method. as contaminants in scampi 36. as spoilage organisms in tropical crustacean shellfish 81 Coryneforms. 37 as spoilage organisms in chilled products 43 as spoilage organisms in tropical crustacean shellfish 81 as spoilage organisms in vacuum-packed dried fish 111 growth and survival in cured. adverse condition on poorly preserved fish 110 EC grading scheme. processing 334 Cockles. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Crayfish. description and uses 59 effects of processing on microflora 60-4 Cold-smoking 104-5. to detect DSP toxins 150-1 Enrobing process 36-8 Enterobacteriaceae. to detect NSP toxins 155 ELISA kit (Biosense). of fish – different processes 94 Dun. 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. source of azaspiracids 159 source of DSP toxins 148-9. forms for purchase 80 Crustacean shellfish 79-92 Cured fish 93-121 Cyclic imines 160-1 detection methods 162 origins and distribution 161 toxicology 162 Degradation. to determine fresh fish spoilage 11 Eels. as autochthonous bacterial flora of fish 3 Crabs. 149-50.
non-B) viruses. of fish at sea 5 Fatty fish. fermented fish product 124 General Pathogen Surveillance Programme 84 Gravad fish. source of PSP toxins 142 Gyoshoyu. to measure fish skin resistance 10 Flavobacterium. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Herring.FISH AND SEAFOOD EU legislation. 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. spoilage 9-11 Freshness grading schemes 9 Frozen prepared fish products 27-51 Frozen raw fish 1-25 Frozen storage. of fresh fish spoilage 911 Evisceration. 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. definition 1 Fermented fish 123-40 Fermented fish/salt products of Thailand (Table) 125 Fish and shellfish toxins 141-74 Fish cakes. as source of inulin in lactic acid fermentation in fish products 134 Garos. fermented fish product 126 Green cured split cod 97. as source of ciguatoxins 156 Garlic. fish sauce 123 HACCP. preserved by marinating 101 Gravlaks. member of cyclic imine group 161 Gymnodinium catenatum. 102 Gymnodimines. caused by Photobacterium phosphoreum 15 Hon kare bushi 107 Hot-smoking 105-7 250 . mould causing spoilage of dried fish 110 Evaluation. of fish – effects on microflora 5-6 Hepatitis (non-A. 130-5 Fish-spoilage bacteria 12 Fishtester. preserved by marinating 100 preserved by dry/wet salting 99 Histamine fish poisoning. causing pink condition on fish surface 110 in fish sauce 127 Halococcus spp. processing 35 Fish component. in crustacean shellfish 86-7 in molluscan shellfish 59-60 Hepatitis A virus. for evaluation of fish quality 9 Eurotium. specific EU hygiene rules 196-224 Fresh fish. of prepared fish products 44 Gambiertoxins. causing pink condition on fish surface 110 in fish sauce 127 Handling. of chilled and frozen prepared fish products – preparation 33-6 Fish sauces 123-4 Fish sauces and pastes. effects of processing 127-30 Fish sauces and pastes of south-east Asia (Table) 124 Fish/salt/carbohydrate products of Asia (Table) 125. concerning crustacean shellfish 87-8 European Grading Scheme.
preserved by marinating 101 Microbiological criteria. 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. 131 Lactic fermented fish products (Table) 126 fish species used in the Philippines (Table) 126 Lactobacillus. 135 I-zushi. as autochthonous bacterial flora of fish 3 Lean fish. smoked and dried fish (Table) 116-7 for shellfish products 70 Microbiological limits. food hygiene 189-224 Lingulodinium polyedrum. enabled by carbohydrate source in fish products 130. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. 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. producer of brevetoxins 154 Katsuobushi 107 Kench curing 97-8 Klipfish 97 Lactic acid bacteria. 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. causing spoilage in vacuum-packed dried salmon 111 Lactic acid fermentation. leading to oxidative rancidity 11 Lipophilic shellfish poisoning toxins 149. preserved by salt-boiling 100 Ionising radiation. as source of botulism 136 International legislation. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 pathogen profile 229-30 Listeria monocytogenes. for fresh and frozen fish (Table) 20 Micrococcus spp. definition 1 Legislation. 151-3 detection methods 153 Liquid smoke 107-8 Listeria spp. initial – of chilled and frozen prepared fish products 31-2 of chilled and frozen raw fish 3-5 251 . hot-smoking 107 preserved by dry/wet salting 99 Marinating 94. as source of yessotoxins 152 Lipid content. 100-1 Masmeen 107 Matjes. in fish fillet 1 Lipid oxidation.INDEX Hygiene regulations for foodstuffs production and distribution (EU) 190-6 Ice storage. cooked – effects of processing on microflora 83 forms for purchase 80 freshness indicators 83 Mackerel. outbreak caused by gravid 115 Lobster. for microbial inactivation in fish products 127. to reduce rate of fish spoilage 12 Ikan kaju.
in crustacean shellfish 867 Norwalk virus. contaminated by azaspiracids 158 description and uses 58 effects of processing on microflora 60-4 hot-smoking 107 Nam-budu. associated with consumption of fermented fish products 137 Oxidative rancidity. of crustacean shellfish – effects of processing 81-3 of cured. shigelloides. reduction in smoking process 108 Norcardiopsis dassonvillei. as autochthonous bacterial flora of fish 3 as spoilage organism in temperate crustacean shellfish 80 in molluscan shellfish 60 Mouse bioassay. growth and survival in chilled and frozen raw fish 17 Paralytic shellfish poisoning (PSP) toxins 142-4 detection methods 144 toxicology 143 252 . contamination leading to illness 68 description and uses 57 effects of processing on microflora 60-4 hot-smoking 107 spoilage organisms 65 P. pathogenic microorganisms in molluscan shellfish 59-60 Opisthorchis viverrini. 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.FISH AND SEAFOOD of crustacean shellfish 80-1 of cured. 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. smoked and dried fish 95 of fermented fish 126-7 of molluscan shellfish 59-60 Microflora. fish sauce 123 Neosaxitoxin (neoSTS) 142-3 Neurotoxin shellfish poisoning (NSP) toxins 153-5 detection methods 155 Nitrosamines. 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. as TTX producer 145 Norovirus.
preserved by salt-boiling 100 Pink. as cause of histamine fish poisoning 15 as spoilage organism in chilled products 43 as spoilage organism in fresh fish 14-5 Pickling 94. contact details 241-2 Prorocentrum spp. 161. as component for seafood ready meals 40 Pathogen profiles. adverse condition on poorly preserved fish 110 Pinnatoxins member of cyclic imine group 164. effects on microflora of fish 5-9 Professional bodies. as source of yessotoxins 152 Protoperidinium cassipes. as source of DSP toxins 148-9 Protein content. as autochthonous bacterial flora of fish 3 as spoilage organism in chilled products 43 as spoilage organism in crustacean shellfish 80. in fish fillet 1 Proteus. mould causing spoilage of dried fish 110 Potato. causing spoilage of dried fish 110-1 Photobacterium phosphoreum. as component for seafood ready meals 40 Prawns. detection methods 165 origins and distribution 165 toxicology 165 Pissala. 225-39 Pathogens. causing spoilage in vacuum-packed dried fish 111 Pseudomonads. 163. 197 Plesiomonas. smoked and dried fish 113-6 growth and survival in fermented fish 136-7 growth and survival in molluscan shellfish 66-70 Pectenotoxins 151. spoilage organism in oysters 65 Protoceratium reticulatum. 69-70 Polycyclic aromatic hydrocarbons. 152-3 Penicillium spp. in molluscan shellfish 60 Pseudomonas spp. reduction in smoking process 103.INDEX Parasites. 81 as spoilage organism in fresh fish 13 as spoilage organism in oysters 65 as TTX producers 145 253 . for detection of PSP toxins 144. 100-1 Pindang. fermented fish product 124 Plankton. 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. pathogen profile 230 in molluscan shellfish 5960. effects of processing on microflora 81-2 freshness indicators 83 Pre-column derivatisation. 108 Polypaecilum pisce. 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. planktonic source of azaspiracids 159 Pseudomonadaceae. smoked and dried fish 115 Pasta. as source of toxins 159.
100 Salting 97-9 definitions 93-4 Sardines. resulting from poorly salted cod 110 Pyrodinium. to measure fish skin resistance 10 Salmon. fermented fish product 126 Raphidophytes. antibody-based kit to detect STX 144 RT Freshmeter. source of PSP toxins 142 Quality Index Method.) 41 ingredients preparation. as carbohydrate source in lactic acid fermentation in fish products 134 as component for seafood ready meals 40 RIDASCREEN. 83 254 . detected in cod 115 Pteriatoxins. as source of tetrotoxin 144-5 Putty fish. for evaluation of fish quality 9 Radiation. producers of brevetoxins 154 Raw fish.) 41 Recipe fish products 38-40 Rhodotorula rubra. initial microflora 31 Seafood ready meals 39-40 Seafood recipe meal centres 38-9 Shaped fish products. vacuum packing 105 Salmonella spp. importance in lactic acid fermentation in fish products 130 importance in salting. assembly and packaging. processing 34. smoking and marinating of fish 96 Salt-boiling 94. as spoilage organism in chilled products 43 as spoilage organism in fresh fish 13-4 as spoilage organism in temperate crustacean shellfish 80. for microbial inactivation in fish products 127. flow diagram (Fig. for fish ready meals – manufacture 40-2 Saxitoxin (STX) 142-3 Scallops. flow diagram (Fig. as autochthonous bacterial flora of fish 3 inhibition of growth by carbon dioxide 6 Shewanella putrefaciens. preserved in saturated brine 99 Sauce. chilled and frozen 1-25 Recipe dish. in molluscan shellfish 60 Rice. processing and effects on microflora 60-1 Shewanella. implicated in amnesic shellfish poisoning 146-7 Pseudoterranova. 36 Seafood. detection methods 165 member of cyclic imine group 164 origins and distribution 165 toxicology 165 Puffer fish. smoked and dried fish 114 pathogen profile 230-1 pathogenic microorganisms in molluscan shellfish 59-60 Salt burn. 135 Rakefisk. growth and survival in chilled and frozen raw fish 19 Pseudoterranova dicipiens. fermented fish product 126 as source of botulism 136 Rakørret. hazard in salting of fish 97 Salt concentration. description and uses 58-9 Scampi. processing 35 Shellstock.FISH AND SEAFOOD inhibition of growth by carbon dioxide 6 Pseudonitzschia multiseries. as hazard in regard to reef-caught fish 28 growth and survival in crustacean shellfish 84 growth and survival in cured.
as contaminant in scampi 36 as contaminant in enrobing process 37 as hazard in dry salting of fish 89 growth and survival in chilled and frozen prepared fish products 46-7 growth and survival in crustacean shellfish 84 growth and survival in cured. of fish after capture 5 Surstrømming. fish and shellfish 141-74 Trade associations. 101 Storage. growth and survival in chilled and frozen prepared fish products 46 growth and survival in cured. for evaluation of fish quality 9 Torrymeter. as measure of fish degradation 10 Sliming. to measure fish skin resistance 10 Total Volatile Base-Nitrogen. role in fish degradation 10. fermented fish product 126 Taste panel evaluation. of dried fish 111 of fish 6-7 of fish – (US) NACMCF recommendations 7-8 of fish – Codex Committee 255 . in poorly salted cod 110 Smoked fish 93-121 Smoking 103-8 different processes 94-5 Snow Mountain virus. definitions of forms for purchase 79-80 effects of processing on microflora 81-2 freshness indicators 83 Shucked shellfish. of cooked fish 9-10 Tetrotoxin 144-6 detection methods 145-6 toxicology 145 TMAO. contact details 241-2 Trematode infections foodborne 137 Trichosporon. of crustacean shellfish 80. 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. processing and effects on microflora 63-4 Skin resistance.INDEX Shigella spp. smoked and dried fish 114 pathogenic microorganisms in molluscan shellfish 59-60 Shottsuru. 12-5 of recipe fish meals 43-4 of shellfish 65 Staphylococcus. 83 of fermented fish 135-6 of fish subjected to different preservation methods 108-10 of fresh fish 9-11. 14 Torry Scale. fish sauce 123 Shrimps. 13. concerning crustacean shellfish 89 Vacuum packaging. detection methods 164 member of cyclic imine group 162-4 origins and distribution 163 toxicology 163-5 Spoilage. as indicator of fresh fish spoilage 11 Toxins. pathogenic microorganisms in molluscan shellfish 59-60 Specific spoilage organisms (SSOs) 12-5 Spirolides. in molluscan shellfish 60 Tuna. grading of 31 hot-smoking 107 US legislation.
growth and survival in chilled and frozen raw fish 17 Vibrio cholerae 01 and 0139. growth and survival in crustacean shellfish 85 pathogen profile 233-4 Vibrio cholerae 01. causing dun condition on dried fish 110 Wet salting 98-9 Wet storage. 68 Vibrio cholerae non-01. growth and survival in chilled and frozen prepared fish products 47 growth and survival in crustacean shellfish 86-7 Wallemia sebi. 152-3 256 . 81 as TTX producers 145 pathogenic microorganisms in molluscan shellfish 59-60 risk in cold-smoking 105 Vibrio cholerae. 62 Yersinia spp. processing for seafood recipe meal centres 39 Vibrio spp. growth and survival in molluscan shellfish 67-8 Vibrio parahaemolyticus. as spoilage organisms in chilled recipe meals 43 Yessotoxins 151. 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. as autochthonous bacterial flora of fish 3 as spoilage organisms in chilled products 43 as spoilage organisms in vacuum-packed dried fish 111 Viruses. 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.FISH AND SEAFOOD guidance 7 of fresh chilled products 42 Vegetables. as hazard in regard to reef-caught fish 28 as spoilage organism in crustacean shellfish 80.
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